Researcher basic information

• 博士（獣医学）, 北海道大学

• https://researchmap.jp/s.murata?lang=en

• https://jglobal.jst.go.jp/detail?JGLOBAL_ID=201301059666482919

• 201301059666482919

• Marek's Disease
• 腫瘍
• ウイルス学
• 感染免疫
• ワクモ

• Life Science, Veterinary medical science

• Bachelor's degree program, School of Veterinary Medicine
• Doctoral (PhD) degree program, Graduate School of Infectious Diseases

Research activity information

• Sep. 2023, The Japanese Society of Veterinary Science, Veterinary Science Award
  Studies on the epidemiological surveys, pathogenesis and strategies for the control of avian diseases

• Clinical efficacy of anti-programmed death ligand 1 antibody HFC-L1/c4G12 in dogs with malignant tumors: an exploratory study.
  Satoshi Takagi; Michihito Tagawa; Naoya Maekawa; Satoru Konnai; Yumiko Kagawa; Kenji Hosoya; Akinori Yamauchi; Ayano Kudo; Shintaro Kamo; Sangho Kim; Ryohei Kinoshita; Tatsuya Deguchi; Ryo Owaki; Yurika Tachibana; Madoka Yokokawa; Hiroto Takeuchi; Hayato Nakamura; Yukinari Kato; Shigeki Kanazawa; Tomoyuki Abe; Takuya Furuta; Keiichi Yamamoto; Yasuhiko Suzuki; Tomohiro Okagawa; Shiro Murata; Kazuhiko Ohashi
  The Journal of veterinary medical science, 87, 12, 1419, 1425, 01 Dec. 2025, [Domestic magazines]
  English, Scientific journal, Cancer in dogs remains a major challenge in modern veterinary medicine. Immunotherapy using immune checkpoint inhibitors (ICIs) is available for various human tumor types, and recent veterinary clinical studies have shown that ICIs are a promising approach for treating canine cancers. A canine chimeric anti-PD-L1 antibody, c4G12 (HFC-L1), has been investigated for canine cancer immunotherapy; however, its clinical benefits have not been well characterized in tumors other than pulmonary metastatic (stage IV) oral malignant melanoma (OMM). To explore the efficacy and safety of HFC-L1, we conducted a clinical study in dogs with stage I-III OMM or other tumor types (n=12). HFC-L1 treatment at a dose of 5 mg/kg every 2 weeks was well tolerated, and no grade 3 or higher treatment-related adverse events were reported. Among the dogs eligible for response evaluation (n=10), a partial response was observed in one dog with squamous cell carcinoma, resulting in an objective response rate of 10%. In addition, in a dog with ceruminous cell carcinoma, clinical evidence of a tumor response was observed in metastatic lung lesions. Together, these results suggest that the HFC-L1 therapy is applicable for the treatment of various tumor types, although its clinical benefits should be further evaluated in clinical studies involving a larger number of dogs with each tumor type.
• Genetic diversity and phylogenetic relationships among western and Asian honey bees, and Eristalis tenax collected in Myanmar
  Saw Bawm, Aye Pyae Pyae Khaing, Hlaing Min Oo, Myint Myint Mu, Ngwe Chi Hmuu, Aye Nyein Sandi Zaw, Nyunt Lwin, Hla Myet Chel, Yadanar Khaing, Shwe Yee Win, Lat Lat Htun, Shiro Murata, Satoru Konnai, Kazuhiko Ohashi, Nariaki Nonaka, Ryo Nakao, Than Naing Tun
  Journal of Asia-Pacific Entomology, 28, 4, 102479, Dec. 2025, [Peer-reviewed]
• Enhancement of Vaccine-Induced T-Cell Responses by Probiotics in Calves.
  Mari Ikehata; Tomohiro Okagawa; Hayato Nakamura; Naoya Maekawa; Yasuhiko Suzuki; Shiro Murata; Kazuhiko Ohashi; Satoru Konnai
  Vaccines, 13, 11, 31 Oct. 2025, [International Magazine]
  English, Scientific journal, BACKGROUND/OBJECTIVES: Calves have immature immune systems, hence immunization with vaccines is essential to protect them from infectious diseases. However, immune responses to vaccines vary widely among individuals. Therefore, strategies for enhancing vaccine efficacy are needed, particularly those targeting low responders to vaccines. Probiotics have attracted attention because of their beneficial immunomodulatory effects on the host. Although probiotics may improve calf immunity, their potential to enhance immune responses to vaccines in calves remains unclear. Thus, we investigated whether immune responses to vaccines, especially T-cell responses, are enhanced when calves receive a combination of probiotic supplementation and vaccination. METHODS: Calves were divided into three feeding groups, as follows: negative control feed, live bacteria-mixed feed (Zeosapo KB), and Clostridium butyricum-only feed (CB). After weaning, all calves received two doses of a live attenuated hexavalent viral vaccine. T-cell responses to a vaccine antigen were evaluated by measuring the expression levels of lymphocyte activation markers CD25 and CD69, as well as Th1 cytokine production, in peripheral blood mononuclear cell culture assays. RESULTS: CD25 expression significantly increased in CD4+ T cells four weeks after the booster vaccination in the Zeosapo KB- and CB-fed groups. In addition, the CD25+CD69+ cell ratio in CD4+ T cells was increased in these groups. The production of Th1 cytokines in the culture supernatant was also increased in the CB-fed group. CONCLUSIONS: This clinical study demonstrates that probiotics activate CD4+ T cells and enhance Th1 cytokine responses in vaccinated calves.
• Two distinct polymorphisms in the basic region of Meq protein of marek's disease virus alter pathological progression and clinical manifestations.
  Jumpei Sato; Aoi Kurokawa; Yoshinosuke Motai; Shunsuke Yamagami; Shwe Yee Win; Fumiya Horio; Hikaru Saeki; Naoya Maekawa; Tomohiro Okagawa; Benedikt B Kaufer; Nikolaus Osterrieder; Mark S Parcells; Satoru Konnai; Kazuhiko Ohashi; Shiro Murata
  Virology journal, 22, 1, 303, 303, 26 Sep. 2025, [Peer-reviewed], [Last author, Corresponding author], [International Magazine]
  English, Scientific journal, BACKGROUND: Marek's disease virus (MDV) causes Marek's disease (MD) in chickens, which is characterized by malignant lymphomas and neurological disorders. Although MD is currently controlled using live vaccines, the virulence of field strains has continuously increased in recent decades. Polymorphisms in the MDV-encoded oncoprotein Meq are shared among field strains according to their virulence. In particular, very virulent MDV strains harbor characteristic amino acid changes in the basic region of Meq at positions 77 and 80; however, the contribution of these polymorphisms to virulence remains unclear. METHODS: To assess the impact of these polymorphisms on MDV virulence, we generated recombinant MDV (rMDV) based on the very virulent RB-1B strain, harboring K77E and D80Y substitutions in Meq found in low-virulent strains (rRB-1B_Meq77/80). Chickens were challenged with rMDVs, and survival rates and tumor incidence were evaluated. Viral loads in major organs were quantified by quantitative PCR, and the dynamics of MDV-infected cells and T cells were analyzed using flow cytometry. In addition, histopathological analysis was performed to further examine differences in pathogenesis in detail. To elucidate the mechanisms underlying pathogenesis, we conducted reporter assays to assess the effect of these polymorphisms in the basic region on its transcriptional regulatory activity. RESULTS: rRB-1B_Meq77/80 exhibited a reduced virulence but unexpectedly caused other clinical signs, including open-mouth breathing, in infected chickens. Quantitative PCR analysis showed consistently lower viral loads across all examined organs in rRB-1B_Meq77/80-infected chickens. Flow cytometric analysis revealed a reduction in MDV-infected cells, accompanied by a notable increase in CD8⁺ T cell populations. Histopathological analysis showed bronchus-associated lymphoid tissue hyperplasia in the lungs. Reporter assays revealed that most amino acid substitutions in the basic region in low-virulence strains reduced transcriptional regulatory activity. CONCLUSION: Our data indicate that polymorphisms at positions 77 and 80 in the Meq of low-virulence strains reduce MDV virulence and Meq-mediated transcription and possibly alter pathogenesis. This study improves our understanding of the mechanisms underlying MDV virulence.
• Exploratory, Randomized, Dose-Response Study of the Anti-PD-L1 Antibody HFC-L1/c4G12 in Dogs with Pulmonary Metastatic Oral Malignant Melanoma.
  Kenji Hosoya; Sangho Kim; Ryohei Kinoshita; Naoya Maekawa; Satoru Konnai; Satoshi Takagi; Michihito Tagawa; Yumiko Kagawa; Tatsuya Deguchi; Ryo Owaki; Yurika Tachibana; Madoka Yokokawa; Hiroto Takeuchi; Hayato Nakamura; Akinori Yamauchi; Ayano Kudo; Shintaro Kamo; Yukinari Kato; Shigeki Kanazawa; Tomoyuki Abe; Takuya Furuta; Keiichi Yamamoto; Yasuhiko Suzuki; Tomohiro Okagawa; Shiro Murata; Kazuhiko Ohashi
  Veterinary sciences, 12, 9, 02 Sep. 2025, [International Magazine]
  English, Scientific journal, Oral malignant melanoma (OMM) is a highly aggressive malignancy in dogs. The development of effective systemic therapies is urgently required to improve the treatment of canine OMM. Immunotherapy using immune checkpoint inhibitors (ICIs) has been investigated in canines following their dramatic success in human cancer treatment; however, there is still a need for extensive veterinary clinical studies to clarify and optimize their clinical benefits. Among the ICIs under development for canine cancer immunotherapy, c4G12 (HFC-L1), a canine chimeric anti-PD-L1 antibody, has shown promising efficacy in dogs with pulmonary metastatic OMM in previous clinical studies. However, the optimal dose of HFC-L1/c4G12 has not yet been determined. To explore the dose-response relationship of HFC-L1, a multicenter, randomized clinical study was conducted using three different doses (2, 5, or 10 mg/kg via intravenous infusion every 2 weeks) to treat dogs with pulmonary metastatic OMM (n = 8-9 per group). The safety profiles were similar among the dose groups, and numerically longer median overall survival was achieved in the higher dose groups (5 and 10 mg/kg) than in the 2 mg/kg group. Although the study was exploratory in nature with a small sample size, 5-10 mg/kg should be considered the preferred dose in future clinical studies using HFC-L1.
• Evaluation of PD-L1 and TIM-3 Pathways in T Cells During Experimental Bovine Leukemia Virus Infection in Sheep.
  Wisa Tiyamanee; Tomohiro Okagawa; Shinji Yamada; Mari Ikehata; Hayato Nakamura; Maho Inoue; Naoya Maekawa; Yukinari Kato; Shiro Murata; Kazuhiko Ohashi; Kenji Murakami; Satoru Konnai
  Veterinary sciences, 12, 9, 26 Aug. 2025, [International Magazine]
  English, Scientific journal, Bovine leukemia virus (BLV) infects B cells in ruminants and causes lymphoma after an extended latency period. Previous studies have demonstrated T-cell exhaustion through the upregulation of immunoinhibitory molecules, including programmed death-ligand 1 (PD-L1) and T-cell immunoglobulin and mucin domain-3 (TIM-3), in BLV-infected cattle. However, studying T-cell exhaustion across all BLV infection stages remains challenging due to the virus's prolonged latency in cattle. Sheep provide a valuable model, as they develop lymphoma more rapidly than cattle. This study examined PD-L1 and TIM-3 expression kinetics and T-cell function in BLV-infected sheep. During persistent infection, PD-L1 expression was correlated with BLV proviral load. TIM-3 expression increased in CD4+, CD8+, and γδTCR+ T cells. Functional analysis revealed that TIM-3 blockade enhanced T-cell activation markers (CD25 and CD69) in cultured PBMCs from infected sheep and increased CD69+IFN-γ+ and CD69+TNF-α+ populations, particularly among CD4+ T cells. Combined PD-L1 and TIM-3 blockade significantly enhanced cytokine production in both CD4+ and CD8+ T cells, while PD-L1 blockade alone showed limited effects. These findings demonstrate the effect of TIM-3 blockade in restoring immune function during chronic BLV infection, effective both alone and in combination. This study validates sheep as a valuable model for investigating immune checkpoint dynamics and evaluating immunotherapies for BLV infection and other chronic diseases.
• Amino acid insertion in the Meq protein of Marek's disease virus, an avian oncogenic herpesvirus, accelerates tumorigenesis.
  Jumpei Sato; Shiro Murata; Yoshinosuke Motai; Shwe Yee Win; Hikari Seo; Shunsuke Yamagami; Naoya Maekawa; Tomohiro Okagawa; Satoru Konnai; Kazuhiko Ohashi
  Microbiology spectrum, 13, 8, e0336824, 05 Aug. 2025, [Peer-reviewed], [Corresponding author], [International Magazine]
  English, Scientific journal, Marek's disease virus (MDV) causes lymphomas (Marek's disease) in chickens. Despite vaccination, MDV field strains exhibit increased virulence, and sporadic outbreaks are still reported. An insertion/deletion in Meq has been identified in several MDV strains, and our previous study using recombinant MDV (rMDV) demonstrated that an insertion in Meq enhanced MDV virulence, whereas a deletion reduced its virulence. However, the mechanisms by which these indels in Meq alter MDV virulence remain elusive. We aimed to clarify the impact of the insertion in Meq on pathogenesis. We compared the effects of Meq and Meq with the insertion, termed L-Meq, on transcriptional regulation, the dynamics of transformed T cells and relevant T-cell subsets, and the patterns of gene expression in tumor lesions of infected chickens. Reporter assays revealed that insertion increased the transactivation activity on the infected-cell protein 4 promoter in the MDV genome and bcl-2/cd30 promoters in the host genome. rMDV encoding L-Meq (vRB-1B_L-Meq) exhibited higher mortality and tumor incidence than rMDV encoding Meq (vRB-1B_Meq), and vRB-1B_L-Meq infection increased the proportion of CD4+ T cells, which are targets for transformation by MDV, in the early post-infection phase, compared with vRB-1B_Meq. RNA-seq analysis revealed that similar genes were modulated in tumor lesions of chickens infected with vRB-1B_Meq or vRB-1B_L-Meq. These findings suggest that although the insertion in Meq does not alter the target genes for transcriptional regulation, it accelerates the process of tumorigenesis by enhancing the transactivation activity.IMPORTANCEMarek's disease, an avian lymphoproliferative disease, is caused by Marek's disease virus (MDV). Meq, an MDV oncoprotein, regulates the expression of viral and host genes. Meq with an insertion, termed L-Meq, has been identified as a factor that enhances MDV virulence. However, the mechanisms by which the insertion in Meq alters MDV virulence remain unknown. Our study clarified that the insertion enhances the transactivation activity of Meq on the host promoters related to tumorigenesis. Notably, the transcriptomes of tumor lesions in chickens infected with recombinant MDV (rMDV) with L-Meq and those infected with rMDV encoding Meq without the insertion were similar; however, chickens infected with rMDV harboring L-Meq exhibited higher proportions of CD4+ T cells and regulatory T cells, which are targets for transformation by MDV, in the early post-infection phase, suggesting accelerated tumorigenesis. This study contributes to the current understanding of the mechanisms underlying MDV virulence.
• Amino Acid Polymorphisms in the Basic Region of Meq of Vaccine Strain CVI988 Drastically Diminish the Virulence of Marek's Disease Virus.
  Jumpei Sato; Yoshinosuke Motai; Shunsuke Yamagami; Aoi Kurokawa; Shwe Yee Win; Fumiya Horio; Hikaru Saeki; Naoya Maekawa; Tomohiro Okagawa; Satoru Konnai; Kazuhiko Ohashi; Shiro Murata
  Viruses, 17, 7, 26 Jun. 2025, [Peer-reviewed], [Last author, Corresponding author], [International Magazine]
  English, Scientific journal, Marek's disease virus (MDV) is the etiological agent of Marek's disease (MD), a lymphoproliferative disorder in chickens. Polymorphisms in the MDV-encoded oncoprotein Meq are shared among field strains and correlate with their virulence. The attenuated vaccine strain CVI988 harbors unique amino acid polymorphisms in Meq, particularly at positions 71, 77, and 326. In this study, we investigated the impact of these polymorphisms on Meq protein function and MDV virulence. Reporter assays revealed that the substitutions, particularly A71S and K77E, markedly impaired the transcriptional regulatory activity of Meq. To evaluate their effect on virulence, we generated a recombinant MDV based on the very virulent RB-1B strain, encoding Meq with A71S and K77E substitutions (rRB-1B_Meq71/77). Chickens infected with rRB-1B_Meq71/77 developed neither clinical signs nor lymphomas. Flow cytometry revealed no expansion of infected cells in this group, but a marked increase in CD8+ T and γδ T cells during early infection. Histopathological analysis also confirmed the absence of MD-associated lesions. These findings demonstrate that the polymorphisms at positions 71 and 77 in the CVI988 strain are sufficient to abolish MDV virulence. This study provides insight into the molecular basis of MDV virulence and informs the strategy for the design of more effective vaccines.
• Comprehensive analysis of immune checkpoint molecules profiles phenotype and function of exhausted T cells in enzootic bovine leukosis.
  Hayato Nakamura; Satoru Konnai; Tomohiro Okagawa; Naoya Maekawa; Wisa Tiyamanee; Mari Ikehata; Koume Matsubara; Kei Watari; Kana Kamitani; Maya Saito; Yukinari Kato; Yasuhiko Suzuki; Shiro Murata; Kazuhiko Ohashi
  Journal of immunology (Baltimore, Md. : 1950), 27 May 2025, [Peer-reviewed], [International Magazine]
  English, Scientific journal, Bovine leukemia virus (BLV) causes enzootic bovine leukosis (EBL), a B-cell lymphoma in cattle. Previous studies have demonstrated that T cells of BLV-infected cattle show increased expression of immune checkpoint molecules, including programmed death-1 (PD-1), lymphocyte-activation gene-3 (LAG-3), cytotoxic T-lymphocyte-associated antigen-4 (CTLA-4), and T-cell immunoglobulin domain and mucin domain-3 (TIM-3), leading to T-cell exhaustion. However, the key immune checkpoint molecules driving T-cell exhaustion in BLV-induced tumorigenesis remained unclear. In this study, we identified the key immune checkpoint molecules by performing comprehensive flow cytometric analyses of T cells from EBL cattle, and elucidated the phenotype and function of exhausted T cells using a transcriptomic analysis by RNA sequencing and cell culture assays. The comprehensive expression analysis revealed that the proportion of CD4+ and CD8+ T cells co-expressing PD-1 and TIM-3 was significantly increased in the peripheral blood and tumor tissues of EBL cattle compared to healthy cattle. Transcriptomic analysis of PD-1+TIM-3+ T cells revealed the upregulation of genes related to terminal exhaustion and the downregulation of genes related to T-cell differentiation and response in this subset. Additionally, PD-1+TIM-3+ T cells exhibited higher expression of CTLA-4, LAG-3, and Eomes, and lower expression of T-bet, suggesting a terminally exhausted phenotype. Cell culture assays revealed a significant impairment in IFN-γ production in PD-1+TIM-3+ T cells upon stimulation, reflecting severe dysfunction. These findings indicate that PD-1+TIM-3+ T cells play a central role in T-cell exhaustion during BLV-induced tumorigenesis. This study provides valuable insights for future therapeutic strategies against BLV infection.
• Programmed Cell Death-1 Expression in T-Cell Subsets in Chickens Infected with Marek's Disease Virus.
  Jumpei Sato; Yoshinosuke Motai; Shunsuke Yamagami; Shwe Yee Win; Fumiya Horio; Hikaru Saeki; Naoya Maekawa; Tomohiro Okagawa; Satoru Konnai; Kazuhiko Ohashi; Shiro Murata
  Pathogens (Basel, Switzerland), 14, 5, 29 Apr. 2025, [Peer-reviewed], [Last author, Corresponding author], [International Magazine]
  English, Scientific journal, Marek's disease virus (MDV) causes Marek's disease (MD) in chickens, characterized by malignant lymphomas and immunosuppression. Sporadic MD outbreaks continue to occur even among vaccinated flocks in certain regions due to the increased virulence of the field strains. However, the mechanisms of tumorigenesis and immunosuppression caused by MDV remain to be fully elucidated. We previously reported that the mRNA expression of programmed cell death 1 (PD-1), an immune checkpoint molecule, was increased in tumor lesions caused by MDV, and its expression was positively correlated with the mRNA expression of Meq, an MDV-specific oncogene. In this study, we characterized PD-1-expressing T-cell subsets in the spleen and tissues of chickens that developed tumors to investigate the association between PD-1 expression and immunosuppression. Flow cytometric analysis revealed that the proportion of PD-1-expressing CD4+ T-cells, which are targets of MDV tumorigenesis, increased in the spleen and tumor tissues of chickens with MD. The proportion of PD-1+ CD4+ T-cells was higher in Meq-expressing cells than in those that were not. In the spleens of chickens with MD, the proportions of PD-1-expressing cells were increased in CD8+ and γδ T-cells, which play pivotal roles in defense against MD pathogenesis, relative to those of spleens from uninfected chickens. Moreover, the proportion of PD-1+ CD8+ T-cells expressing interferon (IFN)-γ did not increase in the spleen of chickens with MD. Additionally, no difference in the proportion of IFN-γ+ γδ T-cells expressing and not expressing PD-1 was observed in the spleens of chickens with MD, although the proportion of IFN-γ+ γδ T-cells expressing PD-1 in the spleens of uninfected chickens was higher. The function of PD-1-expressing CD8+ and γδ T-cells in chickens may be impaired after developing MD, which may cause MDV-induced immunosuppression.
• Microscopic examination of haemoparasites and the first molecular detection of Theileria equi in horses in Myanmar.
  Yadanar Khaing; Lat Lat Htun; Kyaw San Linn; Win Ohnmar Kyaw; Theint Theint Nwae; Hla Myet Chel; Shwe Yee Win; Shiro Murata; Ryo Nakao; Nariaki Nonaka; Saw Bawm
  Parasitology research, 124, 4, 42, 42, 21 Apr. 2025, [Peer-reviewed], [International Magazine]
  English, Scientific journal, The study aimed to determine the prevalence of blood parasites in horses and identify risk factors and molecular detection of piroplasm species (Theileria equi and Babesia caballi) of horses in Myanmar. Blood samples (n = 302) were collected from five regions of Myanmar. Blood smears were screened for presence of piroplasms. Samples positive for piroplasms were subjected to molecular identification using primers specific to the small subunit ribosomal RNA (SSU rRNA) of piroplasms. The overall prevalence of blood parasites (piroplasms, Trypanosoma spp., and microfilaria of Setaria equina) was 30.8% (93/302). The prevalence of piroplasms, Trypanosoma spp., and microfilaria of S. equina, were 22.5% (68/302), 8.3% (25/302), and 2.3% (7/302), respectively. Samples from Yangon (35.0%) and Mandalay (35.0%) showed the highest prevalence, followed by Northern Shan State, Ayeyarwady, and Nay Pyi Taw (33.9%, 25.0% and 24.6%, respectively). The hypothesized factors (age, sex, and breed) showed no significant association (p > 0.05) with the overall occurrence of blood parasites. Although no significant association (p > 0.05) was found between blood parameters (WBCs, RBCs, Hb, and HCT) and the presence of blood parasites, the negative group had a larger WBC count than the positive group. Molecular characterization of piroplasm of two obtained sequences confirmed Theileria equi. This is the first report on microscopic and molecular detection of T. equi in horses in Myanmar, and the findings provide baseline information for blood parasites in horses.
• Genomic characteristics and distinctive genetic polymorphisms of Marek's disease virus strains circulating in Japan.
  Shunsuke Yamagami; Jumpei Sato; Yoshinosuke Motai; Naoya Maekawa; Tomohiro Okagawa; Satoru Konnai; Kazuhiko Ohashi; Shiro Murata
  The Journal of veterinary medical science, 03 Apr. 2025, [Peer-reviewed], [Last author, Corresponding author], [Domestic magazines]
  English, Scientific journal, Marek's disease virus (MDV) is an oncogenic herpesvirus that causes malignant lymphomas in chickens (Marek's disease, MD). Although MD is currently well controlled by vaccination, field strains of MDV tend to increase in virulence, and sporadic outbreaks have been reported worldwide, including in Japan. Previously, we determined the whole-genome sequence of a Japanese MDV strain, Kgs-c1, isolated in 2014, and analyzed its genetic characteristics. However, to date, Kgs-c1 is the only MDV strain that has been subjected to whole-genome sequencing, leaving the possibility that the key genetic features of the Japanese strains remain undiscovered. Therefore, we conducted whole-genome sequencing of two MDV strains, Kgs-c2 and Sg-c1, isolated in 2016 and 2019, respectively, and analyzed their genetic characteristics. Consistent with our previous findings, the genomic sequences of the Japanese strains were classified into the Eurasian cluster, suggesting that the Japanese strains are genetically closer to the European and Chinese strains than they are to the US strains. In contrast, the Japanese strains exhibited genetic features different from those of pathogenic European and Chinese strains. Furthermore, polymorphisms in the meq and infected cell protein 4 (ICP4) genes, which are candidates involved in the enhanced virulence of MDV, were observed among the newly isolated strains and Kgs-c1. Thus, three MDV strains isolated in Japan had different genetic features from those in other countries. These findings emphasize the necessity of periodic monitoring of MDV field strains in order to determine shifts in their characteristics and effectively control MD.
• Performance evaluation of an improved RAISING method for clonality analysis of bovine leukemia virus-infected cells: a collaborative study in Japan.
  Tomohiro Okagawa; Naomi Nojiri; Hazuka Yoshida-Furihata; Naganori Nao; Misono Tominaga; Junko Kohara; Satoshi Gondaira; Hidetoshi Higuchi; Yohei Takeda; Haruko Ogawa; Shinji Yamada; Kenji Murakami; Yasunori Suzuki; Shinji Takai; Masaki Maezawa; Hisashi Inokuma; Kaori Shimizu; Yasuo Inoshima; Tatsufumi Usui; Michihito Tagawa; Mari Yamamoto; Hirohisa Mekata; Mana Esaki; Makoto Ozawa; Takahiro Matsudaira; Naoya Maekawa; Shiro Murata; Kazuhiko Ohashi; Masumichi Saito; Satoru Konnai
  The Journal of veterinary medical science, 28 Mar. 2025, [Peer-reviewed], [Domestic magazines]
  English, Scientific journal, Bovine leukemia virus (BLV), a retrovirus that is widespread worldwide, causes enzootic bovine leukosis (EBL), a B-cell leukemia/lymphoma with a poor prognosis that ultimately results in death. In Japan, the number of cattle infected with this virus is increasing, and it is estimated more than 35% of cattle are currently infected. Since no vaccines or treatments against BLV infection are currently available, it is important to establish a method of early diagnosis for EBL to reduce economic losses caused by the disposal of EBL cattle in Japan, where a large number of expensive beef cattle are raised. We previously developed Rapid Amplification of the Integration Site without Interference by Genomic DNA Contamination (RAISING), a cost-effective, rapid, and sensitive method for the clonality analysis of BLV-infected cells. Despite its usefulness for the early diagnosis of EBL, RAISING had drawbacks preventing its practical application. Here, we report the development of an improved method, RAISING ver.2, and its performance. Compared to BLV clonality analysis using the previous method, RAISING ver.2 was found to maintain high accuracy and reproducibility despite its simplification. Moreover, its performance was also validated in a multicenter validation study. Taken together, our results strongly suggest that RAISING ver.2 can be fully utilized in clinical practice. Successful commercialization of a RAISING test kit could overcome the concerns of livestock farmers suffering from EBL, thereby promoting a stable supply of Japanese beef, both domestically and internationally.
• Characterization of a lipocalin-like molecule from Dermanyssus gallinae as a potential vaccine antigen.
  Fumiya Horio; Hikari Seo; Shwe Yee Win; Jumpei Sato; Yoshinosuke Motai; Shunsuke Yamagami; Takumi Sato; Eiji Ohishi; Naoya Maekawa; Tomohiro Okagawa; Satoru Konnai; Kazuhiko Ohashi; Shiro Murata
  Veterinary immunology and immunopathology, 283, 110921, 110921, 24 Mar. 2025, [Peer-reviewed], [Last author, Corresponding author], [International Magazine]
  English, Scientific journal, Poultry red mites (PRMs, Dermanyssus gallinae) are hematophagous ectoparasites of chickens that pose a significant threat to the egg-laying industry. The emergence of acaricide-resistant PRMs raises the demand for alternative control approaches such as vaccination. However, several vaccine antigens have failed to suppress the growth of PRM populations in field trials due to difficulties in maintaining antibody levels. In ticks, the molecules exposed to the host, such as lipocalins, can facilitate antibody production, and are therefore considered advantageous as vaccine antigens. Therefore, we focused on a lipocalin-like molecule (Dg-Lipocalin) identified from an RNA-seq analysis reported by Fujisawa et al. (2020) and analyzed its exposure to the host and potential as a vaccine antigen. Western blotting using 500-fold diluted plasma of chickens from PRM-contaminated farms revealed the presence of antibodies against Dg-Lipocalin, suggesting its exposure to the host. To evaluate its potential as a vaccine antigen, PRMs were artificially fed immune plasma with 32,000- to 64,000-fold antibody titers or plasma from PBS-inoculated control chickens, and their mortality was observed for 7 days. In experiment 1, the immune plasma significantly increased PRM mortality compared to the control plasma. However, these effects were not observed in experiment 2, although the total mortality was significantly increased in immune plasma-fed PRMs. Thus, the efficacy of Dg-Lipocalin appears to be limited; however, its exposure to the host may result in sustained antibody titers. Further investigation is required to evaluate its feasibility.
• Development of caninized anti-CTLA-4 antibody as salvage combination therapy for anti-PD-L1 refractory tumors in dogs.
  Naoya Maekawa; Satoru Konnai; Kei Watari; Hiroto Takeuchi; Takeshi Nakanishi; Taro Tachibana; Kenji Hosoya; Sangho Kim; Ryohei Kinoshita; Ryo Owaki; Madoka Yokokawa; Yumiko Kagawa; Satoshi Takagi; Tatsuya Deguchi; Hiroshi Ohta; Yukinari Kato; Satoshi Yamamoto; Keiichi Yamamoto; Yasuhiko Suzuki; Tomohiro Okagawa; Shiro Murata; Kazuhiko Ohashi
  Frontiers in immunology, 16, 1570717, 1570717, 2025, [International Magazine]
  English, Scientific journal, Immune checkpoint inhibitors (ICIs) are widely used for cancer immunotherapy; however, the clinical efficacy of anti-PD-1/PD-L1 monotherapy is generally limited, highlighting the need to develop combination therapies. Dogs develop spontaneous tumors in immunocompetent settings, and anti-PD-1/PD-L1 antibodies exert similar clinical benefits. However, no clinically relevant anti-CTLA-4 antibody has been reported, limiting the value of canine tumors as comparative models for human ICI research. Here, canine CTLA-4 was molecularly characterized, and a caninized anti-CTLA-4 antibody (ca1C5) that blocks CTLA-4/ligand binding was developed. Treatment with ca1C5 increased cytokine production in canine immune cell cultures, and the immunostimulatory effect was enhanced when used in combination with the anti-PD-L1 antibody c4G12. As a proof-of-concept, a veterinary clinical study was conducted to demonstrate the safety and clinical efficacy of anti-CTLA-4 antibody as salvage combination therapy in dogs with advanced tumors refractory to prior c4G12 monotherapy. The combination treatment (c4G12 plus ca1C5) was well-tolerated, and evidence of antitumor activity was observed in one dog with oral malignant melanoma. Further studies are warranted to advance veterinary care for dogs and to better characterize canine ICI models for human onco-immunology research.
• Potential of histamine release factor for the utilization as a universal vaccine antigen against poultry red mites, tropical fowl mites, and northern fowl mites.
  Shwe Yee Win; Fumiya Horio; Jumpei Sato; Yoshinosuke Motai; Hikari Seo; Sotaro Fujisawa; Takumi Sato; Eiji Oishi; Lat Lat Htun; Saw Bawm; Tomohiro Okagawa; Naoya Maekawa; Satoru Konnai; Kazuhiko Ohashi; Shiro Murata
  The Journal of veterinary medical science, 20 Nov. 2024, [Peer-reviewed], [Last author, Corresponding author], [Domestic magazines]
  English, Scientific journal, Poultry red mites (PRMs, Dermanyssus gallinae), tropical fowl mites (TFMs, Ornithonyssus bursa), and northern fowl mites (NFMs, Ornithonyssus sylviarum) are hematophagous mites that are distributed worldwide which pose a serious challenge to the poultry industry and negatively impact poultry production and welfare. Vaccines represent a promising approach for controlling avian mites, and the identification of antigens with broad efficacy against multiple avian mite species is advantageous for vaccine control. This study aimed to identify histamine release factor (HRF), which was previously reported as a candidate vaccine antigen against PRMs, from TFMs and NFMs and to analyze its cross-reactivity and acaricidal effects on different avian mite species. The deduced amino acid sequences of the HRFs identified in the TFMs and NFMs were highly homologous to those of the PRMs. We generated recombinant HRF (rHRF) of TFMs, NFMs, and PRMs, and immune plasma against each rHRF was produced by immunization with each antigen. The immune plasma contained antibodies specific to each antigen and showed cross-reactivity with rHRFs from different avian mites. Moreover, PRM nymphs (protonymphs) artificially fed each immune plasma showed higher mortality rates than those fed the control plasma. These results suggest that HRFs can be used as candidate antigens for a universal vaccine with broad efficacy across avian mites.
• In Vivo Characterization of the Anti-Glutathione S-Transferase Antibody Using an In Vitro Mite Feeding Model.
  Shwe Yee Win; Hikari Seo; Fumiya Horio; Sotaro Fujisawa; Jumpei Sato; Yoshinosuke Motai; Takumi Sato; Eiji Oishi; Akira Taneno; Lat Lat Htun; Saw Bawm; Tomohiro Okagawa; Naoya Maekawa; Satoru Konnai; Kazuhiko Ohashi; Shiro Murata
  Vaccines, 12, 2, 148, 148, MDPI AG, 30 Jan. 2024, [Peer-reviewed], [Last author, Corresponding author], [International Magazine]
  English, Scientific journal, Poultry red mites (Dermanyssus gallinae, PRMs), tropical fowl mites (Ornithonyssus bursa, TFMs), and northern fowl mites (O. sylviarum, NFMs) are blood-feeding pests that debilitate poultry worldwide. Glutathione S-transferase (GST) plays an important role in the detoxification and drug metabolism of mites. However, research on avian mite GSTs as vaccine antigens is still lacking. Therefore, we aimed to evaluate the potential of avian mite GSTs for vaccine development. We identified GST genes from TFMs and NFMs. We prepared recombinant GST (rGST) from TFMs, NFMs, and PRMs, and assessed their protein functions. Moreover, we evaluated the cross-reactivity and acaricidal effect of immune plasma against each rGST on TFMs, NFMs, and PRMs. The deduced amino acid sequences of GSTs from TFMs and NFMs were 80% similar to those of the PRMs. The rGSTs exhibited catalytic activity in conjugating glutathione to the 1-chloro-2,4-dinitrobenzene substrate. Immune plasma against each rGST showed cross-reactivity with rGST from different mite species. Moreover, the survival rate of PRMs fed with immune plasma against the rGST of TFMs and NFMs was significantly lower than that of the control plasma. These results demonstrate the potential application of GST as an antigen for the development of a broad-spectrum vaccine against avian mites.
• Characterization of a Very Short Meq Protein Isoform in a Marek's Disease Virus Strain in Japan.
  Yoshinosuke Motai; Shiro Murata; Jumpei Sato; Akihito Nishi; Naoya Maekawa; Tomohiro Okagawa; Satoru Konnai; Kazuhiko Ohashi
  Veterinary sciences, 11, 1, 20 Jan. 2024, [Peer-reviewed], [Corresponding author], [International Magazine]
  English, Scientific journal, Marek's disease virus (MDV) causes malignant lymphoma (Marek's disease; MD) in chickens. The Meq protein is essential for tumorigenesis since it regulates the expression of host and viral genes. Previously, we reported that the deletion of the short isoform of Meq (S-Meq) decreases the pathogenicity of MDV. Recently, we identified a further short isoform of Meq (very short isoform of Meq, VS-Meq) in chickens with MD in Japan. A 64-amino-acid deletion was confirmed at the C-terminus of VS-Meq. We measured the transcriptional regulation by VS-Meq in three gene promoters to investigate the effect of VS-Meq on protein function. Wild-type VS-Meq decreased the transrepression of the pp38 promoter but did not alter the transactivation activity of the Meq and Bcl-2 promoters. The deletion in VS-Meq did not affect the activity of the pp38 promoter but enhanced the transactivation activities of the Meq and Bcl-2 promoters. Collectively, the deletion of VS-Meq potentially enhanced the activity of the Meq promoter, while other amino acid sequences in wild-type VS-Meq seemed to affect the weak transrepression of the pp38 promoter. Further investigation is required to clarify the effects of these changes on pathogenicity.
• Development of a high-affinity anti-bovine PD-1 rabbit-bovine chimeric antibody using an efficient selection and large production system.
  Tomohiro Okagawa; Satoru Konnai; Shinya Goto; Yamato Sajiki; Otgontuya Ganbaatar; Kei Watari; Hayato Nakamura; Cai-Xia Wang; Taro Tachibana; Yukinari Kato; Yayoi Kameda; Junko Kohara; Nobuhiro Terasaki; Manabu Kubota; Akira Takeda; Hirofumi Takahashi; Yasuhiko Suzuki; Naoya Maekawa; Shiro Murata; Kazuhiko Ohashi
  Veterinary research, 54, 1, 82, 82, 27 Sep. 2023, [Peer-reviewed], [International Magazine]
  English, Scientific journal, Immune checkpoint molecules PD-1/PD-L1 cause T-cell exhaustion and contribute to disease progression in chronic infections of cattle. We established monoclonal antibodies (mAbs) that specifically inhibit the binding of bovine PD-1/PD-L1; however, conventional anti-PD-1 mAbs are not suitable as therapeutic agents because of their low binding affinity to antigen. In addition, their sensitivity for the detection of bovine PD-1 is low and their use for immunostaining PD-1 is limited. To address these issues, we established two anti-bovine PD-1 rabbit mAbs (1F10F1 and 4F5F2) and its chimeric form using bovine IgG1 (Boch1D10F1), which exhibit high binding affinity. One of the rabbit mAb 1D10F1 binds more strongly to bovine PD-1 compared with a conventional anti-PD-1 mAb (5D2) and exhibits marked inhibitory activity on the PD-1/PD-L1 interaction. In addition, PD-1 expression in bovine T cells could be detected with higher sensitivity by flow cytometry using 1D10F1. Furthermore, we established higher-producing cells of Boch1D10F1 and succeeded in the mass production of Boch1D10F1. Boch1D10F1 exhibited a similar binding affinity to bovine PD-1 and the inhibitory activity on PD-1/PD-L1 binding compared with 1D10F1. The immune activation by Boch1D10F1 was also confirmed by the enhancement of IFN-γ production. Finally, Boch1D10F1 was administered to bovine leukemia virus-infected cows to determine its antiviral effect. In conclusion, the high-affinity anti-PD-1 antibody developed in this study represents a powerful tool for detecting and inhibiting bovine PD-1 and is a candidate for PD-1-targeted immunotherapy in cattle.
• Enhanced Systemic Antitumour Immunity by Hypofractionated Radiotherapy and Anti-PD-L1 Therapy in Dogs with Pulmonary Metastatic Oral Malignant Melanoma.
  Tatsuya Deguchi; Naoya Maekawa; Satoru Konnai; Ryo Owaki; Kenji Hosoya; Keitaro Morishita; Motoji Nakamura; Tomohiro Okagawa; Hiroto Takeuchi; Sangho Kim; Ryohei Kinoshita; Yurika Tachibana; Madoka Yokokawa; Satoshi Takagi; Yukinari Kato; Yasuhiko Suzuki; Shiro Murata; Kazuhiko Ohashi
  Cancers, 15, 11, 01 Jun. 2023, [Peer-reviewed], [International Magazine]
  English, Scientific journal, Although immune checkpoint inhibitors (ICIs), such as the anti-programmed death-ligand 1 (PD-L1) antibody, have been developed for the treatment of canine malignant melanoma, desirable clinical efficacies have not been achieved. Recent studies in humans have suggested that radiation therapy (RT) combined with ICIs induces robust systemic antitumour immunity in patients with cancer. This study retrospectively examined the therapeutic efficacy of combination therapy (hypofractionated RT and anti-PD-L1 antibody [c4G12]) in dogs with pulmonary metastatic oral malignant melanoma. The intrathoracic clinical benefit rate (CBR)/median overall survival (OS) in the no RT (n = 20, free from the effect of RT), previous RT (n = 9, received RT ≤8 weeks prior to the first c4G12 dose), and concurrent RT (n = 10, c4G12 therapy within ±1 week of the first RT fraction) groups were 10%/185 days, 55.6%/283.5 days (p < 0.05 vs. no RT group), and 20%/129 days (p > 0.05 vs. no RT group), respectively. The adverse events were considered to be tolerable in the combination therapy. Thus, hypofractionated RT before the initiation of c4G12 therapy can be an effective approach for enhancing the therapeutic efficacy of immunotherapy, with acceptable safety profiles. Further prospective clinical studies are required to confirm the findings of this study.
• Molecular characterization of immunoinhibitory factors PD-1/PD-L1 in sheep.
  Wisa Tiyamanee; Satoru Konnai; Tomohiro Okagawa; Yutaro Nojima; Otgontuya Ganbaatar; Naoya Maekawa; Rie Hasebe; Yumiko Kagawa; Yukinari Kato; Yasuhiko Suzuki; Shiro Murata; Kazuhiko Ohashi
  Veterinary immunology and immunopathology, 261, 110609, 110609, 11 May 2023, [Peer-reviewed], [International Magazine]
  English, Scientific journal, Sheep have been used as a large animal experimental model for studying infectious diseases. However, due to a lack of staining antibodies and reagents, immunological studies on sheep have not progressed. The immunoinhibitory receptor programmed death-1 (PD-1) is expressed on T lymphocytes. The interaction of PD-1 with its ligand PD-ligand 1 (PD-L1) delivers inhibitory signals and impairs proliferation, cytokine production, and cytotoxicity of T cells. We previously reported that the PD-1/PD-L1 pathway was closely associated with T-cell exhaustion and disease progression in bovine chronic infections using anti-bovine PD-L1 monoclonal antibodies (mAbs). Furthermore, we found that blocking antibodies against PD-1 and PD-L1 restore T-cell functions and could be used in immunotherapy of cattle. However, the immunological role of the PD-1/PD-L1 pathway in chronic diseases of sheep remains unknown. In this study, we identified cDNA sequences of ovine PD-1 and PD-L1 and examined the cross-activity of anti-bovine PD-L1 mAbs against ovine PD-L1 as well as the expression of PD-L1 in ovine listeriosis. The amino acid sequences of ovine PD-1 and PD-L1 share a high degree of identity and similarity with homologs from ruminants and other mammalian species. Anti-bovine PD-L1 mAb recognized ovine PD-L1 on lymphocytes in the flow cytometric assay. Furthermore, an immunohistochemical staining confirmed the PD-L1 expression on macrophages in the brain lesions of ovine listeriosis. These findings indicated that our anti-PD-L1 mAb would be useful for analyzing the ovine PD-1/PD-L1 pathway. Further research is needed to determine the immunological role of PD-1/PD-L1 in chronic diseases such as BLV infection through experimental infection of sheep.
• Investigation of peripheral blood responses in chickens infested with Dermanyssus gallinae.
  Sotaro Fujisawa; Shiro Murata; Takumi Sato; Eiji Oishi; Akira Taneno; Satoru Konnai; Kazuhiko Ohashi
  Parasitology international, 95, 102754, 102754, 22 Apr. 2023, [Peer-reviewed], [Corresponding author], [International Magazine]
  English, Scientific journal, Among haematophagous ectoparasites that infest chickens, poultry red mite (Dermanyssus gallinae, PRM) is one of the most serious threats to poultry farms. Mass PRM infestation causes various health problems in chickens, resulting in significant productivity reduction in the poultry industry. Despite the efficiency of acaricides for controlling PRMs, the emergence of acaricide-resistant PRMs represents a challenging setback. Infestation with haematophagous ectoparasites, such as PRMs, induces inflammatory and haemostatic reactions in the host. Therefore, we aimed to explore the gene expression in chicken peripheral blood cells to elucidate host responses against PRM infestation in detail. RNA sequencing of blood-fed PRMs was performed, and the levels of the chicken-derived transcripts obtained from the ingested blood cells were analysed. Genes encoding haemoglobin subunits were found to be significantly more expressed, suggesting that PRM infestation causes anaemia in chickens. Additionally, the mRNA and plasma concentrations of CC chemokine ligand 4 and β2 microglobulin among the immune-related molecules were found to be significantly higher in PRM-infested chickens compared with non-infested animals. These results suggest that PRM infestation induce inflammation in chicken. Further studies are warranted to better understand the influence of PRM infestation on the host physiological states, including immunity.
• Analysis of gene expression in poultry red mite, Dermanyssus gallinae, by RNAscope in situ hybridization.
  Hikari Seo; Shiro Murata; Osamu Ichii; Takashi Namba; Shwe Yee Win; Takumi Sato; Eiji Oishi; Akira Taneno; Naoya Maekawa; Tomohiro Okagawa; Satoru Konnai; Kazuhiko Ohashi
  The Journal of veterinary medical science, 85, 5, 532, 535, 14 Mar. 2023, [Peer-reviewed], [Corresponding author], [Domestic magazines]
  English, Scientific journal, The poultry red mite (PRM; Dermanyssus gallinae) is a hematophagous ectoparasite that mainly infests chickens, and its infestation causes significant economic losses to the poultry industry. In this study, we examined the use of RNAscope-based in situ hybridization (ISH) to characterize gene expression in PRM. We analyzed the mRNA expression of Dermanyssus gallinaecathepsin D-1 (Dg-CatD-1) and Dermanyssus gallinae cystatin (Dg-Cys). RNAscope ISH analysis revealed that mRNA expression of Dg-CatD-1 was observed in the digestive tract, and Dg-Cystatin mRNA was expressed in the ovaries in addition to the digestive tract. RNAscope ISH could be applicable for the analysis of gene expression in each tissue of PRM and is an effective method to investigate the characteristics of target genes.
• Enhancement of Vaccine-Induced T-Cell Responses by PD-L1 Blockade in Calves.
  Tomohiro Okagawa; Satoru Konnai; Hayato Nakamura; Otgontuya Ganbaatar; Yamato Sajiki; Kei Watari; Haruka Noda; Mitsuru Honma; Yukinari Kato; Yasuhiko Suzuki; Naoya Maekawa; Shiro Murata; Kazuhiko Ohashi
  Vaccines, 11, 3, 01 Mar. 2023, [Peer-reviewed], [International Magazine]
  English, Scientific journal, Interactions between programmed death 1 (PD-1) and PD-ligand 1 (PD-L1) cause functional exhaustion of T cells by inducing inhibitory signals, thereby attenuating effector functions of T cells. We have developed an anti-bovine PD-L1 blocking antibody (Ab) and have demonstrated that blockade of the interaction between PD-1 and PD-L1 reactivates T-cell responses in cattle. In the present study, we examined the potential utility of PD-1/PD-L1-targeted immunotherapy in enhancing T-cell responses to vaccination. Calves were inoculated with a hexavalent live-attenuated viral vaccine against bovine respiratory infections in combination with treatment with an anti-PD-L1 Ab. The expression kinetics of PD-1 in T cells and T-cell responses to viral antigens were measured before and after vaccination to evaluate the adjuvant effect of anti-PD-L1 Ab. PD-1 expression was upregulated in vaccinated calves after the administration of a booster vaccination. The activation status of CD4+, CD8+, and γδTCR+ T cells was enhanced by the combination of vaccination and PD-L1 blockade. In addition, IFN-γ responses to viral antigens were increased following combinatorial vaccination with PD-L1 blockade. In conclusion, the blockade of the PD-1/PD-L1 interaction enhances T-cell responses induced by vaccination in cattle, indicating the potential utility of anti-PD-L1 Ab in improving the efficacy of current vaccination programs.
• Suppressive modulation of host immune responses by Dermanyssus gallinae infestation.
  Sotaro Fujisawa; Shiro Murata; Masayoshi Isezaki; Shwe Yee Win; Takumi Sato; Eiji Oishi; Akira Taneno; Naoya Maekawa; Tomohiro Okagawa; Satoru Konnai; Kazuhiko Ohashi
  Poultry science, 102, 4, 102532, 102532, 26 Jan. 2023, [Peer-reviewed], [Corresponding author], [International Magazine]
  English, Scientific journal, The poultry red mite (Dermanyssus gallinae, PRM) is a blood-sucking ectoparasite in chickens and is one of the most serious threats to poultry farms. Mass infestation with PRMs causes various health problems in chickens, resulting in significant productivity reduction in the poultry industry. Infestation with hematophagous ectoparasites, such as ticks, induces host inflammatory and hemostatic reactions. On the other hand, several studies have reported that hematophagous ectoparasites secrete various immunosuppressants from their saliva to suppress host immune responses to maintain blood sucking. Here, we examined the expression of cytokines in peripheral blood cells to investigate whether PRM infestation affects immunological states in chickens. In PRM-infested chickens, anti-inflammatory cytokines, IL-10 and TGF-β1, and immune checkpoint molecules, CTLA-4 and PD-1, were highly expressed compared to noninfested chickens. PRM-derived soluble mite extracts (SME) upregulated the gene expression of IL-10 in peripheral blood cells and HD-11 chicken macrophages. In addition, SME suppressed the expression of interferons and inflammatory cytokines in HD-11 chicken macrophages. Moreover, SME induces the polarization of macrophages into anti-inflammatory phenotypes. Collectively, PRM infestation could affect host immune responses, especially suppress the inflammatory responses. Further studies are warranted to fully understand the influence of PRM infestation on host immunity.
• Combined Immune Checkpoint Blockade Enhances Antiviral Immunity against Bovine Leukemia Virus.
  Hayato Nakamura; Satoru Konnai; Tomohiro Okagawa; Naoya Maekawa; Yamato Sajiki; Kei Watari; Kana Kamitani; Maya Saito; Yukinari Kato; Yasuhiko Suzuki; Shiro Murata; Kazuhiko Ohashi
  Journal of virology, 97, 1, e0143022, 04 Jan. 2023, [Peer-reviewed], [International Magazine]
  English, Scientific journal, Bovine leukemia virus (BLV) is a retrovirus that causes enzootic bovine leukosis (EBL) in cattle and is widespread in many countries, including Japan. Recent studies have revealed that the expression of immunoinhibitory molecules, such as programmed death-1 (PD-1) and PD-ligand 1, plays a critical role in immunosuppression and disease progression during BLV infection. In addition, a preliminary study has suggested that another immunoinhibitory molecule, T-cell immunoglobulin domain and mucin domain-3 (TIM-3), is involved in immunosuppression during BLV infection. Therefore, this study was designed to further elucidate the immunoinhibitory role of immune checkpoint molecules in BLV infection. TIM-3 expression was upregulated on peripheral CD4+ and CD8+ T cells in BLV-infected cattle. Interestingly, in EBL cattle, CD4+ and CD8+ T cells infiltrating lymphomas expressed TIM-3. TIM-3 and PD-1 were upregulated and coexpressed in peripheral CD4+ and CD8+ T cells from BLV-infected cattle. Blockade by anti-bovine TIM-3 monoclonal antibody increased CD69 expression on T cells and gamma interferon (IFN-γ) production from peripheral blood mononuclear cells from BLV-infected cattle. A syncytium formation assay also demonstrated the antiviral effects of TIM-3 blockade against BLV infection. The combined inhibition of TIM-3 and PD-1 pathways significantly enhanced IFN-γ production and antiviral efficacy compared to inhibition alone. In conclusion, the combined blockade of TIM-3 and PD-1 pathways shows strong immune activation and antiviral effects and has potential as a novel therapeutic method for BLV infection. IMPORTANCE Enzootic bovine leukosis caused by bovine leukemia virus (BLV) is an important viral disease in cattle, causing severe economic losses to the cattle industry worldwide. The molecular mechanisms of BLV-host interactions are complex. Previously, it was found that immune checkpoint molecules, such as PD-1, suppress BLV-specific Th1 responses as the disease progresses. To date, most studies have focused only on how PD-1 facilitates escape from host immunity in BLV-infected cattle and the antiviral effects of the PD-1 blockade. In contrast, how T-cell immunoglobulin domain and mucin domain-3 (TIM-3), another immune checkpoint molecule, regulates anti-BLV immune responses is rarely reported. It is also unclear why PD-1 inhibition alone was insufficient to exert anti-BLV effects in previous clinical studies. In this study, the expression profile of TIM-3 in T cells derived from BLV-infected cattle suggested that TIM-3 upregulation is a cause of immunosuppression in infected cattle. Based on these results, anti-TIM-3 antibody was used to experimentally evaluate its function in influencing immunity against BLV. Results indicated that TIM-3 upregulation induced by BLV infection suppressed T-cell activation and antiviral cytokine production. Some T cells coexpressed PD-1 and TIM-3, indicating that simultaneous inhibition of PD-1 and TIM-3 with their respective antibodies synergistically restored antiviral immunity. This study could open new avenues for treating bovine chronic infections.
• Safety and clinical efficacy of an anti-PD-L1 antibody (c4G12) in dogs with advanced malignant tumours.
  Naoya Maekawa; Satoru Konnai; Kenji Hosoya; Sangho Kim; Ryohei Kinoshita; Tatsuya Deguchi; Ryo Owaki; Yurika Tachibana; Madoka Yokokawa; Hiroto Takeuchi; Yumiko Kagawa; Satoshi Takagi; Hiroshi Ohta; Yukinari Kato; Satoshi Yamamoto; Keiichi Yamamoto; Yasuhiko Suzuki; Tomohiro Okagawa; Shiro Murata; Kazuhiko Ohashi
  PloS one, 18, 10, e0291727, 2023, [Peer-reviewed], [International Magazine]
  English, Scientific journal, Immune checkpoint inhibitors (ICIs) have been developed for canine tumour treatment, and pilot clinical studies have demonstrated their antitumour efficacy in dogs with oral malignant melanoma (OMM). Although ICIs have been approved for various human malignancies, their clinical benefits in other tumour types remain to be elucidated in dogs. Here, we conducted a clinical study of c4G12, a canine chimeric anti-PD-L1 antibody, to assess its safety and efficacy in dogs with various advanced malignant tumours (n = 12) at the Veterinary Teaching Hospital of Hokkaido University from 2018 to 2023. Dogs with digit or foot pad malignant melanoma (n = 4), osteosarcoma (n = 2), hemangiosarcoma (n = 1), transitional cell carcinoma (n = 1), nasal adenocarcinoma (n = 1), B-cell lymphoma (n = 1), or undifferentiated sarcoma (n = 2) were treated with 2 or 5 mg/kg c4G12 every 2 weeks. Treatment-related adverse events of any grade were observed in eight dogs (66.7%), including elevated aspartate aminotransferase (grade 3) in one dog (8.3%) and thrombocytopenia (grade 4) in another dog (8.3%). Among dogs with target disease at baseline (n = 8), as defined by the response evaluation criteria for solid tumours in dogs (cRECIST), one dog with nasal adenocarcinoma and another with osteosarcoma experienced a partial response (PR), with an objective response rate of 25.0% (2 PR out of 8 dogs; 95% confidence interval: 3.2-65.1%). These results suggest that c4G12 is safe and tolerable and shows antitumor effects in dogs with malignant tumours other than OMM. Further clinical studies are warranted to identify the tumour types that are most likely to benefit from c4G12 treatment.
• Characterization of cysteine proteases from poultry red mite, tropical fowl mite, and northern fowl mite to assess the feasibility of developing a broadly efficacious vaccine against multiple mite species.
  Shwe Yee Win; Shiro Murata; Sotaro Fujisawa; Hikari Seo; Jumpei Sato; Yoshinosuke Motai; Takumi Sato; Eiji Oishi; Akira Taneno; Lat Lat Htun; Saw Bawm; Tomohiro Okagawa; Naoya Maekawa; Satoru Konnai; Kazuhiko Ohashi
  PloS one, 18, 7, e0288565, 2023, [Peer-reviewed], [Corresponding author], [International Magazine]
  English, Scientific journal, Infestation with poultry red mites (PRM, Dermanyssus gallinae) causes anemia, reduced egg production, and death in serious cases, resulting in significant economic losses to the poultry industry. As a novel strategy for controlling PRMs, vaccine approaches have been focused upon and several candidate vaccine antigens against PRMs have been reported. Tropical (TFM, Ornithonyssus bursa) and northern (NFM, Ornithonyssus sylviarum) fowl mites are also hematophagous and cause poultry industry problems similar to those caused by PRM. Therefore, ideal antigens for anti-PRM vaccines are molecules that cross-react with TFMs and NFMs, producing pesticidal effects similar to those against PRMs. In this study, to investigate the potential feasibility of developing vaccines with broad efficacy across mite species, we identified and characterized cysteine proteases (CPs) of TFMs and NFMs, which were previously reported to be effective vaccine antigens of PRMs. The open reading frames of CPs from TFMs and NFMs had the same sequences, which was 73.0% similar to that of PRMs. Phylogenetic analysis revealed that the CPs of TFMs and NFMs clustered in the same clade as CPs of PRMs. To assess protein functionality, we generated recombinant peptidase domains of CPs (rCP-PDs), revealing all rCP-PDs showed CP-like activities. Importantly, the plasma obtained from chickens immunized with each rCP-PD cross-reacted with rCP-PDs of different mites. Finally, all immune plasma of rCP-PDs reduced the survival rate of PRMs, even when the plasma was collected from chickens immunized with rCP-PDs derived from TFM and NFM. Therefore, CP antigen is a promising, broadly efficacious vaccine candidate against different avian mites.
• Potential of ferritin 2 as an antigen for the development of a universal vaccine for avian mites, poultry red mites, tropical fowl mites, and northern fowl mites.
  Shwe Yee Win; Shiro Murata; Sotaro Fujisawa; Hikari Seo; Jumpei Sato; Yoshinosuke Motai; Takumi Sato; Eiji Oishi; Akira Taneno; Lat Lat Htun; Saw Bawm; Tomohiro Okagawa; Naoya Maekawa; Satoru Konnai; Kazuhiko Ohashi
  Frontiers in veterinary science, 10, 1182930, 1182930, 2023, [Peer-reviewed], [Corresponding author], [International Magazine]
  English, Scientific journal, INTRODUCTION: Poultry red mites (PRMs, Dermanyssus gallinae), blood-sucking ectoparasites, are a threat to the poultry industry because of reduced production caused by infestation. In addition, tropical fowl mites (TFMs, Ornithonyssus bursa) and northern fowl mites (NFMs, Ornithonyssus sylviarum) are hematophagous, distributed in various regions, genetically and morphologically close to PRMs, and cause similar problems to the poultry industry. Vaccine approaches have been studied for PRM control, and several molecules have been identified in PRMs as candidates for effective vaccine antigens. The development of an anti-PRM vaccine as a universal vaccine with broad efficacy against avian mites could improve the productivity of poultry farms worldwide. Molecules that are highly conserved among avian mites and have critical functions in the physiology and growth of mites could be ideal antigen candidates for the development of universal vaccines. Ferritin 2 (FER2), an iron-binding protein, is critical for the reproduction and survival of PRMs and has been reported as a useful vaccine antigen for the control of PRMs and a candidate for the universal vaccine antigen in some tick species. METHOD AND RESULTS: Herein, we identified and characterized FER2 in TFMs and NFM. Compared with the sequence of PRM, the ferroxidase centers of the heavy chain subunits were conserved in FER2 of TFMs and NFMs. Phylogenetic analysis revealed that FER2 belongs to clusters of secretory ferritins of mites and other arthropods. Recombinant FER2 (rFER2) proteins from PRMs, TFMs, and NFMs exhibited iron-binding abilities. Immunization with each rFER2 induced strong antibody responses in chickens, and each immune plasma cross-reacted with rFER2 from different mites. Moreover, mortality rates of PRMs fed with immune plasma against rFER2 from TFMs or NFMs, in addition to PRMs, were higher than those of control plasma. DISCUSSION: rFER2 from each avian mite exhibited anti-PRM effects. This data suggests that it has the potential to be used as an antigen candidate for a universal vaccine against avian mites. Further studies are needed to access the usefulness of FER2 as a universal vaccine for the control of avian mites.
• In vitro evaluation of Lactiplantibacillus plantarum HOKKAIDO strain, effective lactic acid bacteria for calf diarrhea.
  Mari Ikehata; Satoru Konnai; Tomohiro Okagawa; Kentaro Abe; Mitsuru Honma; Toru Kitamura; Naoya Maekawa; Yasuhiko Suzuki; Shiro Murata; Kazuhiko Ohashi
  Frontiers in veterinary science, 10, 1145445, 1145445, 2023, [Peer-reviewed], [International Magazine]
  English, Scientific journal, Calf diarrhea adversely affects growth and sometimes results in mortality, leading to severe economic losses to the cattle industry. Antibiotics are useful in the treatment against bacterial diarrhea, but not against viral, protozoan, and antibiotic-resistant bacterial diarrhea. Therefore, there are growing requirements for a novel control method for calf diarrhea. Probiotics have been considered promising candidates for preventive and supportive therapy for calf diarrhea for many years. A recent study has revealed that Lactiplantibacillus plantarum HOKKAIDO strain (Lp-HKD) reduces intestinal pathology and the severity of diarrhea in bovine rotavirus (BRV)-infected calves. Lp-HKD is known to enhance the function of human immune cells and expected to be used as probiotics for humans. Therefore, it is hypothesized that Lp-HKD modulates antiviral immune response in cattle and provide the clinical benefits in BRV-infected calves. However, the detailed mechanism of Lp-HKD-induced immunomodulation remains unknown. Thus, this study aimed to elucidate the immunomodulatory and antiviral effects of Lp-HKD in cattle. Cultivation assay of bovine peripheral blood mononuclear cells (PBMCs) showed that live and heat-killed Lp-HKD stimulates the production of interleukin-1β (IL-1β), IL-6, IL-10, and interferon-γ (IFN-γ) from PBMCs. Stimulation by heat-killed Lp-HKD yielded stronger cytokine production than stimulation by the live Lp-HKD. Additionally, CD14+ monocytes were identified as major producers of IL-1β, IL-6, and IL-10 under Lp-HKD stimulation; however, IFN-γ was mainly produced from immune cells other than CD14+ monocytes. Depletion of CD14+ monocytes from the PBMCs cultivation strongly decreased cytokine production induced by heat-killed Lp-HKD. The inhibition of toll-like receptor (TLR) 2/4 signaling decreased IL-1β and IL-6 production induced by live Lp-HKD and IL-1β, IL-6, and IFN-γ production induced by heat-killed Lp-HKD. Furthermore, live or heat-killed Lp-HKD also activated T cells and their production of IFN-γ and tumor necrosis factor-α. Then, culture supernatants of bovine PBMCs treated with heat-killed Lp-HKD demonstrated antiviral effects against BRV in vitro. In conclusion, this study demonstrated that Lp-HKD activates the functions of bovine immune cells via TLR2/4 signaling and exerts an antiviral effect against BRV through the induction of antiviral cytokines. Lp-HKD could be useful for the prevention and treatment of calf diarrhea through its immune activating effect.
• Molecular characterization of feline immune checkpoint molecules and establishment of PD-L1 immunohistochemistry for feline tumors.
  Naoya Maekawa; Satoru Konnai; Yumie Asano; Takumi Otsuka; Eri Aoki; Hiroto Takeuchi; Yukinari Kato; Mika K Kaneko; Shinji Yamada; Yumiko Kagawa; Maki Nishimura; Satoshi Takagi; Tatsuya Deguchi; Hiroshi Ohta; Takayuki Nakagawa; Yasuhiko Suzuki; Tomohiro Okagawa; Shiro Murata; Kazuhiko Ohashi
  PloS one, 18, 1, e0281143, 2023, [Peer-reviewed], [International Magazine]
  English, Scientific journal, Spontaneous tumors are a major cause of death in cats. Treatment of human tumors has progressed dramatically in the past decade, partly due to the success of immunotherapies using immune checkpoint inhibitors, such as anti-programmed death 1 (PD-1) and anti-PD-ligand 1 (PD-L1) antibodies. However, little is known about the PD-1 pathway and its association with tumor disease in cats. This study investigated the applicability of anti-PD-1/PD-L1 therapy in feline tumors. We first determined the complete coding sequence of feline PD-L1 and PD-L2, and found that the deduced amino acid sequences of feline PD-L1/PD-L2 share high sequence identities (66-83%) with orthologs in other mammalian species. We prepared recombinant feline PD-1, PD-L1, and PD-L2 proteins and confirmed receptor-ligand binding between PD-1 and PD-L1/PD-L2 using flow cytometry. Next, we established an anti-feline PD-L1 monoclonal antibody (clone CL1Mab-7) to analyze the expression of PD-L1. Flow cytometry using CL1Mab-7 revealed the cell surface expression of PD-L1 in a feline macrophage (Fcwf-4) and five mammary adenocarcinoma cell lines (FKNp, FMCm, FYMp, FONp, and FONm), and showed that PD-L1 expression was upregulated by interferon-γ stimulation. Finally, immunohistochemistry using CL1Mab-7 also showed PD-L1 expression in feline squamous cell carcinoma (5/5, 100%), mammary adenocarcinoma (4/5, 80%), fibrosarcoma (5/5, 100%), and renal cell carcinoma (2/2, 100%) tissues. Our results strongly encourage further investigations of the PD-1/PD-L1 pathway as a potential therapeutic target for feline tumors.
• Characterization of SpsQ from Staphylococcus pseudintermedius as an affinity chromatography ligand for canine therapeutic antibodies.
  Hiroto Takeuchi; Chie Nakajima; Satoru Konnai; Naoya Maekawa; Tomohiro Okagawa; Masaru Usui; Yutaka Tamura; Yasuhiko Suzuki; Shiro Murata; Kazuhiko Ohashi
  PloS one, 18, 1, e0281171, 2023, [Peer-reviewed], [International Magazine]
  English, Scientific journal, Coagulase-positive Staphylococci express protein A, which binds to host antibodies, to evade the immune system. Taking advantage of its specific binding to antibodies, protein A from Staphylococcus aureus, which is called SpA, is commonly used as an affinity chromatography ligand for human therapeutic antibodies. However, among four canine IgG subclasses (A, B, C, and D), only IgG-B binds to SpA strongly and establishing an efficient and robust purification scheme for canine therapeutic antibodies whose IgG subclass is A, C, or D remains difficult and depends on finding a suitable substitute to SpA. S. pseudintermedius, a major coagulase-positive Staphylococci found in dogs, expresses spsQ gene which is orthologous to S. aureus spa. We hypothesized that to serve S. pseudintermedius to better adapt to the dog immune system, SpsQ would bind to canine IgGs stronger than SpA, making it a better affinity chromatography ligand for canine therapeutic antibodies. To characterize SpsQ, we first determined the spsQ nucleotide sequence from S. pseudintermedius isolates. Based on the identified sequence, we prepared recombinant proteins containing the immunoglobulin-binding domains of SpA (r-SpA) and SpsQ (r-SpsQ) and determined their binding capacity for each canine IgG subclass. The binding capacity of r-SpsQ for IgG-B was almost as high as that of r-SpA. Interestingly, while both r-SpsQ and r-SpA showed no binding to IgG-C, the binding capacity of r-SpsQ for IgG-A and IgG-D was significantly higher than that of r-SpA. Finally, we performed affinity chromatography using r-SpsQ- or r-SpA-immobilized resin and revealed that the recovery rates of IgG-A and IgG-D using r-SpsQ were significantly higher than those using r-SpA. Our findings indicate that SpsQ has a strong potential to be used as an affinity chromatography ligand for canine therapeutic antibodies of subclass A, B, and D.
• Diagnosis and Early Prediction of Lymphoma Using High-Throughput Clonality Analysis of Bovine Leukemia Virus-Infected Cells.
  Tomohiro Okagawa; Honami Shimakura; Satoru Konnai; Masumichi Saito; Takahiro Matsudaira; Naganori Nao; Shinji Yamada; Kenji Murakami; Naoya Maekawa; Shiro Murata; Kazuhiko Ohashi
  Microbiology spectrum, 10, 6, e0259522, 13 Oct. 2022, [Peer-reviewed], [International Magazine]
  English, Scientific journal, Bovine leukemia virus (BLV), a retrovirus, infects B cells of ruminants and is integrated into the host genome as a provirus for lifelong infection. After a long latent period, 1% to 5% of BLV-infected cattle develop aggressive lymphoma, enzootic bovine leukosis (EBL). Since the clonal expansion of BLV-infected cells is essential for the development of EBL, the clonality of proviral integration sites could be a molecular marker for diagnosis and early prediction of EBL. Recently, we developed Rapid Amplification of the Integration Site without Interference by Genomic DNA Contamination (RAISING) and an analysis software of clonality value (CLOVA) to analyze the clonality of transgene-integrated cells. RAISING-CLOVA is capable of assessing the risk of adult T-cell leukemia/lymphoma development in human T-cell leukemia virus-I-infected individuals through the clonality analysis of proviral integration sites. Thus, we herein examined the performance of RAISING-CLOVA for the clonality analysis of BLV-infected cells and conducted a comprehensive clonality analysis by RAISING-CLOVA in EBL and non-EBL cattle. RAISING-CLOVA targeting BLV was a highly accurate and reproducible method for measuring the clonality value. The comprehensive clonality analysis successfully distinguished EBL from non-EBL specimens with high sensitivity and specificity. A longitudinal clonality analysis in BLV-infected sheep, an experimental model of lymphoma, also confirmed the effectiveness of RAISING-CLOVA for early detection of EBL development. Therefore, our study emphasizes the usefulness of RAISING-CLOVA as a routine clinical test for monitoring virus-related cancers. IMPORTANCE Bovine leukemia virus (BLV) infection causes aggressive B-cell lymphoma in cattle and sheep. The virus has spread to farms around the world, causing significant economic damage to the livestock industry. Thus, the identification of high-risk asymptomatic cattle before they develop lymphoma can be effective in reducing the economic damage. Clonal expansion of BLV-infected cells is a promising marker for the development of lymphoma. Recently, we have developed a high-throughput method to amplify random integration sites of transgenes in host genomes and analyze their clonality, named as RAISING-CLOVA. As a new application of our technology, in this study, we demonstrate the value of the RAISING-CLOVA method for the diagnosis and early prediction of lymphoma development by BLV infection in cattle. RAISING-CLOVA is a reliable technology for monitoring the clonality of BLV-infected cells and would contribute to reduce the economic losses by EBL development.
• Prostaglandin E-2-Induced Immune Suppression via Cytotoxic T-Lymphocyte Antigen 4 in Paratuberculosis
  Yamato Sajiki; Satoru Konnai; Kei Watari; Tomohiro Okagawa; Akina Tanaka; Satoko Kawaji; Reiko Nagata; Naoya Maekawa; Yasuhiko Suzuki; Yukinari Kato; Shiro Murata; Yasuyuki Mori; Kazuhiko Ohashi
  INFECTION AND IMMUNITY, 90, 10, e0021022, Sep. 2022, [Peer-reviewed], [International Magazine]
  English, Scientific journal
• Suppressive effects of Ixodes persulcatus sialostatin L2 against Borrelia miyamotoi-stimulated immunity
  Yamato Sajiki; Satoru Konnai; Tomohiro Okagawa; Naoya Maekawa; Masayoshi Isezaki; Shinji Yamada; Takuya Ito; Kozue Sato; Hiroki Kawabata; Carlos Logullo; Itabajara da Silva Vaz Jr; Shiro Murata; Kazuhiko Ohashi
  TICKS AND TICK-BORNE DISEASES, 13, 4, 101963, 101963, Jul. 2022, [Peer-reviewed], [International Magazine]
  English, Scientific journal
• Exploration of serum biomarkers in dogs with malignant melanoma receiving anti-PD-L1 therapy and potential of COX-2 inhibition for combination therapy.
  Naoya Maekawa; Satoru Konnai; Yumie Asano; Yamato Sajiki; Tatsuya Deguchi; Tomohiro Okagawa; Kei Watari; Hiroto Takeuchi; Satoshi Takagi; Kenji Hosoya; Sangho Kim; Hiroshi Ohta; Yukinari Kato; Yasuhiko Suzuki; Shiro Murata; Kazuhiko Ohashi
  Scientific reports, 12, 1, 9265, 9265, 03 Jun. 2022, [Peer-reviewed], [International Magazine]
  English, Scientific journal, Immune checkpoint inhibitors (ICIs) such as anti-PD-L1 antibodies are widely used to treat human cancers, and growing evidence suggests that ICIs are promising treatments for canine malignancies. However, only some canine oral malignant melanoma (OMM) cases respond to ICIs. To explore biomarkers predictive of survival in dogs with pulmonary metastatic OMM receiving the anti-PD-L1 antibody c4G12 (n = 27), serum concentrations of prostaglandin E2 (PGE2), cytokines, chemokines, and growth factors were measured prior to treatment initiation. Among 12 factors tested, PGE2, interleukin (IL)-12p40, IL-8, monocyte chemotactic protein-1 (MCP-1), and stem cell factor (SCF) were higher in OMM dogs compared to healthy dogs (n = 8). Further, lower baseline serum PGE2, MCP-1, and vascular endothelial growth factor (VEGF)-A concentrations as well as higher IL-2, IL-12, and SCF concentrations predicted prolonged overall survival. These observations suggest that PGE2 confers resistance against anti-PD-L1 therapy through immunosuppression and thus is a candidate target for combination therapy. Indeed, PGE2 suppressed IL-2 and interferon (IFN)-γ production by stimulated canine peripheral blood mononuclear cells (PBMCs), while inhibition of PGE2 biosynthesis using the COX-2 inhibitor meloxicam in combination with c4G12 enhanced Th1 cytokine production by PBMCs. Thus, serum PGE2 may be predictive of c4G12 treatment response, and concomitant use of COX-2 inhibitors may enhance ICI antitumor efficacy.
• Estradiol-induced immune suppression via prostaglandin E-2 during parturition in bovine leukemia virus-infected cattle
  Yamato Sajiki; Satoru Konnai; Tomohiro Okagawa; Naoya Maekawa; Shinya Goto; Junko Kohara; Atsushi Nitanai; Hirofumi Takahashi; Kentaro Kubota; Hiroshi Takeda; Shiro Murata; Kazuhiko Ohashi
  PLOS ONE, 17, 3, e0263660, e0263660, 09 Mar. 2022, [Peer-reviewed], [International Magazine]
  English, Scientific journal
• In vitro evaluation of a cysteine protease from poultry red mites, Demanyssus gallinae, as a vaccine antigen for chickens.
  Takuma Ariizumi; Shiro Murata; Sotaro Fujisawa; Masayoshi Isezaki; Takumi Sato; Eiji Oishi; Akira Taneno; Osamu Ichii; Naoya Maekawa; Tomohiro Okagawa; Satoru Konnai; Kazuhiko Ohashi
  Poultry science, 101, 3, 101638, 101638, Elsevier BV, Mar. 2022, [Peer-reviewed], [Corresponding author], [International Magazine]
  English, Scientific journal, Poultry red mites (PRMs, Dermanyssus gallinae) are hematophagous ectoparasites that negatively affect egg production, which causes serious economic losses to the poultry industry worldwide. Currently, the emergence of acaricide-resistant PRMs has impeded PRM control in poultry farms. Several alternatives for acaricide use have been described for managing PRM-caused problems. Vaccination is among the methods for controlling PRMs in poultry houses. Currently, several candidates for vaccine antigens have been identified. This study identified a cysteine protease, Deg-CPR-2, which differs from 2 other previously reported cysteine proteases in PRMs, from previously obtained data from RNA-sequencing (RNA-seq) analysis. We investigated the characteristics of Deg-CPR-2 and assessed its efficacy as a vaccine antigen in vitro. Phylogenetic analysis revealed that Deg-CPR-2 belonged to a different cluster from those of other cysteine proteases in PRMs. This cluster also included cathepsin L-like proteases, enzymes thought to be involved in hemoglobin digestion in ticks. Expression analysis revealed Deg-CPR-2 expression in midguts and all the life-stages; however, there were differences in the expression levels across the life-stages. The enzyme activity of recombinant Deg-CPR-2 was inhibited in the presence of a cysteine protease inhibitor, which suggests that Deg-CPR-2 functions as a cysteine protease in PRMs. Finally, there was an in vitro increase in the mortality of PRMs, mainly protonymphs that were artificially fed with plasma from chickens immunized with Deg-CPR-2. These findings suggest that Deg-CPR-2 may contribute to protein digestion in the midgut of PRMs and is crucially involved in physiological processes in PRMs. Additionally, immunization with Deg-CPR-2 may reduce the number of protonymphs, and Deg-CPR-2 should be considered as a candidate antigen for anti-PRM vaccine development.
• Effect of Insertion and Deletion in the Meq Protein Encoded by Highly Oncogenic Marek's Disease Virus on Transactivation Activity and Virulence
  Jumpei Sato; Shiro Murata; Zhiyuan Yang; Benedikt B. Kaufer; Sotaro Fujisawa; Hikari Seo; Naoya Maekawa; Tomohiro Okagawa; Satoru Konnai; Nikolaus Osterrieder; Mark S. Parcells; Kazuhiko Ohashi
  VIRUSES-BASEL, 14, 2, 382, 382, 14 Feb. 2022, [Peer-reviewed], [Corresponding author], [International Magazine]
  English, Scientific journal
• Enhancement of interleukin-2 production by bovine peripheral blood mononuclear cells treated with the combination of anti- programmed death-ligand 1 and cytotoxic T lymphocyte antigen 4 chimeric monoclonal antibodies
  Kei Watari; Satoru Konnai; Tomohiro Okagawa; Naoya Maekawa; Yamato Sajiki; Yukinari Kato; Yasuhiko Suzuki; Shiro Murata; Kazuhiko Ohashi
  JOURNAL OF VETERINARY MEDICAL SCIENCE, 84, 1, 6, 15, 07 Jan. 2022, [Peer-reviewed], [Domestic magazines]
  English, Scientific journal
• Characterization of a copper transporter 1 from Dermanyssus gallinae as a vaccine antigen.
  Sotaro Fujisawa; Shiro Murata; Masayoshi Isezaki; Takuma Ariizumi; Takumi Sato; Eiji Oishi; Akira Taneno; Naoya Maekawa; Tomohiro Okagawa; Osamu Ichii; Satoru Konnai; Kazuhiko Ohashi
  Parasitology, 149, 1, 105, 115, Jan. 2022, [Peer-reviewed], [Corresponding author], [International Magazine]
  English, Scientific journal
• Characterization of a Novel Cysteine Protease Inhibitor from Poultry Red Mites: Potential Vaccine for Chickens.
  Sotaro Fujisawa; Shiro Murata; Masayoshi Isezaki; Takuma Ariizumi; Takumi Sato; Eiji Oishi; Akira Taneno; Naoya Maekawa; Tomohiro Okagawa; Osamu Ichii; Satoru Konnai; Kazuhiko Ohashi
  Vaccines, 9, 12, 1472, 1472, MDPI AG, 13 Dec. 2021, [Peer-reviewed], [Corresponding author], [International Magazine]
  English, Scientific journal, Poultry red mite (PRM; Dermanyssus gallinae) is a hazardous, blood-sucking ectoparasite of birds that constitutes a threat to poultry farming worldwide. Acaricides, commonly used in poultry farms to prevent PRMs, are not effective because of the rapid emergence of acaricide-resistant PRMs. However, vaccination may be a promising strategy to control PRM. We identified a novel cystatin-like molecule in PRMs: Dg-Cys. Dg-Cys mRNA expression was detected in the midgut and ovaries, in all stages of life. The PRM nymphs that were artificially fed with the plasma from chickens that were immunized with Dg-Cys in vitro had a significantly reduced reproductive capacity and survival rate. Moreover, combination of Dg-Cys with other antigen candidates, like copper transporter 1 or adipocyte plasma membrane-associated protein, enhanced vaccine efficacies. vaccination and its application as an antigen for cocktail vaccines could be an effective strategy to reduce the damage caused by PRMs in poultry farming.
• Programmed death-ligand 1 expression in swine chronic infections and enhancement of interleukin-2 production via programmed death-1/programmed death-ligand 1 blockade.
  Otgontuya Ganbaatar; Satoru Konnai; Tomohiro Okagawa; Yutaro Nojima; Naoya Maekawa; Yoshiki Ichikawa; Atsushi Kobayashi; Tomoyuki Shibahara; Yojiro Yanagawa; Hidetoshi Higuchi; Yukinari Kato; Yasuhiko Suzuki; Shiro Murata; Kazuhiko Ohashi
  Immunity, inflammation and disease, 9, 4, 1573, 1583, Dec. 2021, [Peer-reviewed], [International Magazine]
  English, Scientific journal, INTRODUCTION: Chronic infections lead to the functional exhaustion of T cells. Exhausted T cells are phenotypically differentiated by the surface expression of the immunoinhibitory receptor, such as programmed death-1 (PD-1). The inhibitory signal is produced by the interaction between PD-1 and its PD-ligand 1 (PD-L1) and impairs the effector functions of T cells. However, the expression dynamics of PD-L1 and the immunological functions of the PD-1/PD-L1 pathway in chronic diseases of pigs are still poorly understood. In this study, we first analyzed the expression of PD-L1 in various chronic infections in pigs, and then evaluated the immune activation by the blocking assay targeting the swine PD-1/PD-L1 pathway. METHODS: In the initial experiments, anti-bovine PD-L1 monoclonal antibodies (mAbs) were tested for cross-reactivity with swine PD-L1. Subsequently, immunohistochemical analysis was conducted using the anti-PD-L1 mAb. Finally, we assessed the immune activation of swine peripheral blood mononuclear cells (PBMCs) by the blockade with anti-PD-L1 mAb. RESULTS: Several anti-PD-L1 mAbs tested recognized swine PD-L1-expressing cells. The binding of swine PD-L1 protein to swine PD-1 was inhibited by some of these cross-reactive mAbs. In addition, immunohistochemical analysis revealed that PD-L1 was expressed at the site of infection in chronic infections of pigs. The PD-L1 blockade increased the production of interleukin-2 from swine PBMCs. CONCLUSIONS: These findings suggest that the PD-1/PD-L1 pathway could be also involved in immunosuppression in chronic infections in pigs. This study provides a new perspective on therapeutic strategies for chronic diseases in pigs by targeting immunosuppressive pathways.
• Research Note: Characterization of S-Meq containing the deletion in Meq protein's transactivation domain in a Marek's disease virus strain in Japan.
  Shiro Murata; Eiji Yamamoto; Natsumi Sakashita; Naoya Maekawa; Tomohiro Okagawa; Satoru Konnai; Kazuhiko Ohashi
  Poultry science, 100, 11, 101461, 101461, Nov. 2021, [Peer-reviewed], [Lead author, Corresponding author], [International Magazine]
  English, Scientific journal, Marek's disease virus (MDV) causes malignant lymphomas in chickens (Marek's disease; MD). Although MD has caused significant economic losses to the poultry industry, currently, its occurrence in the field is effectively controlled by vaccination. However, the genetic characteristics of MDV strains have changed, and the poultry industry has experienced MD outbreaks in vaccinated chickens because of enhanced virulence. Meq, an oncoprotein of MDV, is a key transcription factor correlated with the tumorigenesis in MD. Animal experiments using recombinant MDV revealed that distinct polymorphisms in Meq affect the virulence of MDV strains. Meq containing an insertion or deletion is present in some MDV strains. In the 2010s, field strains that encode Meq containing the deletion (S-Meq) were reported. In this study, we characterized the genetic features of S-Meq detected in a Japanese MDV strain and analyzed its transactivation activity to investigate S-Meq's protein function. S-Meq lacked 41 amino acids, and the deletion was at the same position as those observed in other countries. In addition, S-Meq exhibited higher transactivation activity than Meq from other MDV strains circulating in Japan. These results suggest that the deletion in the transactivation domain may enhance the Meq protein's function. Further investigation is needed to clarify whether the deletion in the transactivation domain of Meq affects MDV's virulence.
• In vitro characterization of adipocyte plasma membrane-associated protein from poultry red mites, Dermanyssus gallinae, as a vaccine antigen for chickens.
  Sotaro Fujisawa; Shiro Murata; Masaki Takehara; Julia Aoyama; Ayu Morita; Masayoshi Isezaki; Shwe Yee Win; Takuma Ariizumi; Takumi Sato; Eiji Oishi; Akira Taneno; Naoya Maekawa; Tomohiro Okagawa; Osamu Ichii; Satoru Konnai; Kazuhiko Ohashi
  Vaccine, 39, 41, 6057, 6066, 01 Oct. 2021, [Peer-reviewed], [Corresponding author], [International Magazine]
  English, Scientific journal, The poultry red mite (Dermanyssus gallinae; PRM) is a blood-sucking ectoparasite of chickens that is a threat to poultry farming worldwide and significantly reduces productivity in the egg-laying industry. Chemical acaricides that are widely used in poultry farms for the prevention of PRMs are frequently ineffective due to the emergence of acaricide-resistant PRMs. Therefore, alternative control methods are needed, and vaccination is a promising strategy for controlling PRMs. A novel adipocyte-plasma membrane-associated protein-like molecule (Dg-APMAP) is highly expressed in blood-fed PRMs according to a previous RNA sequencing analysis. Here, we attempted to identify the full sequence of Dg-APMAP, study its expression in different life stages of PRMs, and evaluate its potential as a vaccine antigen. Dg-APMAP mRNA was expressed in the midgut and ovaries, and in all life stages regardless of feeding states. Importantly, in vitro feeding of PRMs with plasma derived from chickens immunized with the recombinant protein of the extracellular region of Dg-APMAP significantly reduced their survival rate in nymphs and adults, which require blood meals. Our data suggest that the host immune responses induced by vaccination with Dg-APMAP could be an effective strategy to reduce the suffering caused by PRMs in the poultry industry.
• Prostaglandin-related immune suppression in cattle
  Y. Sajiki; S. Konnai; Y. Ikenaka; T. Okagawa; N. Maekawa; C. Logullo; I. da Silva Vaz; S. Murata; K. Ohashi
  Veterinary Immunology and Immunopathology, 236, 110238, 110238, Elsevier BV, Jun. 2021, [Peer-reviewed]
  Scientific journal
• Selection of reference genes for quantitative PCR analysis in poultry red mite (Dermanyssus gallinae).
  Takuma Ariizumi; Shiro Murata; Sotaro Fujisawa; Masayoshi Isezaki; Naoya Maekawa; Tomohiro Okagawa; Takumi Sato; Eiji Oishi; Akira Taneno; Satoru Konnai; Kazuhiko Ohashi
  The Journal of veterinary medical science, 83, 4, 558, 565, 09 Apr. 2021, [Peer-reviewed], [Corresponding author], [Domestic magazines]
  English, Scientific journal, Poultry red mites (PRMs, Dermanyssus gallinae) are harmful ectoparasites that affect farmed chickens and cause serious economic losses in the poultry industry worldwide. Acaricides are used for PRM control; however, some PRMs have developed acaricide-resistant properties, which have indicated the need for different approaches for PRM control. Therefore, it is necessary to elucidate the biological status of PRMs to develop alternative PRM control strategies. Quantitative polymerase chain reaction (qPCR) allows analysis of the biological status at the transcript level. However, reference genes are preferable for accurate comparison of expression level changes given the large variation in the quality of the PRM samples collected in each farm. This study aimed to identify candidate reference genes with stable expression levels in the different blood feeding states and life stages of PRMs. First, we selected candidates based on the following criteria: sufficient expression intensity and no significant expression difference between fed and starved states. We selected and characterized seven candidate reference genes. Among them, we evaluated the gene expression stability between the starved and fed states using RefFinder; moreover, we compared their expression levels in each life-stage and identified two reference genes, Elongation factor 1-alpha (ELF1A)-like and apolipophorins-like. Finally, we evaluated the utility of the candidates as reference genes, and the use of ELF1A-like and apolipophorins-like successfully normalized ATP synthase subunit g -like gene expression. Thus, ELF1A-like and apolipophorins-like could be suitable reference genes in PRMs.
• Direct evidence of the preventive effect of milk replacer-based probiotic feeding in calves against severe diarrhea.
  Fumi Kayasaki; Tomohiro Okagawa; Satoru Konnai; Junko Kohara; Yamato Sajiki; Kei Watari; Otgontuya Ganbaatar; Shinya Goto; Hayato Nakamura; Honami Shimakura; Erina Minato; Atsushi Kobayashi; Manabu Kubota; Nobuhiro Terasaki; Akira Takeda; Haruka Noda; Mitsuru Honma; Naoya Maekawa; Shiro Murata; Kazuhiko Ohashi
  Veterinary microbiology, 254, 108976, 108976, Mar. 2021, [Peer-reviewed], [International Magazine]
  English, Scientific journal, Diarrhea is a major cause of death in calves and this is linked directly to economic loss in the cattle industry. Fermented milk replacer (FMR) has been used widely in clinical settings for calf feeding to improve its health and growth. However, the protective efficacy of FMR on calf diarrhea remains unclear. In this study, we verified the preventive effects of FMR feeding on calf diarrhea using an experimental infection model of bovine rotavirus (BRV) in newborn calves and a field study in dairy farms with calf diarrhea. In addition, we evaluated the protective efficacy of lactic acid bacteria-supplemented milk replacer (LAB-MR) in an experimental infection model. In the experimental infection, calves fed FMR or high-concentrated LAB-MR had diarrhea, but the water content of feces was lower and more stable than that of calves fed normal milk replacer. The amount of milk intake also decreased temporarily, but recovered immediately in the FMR- and LAB-MR-fed calves. As compared with the control calves, FMR- or LAB-MR-fed calves showed less severe or reduced histopathological lesions of enteritis in the intestinal mucosa. In a field study using dairy calves, FMR feeding significantly reduced the incidence of enteritis, mortality from enteritis, duration of a series of treatment for enteritis, number of consultations, and cost of medical care for the disease. These results suggest that feeding milk replacer-based probiotics to calves reduces the severity of diarrhea and tissue damage to the intestinal tract caused by BRV infection and provides significant clinical benefits to the prevention and treatment of calf diarrhea.
• PD-L1 immunohistochemistry for canine cancers and clinical benefit of anti-PD-L1 antibody in dogs with pulmonary metastatic oral malignant melanoma.
  Naoya Maekawa; Satoru Konnai; Maki Nishimura; Yumiko Kagawa; Satoshi Takagi; Kenji Hosoya; Hiroshi Ohta; Sangho Kim; Tomohiro Okagawa; Yusuke Izumi; Tatsuya Deguchi; Yukinari Kato; Satoshi Yamamoto; Keiichi Yamamoto; Mikihiro Toda; Chie Nakajima; Yasuhiko Suzuki; Shiro Murata; Kazuhiko Ohashi
  NPJ precision oncology, 5, 1, 10, 10, 12 Feb. 2021, [Peer-reviewed], [International Magazine]
  English, Scientific journal, Immunotherapy targeting programmed cell death 1 (PD-1) and PD-ligand 1 (PD-L1) represents promising treatments for human cancers. Our previous studies demonstrated PD-L1 overexpression in some canine cancers, and suggested the therapeutic potential of a canine chimeric anti-PD-L1 monoclonal antibody (c4G12). However, such evidence is scarce, limiting the clinical application in dogs. In the present report, canine PD-L1 expression was assessed in various cancer types, using a new anti-PD-L1 mAb, 6C11-3A11, and the safety and efficacy of c4G12 were explored in 29 dogs with pulmonary metastatic oral malignant melanoma (OMM). PD-L1 expression was detected in most canine malignant cancers including OMM, and survival was significantly longer in the c4G12 treatment group (median 143 days) when compared to a historical control group (n = 15, median 54 days). In dogs with measurable disease (n = 13), one dog (7.7%) experienced a complete response. Treatment-related adverse events of any grade were observed in 15 dogs (51.7%). Here we show that PD-L1 is a promising target for cancer immunotherapy in dogs, and dogs could be a useful large animal model for human cancer research.
• Intrauterine infection with Mycobacterium avium subsp. paratuberculosis in pregnant cattle diagnosed with Johne's disease
  Yukiko Taniguchi; Satoru Konnai; Shinichi Sakakibara; Ayano Yamamoto; Tomohiro Okagawa; Naoya Maekawa; Shiro Murata; Kazuhiko Ohashi
  JAPANESE JOURNAL OF VETERINARY RESEARCH, 69, 1, 51, 55, Feb. 2021, [Peer-reviewed]
  English, Scientific journal
• Cross-species reactivity of antibodies against Ixodes persulcatus ferritin 2 to Rhipicephalus microplus
  Marina Amaral Xavier; Satoru Konnai; Luis Fernando Parizi; Naftaly Wangombe Githaka; Masayoshi Isezaki; Shinya Goto; Sotaro Fujisawa; Shinji Yamada; Tomohiro Okagawa; Naoya Maekawa; Carlos Logullo; Itabajara da Silva Vaz Jr; Shiro Murata; Kazuhiko Ohashi
  JAPANESE JOURNAL OF VETERINARY RESEARCH, 69, 1, 57, 65, Feb. 2021
  English, Scientific journal
• The enhancement of Th1 immune response by anti-PD-L1 antibody in cattle infected with Mycobacterium avium subsp. paratuberculosis
  Yamato Sajiki; Satoru Konnai; Reiko Nagata; Satoko Kawaji; Hayato Nakamura; Sotaro Fujisawa; Tomohiro Okagawa; Naoya Maekawa; Yukinari Kato; Yasuhiko Suzuki; Shiro Murata; Yasuyuki Mori; Kazuhiko Ohashi
  JOURNAL OF VETERINARY MEDICAL SCIENCE, 83, 2, 162, 166, 25 Feb. 2021, [Peer-reviewed], [Domestic magazines]
  English, Scientific journal
• A TLR7 agonist activates bovine Th1 response and exerts antiviral activity against bovine leukemia virus.
  Yamato Sajiki; Satoru Konnai; Tomohiro Okagawa; Naoya Maekawa; Hayato Nakamura; Yukinari Kato; Yasuhiko Suzuki; Shiro Murata; Kazuhiko Ohashi
  Developmental and comparative immunology, 114, 103847, 103847, Jan. 2021, [Peer-reviewed], [International Magazine]
  English, Scientific journal, Bovine leukemia virus (BLV) infection is a bovine chronic infection caused by BLV, a member of the genus Deltaretrovirus. In this study, we examined the immunomodulatory effects of GS-9620, a toll-like receptor (TLR) 7 agonist, in cattle (Bos taurus) and its therapeutic potential for treating BLV infection. GS-9620 induced cytokine production in peripheral blood mononuclear cells (PBMCs) as well as CD80 expression in CD11c+ cells and increased CD69 and interferon (IFN)-γ expressions in T cells. Removing CD11c+ cells from PBMCs decreased CD69 expression in T cells in the presence of GS-9620. These results suggest that TLR7 agonism promotes T-cell activation via CD11c+ cells. Analyses using PBMCs from BLV-infected cattle revealed that TLR7 expression in CD11c+ cells was upregulated during late-stage BLV infection. Furthermore, GS-9620 increased IFN-γ and TNF-α production and inhibited syncytium formation in vitro, suggesting that GS-9620 may be used to treat BLV infection.
• Tick saliva-induced programmed death-1 and PD-ligand 1 and its related host immunosuppression
  Yamato Sajiki; Satoru Konnai; Yoshinori Ikenaka; Kevin Christian Montecillo Gulay; Atsushi Kobayashi; Luis Fernando Parizi; Benvindo Capela Joao; Kei Watari; Sotaro Fujisawa; Tomohiro Okagawa; Naoya Maekawa; Carlos Logullo; Itabajara da Silva Vaz; Shiro Murata; Kazuhiko Ohashi
  SCIENTIFIC REPORTS, 11, 1, 1063, 1063, 13 Jan. 2021, [Peer-reviewed], [International Magazine]
  English, Scientific journal
• Characterisation of a cysteine protease from poultry red mites and its potential use as a vaccine for chickens.
  Shiro Murata; Ayaka Taniguchi; Masayoshi Isezaki; Sotaro Fujisawa; Eishi Sakai; Akira Taneno; Osamu Ichii; Takuya Ito; Naoya Maekawa; Tomohiro Okagawa; Satoru Konnai; Kazuhiko Ohashi
  Parasite (Paris, France), 28, 9, 9, 2021, [Peer-reviewed], [Lead author, Corresponding author], [International Magazine]
  English, Scientific journal, Poultry red mites (PRMs, Dermanyssus gallinae) are ectoparasites that negatively affect farmed chickens, leading to serious economic losses worldwide. Acaricides have been used to control PRMs in poultry houses. However, some PRMs have developed resistance to acaricides, and therefore different approaches are required to manage the problems caused by PRMs. Vaccination of chickens is one of the methods being considered to reduce the number of PRMs in poultry houses. In a previous study, a cysteine protease, Deg-CPR-1, was identified as a candidate vaccine against PRMs distributed in Europe. In this study, we investigated the characteristics of Deg-CPR-1. A phylogenetic analysis revealed that Deg-CPR-1 is closely related to the digestive cysteine proteases of other mite species, and it was classified into a cluster different from that of chicken cathepsins. Deg-CPR-1 of PRMs in Japan has an amino acid substitution compared with that of PRMs in Europe, but it showed efficacy as a vaccine, consistent with previous findings. Deg-CPR-1 exhibited cathepsin L-like enzyme activity. In addition, the Deg-CPR-1 mRNA was expressed in the midgut and in all stages of PRMs that feed on blood. These results imply that Deg-CPR-1 in the midgut may have important functions in physiological processes, and the inhibition of its expression may contribute to the efficacy of a Deg-CPR-1-based vaccine. Further research is required to fully understand the mechanisms of vaccine efficacy.
• Enhanced Immunotherapeutic Efficacy of Anti-PD-L1 Antibody in Combination with an EP4 Antagonist.
  Yamato Sajiki; Satoru Konnai; Zimeng Cai; Kensuke Takada; Tomohiro Okagawa; Naoya Maekawa; Sotaro Fujisawa; Yukinari Kato; Yasuhiko Suzuki; Shiro Murata; Kazuhiko Ohashi
  ImmunoHorizons, 4, 12, 837, 850, 21 Dec. 2020, [Peer-reviewed], [International Magazine]
  English, Scientific journal, Combination treatment approaches are increasingly considered to overcome resistance to immunotherapy targeting immunoinhibitory molecules such as programmed death (PD)-1 and PD-ligand 1 (PD-L1). Previous studies have demonstrated that the therapeutic efficacy of anti-PD-L1 Abs is enhanced by combination treatment with cyclooxygenase-2 inhibitors, through downregulation of the immunosuppressive eicosanoid PGE2, although the underlying mechanism remains unclear. In this study, we show that serum PGE2 levels are upregulated after anti-PD-L1 Ab administration in a bovine model of immunotherapy and that PGE2 directly inhibits T cell activation via its receptor E prostanoid (EP) 4. Additionally, anti-PD-L1 Ab induces TNF-α production and TNF-α blockade reduces PGE2 production in the presence of anti-PD-L1 Ab, suggesting that anti-PD-L1 Ab-induced TNF-α impairs T cell activation by PGE2 upregulation. Our studies examining the therapeutic potential of the dual blockade of PD-L1 and EP4 in bovine and murine immune cells reveal that the dual blockade of PD-L1 and EP4 significantly enhances Th1 cytokine production in vitro. Finally, we show that the dual blockade decreases tumor volume and prolongs survival in mice inoculated with the murine lymphoma cell line EG7. Altogether, these results suggest that TNF-α induced by anti-PD-L1 Ab treatment is associated with T cell dysfunction via PGE2/EP4 pathway and that the dual blockade of PD-L1 and EP4 should be considered as a novel immunotherapy for cancer.
• Detection and molecular identification of Leucocytozoon and Plasmodium species from village chickens in different areas of Myanmar
  Shwe Yee Win; Hla Myet Chel; Myint Myint Hmoon; Lat Lat Htun; Saw Bawm; Mar Mar Win; Shiro Murata; Nariaki Nonaka; Ryo Nakao; Ken Katakura
  ACTA TROPICA, 212, 105719, 105719, Dec. 2020, [Peer-reviewed], [International Magazine]
  English, Scientific journal
• Distinct polymorphisms in a single herpesvirus gene are capable of enhancing virulence and mediating vaccinal resistance.
  Andelé M Conradie; Luca D Bertzbach; Jakob Trimpert; Joseph N Patria; Shiro Murata; Mark S Parcells; Benedikt B Kaufer
  PLoS pathogens, 16, 12, e1009104, Dec. 2020, [Peer-reviewed], [International Magazine]
  English, Scientific journal, Modified-live herpesvirus vaccines are widely used in humans and animals, but field strains can emerge that have a higher virulence and break vaccinal protection. Since the introduction of the first vaccine in the 1970s, Marek's disease virus overcame the vaccine barrier by the acquisition of numerous genomic mutations. However, the evolutionary adaptations in the herpesvirus genome responsible for the vaccine breaks have remained elusive. Here, we demonstrate that point mutations in the multifunctional meq gene acquired during evolution can significantly alter virulence. Defined mutations found in highly virulent strains also allowed the virus to overcome innate cellular responses and vaccinal protection. Concomitantly, the adaptations in meq enhanced virus shedding into the environment, likely providing a selective advantage for the virus. Our study provides the first experimental evidence that few point mutations in a single herpesviral gene result in drastically increased virulence, enhanced shedding, and escape from vaccinal protection.
• Genetic characterization of a Marek's disease virus strain isolated in Japan.
  Shiro Murata; Yuka Machida; Masayoshi Isezaki; Naoya Maekawa; Tomohiro Okagawa; Satoru Konnai; Kazuhiko Ohashi
  Virology journal, 17, 1, 186, 186, 23 Nov. 2020, [Peer-reviewed], [Lead author, Corresponding author], [International Magazine]
  English, Scientific journal, BACKGROUND: Marek's disease virus (MDV) causes malignant lymphomas in chickens (Marek's disease, MD). MD is currently controlled by vaccination; however, MDV strains have a tendency to develop increased virulence. Distinct diversity and point mutations are present in the Meq proteins, the oncoproteins of MDV, suggesting that changes in protein function induced by amino acid substitutions might affect MDV virulence. We previously reported that recent MDV isolates in Japan display distinct mutations in Meq proteins from those observed in traditional MDV isolates in Japan, but similar to those in MDV strains isolated from other countries. METHODS: To further investigate the genetic characteristics in Japanese field strains, we sequenced the whole genome of an MDV strain that was successfully isolated from a chicken with MD in Japan. A phylogenetic analysis of the meq gene was also performed. RESULTS: Phylogenetic analysis revealed that the Meq proteins in most of the Japanese isolates were similar to those of Chinese and European strains, and the genomic sequence of the Japanese strain was classified into the Eurasian cluster. Comparison of coding region sequences among the Japanese strain and MDV strains from other countries revealed that the genetic characteristics of the Japanese strain were similar to those of Chinese and European strains. CONCLUSIONS: The MDV strains distributed in Asian and European countries including Japan seem to be genetically closer to each other than to MDV strains from North America. These findings indicate that the genetic diversities of MDV strains that emerged may have been dependent on the different vaccination-based control approaches.
• Identification and functional analysis of ferritin 2 from the Taiga tick Ixodes persulcatus Schulze.
  Naftaly Wang'ombe Githaka; Satoru Konnai; Masayoshi Isezaki; Shinya Goto; Marina Amaral Xavier; Sotaro Fujisawa; Shinji Yamada; Tomohiro Okagawa; Naoya Maekawa; Carlos Logullo; Itabajara da Silva Vaz Jr; Shiro Murata; Kazuhiko Ohashi
  Ticks and tick-borne diseases, 11, 6, 101547, 101547, Nov. 2020, [Peer-reviewed], [International Magazine]
  English, Scientific journal, Ferritin 2 (FER2) is an iron storage protein, which has been shown to be critical for iron homeostasis during blood feeding and reproduction in ticks and is therefore suitable as a component for anti-tick vaccines. In this study, we identified the FER2 of Ixodes persulcatus, a major vector for zoonotic diseases such as Lyme borreliosis and tick-borne relapsing fever in Japan, and investigated its functions. Ixodes persulcatus-derived ferritin 2 (Ip-FER2) showed concentration-dependent iron-binding ability and high amino acid conservation, consistent with FER2s of other tick species. Vaccines containing the recombinant Ip-FER2 elicited a significant reduction of the engorgement weight of adult I. persulcatus. Interestingly, the reduction of engorgement weight was also observed in Ixodes ovatus, a sympatric species of I. persulcatus. In silico analyses of FER2 sequences of I. persulcatus and other ticks showed a greater similarity with I. scapularis and I. ricinus and lesser similarity with Hyalomma anatolicum, Haemaphysalis longicornis, Rhipicephalus microplus, and R. appendiculatus. Moreover, it was observed that the tick FER2 sequences possess conserved regions within the primary structures, and in silico epitope mapping analysis revealed that antigenic regions were also conserved, particularly among Ixodes spp ticks. In conclusion, the data support further protective tick vaccination applications using the Ip-FER2 antigens identified herein.
• Molecular detection and genetic characterization of infectious laryngotracheitis virus in poultry in Myanmar
  Zhiyuan Yang; Shiro Murata; Sotaro Fujisawa; Masaki Takehara; Ken Katakura; Myint Myint Hmoon; Shwe Yee Win; Saw Bawm; Satoru Konnai; Kazuhiko Ohashi
  BMC VETERINARY RESEARCH, 16, 1, 453, 453, 23 Nov. 2020, [Peer-reviewed], [Corresponding author], [International Magazine]
  English, Scientific journal
• Transcriptome dynamics of blood-fed and starved poultry red mites, Dermanyssus gallinae.
  Sotaro Fujisawa; Shiro Murata; Masayoshi Isezaki; Eiji Oishi; Akira Taneno; Naoya Maekawa; Tomohiro Okagawa; Satoru Konnai; Kazuhiko Ohashi
  Parasitology international, 78, 102156, 102156, Oct. 2020, [Peer-reviewed], [Corresponding author], [International Magazine]
  English, Scientific journal
• Expression Analysis of Canine CMTM6 and CMTM4 as Potential Regulators of the PD-L1 Protein in Canine Cancers
  Hiroto Takeuchi; Satoru Konnai; Naoya Maekawa; Erina Minato; Yoshiki Ichikawa; Atsushi Kobayashi; Tomohiro Okagawa; Shiro Murata; Kazuhiko Ohashi
  FRONTIERS IN VETERINARY SCIENCE, 7, 330, 330, Jun. 2020, [Peer-reviewed], [International Magazine]
  English, Scientific journal
• Immunosuppressive effects of sialostatin L1 and L2 isolated from the taiga tick Ixodes persulcatus Schulze.
  Yamato Sajiki; Satoru Konnai; Akie Ochi; Tomohiro Okagawa; Naftaly Githaka; Masayoshi Isezaki; Shinji Yamada; Takuya Ito; Shuji Ando; Hiroki Kawabata; Carlos Logullo; Itabajara da Silva Vaz Jr; Naoya Maekawa; Shiro Murata; Kazuhiko Ohashi
  Ticks and tick-borne diseases, 11, 2, 101332, 101332, Mar. 2020, [Peer-reviewed], [International Magazine]
  English, Scientific journal, Tick saliva contains immunosuppressants which are important to obtain a blood meal and enhance the infectivity of tick-borne pathogens. In Japan, Ixodes persulcatus is a major vector for Lyme borreliosis pathogens, such as Borrelia garinii, as well as for those causing relapsing fever, such as B. miyamotoi. To date, little information is available on bioactive salivary molecules, produced by this tick. Thus, in this study, we identified two proteins, I. persulcatus derived sialostatin L1 (Ip-sL1) and sL2 (Ip-sL2), as orthologs of I. scapularis derived sL1 and sL2. cDNA clones of Ip-sL1 and Ip-sL2 shared a high identity with sequences of sL1 and sL2 isolated from the salivary glands of I. scapularis. Semi-quantitative PCR revealed that Ip-sL1 and Ip-sL2 were expressed in the salivary glands throughout the life of the tick. In addition, Ip-sL1 and Ip-sL2 were expressed even before the ticks started feeding, and their expression continued during blood feeding. Recombinant Ip-sL1 and Ip-sL2 were developed to characterize the proteins via biological and immunological analyses. These analyses revealed that both Ip-sL1 and Ip-sL2 had inhibitory effects on cathepsins L and S. Ip-sL1 and Ip-sL2 inhibited the production of IP-10, TNFα, and IL-6 by LPS-stimulated bone-marrow-derived dendritic cells (BMDCs). Additionally, Ip-sL1 significantly impaired BMDC maturation. Taken together, these results suggest that Ip-sL1 and Ip-sL2 confer immunosuppressive functions and appear to be involved in the transmission of pathogens by suppressing host immune responses, such as cytokine production and dendritic cell maturation. Therefore, further studies are warranted to investigate the immunosuppressive functions of Ip-sL1 and Ip-sL2 in detail to clarify their involvement in pathogen transmission via I. persulcatus.
• PD-L1 expression in equine malignant melanoma and functional effects of PD-L1 blockade.
  Otgontuya Ganbaatar; Satoru Konnai; Tomohiro Okagawa; Yutaro Nojima; Naoya Maekawa; Erina Minato; Atsushi Kobayashi; Ryo Ando; Nobuya Sasaki; Daisuke Miyakoshi; Osamu Ichii; Yukinari Kato; Yasuhiko Suzuki; Shiro Murata; Kazuhiko Ohashi
  PloS one, 15, 11, e0234218, 2020, [Peer-reviewed], [International Magazine]
  English, Scientific journal, Programmed death-1 (PD-1) is an immunoinhibitory receptor expressed on lymphocytes. Interaction of PD-1 with its ligand PD-ligand 1 (PD-L1) delivers inhibitory signals and impairs proliferation, cytokine production, and cytotoxicity of T cells. In our previous studies, we have developed anti-bovine PD-L1 monoclonal antibodies (mAbs) and reported that the PD-1/PD-L1 pathway was closely associated with T-cell exhaustion and disease progression in bovine chronic infections and canine tumors. Furthermore, we found that blocking antibodies that target PD-1 and PD-L1 restore T-cell functions and could be used in immunotherapy in cattle and dogs. However, the immunological role of the PD-1/PD-L1 pathway for chronic equine diseases, including tumors, remains unclear. In this study, we identified cDNA sequences of equine PD-1 (EqPD-1) and PD-L1 (EqPD-L1) and investigated the role of anti-bovine PD-L1 mAbs against EqPD-L1 using in vitro assays. In addition, we evaluated the expression of PD-L1 in tumor tissues of equine malignant melanoma (EMM). The amino acid sequences of EqPD-1 and EqPD-L1 share a considerable identity and similarity with homologs from non-primate species. Two clones of the anti-bovine PD-L1 mAbs recognized EqPD-L1 in flow cytometry, and one of these cross-reactive mAbs blocked the binding of equine PD-1/PD-L1. Of note, immunohistochemistry confirmed the PD-L1 expression in EMM tumor tissues. A cultivation assay revealed that PD-L1 blockade enhanced the production of Th1 cytokines in equine immune cells. These findings showed that our anti-PD-L1 mAbs would be useful for analyzing the equine PD-1/PD-L1 pathway. Further research is warranted to discover the immunological role of PD-1/PD-L1 in chronic equine diseases and elucidate a future application in immunotherapy for horses.
• The Suppression of Th1 Response by Inducing TGF-β1 From Regulatory T Cells in Bovine Mycoplasmosis.
  Yamato Sajiki; Satoru Konnai; Shinya Goto; Tomohiro Okagawa; Kosuke Ohira; Honami Shimakura; Naoya Maekawa; Satoshi Gondaira; Hidetoshi Higuchi; Motoshi Tajima; Yuki Hirano; Junko Kohara; Shiro Murata; Kazuhiko Ohashi
  Frontiers in veterinary science, 7, 609443, 609443, 2020, [Peer-reviewed], [International Magazine]
  English, Scientific journal, Regulatory T cells (Tregs) regulate immune responses and maintain host immune homeostasis. Tregs contribute to the disease progression of several chronic infections by oversuppressing immune responses via the secretion of immunosuppressive cytokines, such as transforming growth factor (TGF)-β and interleukin-10. In the present study, we examined the association of Tregs with Mycoplasma bovis infection, in which immunosuppression is frequently observed. Compared with uninfected cattle, the percentage of Tregs, CD4+CD25highFoxp3+ T cells, was increased in M. bovis-infected cattle. Additionally, the plasma of M. bovis-infected cattle contained the high concentrations of TGF-β1, and M. bovis infection induced TGF-β1 production from bovine immune cells in in vitro cultures. Finally, we analyzed the immunosuppressive effects of TGF-β1 on bovine immune cells. Treatment with TGF-β1 significantly decreased the expression of CD69, an activation marker, in T cells, and Th1 cytokine production in vitro. These results suggest that the increase in Tregs and TGF-β1 secretion could be one of the immunosuppressive mechanisms and that lead to increased susceptibility to other infections in terms of exacerbation of disease during M. bovis infection.
• Upregulation of PD-L1 Expression by Prostaglandin E2 and the Enhancement of IFN-γ by Anti-PD-L1 Antibody Combined With a COX-2 Inhibitor in Mycoplasma bovis Infection.
  Shinya Goto; Satoru Konnai; Yuki Hirano; Junko Kohara; Tomohiro Okagawa; Naoya Maekawa; Yamato Sajiki; Kei Watari; Erina Minato; Atsuhi Kobayashi; Satoshi Gondaira; Hidetoshi Higuchi; Masateru Koiwa; Motoshi Tajima; Eiji Taguchi; Ryoko Uemura; Shinji Yamada; Mika K Kaneko; Yukinari Kato; Keiichi Yamamoto; Mikihiro Toda; Yasuhiko Suzuki; Shiro Murata; Kazuhiko Ohashi
  Frontiers in veterinary science, 7, 12, 12, 2020, [Peer-reviewed], [International Magazine]
  English, Scientific journal, Bovine mycoplasmosis caused by Mycoplasma bovis results in pneumonia and mastitis in cattle. We previously demonstrated that the programmed death 1 (PD-1)/PD-ligand 1 (PD-L1) pathway is involved in immune dysfunction during M. bovis infection and that prostaglandin E2 (PGE2) suppressed immune responses and upregulated PD-L1 expression in Johne's disease, a bacterial infection in cattle. In this study, we investigated the role of PGE2 in immune dysfunction and the relationship between PGE2 and the PD-1/PD-L1 pathway in M. bovis infection. In vitro stimulation with M. bovis upregulated the expressions of PGE2 and PD-L1 presumably via Toll-like receptor 2 in bovine peripheral blood mononuclear cells (PBMCs). PGE2 levels of peripheral blood in infected cattle were significantly increased compared with those in uninfected cattle. Remarkably, plasma PGE2 levels were positively correlated with the proportions of PD-L1+ monocytes in M. bovis-infected cattle. Additionally, plasma PGE2 production in infected cattle was negatively correlated with M. bovis-specific interferon (IFN)-γ production from PBMCs. These results suggest that PGE2 could be one of the inducers of PD-L1 expression and could be involved in immunosuppression during M. bovis infection. In vitro blockade assays using anti-bovine PD-L1 antibody and a cyclooxygenase 2 inhibitor significantly upregulated the M. bovis-specific IFN-γ response. Our study findings might contribute to the development of novel therapeutic strategies for bovine mycoplasmosis that target PGE2 and the PD-1/PD-L1 pathway.
• Clinical efficacy of the combined treatment of anti-PD-L1 rat-bovine chimeric antibody with a COX-2 inhibitor in calves infected with Mycoplasma bovis
  Goto S; Konnai S; Hirano Y; Kohara J; Okagawa T; Maekawa N; Sajiki Y; Watari K; Minato E; Kobayashi A; Gondaira S; Higuchi H; Koiwa M; Tajima M; Taguchi E; Ishida M; Uemura R; Yamada S; Kaneko M; Kato Y; Yamamoto K; Toda M; Suzuki Y; Murata S; Ohashi K
  Jpn J Vet Res, 68, 2, 77, 90, 2020, [Peer-reviewed]
  English, Scientific journal
• Immune inhibitory function of bovine CTLA-4 and the effects of its blockade in IFN-gamma production
  Kei Watari; Satoru Konnai; Naoya Maekawa; Tomohiro Okagawa; Yasuhiko Suzuki; Shiro Murata; Kazuhiko Ohashi
  BMC VETERINARY RESEARCH, 15, 1, 380, 380, 29 Oct. 2019, [Peer-reviewed], [International Magazine]
  English, Scientific journal
• Prostaglandin E2-Induced Immune Exhaustion and Enhancement of Antiviral Effects by Anti-PD-L1 Antibody Combined with COX-2 Inhibitor in Bovine Leukemia Virus Infection.
  Yamato Sajiki; Satoru Konnai; Tomohiro Okagawa; Asami Nishimori; Naoya Maekawa; Shinya Goto; Kei Watari; Erina Minato; Atsushi Kobayashi; Junko Kohara; Shinji Yamada; Mika K Kaneko; Yukinari Kato; Hirofumi Takahashi; Nobuhiro Terasaki; Akira Takeda; Keiichi Yamamoto; Mikihiro Toda; Yasuhiko Suzuki; Shiro Murata; Kazuhiko Ohashi
  Journal of immunology (Baltimore, Md. : 1950), 203, 5, 1313, 1324, 01 Sep. 2019, [Peer-reviewed], [International Magazine]
  English, Scientific journal, Bovine leukemia virus (BLV) infection is a chronic viral infection of cattle and endemic in many countries, including Japan. Our previous study demonstrated that PGE2, a product of cyclooxygenase (COX) 2, suppresses Th1 responses in cattle and contributes to the progression of Johne disease, a chronic bacterial infection in cattle. However, little information is available on the association of PGE2 with chronic viral infection. Thus, we analyzed the changes in plasma PGE2 concentration during BLV infection and its effects on proviral load, viral gene transcription, Th1 responses, and disease progression. Both COX2 expression by PBMCs and plasma PGE2 concentration were higher in the infected cattle compared with uninfected cattle, and plasma PGE2 concentration was positively correlated with the proviral load. BLV Ag exposure also directly enhanced PGE2 production by PBMCs. Transcription of BLV genes was activated via PGE2 receptors EP2 and EP4, further suggesting that PGE2 contributes to disease progression. In contrast, inhibition of PGE2 production using a COX-2 inhibitor activated BLV-specific Th1 responses in vitro, as evidenced by enhanced T cell proliferation and Th1 cytokine production, and reduced BLV proviral load in vivo. Combined treatment with the COX-2 inhibitor meloxicam and anti-programmed death-ligand 1 Ab significantly reduced the BLV proviral load, suggesting a potential as a novel control method against BLV infection. Further studies using a larger number of animals are required to support the efficacy of this treatment for clinical application.
• Vector transmission of bovine leukemia virus during summer season in Northern Hokkaido
  Inagaki H; Konnai S; Kaburagi H; Murota H; Takabatake N; Watari K; Okagawa T; Maekawa N; Murata S; Ohashi K
  Jpn. J. Vet. Res, 67, 3, 235, 239, Aug. 2019, [Peer-reviewed]
  English, Scientific journal
• Molecular detection and genetic characterization of Mycoplasma gallisepticum, Mycoplama synoviae, and infectious bronchitis virus in poultry in Myanmar.
  Fujisawa S; Murata S; Takehara M; Katakura K; Hmoon MM; Win SY; Ohashi K
  BMC veterinary research, 15, 1, 261, Springer Science and Business Media LLC, Jul. 2019, [Peer-reviewed], [Corresponding author]
  Scientific journal
• Haematophagous mites on poultry farms in the Republic of the Union of Myanmar
  Masaki Takehara; Shiro Murata; Ken Katakura; Sotaro Fujisawa; Myint Myint Hmoon; Shwe Yee Win; Saw Bawm; Lat Lat Htun; Ye Htut Aung; Mar Mar Win; Masayoshi Isezaki; Naoya Maekawa; Tomohiro Okagawa; Satoru Konnai; Kazuhiko Ohashi
  HELIYON, 5, 4, e01544, Apr. 2019, [Peer-reviewed], [Corresponding author], [International Magazine]
  English, Scientific journal
• Effects of bovine tumor necrosis factor alpha decoy receptors on cell death and inflammatory cytokine kinetics: potential for bovine inflammation therapy.
  Fujisawa S; Konnai S; Okagawa T; Maekawa N; Tanaka A; Suzuki Y; Murata S; Ohashi K
  BMC veterinary research, 15, 1, 68, 68, Feb. 2019, [Peer-reviewed], [International Magazine]
  English, Scientific journal, BACKGROUND: Refractory diseases, including bacterial infections, are causing huge economic losses in dairy farming. Despite efforts to prevent and treat those diseases in cattle, including the use of antimicrobials, it is not well controlled in the field. Several inflammatory cytokines, including tumor necrosis factor alpha (TNF-α), play important roles in disease progression; thus, blocking these cytokines can attenuate the acute and sever inflammation and may be a novel strategy for treatment. However, biological drugs targeting inflammatory cytokines have not been used in cattle. Therefore, in this study, bovine sTNFR1 and sTNFR2 IgG1 Fc-fusion proteins (TNFR1-Ig and TNFR2-Ig) were produced, and their anti-inflammatory functions were analyzed in vitro, to develop decoy receptors for bovine TNF-α. RESULTS: Both TNFR1-Ig and TNFR2-Ig were shown to bind with TNF-α, and TNFR2-Ig showed higher affinity toward TNF-α than TNFR1-Ig. We next stimulated murine fibroblast-derived cells (L929 cells) with TNF-α to induce cell death and analyzed cell viability in the presence of TNFR-Ig proteins. Both TNFR1-Ig and TNFR2-Ig suppressed TNF-α-induced cell death, significantly improving cell viability. In addition, cell death induced by TNF-α was suppressed, even at low TNFR2-Ig concentrations, suggesting TNFR2-Ig has higher activity to suppress TNF-α functions than TNFR1-Ig. Finally, to examine TNFR2-Ig's anti-inflammatory, we cultured peripheral blood mononuclear cells from cattle with TNF-α in the presence of TNFR2-Ig and analyzed the gene expression and protein production of the inflammatory cytokines IL-1β and TNF-α. TNFR2-Ig significantly reduced the gene expression and protein production of these cytokines. Our results suggest that TNFR2-Ig inhibits inflammatory cytokine kinetics by blocking TNF-α to transmembrane TNFR, thereby attenuating excessive inflammation induced by TNF-α. CONCLUSIONS: Collectively, the findings of this study demonstrated the potential of TNFR2-Ig as a novel therapeutic for inflammatory diseases, such as bovine clinical mastitis. Further investigation is required for future clinical application.
• Identification of immuno-inhibitory molecules in Mongolian native cattle and yak
  Ochirkhuu N; Konnai S; Odbileg R; Okagawa T; Maekawa N; Murata S; Ohashi K
  Jpn J Vet Res, 66, 3, 177, 192, Aug. 2018, [Peer-reviewed]
  English, Scientific journal
• Cooperation of PD-1 and LAG-3 in the exhaustion of CD4+ and CD8+ T cells during bovine leukemia virus infection.
  Tomohiro Okagawa; Satoru Konnai; Asami Nishimori; Naoya Maekawa; Shinya Goto; Ryoyo Ikebuchi; Junko Kohara; Yasuhiko Suzuki; Shinji Yamada; Yukinari Kato; Shiro Murata; Kazuhiko Ohashi
  Veterinary research, 49, 1, 50, 50, 19 Jun. 2018, [Peer-reviewed], [International Magazine]
  English, Scientific journal, Bovine leukemia virus (BLV) is a retrovirus that infects B cells in cattle and causes bovine leukosis after a long latent period. Progressive exhaustion of T cell functions is considered to facilitate disease progression of BLV infection. Programmed death-1 (PD-1) and lymphocyte activation gene-3 (LAG-3) are immunoinhibitory receptors that contribute to T-cell exhaustion caused by BLV infection in cattle. However, it is unclear whether the cooperation of PD-1 and LAG-3 accelerates disease progression of BLV infection. In this study, multi-color flow cytometric analyses of PD-1- and LAG-3-expressing T cells were performed in BLV-infected cattle at different stages of the disease. The frequencies of PD-1+LAG-3+ heavily exhausted T cells among CD4+ and CD8+ T cells was higher in the blood of cattle with B-cell lymphoma over that of BLV-uninfected and BLV-infected cattle without lymphoma. In addition, blockade assays of peripheral blood mononuclear cells were performed to examine whether inhibition of the interactions between PD-1 and LAG-3 and their ligands by blocking antibodies could restore T-cell function during BLV infection. Single or dual blockade of the PD-1 and LAG-3 pathways reactivated the production of Th1 cytokines, interferon-γ and tumor necrosis factor-α, from BLV-specific T cells of the infected cattle. Taken together, these results indicate that PD-1 and LAG-3 cooperatively mediate the functional exhaustion of CD4+ and CD8+ T cells and are associated with the development of B-cell lymphoma in BLV-infected cattle.
• Prostaglandin E2 Induction Suppresses the Th1 Immune Responses in Cattle with Johne's Disease.
  Yamato Sajiki; Satoru Konnai; Tomohiro Okagawa; Asami Nishimori; Naoya Maekawa; Shinya Goto; Ryoyo Ikebuchi; Reiko Nagata; Satoko Kawaji; Yumiko Kagawa; Shinji Yamada; Yukinari Kato; Chie Nakajima; Yasuhiko Suzuki; Shiro Murata; Yasuyuki Mori; Kazuhiko Ohashi
  Infection and immunity, 86, 5, May 2018, [Peer-reviewed], [International Magazine]
  English, Scientific journal, Johne's disease, caused by Mycobacterium avium subsp. paratuberculosis, is a bovine chronic infection that is endemic in Japan and many other countries. The expression of immunoinhibitory molecules is upregulated in cattle with Johne's disease, but the mechanism of immunosuppression is poorly understood. Prostaglandin E2 (PGE2) is immunosuppressive in humans, but few veterinary data are available. In this study, functional and kinetic analyses of PGE2 were performed to investigate the immunosuppressive effect of PGE2 during Johne's disease. In vitro PGE2 treatment decreased T-cell proliferation and Th1 cytokine production and upregulated the expression of immunoinhibitory molecules such as interleukin-10 and programmed death ligand 1 (PD-L1) in peripheral blood mononuclear cells (PBMCs) from healthy cattle. PGE2 was upregulated in sera and intestinal lesions of cattle with Johne's disease. In vitro stimulation with Johnin purified protein derivative (J-PPD) induced cyclooxygenase-2 (COX-2) transcription, PGE2 production, and upregulation of PD-L1 and immunoinhibitory receptors in PBMCs from cattle infected with M. avium subsp. paratuberculosis Therefore, Johnin-specific Th1 responses could be limited by the PGE2 pathway in cattle. In contrast, downregulation of PGE2 with a COX-2 inhibitor promoted J-PPD-stimulated CD8+ T-cell proliferation and Th1 cytokine production in PBMCs from the experimentally infected cattle. PD-L1 blockade induced J-PPD-stimulated CD8+ T-cell proliferation and interferon gamma production in vitro Combined treatment with a COX-2 inhibitor and anti-PD-L1 antibodies enhanced J-PPD-stimulated CD8+ T-cell proliferation in vitro, suggesting that the blockade of both pathways is a potential therapeutic strategy to control Johne's disease. The effects of COX-2 inhibition warrant further study as a novel treatment of Johne's disease.
• Analysis of Porcine Circovirus Type 2 Load in Serum from Pigs Recently Imported into Japan Using Quantitative Real-Time PCR
  柴田明弘; 村田史郎; 中尾哲也; 井関博
  日本獣医師会雑誌, 71, 3, 135‐139, 20 Mar. 2018, [Peer-reviewed]
  Japanese
• Intrauterine infection with bovine leukemia virus in pregnant dam with high viral load.
  Yamato Sajiki; Satoru Konnai; Asami Nishimori; Tomohiro Okagawa; Naoya Maekawa; Shinya Goto; Masashi Nagano; Junko Kohara; Nana Kitano; Toshihiko Takahashi; Motoshi Tajima; Hirohisa Mekata; Yoichiro Horii; Shiro Murata; Kazuhiko Ohashi
  The Journal of veterinary medical science, 79, 12, 2036, 2039, 22 Dec. 2017, [Peer-reviewed], [Domestic magazines]
  English, Scientific journal, Enzootic bovine leukemia is caused by the bovine leukemia virus (BLV). BLV is transmitted vertically or horizontally through the transfer of infected cells via direct contact, through milk, insect bites and contaminated iatrogenic procedures. However, we lacked direct evidence of intrauterine infection. The purpose of this study was to confirm intrauterine BLV infection in two pregnant dams with high viral load by cesarean delivery. BLV was detected in cord and placental blood, and the BLV in the newborns showed 100% nucleotide identity with the BLV-env sequence from the dams. Notably, a newborn was seropositive for BLV but had no colostral antibodies. In this study, we presented a direct evidence of intrauterine BLV transmission in pregnant dam with a high proviral load. These results could aid the development of BLV control measures targeting viral load.
• Correction for Nishimori et al., “Identification of an Atypical Enzootic Bovine Leukosis in Japan by Using a Novel Classification of Bovine Leukemia Based on Immunophenotypic Analysis”
  Asami Nishimori; Satoru Konnai; Tomohiro Okagawa; Naoya Maekawa; Shinya Goto; Ryoyo Ikebuchi; Ayako Nakahara; Yuzumi Chiba; Masaho Ikeda; Shiro Murata; Kazuhiko Ohashi
  Clinical and vaccine immunology : CVI, 24, 12, Dec. 2017, [International Magazine]
  English, Scientific journal
• Detection and molecular characterization of equine infectious anemia virus in Mongolian horses
  Tumenjargal Sharav; Satoru Konnai; Nyamsuren Ochirkhuu; Erdene T. S. Ochir; Hirohisa Mekata; Yoshihiro Sakoda; Takashi Umemura; Shiro Murata; Tungalag Chultemdorj; Kazuhiko Ohashi
  JOURNAL OF VETERINARY MEDICAL SCIENCE, 79, 11, 1884, 1888, Nov. 2017, [Peer-reviewed]
  English, Scientific journal
• Identification of an Atypical Enzootic Bovine Leukosis in Japan by Using a Novel Classification of Bovine Leukemia Based on Immunophenotypic Analysis
  Asami Nishimori; Satoru Konnai; Tomohiro Okagawa; Naoya Maekawa; Shinya Goto; Ryoyo Ikebuchi; Ayako Nakahara; Yuzumi Chiba; Masaho Ikeda; Shiro Murata; Kazuhiko Ohashi
  CLINICAL AND VACCINE IMMUNOLOGY, 24, 9, Sep. 2017, [Peer-reviewed]
  English, Scientific journal
• Molecular detection and phylogenetic analysis of Ehrlichia canis in a Philippine dog.
  Maekawa N; Konnai S; Balbin MM; Mingala CN; Gicana KRB; Bernando FAEM; Murata S; Ohashi K
  Ticks and tick-borne diseases, 9, 2, 266, 269, Sep. 2017, [Peer-reviewed], [International Magazine]
  English, Scientific journal, Canine monocytic ehrlichiosis (CME), caused by a rickettsial bacterium, Ehrlichia canis, is distributed worldwide, particularly in tropical and subtropical regions. Transmission of E. canis is primarily mediated by the vector tick, Rhipicephalus sanguineus sensu lato and the bacteria then infect and replicate in monocytes and macrophages. Many cases are seen in veterinary hospitals and treated routinely; however, the genetic variation of E. canis strains found in the Philippines has been poorly investigated to date. In this study, the 16S rRNA gene and the gp200 gene of E. canis were detected by polymerase chain reaction from an infected dog in the Philippines, and the deduced amino acid sequence of the gp200 gene was subjected to a phylogenetic analysis. The Philippine genotype formed a cluster with the Taiwan genotype, and was somewhat divergent from the USA and Brazil strains. This suggested that E. canis underwent evolution in East and Southeast Asia, confirming the utility of the gp200 gene for the assessment of genetic relationships among strains.
• Molecular Epidemiological Survey and Genetic Characterization of Anaplasma Species in Mongolian Livestock
  Nyamsuren Ochirkhuu; Satoru Konnai; Raadan Odbileg; Shiro Murata; Kazuhiko Ohashi
  VECTOR-BORNE AND ZOONOTIC DISEASES, 17, 8, 539, 549, Aug. 2017, [Peer-reviewed]
  English, Scientific journal
• A canine chimeric monoclonal antibody targeting PD-L1 and its clinical efficacy in canine oral malignant melanoma or undifferentiated sarcoma
  Naoya Maekawa; Satoru Konnai; Satoshi Takagi; Yumiko Kagawa; Tomohiro Okagawa; Asami Nishimori; Ryoyo Ikebuchi; Yusuke Izumi; Tatsuya Deguchi; Chie Nakajima; Yukinari Kato; Keiichi Yamamoto; Hidetoshi Uemura; Yasuhiko Suzuki; Shiro Murata; Kazuhiko Ohashi
  SCIENTIFIC REPORTS, 7, 1, 8951, Aug. 2017, [Peer-reviewed]
  English, Scientific journal
• Anti-Bovine Programmed Death-1 Rat-Bovine Chimeric Antibody for Immunotherapy of Bovine Leukemia Virus Infection in Cattle
  Tomohiro Okagawa; Satoru Konnai; Asami Nishimori; Naoya Maekawa; Ryoyo Ikebuchi; Shinya Goto; Chie Nakajima; Junko Kohara; Satoshi Ogasawara; Yukinari Kato; Yasuhiko Suzuki; Shiro Murata; Kazuhiko Ohashi
  FRONTIERS IN IMMUNOLOGY, 8, 650, Jun. 2017, [Peer-reviewed]
  English, Scientific journal
• In vitro and in vivo antivirus activity of an anti-programmed death-ligand 1 (PD-L1) ratbovine chimeric antibody against bovine leukemia virus infection
  Asami Nishimori; Satoru Konnai; Tomohiro Okagawa; Naoya Maekawa; Ryoyo Ikebuchi; Shinya Goto; Yamato Sajiki; Yasuhiko Suzuki; Junko Kohara; Satoshi Ogasawara; Yukinari Kato; Shiro Murata; Kazuhiko Ohashi
  PLOS ONE, 12, 4, e0174916, Apr. 2017, [Peer-reviewed]
  English, Scientific journal
• Increase of cells expressing PD-1 and PD-L1 and enhancement of IFN-γ production via PD-1/PD-L1 blockade in bovine mycoplasmosis.
  Goto, S; Konnai, S; Okagawa, T; Nishimori, A; Maekawa, N; Gondaira, S; Higuchi, H; Koiwa, M; Tajima, M; Kohara, J; Ogasawara, S; Kato, Y; Suzuki, Y; Murata, S; Ohashi, K
  Immunity, Inflammation and Disease, 10, 3, 1, 9, 2017, [Peer-reviewed], [International Magazine]
  English, Scientific journal, INTRODUCTION: Bovine mycoplasma, chiefly Mycoplasma bovis, is a pathogen that causes pneumonia, mastitis, arthritis, and otitis media in cattle. This pathogen exerts immunosuppressive effects, such as the inhibition of interferon production. However, the mechanisms involved in bovine mycoplasmosis have not been fully elucidated. In this study, we investigated the role of the programmed death-1 (PD-1)/programmed death-ligand 1 (PD-L1) pathway in immunosuppression in bovine mycoplasmosis. METHODS: In the initial experiments, we used enzyme-linked immunosorbent assay to measure interferon-γ (IFN-γ) from peripheral blood mononuclear cells (PBMCs) isolated from cattle with mycoplasmosis. RESULTS: Expectedly, IFN-γ production significantly decreased in cattle with mycoplasmosis compared with that in clinically healthy cattle. Concomitantly, flow cytometric analysis revealed that the proportions of PD-1+ CD4+ and PD-L1+ CD14+ cells significantly increased in peripheral blood of the infected cattle. Interestingly, the number of PD-1+ CD4+ and PD-1+ CD8+ T cells were negatively correlated with IFN-γ production from PBMCs in bovine mycoplasmosis. Additionally, blockade of the PD-1/PD-L1 pathway in vitro by anti-bovine PD-1- and anti-bovine PD-L1 antibodies significantly upregulated the production of IFN-γ from anti-mycoplasma-specific cells. CONCLUSIONS: These results suggest that the PD-1/PD-L1 pathway could be involved in immune exhaustion of bovine mycoplasma-specific T cells. In conclusion, our study opens up a new perspective in the therapeutic strategy for bovine mycoplasmosis by targeting the immunoinhibitory receptor pathways.
• Isolation and purification of Gallid herpesvirus 2 strains currently distributed in Japan
  Yuka Machida; Shiro Murata; Ayumi Matsuyama-Kato; Masayoshi Isezaki; Akira Taneno; Eishi Sakai; Satoru Konnai; Kazuhiko Ohashi
  JOURNAL OF VETERINARY MEDICAL SCIENCE, 79, 1, 115, 122, Jan. 2017, [Peer-reviewed]
  English, Scientific journal
• Immune exhaustion during chronic infections in cattle
  Satoru Konnai; Shiro Murata; Kazuhiko Ohashi
  JOURNAL OF VETERINARY MEDICAL SCIENCE, 79, 1, 1, 5, Jan. 2017, [Peer-reviewed]
  English
• Molecular epidemiological survey and genetic characterization of ovine gammaherpesvirus-2 in Mongolian livestock
  Nyamsuren Ochirkhuu; Satoru Konnai; Raadan Odbileg; Shiro Murata; Kazuhiko Ohashi
  Journal of Veterinary Medical Science, 79, 12, 2040, 2042, Japanese Society of Veterinary Science, 2017, [Peer-reviewed]
  English
• Cooperation of PD-1 and LAG-3 Contributes to T-Cell Exhaustion in Anaplasma marginale-Infected Cattle
  Tomohiro Okagawa; Satoru Konnai; James R. Deringer; Massaro W. Ueti; Glen A. Scoles; Shiro Murata; Kazuhiko Ohashi; Wendy C. Brown
  INFECTION AND IMMUNITY, 84, 10, 2779, 2790, Oct. 2016, [Peer-reviewed]
  English, Scientific journal
• Molecular detection and characterization of bovine viral diarrhea virus in Mongolian cattle and yaks
  Nyamsuren Ochirkhuu; Satoru Konnai; Raadan Odbileg; Battogtokh Odzaya; Shura Gansukh; Shiro Murata; Kazuhiko Ohashi
  ARCHIVES OF VIROLOGY, 161, 8, 2279, 2283, Aug. 2016, [Peer-reviewed]
  English, Scientific journal
• Immunohistochemical Analysis of PD-L1 Expression in Canine Malignant Cancers and PD-1 Expression on Lymphocytes in Canine Oral Melanoma
  Naoya Maekawa; Satoru Konnai; Tomohiro Okagawa; Asami Nishimori; Ryoyo Ikebuchi; Yusuke Izumi; Satoshi Takagi; Yumiko Kagawa; Chie Nakajima; Yasuhiko Suzuki; Yukinari Kato; Shiro Murata; Kazuhiko Ohashi
  PLOS ONE, 11, 6, e0157176, Jun. 2016, [Peer-reviewed]
  English, Scientific journal
• Direct polymerase chain reaction from blood and tissue samples for rapid diagnosis of bovine leukemia virus infection
  Asami Nishimori; Satoru Konnai; Ryoyo Ikebuchi; Tomohiro Okagawa; Ayako Nakahara; Shiro Murata; Kazuhiko Ohashi
  JOURNAL OF VETERINARY MEDICAL SCIENCE, 78, 5, 791, 796, May 2016, [Peer-reviewed]
  English, Scientific journal
• Detection of bovine leukemia virus and identification of its genotype in Mongolian cattle
  Nyamsuren Ochirkhuu; Satoru Konnai; Raadan Odbileg; Asami Nishimori; Tomohiro Okagawa; Shiro Murata; Kazuhiko Ohashi
  ARCHIVES OF VIROLOGY, 161, 4, 985, 991, Apr. 2016, [Peer-reviewed]
  English, Scientific journal
• Bovine leukemia virus reduces anti-viral cytokine activities and NK cytotoxicity by inducing TGF-β secretion from regulatory T cells.
  Ohira K; Nakahara A; Konnai S; Okagawa T; Nishimori A; Maekawa N; Ikebuchi R; Kohara J; Murata S; Ohashi K
  Immunity, inflammation and disease, 4, 1, 52, 63, Mar. 2016, [Peer-reviewed], [International Magazine]
  English, Scientific journal, CD4(+)CD25(high)Foxp3(+) T cells suppress excess immune responses that lead to autoimmune and/or inflammatory diseases, and maintain host immune homeostasis. However, CD4(+)CD25(high)Foxp3(+) T cells reportedly contribute to disease progression by over suppressing immune responses in some chronic infections. In this study, kinetic and functional analyses of CD4(+)CD25(high)Foxp3(+) T cells were performed in cattle with bovine leukemia virus (BLV) infections, which have reported immunosuppressive characteristics. In initial experiments, production of the Th1 cytokines IFN-γ and TNF-α was reduced in BLV-infected cattle compared with uninfected cattle, and numbers of IFN-γ or TNF-α producing CD4(+) T cells decreased with disease progression. In contrast, IFN-γ production by NK cells was inversely correlated with BLV proviral loads in infected cattle. Additionally, during persistent lymphocytosis disease stages, NK cytotoxicity was depressed as indicated by low expression of the cytolytic protein perforin. Concomitantly, total CD4(+)CD25(high)Foxp3(+) T cell numbers and percentages of TGF-β(+) cells were increased, suggesting that TGF-β plays a role in the functional declines of CD4(+) T cells and NK cells. In further experiments, recombinant bovine TGF-β suppressed IFN-γ and TNF-α production by CD4(+) T cells and NK cytotoxicity in cultured cells. These data suggest that TGF-β from CD4(+)CD25(high)Foxp3(+) T cells is immunosuppressive and contributes to disease progression and the development of opportunistic infections during BLV infection.
• Identification and the preliminary in vitro characterization of IRIS homologue from salivary glands of Ixodes persulcatus Schulze
  Kochi Toyomane; Satoru Konnai; Ayano Niwa; Naftaly Githaka; Masayoshi Isezaki; Shinji Yamada; Takuya Ito; Ai Takano; Shuji Ando; Hiroki Kawabata; Shiro Murata; Kazuhiko Ohashi
  TICKS AND TICK-BORNE DISEASES, 7, 1, 119, 125, 2016, [Peer-reviewed]
  English, Scientific journal
• Bovine Immunoinhibitory Receptors Contribute to Suppression of Mycobacterium avium subsp paratuberculosis-Specific T-Cell Responses
  Tomohiro Okagawa; Satoru Konnai; Asami Nishimori; Ryoyo Ikebuchi; Seiko Mizorogi; Reiko Nagata; Satoko Kawaji; Shogo Tanaka; Yumiko Kagawa; Shiro Murata; Yasuyuki Mori; Kazuhiko Ohashi
  INFECTION AND IMMUNITY, 84, 1, 77, 89, Jan. 2016, [Peer-reviewed]
  English, Scientific journal
• A preliminary survey of the seroprevalence of Mycobacterium avium subspecies paratuberculosis in Mongolian cattle
  Nyamsuren Ochirkhuu; Satoru Konnai; Raadan Odbileg; Shiro Murata; Kazuhiko Ohashi
  JAPANESE JOURNAL OF VETERINARY RESEARCH, 63, 4, 191, 194, Nov. 2015, [Peer-reviewed]
  English, Scientific journal
• Molecular epidemiological survey and genetic analysis of vector-borne infections of cattle in Luzon Island, the Philippines
  Nyamsuren Ochirkhuu; Satoru Konnai; Claro N. Mingala; Tomohiro Okagawa; Marvin Villanueva; Flor Marie Immanuelle R. Pilapil; Shiro Murata; Kazuhiko Ohashi
  VETERINARY PARASITOLOGY, 212, 3-4, 161, 167, Sep. 2015, [Peer-reviewed]
  English, Scientific journal
• Expression of regulatory dendritic cell-related cytokines in cattle experimentally infected with Trypanosoma evansi
  Hirohisa Mekata; Shiro Murata; Claro Niegos Mingala; Kazuhiko Ohashi; Satoru Konnai
  JOURNAL OF VETERINARY MEDICAL SCIENCE, 77, 8, 1017, 1019, Aug. 2015, [Peer-reviewed]
  English, Scientific journal
• Increased expression of the regulatory T cell-associated marker CTLA-4 in bovine leukemia virus infection
  Saori Suzuki; Satoru Konnai; Tomohiro Okagawa; Ryoyo Ikebuchi; Asami Nishimori; Junko Kohara; Claro N. Mingala; Shiro Murata; Kazuhiko Ohashi
  VETERINARY IMMUNOLOGY AND IMMUNOPATHOLOGY, 163, 3-4, 115, 124, Feb. 2015, [Peer-reviewed]
  English, Scientific journal
• An investigation of binding ability of Ixodes persulcatus Schulze Salp15 with Lyme disease spirochetes.
  Murase Y; Konnai S; Yamada S; Githaka N; Isezaki M; Ito T; Takano A; Ando S; Kawabata H; Murata S; Ohashi K
  Insect Biochemistry and Molecular Biology, 60, 59, 67, 2015, [Peer-reviewed]
  English, Scientific journal
• Influence of PD-L1 cross-linking on cell death in PD-L1-expressing cell lines and bovine lymphocytes
  Ryoyo Ikebuchi; Satoru Konnai; Tomohiro Okagawa; Kazumasa Yokoyama; Chie Nakajima; Yasuhiko Suzuki; Shiro Murata; Kazuhiko Ohashi
  IMMUNOLOGY, 142, 4, 551, 561, Aug. 2014, [Peer-reviewed]
  English, Scientific journal
• Expression analysis of programmed death ligand 2 in tumors caused by the avian oncovirus Marek's disease virus
  Ayumi Matsuyama-Kato; Shiro Murata; Masayoshi Isezaki; Sarah Takasaki; Rika Kano; Satoru Konnai; Kazuhiko Ohashi
  ARCHIVES OF VIROLOGY, 159, 8, 2123, 2126, Aug. 2014, [Peer-reviewed]
  English, Scientific journal
• Differences in cellular function and viral protein expression between IgM(high) and IgM(low) B-cells in bovine leukemia virus-infected cattle
  Ryoyo Ikebuchi; Satoru Konnai; Tomohiro Okagawa; Asami Nishimori; Ayako Nakahara; Shiro Murata; Kazuhiko Ohashi
  JOURNAL OF GENERAL VIROLOGY, 95, Pt 8, 1832, 1842, Aug. 2014, [Peer-reviewed]
  English, Scientific journal
• Identification and characterization of bovine programmed death-ligand 2
  Asami Nishimori; Satoru Konnai; Ryoyo Ikebuchi; Tomohiro Okagawa; Chie Nakajima; Yasuhiko Suzuki; Claro N. Mingala; Shiro Murata; Kazuhiko Ohashi
  MICROBIOLOGY AND IMMUNOLOGY, 58, 7, 388, 397, Jul. 2014, [Peer-reviewed]
  English, Scientific journal
• Expression of PD-L1 on Canine Tumor Cells and Enhancement of IFN-gamma Production from Tumor-Infiltrating Cells by PD-L1 Blockade
  Naoya Maekawa; Satoru Konnai; Ryoyo Ikebuchi; Tomohiro Okagawa; Mami Adachi; Satoshi Takagi; Yumiko Kagawa; Chie Nakajima; Yasuhiko Suzuki; Shiro Murata; Kazuhiko Ohashi
  PLOS ONE, 9, 6, e98415, Jun. 2014, [Peer-reviewed]
  English, Scientific journal
• Identification and sequence characterization of novel Theileria genotypes from the waterbuck (Kobus defassa) in a Theileria parva-endemic area in Kenya
  Naftaly Githaka; Satoru Konnai; Richard Bishop; David Odongo; Isaac Lekolool; Edward Kariuki; Francis Gakuya; Lucy Kamau; Masayoshi Isezaki; Shiro Murata; Kazuhiko Ohashi
  VETERINARY PARASITOLOGY, 202, 3-4, 180, 193, May 2014, [Peer-reviewed]
  English, Scientific journal
• Suppressive effects of neutrophil by Salp16-like salivary gland proteins from Ixodes persulcatus Schulze tick
  Hidano A; Konnai S; Yamada S; Githaka N; Isezaki M; Higuchi H; Nagahata H; Ito T; Takano A; Ando S; Kawabata H; Murata S; Ohashi K
  Insect Molecular Biology, 23, 4, 466, 474, 2014, [Peer-reviewed]
  English, Scientific journal
• Horizontal transmission of bovine leukemia virus from lymphocytotic cattle, and beneficial effects of insect vector control
  Ooshiro M; Konnai S; Katagiri Y; Afuso M; Arakaki N; Tsuha O; Murata S; Ohashi K
  Vet Rec., 173, 21, 527, Nov. 2013, [Peer-reviewed]
  English, Scientific journal
• Genotypic variations in field isolates of Theileria species infecting giraffes (Giraffa camelopardalis tippelskirchi and Giraffa camelopardalis reticulata) in Kenya
  Naftaly Githaka; Satoru Konnai; Robert Skilton; Edward Kariuki; Esther Kanduma; Shiro Murata; Kazuhiko Ohashi
  Parasitology International, 62, 5, 448, 453, Oct. 2013, [Peer-reviewed]
  English, Scientific journal
• Expression analysis of Foxp3 in T cells from bovine leukemia virus infected cattle
  Saori Suzuki; Satoru Konnai; Tomohiro Okagawa; Ryoyo Ikebuchi; Tatsuya Shirai; Yuji Sunden; Claro N. Mingala; Shiro Murata; Kazuhiko Ohashi
  MICROBIOLOGY AND IMMUNOLOGY, 57, 8, 600, 604, Aug. 2013, [Peer-reviewed]
  English, Scientific journal
• Characterization of Meq proteins from field isolates of Marek's disease virus in Japan
  Shiro Murata; Tomoyuki Hashiguchi; Yuko Hayashi; Yuki Yamamoto; Ayumi Matsuyama-Kato; Sarah Takasaki; Masayoshi Isezaki; Misao Onuma; Satoru Konnai; Kazuhiko Ohashi
  INFECTION GENETICS AND EVOLUTION, 16, 137, 143, Jun. 2013, [Peer-reviewed], [Lead author, Corresponding author]
  English, Scientific journal
• Expression analysis of Foxp3 in T-cells from bovine leukemia virus infected cattle.
  Suzuki S; Konnai S; Okagawa T; Ikebuchi R; Shirai T; Sunden Y; Mingala CN; Murata S; Ohashi K
  Microbiology and immunology, Jun. 2013, [Peer-reviewed]
• Bovine whole-blood culture as a tool for the measurement of endotoxin activities in Gram-negative bacterial vaccines
  Saiki Imamura; Mari Nakamizo; Michiko Kawanishi; Nao Nakajima; Kinya Yamamoto; Mariko Uchiyama; Fumiya Hirano; Hidetaka Nagai; Mayumi Kijima; Ryoyo Ikebuchi; Hirohisa Mekata; Shiro Murata; Satoru Konnai; Kazuhiko Ohashi
  VETERINARY IMMUNOLOGY AND IMMUNOPATHOLOGY, 153, 1-2, 153, 158, May 2013, [Peer-reviewed]
  English, Scientific journal
• Isolation, cloning, and pathologic analysis of Trypanosoma evansi field isolates
  Hirohisa Mekata; Satoru Konnai; Claro N. Mingala; Nancy S. Abes; Charito A. Gutierrez; Alan P. Dargantes; William H. Witola; Noboru Inoue; Misao Onuma; Shiro Murata; Kazuhiko Ohashi
  PARASITOLOGY RESEARCH, 112, 4, 1513, 1521, Apr. 2013, [Peer-reviewed]
  English, Scientific journal
• Prevalence of Lyme Borrelia in Ixodes persulcatus Ticks from an Area with a Confirmed Case of Lyme Disease
  Yusuke Murase; Satoru Konnai; Naftaly Githaka; Arata Hidano; Kyle Taylor; Takuya Ito; Ai Takano; Shuji Ando; Hiroki Kawabata; Toshio Tsubota; Shiro Murata; Kazuhiko Ohashi
  JOURNAL OF VETERINARY MEDICAL SCIENCE, 75, 2, 215, 218, Feb. 2013, [Peer-reviewed]
  English, Scientific journal
• Recent viral diseases of Japanese anatid with a fatal case of Marek's disease in an endangered species, white-fronted goose (Anser Albifrons)
  Mitsuhiko Asakawa; Tetsuya Nakade; Shiro Murata; Kazuhiko Ohashi; Yuichi Osa; Hiroyuki Taniyama
  Ducks: Habitat, Behavior and Diseases, 37, 47, Nova Science Publishers, Inc., Jan. 2013
  English, In book
• Enhanced expression of LAG-3 on lymphocyte subpopulations from persistently lymphocytotic cattle infected with bovine leukemia virus
  Satoru Konnai; Saori Suzuki; Tatsuya Shirai; Ryoyo Ikebuchi; Tomohiro Okagawa; Yuji Sunden; Claro N. Mingala; Misao Onuma; Shiro Murata; Kazuhiko Ohashi
  COMPARATIVE IMMUNOLOGY MICROBIOLOGY AND INFECTIOUS DISEASES, 36, 1, 63, 69, Jan. 2013, [Peer-reviewed]
  English, Scientific journal
• Identification and partial characterization of a gut Rhipicephalus appendiculatus cystatin
  Saiki Imamura; Satoru Konnai; Shinji Yamada; Luis F. Parizi; Naftaly Githaka; Itabajara da S. Vaz; Shiro Murata; Kazuhiko Ohashi
  TICKS AND TICK-BORNE DISEASES, 4, 1-2, 138, 144, 2013, [Peer-reviewed]
  English, Scientific journal
• Blockade of bovine PD-1 increases T cell function and inhibits bovine leukemia virus expression in B cells in vitro
  Ryoyo Ikebuchi; Satoru Konnai; Tomohiro Okagawa; Kazumasa Yokoyama; Chie Nakajima; Yasuhiko Suzuki; Shiro Murata; Kazuhiko Ohashi
  Veterinary Research, 44, 1, 59, 2013, [Peer-reviewed]
  English, Scientific journal
• Molecular detection and characterization of potentially new Babesia and Theileria species/variants in wild felids from Kenya
  Naftaly Githaka; Satoru Konnai; Edward Kariuki; Esther Kanduma; Shiro Murata; Kazuhiko Ohashi
  ACTA TROPICA, 124, 1, 71, 78, Oct. 2012, [Peer-reviewed]
  English, Scientific journal
• Transcriptional profiling of inflammatory cytokine genes in African, buffaloes (Syncerus caffer) infected with Theileria parva
  Tomohiro Okagawa; Satoru Konnai; Hirohisa Mekata; Naftaly Githaka; Saori Suzuki; Edward Kariuki; Francis Gakuya; Esther Kanduma; Tatsuya Shirai; Ryoyo Ikebuchi; Yoshinori Ikenaka; Mayumi Ishizuka; Shiro Murata; Kazuhiko Ohashi
  VETERINARY IMMUNOLOGY AND IMMUNOPATHOLOGY, 148, 3-4, 373, 379, Aug. 2012, [Peer-reviewed]
  English, Scientific journal
• Kinetics of regulatory dendritic cells in inflammatory responses during Trypanosoma evansi infection
  H. Mekata; S. Konnai; C. N. Mingala; N. S. Abes; C. A. Gutierrez; A. P. Dargantes; W. H. Witola; N. Inoue; M. Onuma; S. Murata; K. Ohashi
  PARASITE IMMUNOLOGY, 34, 6, 318, 329, Jun. 2012, [Peer-reviewed]
  English, Scientific journal
• Surveillance of Marek's disease virus in migratory and sedentary birds in Hokkaido, Japan
  Shiro Murata; Yuko Hayashi; Ayumi Kato; Masayoshi Isezaki; Sarah Takasaki; Misao Onuma; Yuichi Osa; Mitsuhiko Asakawa; Satoru Konnai; Kazuhiko Ohashi
  VETERINARY JOURNAL, 192, 3, 538, 540, Jun. 2012, [Peer-reviewed], [Lead author, Corresponding author]
  English, Scientific journal
• Increased bovine Tim-3 and its ligand expressions during bovine leukemia virus infection
  Tomohiro Okagawa; Satoru Konnai; Ryoyo Ikebuchi; Saori Suzuki; Tatsuya Shirai; Yuji Sunden; Misao Onuma; Shiro Murata; Kazuhiko Ohashi
  VETERINARY RESEARCH, 43, 45, May 2012, [Peer-reviewed]
  English, Scientific journal
• Molecular characterization of immunoinhibitory factors PD-1/PD-L1 in chickens infected with Marek's disease virus
  Ayumi Matsuyama-Kato; Shiro Murata; Masayoshi Isezaki; Rika Kano; Sara Takasaki; Osamu Ichii; Satoru Konnai; Kazuhiko Ohashi
  VIROLOGY JOURNAL, 9, 94, May 2012, [Peer-reviewed]
  English, Scientific journal
• Molecular cloning and characterization of Th1 and Th2 cytokines of African buffalo (Syncerus caffer)
  S. Suzuki; S. Konnai; T. Okagawa; N. W. Githaka; E. Kariuki; F. Gakuya; E. Kanduma; T. Shirai; R. Ikebuchi; Y. Ikenaka; M. Ishizuka; S. Murata; K. Ohashi
  INTERNATIONAL JOURNAL OF IMMUNOGENETICS, 39, 2, 170, 182, Apr. 2012, [Peer-reviewed]
  English, Scientific journal
• Identification of TROSPA homologue in Ixodes persulcatus Schulze, the specific vector for human Lyme borreliosis in Japan
  Satoru Konnai; Shinji Yamada; Saiki Imamura; Hideto Nishikado; Naftaly Githaka; Takuya Ito; Ai Takano; Hiroki Kawabata; Shiro Murata; Kazuhiko Ohashi
  TICKS AND TICK-BORNE DISEASES, 3, 2, 75, 77, 2012, [Peer-reviewed]
  English, Scientific journal
• Molecular cloning of bovine lymphocyte activation gene-3 and its expression characteristics in bovine leukemia virus-infected cattle
  Tatsuya Shirai; Satoru Konnai; Ryoyo Ikebuchi; Tomohiro Okagawa; Saori Suzuki; Yuji Sunden; Misao Onuma; Shiro Murata; Kazuhiko Ohashi
  VETERINARY IMMUNOLOGY AND IMMUNOPATHOLOGY, 144, 3-4, 462, 467, Dec. 2011, [Peer-reviewed]
  English, Scientific journal
• Increase of cells expressing PD-L1 in bovine leukemia virus infection and enhancement of anti-viral immune responses in vitro via PD-L1 blockade
  Ryoyo Ikebuchi; Satoru Konnai; Tatsuya Shirai; Yuji Sunden; Shiro Murata; Misao Onuma; Kazuhiko Ohashi
  VETERINARY RESEARCH, 42, 103, Sep. 2011, [Peer-reviewed]
  English, Scientific journal
• Analysis of transcriptional activities of the Meq proteins present in highly virulent Marek's disease virus strains, RB1B and Md5
  Shiro Murata; Tsukasa Okada; Rika Kano; Yuko Hayashi; Tomoyuki Hashiguchi; Misao Onuma; Satoru Konnai; Kazuhiko Ohashi
  VIRUS GENES, 43, 1, 66, 71, Aug. 2011, [Peer-reviewed], [Lead author, Corresponding author]
  English, Scientific journal
• Molecular detection of Anaplasma phagocytophilum in cattle and Ixodes persulcatus ticks
  Yusuke Murase; Satoru Konnai; Arata Hidano; Naftali W. Githaka; Takuya Ito; Ai Takano; Hiroki Kawabata; Manabu Ato; Tomoko Tajima; Motoshi Tajima; Misao Onuma; Shiro Murata; Kazuhiko Ohashi
  VETERINARY MICROBIOLOGY, 149, 3-4, 504, 507, May 2011, [Peer-reviewed]
  English, Scientific journal
• Molecular identification and expression analysis of lipocalins from blood feeding taiga tick, Ixodes persulcatus Schulze
  Satoru Konnai; Hideto Nishikado; Shinji Yamada; Saiki Imamura; Takuya Ito; Misao Onuma; Shiro Murata; Kazuhiko Ohashi
  EXPERIMENTAL PARASITOLOGY, 127, 2, 467, 474, Feb. 2011, [Peer-reviewed]
  English, Scientific journal
• Serological survey of Toxoplasma gondii in wild waterfowl in Chukotka, Kamchatka, Russia and Hokkaido, Japan
  Takehiro Murao; Yoshitaka Omata; Rika Kano; Shiro Murata; Tsukasa Okada; Satoru Konnai; Mitsuhiko Asakawa; Kazuhiko Ohashi; Misao Onuma
  JOURNAL OF PARASITOLOGY, 94, 4, 830, 833, Aug. 2008, [Peer-reviewed]
  English, Scientific journal
• Development of a nested polymerase chain reaction method to detect oncogenic Marek's disease virus from feather tips
  Shiro Murata; Kyung-Soo Chang; Sung-Il Lee; Satoru Konnai; Misao Onuma; Kazuhiko Ohashi
  JOURNAL OF VETERINARY DIAGNOSTIC INVESTIGATION, 19, 5, 471, 478, Sep. 2007, [Peer-reviewed], [Lead author]
  English, Scientific journal
• Detection of the virulent Marek's disease virus genome from feather tips of wild geese in Japan and the Far East region of Russia
  S. Murata; K.-S. Chang; Y. Yamamoto; T. Okada; S.-I. Lee; S. Konnai; M. Onuma; Y. Osa; M. Asakawa; K. Ohashi
  ARCHIVES OF VIROLOGY, 152, 8, 1523, 1526, Aug. 2007, [Peer-reviewed], [Lead author]
  English, Scientific journal
• Identification and characterization of a novel spliced form of the meq transcript in lymphoblastoid cell lines derived from Marek's disease tumours
  Tsukasa Okada; Michihiro Takagi; Shiro Murata; Misao Onuma; Kazuhiko Ohashi
  JOURNAL OF GENERAL VIROLOGY, 88, 8, 2111, 2120, Aug. 2007, [Peer-reviewed]
  English, Scientific journal
• Rapid detection and differentiation of Newcastle disease virus by real-time PCR with melting-curve analysis
  HM Pham; S Konnai; T Usui; KS Chang; S Murata; M Mase; K Ohashi; M Onuma
  ARCHIVES OF VIROLOGY, 150, 12, 2429, 2438, Dec. 2005, [Peer-reviewed]
  English, Scientific journal

• 馬精上皮腫に対する免疫チェックポイント阻害剤を用いた臨床試験研究の1例
  堀川壮琉; 岡川朋弘; 岡川朋弘; 前川直也; 北内諒; 山家崇史; 村田史郎; 大橋和彦; 加藤史樹; 今内覚; 今内覚, 北海道獣医師会雑誌, 69, 8, 2025
• Effect of polymorphisms in the meq gene of Marek’s disease virus distributed in Japan on the pathogenicity and transcriptional regulation
  もたい吉之介; 村田史郎; 村田史郎; 佐藤純平; 山上竣裕; SHWE Win Yee; 堀尾文彌; 前川直也; 岡川朋弘; 今内覚; 今内覚; 今内覚; 大橋和彦; 大橋和彦; 大橋和彦, 日本ウイルス学会学術集会プログラム・予稿集(Web), 71st, 2024
• Effect of polymorphisms in the basic region of Meq protein of Marek’s disease, an avian oncogenic herpesvirus, on virulence
  佐藤純平; 村田史郎; 村田史郎; もたい吉之介; 山上竣裕; 堀尾文彌; SHWE Win Yee; 前川直也; 岡川朋弘; 今内覚; 今内覚; 今内覚; 大橋和彦; 大橋和彦; 大橋和彦, 日本ウイルス学会学術集会プログラム・予稿集(Web), 71st, 2024
• 非典型的な地方病性牛伝染性リンパ腫症例のRAISING法を用いた性状解析
  松原幸芽; 岡川朋弘; 斎藤益満; 松平崇弘; 直亨則; 神谷可菜; 齋藤麻矢; 前川直也; 村田史郎; 大橋和彦; 今内覚, 日本獣医学会学術集会講演要旨集, 167th, 2024
• 抗PD-L1抗体により治療した悪性黒色腫の犬における血清バイオマーカーの探索
  飯高早紀子; 前川直也; 今内覚; 出口辰弥; 岡川朋弘; 高木哲; 高木哲; 鈴木定彦; 鈴木定彦; 村田史郎; 大橋和彦, 日本獣医学会学術集会講演要旨集, 167th, 2024
• プロバイオティクスによる子牛のワクチン効果増強作用の検証
  池端麻里; 岡川朋弘; 中村隼人; 前川直也; 村田史郎; 大橋和彦; 今内覚; 今内覚, 日本獣医学会学術集会講演要旨集, 167th, 2024
• 日本に分布するマレック病ウイルスmeq遺伝子による病原性への影響
  もたい吉之介; 村田史郎; 佐藤純平; 山上竣裕; WIN Shwe Yee; 堀尾文彌; 前川直也; 岡川朋弘; 今内覚; 今内覚; 大橋和彦, 日本獣医学会学術集会講演要旨集, 167th, 2024
• ワクモ曝露型抗原による抗ワクモワクチン抗原としての評価
  堀尾文彌; 村田史郎; ウィン シュウェ・イー; 佐藤純平; もたい吉之介; 山上竣裕; 佐藤匠; 大石英司; 前川直也; 岡川朋弘; 今内覚; 大橋和彦, 日本獣医学会学術集会講演要旨集, 167th, 2024
• マレック病ウイルスmeq遺伝子の転写活性化調節ドメインの多型と病原性との関連
  山上竣裕; 村田史郎; 佐藤純平; もたい吉之介; WIN Shwe Yee; 堀尾文彌; 前川直也; 岡川朋弘; 今内覚; 大橋和彦, 日本獣医学会学術集会講演要旨集, 167th, 2024
• 抗ワクモワクチンを用いたワクモの制御法の開発(1)
  大橋和彦; 村田史郎, 鶏の研究, 99, 6, 2024
• 抗ワクモワクチンを用いたワクモの制御法の開発(2)
  大橋和彦; 村田史郎, 鶏の研究, 99, 8, 2024
• 抗ワクモワクチンを用いたワクモの制御法の開発(3)
  大橋和彦; 村田史郎, 鶏の研究, 99, 10, 2024
• 子牛における地方病性牛伝染性リンパ腫発症は母牛由来の腫瘍細胞が原因か?
  廣瀬海凪; 今内覚; 岡川朋弘; 富永みその; 松原幸芽; 前川直也; 村田史郎; 大橋和彦, 北海道獣医師会雑誌, 68, 8, 2024
• 抗ワクモワクチン開発に向けた試み
  村田史郎, 衛生動物, 75, Supplement, 2024
• 牛伝染性リンパ腫の診断と早期発症予測法の開発
  富永みその; 今内覚; 岡川朋弘; 神谷可菜; 齋藤麻矢; 安富一郎; 目堅博久; 前川直也; 村田史郎; 大橋和彦, 北海道獣医師会雑誌, 67, 8, 2023
• Chimeric provirus of bovine leukemia virus/ SMAD family member 3 in cattle with enzootic bovine leukosis
  直亨則; 直亨則; 岡川朋弘; 野尻直未; 今内覚; 今内覚; 嶋倉穂南; 富永みその; 吉田初佳; 西山依里; 松平崇弘; 前川直也; 村田史郎; 村田史郎; 村松正道; 村松正道; 大橋和彦; 大橋和彦; 斎藤益満, 日本ウイルス学会学術集会プログラム・予稿集(Web), 70th, 3, 47, 47, 2023, [International Magazine]
  Bovine leukemia virus (BLV) is a member of the family Retroviridae that causes enzootic bovine leukemia (EBL). However, the association between BLV infection and EBL development remains unclear. In this study, we identified a BLV/SMAD3 chimeric provirus within CC2D2A intron 30 in monoclonal expanded malignant cells from a cow with EBL. The chimeric provirus harbored a spliced SMAD3 sequence composed of exons 3-9, encoding the short isoform protein, and the BLV-SMAD3 chimeric transcript was detectable in cattle with EBL. This is the first report of a BLV chimeric provirus that might be involved in EBL tumorigenesis., English
• 免疫チェックポイント分子Programmed death-ligand1(PD-L1)を標的とした抗体薬による犬の鼻腔内腺癌に対する免疫療法の検討
  多田佳史; 前川直也; 今内覚; 細谷謙次; 大脇稜; 竹内寛人; 賀川由美子; 高木哲; 高木哲; 鈴木定彦; 鈴木定彦; 岡川朋弘; 村田史郎; 大橋和彦, 日本獣医学会学術集会講演要旨集, 166th, 2023
• 家禽疾病の発生防止に向けた疫学調査,病態解析および制御法に関する研究
  村田史郎, 日本獣医学会学術集会講演要旨集, 166th, 2023
• 牛伝染性リンパ腫ウイルス感染症における免疫チェックポイント因子TIM-3およびPD-L1を標的とした抗ウイルス効果の検討
  中村隼人; 今内覚; 岡川朋弘; 前川直也; 村田史郎; 大橋和彦, 日本獣医学会学術集会講演要旨集, 166th, 2023
• クローナリティ解析による地方病性牛伝染性リンパ腫に対する発症予測法の開発
  富永みその; 岡川朋弘; 嶋倉穂南; 斎藤益満; 松平崇弘; 直亨則; 山田慎二; 村上賢二; 前川直也; 村田史郎; 大橋和彦; 今内覚, 日本獣医学会学術集会講演要旨集, 166th, 2023
• マレック病ウイルスMeqタンパク質における挿入配列による病原性増強機構の解明
  佐藤純平; 村田史郎; YEE Win Shwe; 瀬尾光里; もたい吉之介; 前川直也; 岡川朋弘; 今内覚; 大橋和彦, 日本獣医学会学術集会講演要旨集, 166th, 2023
• 新たに認められた欠損配列を持つマレック病ウイルスmeq遺伝子の性状解析
  もたい吉之介; 村田史郎; 佐藤純平; 西明仁; 前川直也; 岡川朋弘; 今内覚; 大橋和彦, 日本獣医学会学術集会講演要旨集, 166th, 2023
• ネコ腫瘍組織におけるprogrammed death ligand1(PD-L1)の発現解析
  青木絵理; 今内覚; 今内覚; 前川直也; 岡川朋弘; 竹内寛人; 佐藤純平; 浅野裕美恵; 大塚拓海; 賀川由美子; 加藤幸成; 高木哲; 鈴木定彦; 村田史郎; 村田史郎; 大橋和彦; 大橋和彦; 大橋和彦, 日本獣医学会学術集会講演要旨集, 165th (CD-ROM), 2022
• ワクモ由来の2種類のアスパラギン酸ペプチダーゼの性状解析
  瀬尾光里; 村田史郎; 村田史郎; WIN Shwe Yee; 藤澤宗太郎; 北條巧; 伊勢崎政美; 佐藤匠; 大石英司; 種子野章; 市居修; 市居修; 前川直也; 岡川朋弘; 今内覚; 今内覚; 大橋和彦; 大橋和彦; 大橋和彦, 日本獣医学会学術集会講演要旨集, 165th (CD-ROM), 2022
• 鼻疽の診断抗原の探索
  市川世識; 飯沼由希帆; 岡川朋弘; 前川直也; 村田史郎; 今内覚; 木下優太; 丹羽秀和; 青島圭佑; 小林篤史; 大橋和彦; 木村享史, 日本獣医学会学術集会講演要旨集, 165th (CD-ROM), 2022
• 牛伝染性リンパ腫ウイルス感染牛における分娩が及ぼす免疫抑制メカニズムの解析
  富永みその; 今内覚; 佐治木大和; 岡川朋弘; 小原潤子; 似内厚之; 高橋博文; 窪田健太郎; 武田休史; 前川直也; 村田史郎; 大橋和彦, 北海道獣医師会雑誌, 66, 8, 2022
• 牛伝染性リンパ腫の病態発生機序解析を基盤とする新制御法の試み
  今内覚; 岡川朋弘; 前川直也; 村田史郎; 大橋和彦, 家畜衛生学雑誌, 47, 3, 2021
• Expression and functional analysis of swine PD-1 /PD-L1 pathway
  OTGONTUYA Ganbaatar; 今内覚; 岡川朋弘; 野島裕太郎; 前川直也; 市川世識; 小林篤史; 芝原友幸; 柳川洋二郎; 鈴木定彦; 村田史郎; 大橋和彦, 日本獣医学会学術集会講演要旨集, 164th (CD-ROM), 2021
• ワクモ(Dermanyssus gallinae)の吸血に対する宿主免疫応答の解明
  藤澤宗太郎; 村田史郎; 伊勢崎政美; 佐藤匠; 大石英司; 種子野章; 前川直也; 岡川朋弘; 今内覚; 大橋和彦, 日本獣医学会学術集会講演要旨集, 164th (CD-ROM), 2021
• 【分子標的薬2020 いま知っておくべき獣医療における分子標的療法の現在と未来】将来の展望 免疫チェックポイント阻害薬を用いた腫瘍免疫療法
  前川 直也; 今内 覚; 岡川 朋弘; 村田 史郎; 大橋 和彦, Veterinary Oncology, 7, 2, 89, 94, Apr. 2020
  (株)エデュワードプレス, Japanese
• ウシマイコプラズマ感染症における免疫疲弊化
  今内 覚; 後藤 伸也; 岡川 朋弘; 前川 直也; 村田 史郎; 大橋 和彦, 日本マイコプラズマ学会雑誌, 46, 46, 31, 33, Mar. 2020, [Invited]
  日本マイコプラズマ学会, Japanese, Report scientific journal
• 動物用抗体医薬品の開発および臨床研究
  今内 覚; 岡川 朋弘; 前川 直也; 中島 千絵; 鈴木 定彦; 山本 啓一; 戸田 幹洋; 村田 史郎; 大橋 和彦, JVPA digest : 日本動物用医薬品協会会報, 67, 1, 10, Feb. 2020
  日本動物薬事協会, Japanese
• 牛白血病の病態発生機序解析を基盤とする新制御法の試み
  今内覚; 岡川朋弘; 前川直也; 村田史郎; 大橋和彦, 畜産技術, 784, 2020
• 牛白血病ウイルス感染症における免疫抑制受容体TIM-3の発現解析および機能解析
  中村隼人; 今内覚; 岡川朋弘; 佐治木大和; 渡慧; 神谷可菜; 齋藤麻矢; 前川直也; 村田史郎; 大橋和彦, 日本獣医学会学術集会講演要旨集, 163rd, 221, 221, 2020
  (公社)日本獣医学会, Japanese
• ネコCytotoxic T-lymphocyte antigen-4(CTLA-4)を標的とした新規免疫抑制剤の開発における基礎的検討
  大塚拓海; 今内覚; 前川直也; 渡慧; 岡川朋弘; 村田史郎; 大橋和彦, 日本獣医学会学術集会講演要旨集, 163rd, 221, 221, 2020
  (公社)日本獣医学会, Japanese
• ワクモにおける定量的PCR法確立のための内在性コントロール遺伝子の探索
  有泉拓馬; 村田史郎; 藤澤宗太郎; 伊勢崎政美; 前川直也; 岡川朋弘; 種子野章; 大石英司; 今内覚; 大橋和彦, 日本獣医学会学術集会講演要旨集, 163rd, 248, 248, 2020
  (公社)日本獣医学会, Japanese
• Tick factors associated pathogen transmission~Tick immunosuppressants in saliva~
  今内覚; 今内覚; 岡川朋弘; 前川直也; 村田史郎; 村田史郎; 大橋和彦; 大橋和彦, 衛生動物, 70, 4, 189, 197, Dec. 2019
  マダニの唾液の役割と、唾液に含まれる免疫抑制因子を中心に取り上げ、同定され始めた病原体伝播に関わるマダニ因子について紹介した。ダニの唾液または唾液を産生する唾液腺を用いた研究から、マダニは宿主免疫を撹乱したり抑制したりすることが報告されており、唾液は直接的または間接的に病原体の伝播を補助・促進することも明らかとなっている。同定された免疫抑制因子のうちSalp15は宿主CD4+T細胞のCD4に結合し、IL-2産生の抑制によるCD4+T細胞の増殖および活性化を阻害する。さらに近年になって、マダニの吸血補助や維持に関わる因子の他に、ダニ体内において病原体の伝播や定着に関与する特異的な分子の存在が明らかとなってきた。, 日本衛生動物学会, Japanese
• イヌ腫瘍組織およびイヌ腫瘍由来細胞株におけるイヌHER2の発現解析
  吉武 志江奈; 今内 覚; 前川 直也; 賀川 由美子; 西村 麻紀; 岡川 朋弘; 鈴木 定彦; 高木 哲; 中川 貴之; 村田 史郎; 大橋 和彦, 日本獣医学会学術集会講演要旨集, 162回, 380, 380, Aug. 2019
  (公社)日本獣医学会, Japanese
• イヌ腫瘍由来細胞株におけるCKLF-like MARVEL transmembrane domain containing protein 6(CMTM6)および4(CMTM4)の発現解析
  竹内 寛人; 今内 覚; 岡川 朋弘; 前川 直也; 村田 史郎; 大橋 和彦, 日本獣医学会学術集会講演要旨集, 162回, 380, 380, Aug. 2019
  (公社)日本獣医学会, Japanese
• BLV感染症に対するCOX-2阻害剤と抗PD-L1抗体併用法の抗ウイルス効果の検討
  佐治木 大和; 今内 覚; 岡川 朋弘; 前川 直也; 後藤 伸也; 小原 潤子; 山田 慎二; 加藤 幸成; 鈴木 定彦; 村田 史郎; 大橋 和彦, 日本獣医学会学術集会講演要旨集, 162回, 381, 381, Aug. 2019
  (公社)日本獣医学会, Japanese
• Mycoplasma bovis感染症に対する抗PD-L1キメラ抗体を用いた臨床試験
  後藤 伸也; 今内 覚; 平野 佑気; 小原 潤子; 岡川 朋弘; 前川 直也; 鈴木 定彦; 加藤 幸成; 村田 史郎; 大橋 和彦, 日本獣医学会学術集会講演要旨集, 162回, 381, 381, Aug. 2019
  (公社)日本獣医学会, Japanese
• ミャンマーの養鶏場におけるマイコプラズマ、鶏伝染性気管支炎ウイルス、鶏伝染性喉頭気管炎ウイルスの分子学的検出と遺伝学的性状解析(Molecular detection and genetic characterization of mycoplasmas, infectious bronchitis virus and infectious laryngotracheitis virus in poultry farms in Myanmar)
  Yang Zhiyuan; 藤澤 宗太郎; 村田 史郎; 竹原 昌生; 片倉 賢; Myint Myint Hmoon; Shwe Yee Win; Saw Bawm; Lat Lat Htun; Ye Htut Aung; Mar Mar Win; 今内 覚; 大橋 和彦, 日本獣医学会学術集会講演要旨集, 162回, 411, 411, Aug. 2019
  (公社)日本獣医学会, English
• RNA-Seqを用いたワクモ(Dermanyssus gallinae)の吸血状態別における遺伝子発現解析
  藤澤 宗太郎; 村田 史郎; 竹原 昌生; 伊勢崎 政美; 小川 遼; 宇野 有紀子; 種子野 章; 前川 直也; 岡川 朋弘; 今内 覚; 大橋 和彦, 日本獣医学会学術集会講演要旨集, 162回, 412, 412, Aug. 2019
  (公社)日本獣医学会, Japanese
• COX-2阻害剤の併用による抗PD-L1抗体を用いたイヌ腫瘍免疫療法の効果増強に向けた基礎的検討
  前川 直也; 今内 覚; 浅野 裕美恵; 岡川 朋弘; 高木 哲; 村田 史郎; 大橋 和彦, 日本獣医学会学術集会講演要旨集, 162回, 460, 460, Aug. 2019
  (公社)日本獣医学会, Japanese
• 免疫チェックポイント分子Programmed death‐ligand 1(PD‐L1)を標的とする抗体薬による免疫療法が奏功した肺転移のある口腔内悪性黒色腫の犬の1例
  竹内寛人; 前川直也; 今内覚; 高木哲; 細谷謙次; 賀川由美子; 岡川朋弘; 鈴木定彦; 鈴木定彦; 山本啓一; 山本啓一; 村田史郎; 大橋和彦, 北海道獣医師会雑誌, 63, 8, 343, 343, Aug. 2019
  (公社)北海道獣医師会, Japanese
• ロタウイルス実験感染子牛モデルを用いた3.5倍発酵代用乳の腸炎抑制効果の検証
  茅先史; 今内覚; 久保田学; 岡川朋弘; 佐治木大和; 渡慧; 小原潤子; 前川直也; 村田史郎; 大橋和彦, 北海道獣医師会雑誌, 63, 8, 302, 302, Aug. 2019
  (公社)北海道獣医師会, Japanese
• 免疫学的解析を基盤とした動物用創薬研究
  今内覚; 岡川朋弘; 前川直也; 中島千絵; 中島千絵; 鈴木定彦; 鈴木定彦; 山本啓一; 山本啓一; 戸田幹洋; 戸田幹洋; 村田史郎; 大橋和彦, 臨床獣医, 37, 3, 37, 41, Mar. 2019
  (株)緑書房, Japanese
• 抗ワクモワクチン開発に向けた試み
  村田史郎, 鶏病研究会報, 54, 4, 209‐210, 25 Feb. 2019, [Invited]
  Japanese
• ワクモ由来Adipocyte plasma membrane-associated proteinの抗ワクモワクチン抗原としての評価
  森田鮎; 竹原昌生; 村田史郎; 伊勢崎政美; 藤澤宗太郎; 種子野章; 酒井英史; 宇野有紀子; 小川遼; 市居修; 前川直也; 岡川朋弘; 今内覚; 大橋和彦, 日本獣医学会学術集会講演要旨集, 162nd, 2019
• 動物難治性疾病に対する創薬研究
  今内 覚; 岡川朋弘; 前川直也; 中島千絵; 鈴木定彦; 山本啓一; 戸田幹洋; 村田史郎; 大橋和彦, MPアグロジャーナル, 35, 10, 22, 25, Oct. 2018, [Invited]
  MPアグロ, Japanese, Introduction scientific journal
• 小動物医療における腫瘍の免疫療法 2 治療篇 免疫チェックポイント分子を標的とした腫瘍免疫療法―抗PD‐1/PD‐L1抗体療法
  前川直也; 今内覚; 村田史郎; 大橋和彦; 高木哲; 高木哲, 獣医畜産新報, 71, 9, 659, 663, Sep. 2018
  文永堂出版(株), Japanese
• 牛白血病における新規病態進行評価マーカーの検索
  渡慧; 今内覚; 佐治木大和; 岡川朋弘; 前川直也; 後藤伸也; 村田史郎; 大橋和彦, 北海道獣医師会雑誌, 62, 8, 282, 282, Aug. 2018
  (公社)北海道獣医師会, Japanese
• マレック病ウイルスの病原性進化機構の解明―病原性試験や全ゲノム解析の試み
  大橋和彦; 村田史郎; 町田柚香; 伊勢崎政美; 今内覚, 日本獣医学会学術集会講演要旨集, 161回, 216, 216, Aug. 2018
  (公社)日本獣医学会, Japanese
• 免疫抑制因子PD‐1およびPD‐L1を標的としたイヌ腫瘍に対する新規免疫療法の検討
  前川直也; 今内覚; 村田史郎; 大橋和彦, 日本獣医学会学術集会講演要旨集, 161回, 233, 233, Aug. 2018
  (公社)日本獣医学会, Japanese
• 動物難治性疾病の免疫学的解析を基盤とした新規制御法の開発
  今内覚; 村田史郎; 大橋和彦, 日本獣医学会学術集会講演要旨集, 161回, 202, 202, Aug. 2018
  (公社)日本獣医学会, Japanese
• 地方病性牛白血病の簡易鑑別診断基準の確立と若齢発症牛の実態調査
  田中晶菜; 今内覚; 岡川朋弘; 西森朝美; 前川直也; 戸塚知恵; 千葉由純; 池田昌穂; 村田史郎; 大橋和彦, 日本獣医学会学術集会講演要旨集, 161回, 353, 353, Aug. 2018
  (公社)日本獣医学会, Japanese
• ヨーネ病におけるプロスタグランジンE₂(PGE₂)動態解析及びCOX‐2阻害剤の免疫活性化効果の検討
  佐治木大和; 今内覚; 岡川朋弘; 西森朝美; 前川直也; 後藤伸也; 永田礼子; 川治聡子; 森康行; 村田史郎; 大橋和彦, 日本獣医学会学術集会講演要旨集, 161回, 341, 341, Aug. 2018
  (公社)日本獣医学会, Japanese
• ウシCytotoxic T‐lymphocyte antigen‐4(CTLA‐4)の機能解析および慢性感染症における発現解析
  渡慧; 今内覚; 岡川朋弘; 前川直也; 後藤伸也; 佐治木大和; 村田史郎; 鈴木定彦; 大橋和彦, 日本獣医学会学術集会講演要旨集, 161回, 340, 340, Aug. 2018
  (公社)日本獣医学会, Japanese
• 肺転移のある悪性黒色腫罹患犬に対するPD‐L1を標的とした抗体薬の治療効果
  前川直也; 今内覚; 高木哲; 細谷謙次; 賀川由美子; 岡川朋弘; 和泉雄介; 出口辰弥; 鈴木定彦; 山本啓一; 村田史郎; 大橋和彦, 日本獣医学会学術集会講演要旨集, 161回, 429, 429, Aug. 2018
  (公社)日本獣医学会, Japanese
• イヌCytotoxic T‐lymphocyte antigen‐4(CTLA‐4)を標的としたバイオ医薬品の開発に向けた基礎的検討
  石原悠太郎; 今内覚; 岡川朋弘; 前川直也; 鈴木定彦; 大田寛; 村田史郎; 大橋和彦, 日本獣医学会学術集会講演要旨集, 161回, 430, 430, Aug. 2018
  (公社)日本獣医学会, Japanese
• Mycoplasma bovis感染症における免疫抑制機序の解析
  後藤伸也; 今内覚; 岡川朋弘; 前川直也; 佐治木大和; 渡慧; 樋口豪紀; 小岩政照; 田島誉士; 鈴木定彦; 村田史郎; 大橋和彦, 日本獣医学会学術集会講演要旨集, 161回, 342, 342, Aug. 2018
  (公社)日本獣医学会, Japanese
• 地方病性牛白血病若齢発症牛におけるプロウイルス挿入部位の網羅的解析
  岡川朋弘; 今内覚; 西森朝美; 田中晶菜; 前川直也; 戸塚知恵; 千葉由純; 池田昌穂; 村田史郎; 大橋和彦, 日本獣医学会学術集会講演要旨集, 161回, 351, 351, Aug. 2018
  (公社)日本獣医学会, Japanese
• ミャンマー連邦共和国における鶏の吸血性外部寄生虫の分布状況調査
  竹原昌生; 村田史郎; 片倉賢; MYINT MYINT Hmoon; SHWE YEE Win; SAW Bawm; LAT LAT Htun; YE HTUT Aung; MAR MAR Win; 伊勢崎政美; 今内覚; 大橋和彦, 日本獣医学会学術集会講演要旨集, 161回, 375, 375, Aug. 2018
  (公社)日本獣医学会, Japanese
• ワクモ(Dermanyssus gallinae)由来カテプシンL用タンパク質の性状解析とワクチン抗原としての評価
  村田史郎; 伊勢崎政美; 谷口綾香; 北條巧; 種子野章; 酒井英史; 宇野有紀子; 矢吹卓也; 市居修; 伊東拓也; 今内覚; 大橋和彦, 衛生動物, 69, 2, 103, 103, Jun. 2018
  日本衛生動物学会, Japanese
• 獣医療における抗体医薬の現状
  今内覚; 村田史郎; 大橋和彦, 家畜診療, 658, 233‐240, 01 Apr. 2018
  Japanese
• 日本に分布するマレック病ウイルス野外株の病原性の検討
  村田史郎; 種子野章; 酒井英史; 町田柚香; 松山あゆ美; 伊勢崎政美; 今内覚; 大橋和彦, 日本獣医学会学術集会講演要旨集, 160th, 405, 30 Aug. 2017
  Japanese
• マレック病ウイルス弱毒株を用いた新規抗ワクモワクチン開発に向けた基礎研究
  青山珠里愛; 村田史郎; 伊勢崎政美; 種子野章; 酒井英史; 今内覚; 大橋和彦, 日本獣医学会学術集会講演要旨集, 160th, 405, 30 Aug. 2017
  Japanese
• 牛マイコプラズマ感染症における免疫抑制機序の解明
  後藤伸也; 今内覚; 岡川朋弘; 西森朝美; 前川直也; 樋口豪紀; 小岩政照; 田島誉士; 小原潤子; 加藤幸成; 鈴木定彦; 村田史郎; 大橋和彦, 日本獣医学会学術集会講演要旨集, 160th, 370, 370, 30 Aug. 2017
  (公社)日本獣医学会, Japanese
• Theileria parva感染症における免疫チェックポイント因子の動態解析
  今内覚; 岡山朋弘; 山田慎二; SIMUUNZA Martin; CONNELLEY Timothy; MORRISON Ivan; 村田史郎; 大橋和彦, 日本獣医学会学術集会講演要旨集, 160th, 338, 30 Aug. 2017
  Japanese
• PD‐L1を標的としたイヌ用キメラ抗体医薬の作製と悪性黒色腫および未分化肉腫罹患犬における抗腫瘍効果の検討
  前川直也; 今内覚; 高木哲; 賀川由美子; 岡川朋弘; 西森朝美; 池渕良洋; 和泉雄介; 出口辰弥; 加藤幸成; 鈴木定彦; 村田史郎; 大橋和彦, 日本獣医学会学術集会講演要旨集, 160th, 460, 460, 30 Aug. 2017
  (公社)日本獣医学会, Japanese
• 牛白血病ウイルス(BLV)感染症においてプロスタグランジンE₂(PGE₂)が及ぼす免疫抑制機序の解明
  佐治木大和; 今内覚; 西森朝美; 岡川朋弘; 村田史郎; 大橋和彦, 日本獣医学会学術集会講演要旨集, 160th, 370, 30 Aug. 2017
  Japanese
• BLV感染症をモデルとした抗ウシPD‐L1キメラ抗体の臨床応用試験
  西森朝美; 今内覚; 岡川朋弘; 前川直也; 池渕良洋; 後藤伸也; 佐治木大和; 鈴木定彦; 小原潤子; 小笠原諭; 加藤幸成; 村田史郎; 大橋和彦, 日本獣医学会学術集会講演要旨集, 160th, 370, 370, 30 Aug. 2017
  (公社)日本獣医学会, Japanese
• COX‐2阻害薬と抗PD‐L1抗体の併用によるイヌ腫瘍疾患の制御法の検討
  浅野裕美恵; 今内覚; 岡川朋弘; 前川直也; 西森朝美; 後藤伸也; 高木哲; 村田史郎; 大橋和彦, 日本獣医学会学術集会講演要旨集, 160th, 371, 371, 30 Aug. 2017
  (公社)日本獣医学会, Japanese
• 牛白血病ウイルス(BLV)感染ハイリスク牛における初めての子宮内感染直接証明
  佐治木大和; 今内覚; 西森朝美; 岡川朋弘; 永野昌志; 小原潤子; 村田史郎; 大橋和彦, 北海道獣医師会雑誌, 61, 8, 280, 10 Aug. 2017
  Japanese
• マダニ唾液由来因子の病原体伝播における役割
  今内覚; 伊東拓也; 高野愛; 川端寛樹; 安藤秀二; 村田史郎; 大橋和彦, 日本細菌学雑誌(Web), 72, 1, 35(J‐STAGE), 2017
  Japanese
• ウシCytotoxic T‐lymphocyte antigen‐4(CTLA‐4)の機能解析と特異抗体の樹立
  渡慧; 今内覚; 前川直也; 岡川朋弘; 西森朝美; 鈴木定彦; 村田史郎; 大橋和彦, 日本獣医学会学術集会講演要旨集, 159th, 365, 365, 30 Aug. 2016
  (公社)日本獣医学会, Japanese
• 家畜における免疫チェックポイント関連因子の遺伝子比較解析とウシ用抗体を用いた動物種横断的研究
  野島裕太郎; 今内覚; 西森朝美; 前川直也; 岡川朋弘; 森康行; 賀川由美子; 鈴木定彦; 村田史郎; 大橋和彦, 日本獣医学会学術集会講演要旨集, 159th, 364, 364, 30 Aug. 2016
  (公社)日本獣医学会, Japanese
• シュルツェマダニ(Ixodes persulcatus)由来sialostatinL2の機能解析
  越智晶絵; 今内覚; 伊東拓也; 川端寛樹; 高野愛; 安藤秀二; 村田史郎; 大橋和彦, 日本獣医学会学術集会講演要旨集, 159th, 339, 30 Aug. 2016
  Japanese
• ウシ腫瘍壊死因子(TNF‐α)デコイレセプターによる新規抗炎症剤の開発
  藤澤宗太郎; 今内覚; 岡川朋弘; 西森朝美; 前川直也; 村田史郎; 鈴木定彦; 大橋和彦, 日本獣医学会学術集会講演要旨集, 159th, 364, 364, 30 Aug. 2016
  (公社)日本獣医学会, Japanese
• マレック病ウイルス日本分離株の全ゲノム解析
  村田史郎; 町田柚香; 伊勢崎政美; 今内覚; 大橋和彦, 日本獣医学会学術集会講演要旨集, 159th, 399, 30 Aug. 2016
  Japanese
• 抗ワクモ(Dermanyssus gallinae)ワクチン開発に向けたワクモ由来フェリチン2の性状解析
  伊勢崎政美; 村田史郎; 酒井英史; 矢吹卓也; 種子野章; 市居修; 伊東拓也; 今内覚; 大橋和彦, 日本獣医学会学術集会講演要旨集, 159回, 399, 399, Aug. 2016
  (公社)日本獣医学会, Japanese
• 牛白血病への対策を練る 牛白血病について
  今内覚; 村田史郎; 大橋和彦, 養牛の友, 484, 32‐36, 36, 01 Jul. 2016
  日本畜産振興会, Japanese
• 増加する牛白血病とその対策
  今内覚; 村田史郎; 大橋和彦, 農家の友, 68, 2, 108‐110, 110, 01 Feb. 2016
  北海道農業改良普及協会, Japanese
• 牛白血病ウイルス感染症における疲弊化T細胞の動態とその再活性化
  岡川朋弘; 今内覚; 西森朝美; 前川直也; 池渕良洋; 村田史郎; 大橋和彦, 日本獣医学会学術集会講演要旨集, 158th, 324, 324, 30 Aug. 2015
  (公社)日本獣医学会, Japanese
• イヌ腫瘍浸潤リンパ球におけるPD‐1発現解析と免疫チェックポイント分子を標的とした新規腫瘍免疫療法の検討
  前川直也; 今内覚; 池渕良洋; 岡川朋弘; 足立真実; 高木哲; 賀川由美子; 賀川由美子; 村田史郎; 大橋和彦, 日本獣医学会学術集会講演要旨集, 158th, 405, 405, 30 Aug. 2015
  (公社)日本獣医学会, Japanese
• 日本に分布するマレック病ウイルス野外株の分離
  町田柚香; 村田史郎; 伊勢崎政美; 酒井英史; 種子野章; 今内覚; 大橋和彦, 日本獣医学会学術集会講演要旨集, 158th, 354, 30 Aug. 2015
  Japanese
• シュルツェマダニ(Ixodes persulcatus)唾液腺由来免疫抑制因子sL2の同定および機能解析
  越智晶絵; 今内覚; 岡川朋弘; 西森朝美; 前川直也; 村田史郎; 大橋和彦, 日本獣医学会学術集会講演要旨集, 158th, 301, 301, 30 Aug. 2015
  (公社)日本獣医学会, Japanese
• 日本国内における牛白血病若齢発症牛の実態調査と表現型解析
  西森朝美; 今内覚; 池渕良洋; 岡川朋弘; 中原綾子; 千葉由純; 戸塚知恵; 村田史郎; 大橋和彦, 日本獣医学会学術集会講演要旨集, 158th, 336, 30 Aug. 2015
  Japanese
• ワクモ(Dermanyssus gallinae)由来カテプシンL様タンパク質の性状解析とワクチン抗原としての評価
  北條巧; 谷口綾香; 村田史郎; 伊勢崎政美; 酒井英史; 矢吹卓也; 寺田晴菜; 種子野章; 今内覚; 大橋和彦, 日本獣医学会学術集会講演要旨集, 158th, 353, 30 Aug. 2015
  Japanese
• マレック病ウイルスmeq遺伝子に認められた欠損配列
  村田史郎; 山本英次; 坂下奈津美; 松山あゆ美; 伊勢崎政美; 町田柚香; 今内覚; 大橋和彦, 日本獣医学会学術集会講演要旨集, 158th, 354, 30 Aug. 2015
  Japanese
• シュルツェマダニ(Ixodes persulcatus)由来Salp15因子によるライム病ボレリア伝播促進機能の解析
  村瀬優介; 今内覚; 伊東拓也; 高野愛; 川端寛樹; 安藤秀二; 村田史郎; 大橋和彦, 日本獣医学会学術集会講演要旨集, 158th, 301, 30 Aug. 2015
  Japanese
• マレック病ウイルス癌遺伝子(Meq)と病原性との関連
  高木道浩; 村田史郎; 今内覚; 大橋和彦, 鶏病研究会報, 50, 4, 199, 205, 25 Feb. 2015
  マレック病(MD)は鶏のウイルス感染症であり，家畜伝染病予防法で届出伝染病に指定されている。MDは翼麻痺や脚麻痺などの神経症状を示し，末梢神経の腫大，悪性リンパ腫形成(CD4+T細胞を標的とする)が見られ，死亡率は様々である。原因ウイルスであるMDウイルス(MDV)は直鎖状の二本鎖DNAウイルスでヘルペスウイルス科(Herpesviridae)アルファヘルペスウイルス亜科(Alphaherpesvirinae)，マルディウイルス(Mardivirus)に属する。これまではその病原性に基づきMDVは血清型1から3型(MDV1～3)に分類されていたが，最近，MDV1はGallid herpesvirus 2(GaHV-2)，MDV2はGallid herpesvirus 3(GaHV-3)およびMDV3はMeleagrid herpesvirus 1(MeHV-1)と新たに区分された。MDはワクチンによって効果的にコントロールできる疾病であり，ワクチンとしてすべての血清型があり，GaHV-2では弱毒株が用いられている。しかし，国内ではワクチンブレークによる発生，国外では強毒株の発生が見られている。また，ワクチンの作用機序およびMDVの腫瘍化機序も未だに解明されていない。ここではMDVの癌遺伝子と考えられているmeqについて述べると共に最近のMDV腫瘍化機序の知見について述べる。, 鶏病研究会, Japanese
• ウシの免疫抑制受容体を介したヨーネ菌特異的T細胞の応答の疲弊化
  岡川朋弘; 今中覚; 池渕良洋; 西森朝美; 溝呂木聖子; 永田礼子; 川治聡子; 田中省吾; 森康行; 村田史郎; 大橋和彦, 日本獣医学会学術集会講演要旨集, 157th, 385, 11 Aug. 2014
  Japanese
• 牛白血病ウイルス感染牛におけるナチュラル・キラー細胞の動態および機能解析
  中原綾子; 今内覚; 池渕良洋; 岡川朋弘; 西森朝美; 村田史郎; 大橋和彦, 日本獣医学会学術集会講演要旨集, 157th, 388, 11 Aug. 2014
  Japanese
• ウシ制御性T細胞の機能解析および慢性感染症における動態解析
  大平浩輔; 今内覚; 岡川朋弘; 西森朝美; 前川直也; 中原綾子; 村田史郎; 大橋和彦, 日本獣医学会学術集会講演要旨集, 157th, 388, 388, 11 Aug. 2014
  (公社)日本獣医学会, Japanese
• イヌ免疫抑制因子PD‐1およびPD‐L1を標的とした新規腫瘍免疫療法の検討
  前川直也; 今内覚; 池渕良洋; 岡川朋弘; 足立真実; 高木哲; 賀川由美子; 村田史郎; 大橋和彦, 日本獣医学会学術集会講演要旨集, 157th, 483, 483, 11 Aug. 2014
  (公社)日本獣医学会, Japanese
• Direct PCR法を用いた全血からの牛白血病ウイルス迅速簡易診断法の検討
  西森朝美; 今内覚; 池渕良洋; 岡川朋弘; 大平浩輔; 中原綾子; 村田史郎; 大橋和彦, 日本獣医学会学術集会講演要旨集, 157th, 424, 11 Aug. 2014
  Japanese
• 野生鳥類―畜産防疫の手引き~牛舎に侵入する野生鳥類~
  長雄一; 大越安吾; 平井綱雄; 藤井啓; 大橋和彦; 村田史郎; 遠藤大二; 金子正美; 田中克佳; 浅川満彦, 酪農ジャーナル, 67, 8, 29, 31, 01 Aug. 2014
  酪農学園大学エクステンションセンター, Japanese
• 野生鳥類由来感染症の家畜への伝播リスク評価手法の検討
  長雄一; 藤井啓; 大越安吾; 大橋和彦; 村田史郎; 千嶋淳; 久保清司, 日本野生動物医学会大会・講演要旨集, 20th, 42, 2014
  Japanese
• シュルツェマダニ(lxodes persulcatus)由来因子の宿主免疫抑制機能の解析
  今内覚; 伊東拓也; 川端寛樹; 高野愛; 安藤秀二; 村田史郎; 大橋和彦, 大原綜合病院年報, 53, 65, 15 Dec. 2013
  Japanese
• 免疫抑制受容体PD-1のリガンドPD-L2の機能的特徴と臨床応用研究
  西森朝美; 今内覚; 池渕良洋; 岡川朋弘; 村田史郎; 大橋和彦, 動物ワクチン-バイオ医薬品研究会ニュースレター, 8, 18, 19, Dec. 2013, [Invited]
  Japanese, Introduction scientific journal
• 牛白血病ウイルスの感染予防対策 潜伏期間は3~10年,若齢牛の発症例も多い
  今内覚; 村田史郎; 大橋和彦, Dairyman, 63, 9, 42, 43, 01 Sep. 2013
  Japanese
• イヌ免疫抑制因子PD‐1およびPD‐L1の同定と新規免疫療法樹立への基礎研究
  前川直也; 今内覚; 池渕良洋; 岡川朋弘; 高木哲; 村田史郎; 大橋和彦, 日本獣医学会学術集会講演要旨集, 156th, 257, 257, 30 Aug. 2013
  (公社)日本獣医学会, Japanese
• 牛白血病ウイルス感染牛由来IgM^high B細胞とIgM^low B細胞のウイルス抗原発現解析及び性状解析
  池渕良洋; 今内覚; 岡川朋弘; 西森朝美; 中原綾子; 村田史郎; 大橋和彦, 日本獣医学会学術集会講演要旨集, 156th, 271, 30 Aug. 2013
  Japanese
• マレック病腫瘍病変形成における免疫抑制因子発現機序の検討
  松山あゆ美; 村田史郎; 伊勢崎政美; 今内覚; 大橋和彦, 日本獣医学会学術集会講演要旨集, 156th, 257, 30 Aug. 2013
  Japanese
• シュルツェマダニ由来免疫抑制因子の機能解析
  豊間根耕地; 今内覚; 伊東拓也; 川端寛樹; 高野愛; 安藤秀二; 村田史郎; 大橋和彦, 日本獣医学会学術集会講演要旨集, 156th, 235, 30 Aug. 2013
  Japanese
• 全血培養法によるエンドトキシン活性測定の有用性の検討
  今村彩貴; 中溝万里; 川西路子; 池渕良洋; 目堅博久; 村田史郎; 今内覚; 大橋和彦; 中島奈緒; 山本欣也; 内山万利小; 平野文哉; 永井英貴; 木島まゆみ, 日本獣医学会学術集会講演要旨集, 156th, 260, 30 Aug. 2013
  Japanese
• ウシ免疫抑制因子PD‐L1およびPD‐L2の組換えタンパク質作製と免疫機能解析
  西森朝美; 今内覚; 池渕良洋; 岡川朋弘; 村田史郎; 大橋和彦, 日本獣医学会学術集会講演要旨集, 156th, 257, 30 Aug. 2013
  Japanese
• 牛白血病発生農場におけるサシバエ種の同定とウイルスの検出
  今内覚; 豊間根耕地; 小池菜々子; 伊東拓也; 村田史郎; 大橋和彦, 日本獣医学会学術集会講演要旨集, 156th, 231, 30 Aug. 2013
  Japanese
• 国内に潜在すると考えられる新興回帰熱に関する疫学調査研究
  高野愛; 川端寛樹; 大久保(佐藤)梢; 中尾稔; 伊東拓也; TAYLOR Kyle; 李景利; 坪田敏男; 今内覚; 吉村英紘; 豊間根耕地; 村田史郎; 大橋和彦, 日本獣医学会学術集会講演要旨集, 156回, 304, 304, Aug. 2013
  (公社)日本獣医学会, Japanese
• シュルツェマダニにおける病原体伝播関連因子の同定.
  今内覚; 川端寛樹; 伊東拓也; 高野愛; 安藤秀二; 村田史郎; 大橋和彦, 衛生動物, 64, 2, 112, 15 Jun. 2013
  Japanese
• スーラ病の分子疫学調査および野外分離株を用いた比較病原性解析と病態発生機序の解明
  目堅博久; 今内覚; MINGALA Claro N; 井上昇; 村田史郎; 小沼操; 大橋和彦, 日本獣医学会学術集会講演要旨集, 155th, 81, 04 Mar. 2013
  Japanese
• RNA用試料抽出・保存用濾紙のRNAウイルスPCR検出への利用
  松山智慧麿; 遠藤大二; 大橋和彦; 今内覚; 村田史郎; 有川二郎; 吉松組子; 水谷哲也; 林正信, 日本獣医学会学術集会講演要旨集, 155th, 229, 04 Mar. 2013
  Japanese
• シュルツェマダニ由来因子の病原体伝播における機能解析
  今内覚; 川端寛樹; 伊東拓也; 高野愛; 安藤秀二; 村田史郎; 大橋和彦, 大原綜合病院年報, 52, 105, 106, 15 Dec. 2012
  Japanese
• 感染症と免疫の基礎 最終回 臨床現場で役立つ免疫の知識
  大橋和彦; 村田史郎; 今内覚, 臨床獣医, 30, 11, 50, 52, 01 Nov. 2012
  Japanese
• 感染症と免疫の基礎 第3回 細胞性免疫
  今内覚; 村田史郎; 大橋和彦, 臨床獣医, 30, 10, 42, 44, 01 Oct. 2012
  Japanese
• 感染症と免疫の基礎 第2回 液性免疫
  村田史郎; 今内覚; 大橋和彦, 臨床獣医, 30, 9, 60, 62, 01 Sep. 2012
  Japanese
• ワクモDermanyssus gallinae由来発現遺伝子の網羅的解析
  伊勢崎政美; 村田史郎; 今内覚; 大橋和彦, 日本獣医学会学術集会講演要旨集, 154th, 203, 31 Aug. 2012
  Japanese
• フィリピン共和国におけるTrypanosoma evansi野外流行株の分離および比較病原性解析
  今内覚; 目堅博久; CLARO Mingala; NANCY Abes; ALAN Dargantes; 井上昇; 村田史郎; 大橋和彦, 日本獣医学会学術集会講演要旨集, 154th, 213, 31 Aug. 2012
  Japanese
• マレック病ウイルスワクチン接種鶏における免疫抑制因子PD‐1およびPD‐L1の発現解析
  高崎紗蘭; 村田史郎; 松山あゆ美; 伊勢崎政美; 今内覚; 大橋和彦, 日本獣医学会学術集会講演要旨集, 154th, 234, 31 Aug. 2012
  Japanese
• 牛白血病ウイルス感染牛における免疫抑制因子Tim‐3/Gal‐9の発現解析および機能解析
  岡川朋弘; 今内覚; 池渕良洋; 鈴木紗織; 寸田祐嗣; 村田史郎; 大橋和彦, 日本獣医学会学術集会講演要旨集, 154th, 233, 31 Aug. 2012
  Japanese
• シュルツェマダニ(Ixodes persulcatus)唾液腺由来免疫抑制因子sL2の同定および発現解析
  高田春奈; 今内覚; NAFTALY Githaka; 伊勢崎政美; 伊東拓也; 安藤秀二; 川端寛樹; 村田史郎; 大橋和彦, 日本獣医学会学術集会講演要旨集, 154th, 204, 31 Aug. 2012
  Japanese
• ウシの免疫抑制受容体PD‐1に対するモノクローナル抗体の作製および免疫活性化能の検討
  池渕良洋; 今内覚; 岡川朋弘; 横山和正; 中島千絵; 鈴木定彦; 村田史郎; 大橋和彦, 日本獣医学会学術集会講演要旨集, 154th, 233, 31 Aug. 2012
  Japanese
• 感染症と免疫の基礎 免疫とは何か
  大橋 和彦; 村田史郎; 今内 覚, 臨床獣医, 8, 60, 62, Aug. 2012, [Invited]
  Japanese, Introduction scientific journal
• マレック病感染鶏由来腫瘍病変部における免疫抑制因子PD‐1/PD‐L1発現の形態学的解析
  村田史郎; 松山あゆ美; 伊勢崎政美; 高崎紗蘭; 市居修; 今内覚; 大橋和彦, 日本獣医学会学術集会講演要旨集, 154回, 234, 234, Aug. 2012
  (公社)日本獣医学会, Japanese
• 増加している牛白血病―北海道での現状と対策について―
  今内覚; 村田史郎; 大橋和彦, 北海道獣医師会雑誌, 56, 7, 245, 251, 10 Jul. 2012
  Japanese
• ウシ難治性疾病に対する多機能型新規治療法の開発
  岡川朋弘; 今内覚; 池渕良洋; 鈴木紗織; 村田史郎; 大橋和彦, 動物ワクチン-バイオ医薬品研究会ニュースレター, 5, 21, 23, Jun. 2012, [Invited]
  Japanese, Introduction scientific journal
• A comparative analysis of cytokine profile of Theileria parva infection in Bovidae
  Okagawa T; Konnai S; Githaka N; Mekata H; Suzuki S; Kariuki E; Skilton R; Ishizuka M; Murata S; Ohashi K, Japanese Journal of Veterinary Parasitology, 10, 1-2, 37, Mar. 2012
  Japanese, Report scientific journal
• Babesia ovata感染宿主有核細胞の培養系産物を抗原としたELISAによる抗体検出系
  児玉道; 荻原喜久美; 納谷裕子; 村田史郎; 今内覚; 大橋和彦; 岸川正剛, 日本獣医学会学術集会講演要旨集, 153rd, 223, 01 Mar. 2012
  Japanese
• 牛難治性疾病に対する免疫抑制因子を標的とした治療型ワクチンの開発
  今内覚; 池渕良洋; 岡川朋弘; 鈴木紗織; 白井達哉; 村田史郎; 大橋和彦, 日本獣医学会学術集会講演要旨集, 153rd, 117, 01 Mar. 2012
  Japanese
• 牛白血病ウイルス感染牛における免疫抑制因子PD‐1/PL‐L1の発現解析および機能解析
  池渕良洋; 今内覚; 村田史郎; 大橋和彦, 日本獣医師会獣医学術学会年次大会講演要旨集, 2011, 45, 20 Jan. 2012
  Japanese
• 牛白血病の感染予防対策 最終回 4 ある農場における清浄化の取り組み事例
  今内覚; 安富一郎; 田島誉士; 村田史郎; 大橋和彦, Dairy Jpn, 56, 15, 63, 65, 01 Dec. 2011
  Japanese
• ウシTim-3の発現・機能解析と新規疾病制御法への応用展開
  岡川朋弘; 今内覚; 池渕良洋; 鈴木紗織; 村田史郎; 大橋和彦, 動物ワクチン-バイオ医薬品研究会ニュースレター, 4, 27, 29, Dec. 2011, [Invited]
  Japanese, Introduction scientific journal
• 牛白血病の感染予防対策 —診療・生産現場からの声に対して—
  今内 覚; 安富一郎; 田島誉士; 村田史郎; 大橋和彦, Dairy Japan, 12, 62, 64, Dec. 2011, [Invited]
  Japanese, Introduction scientific journal
• 牛白血病の感染予防対策 3 牛白血病に対する風評被害と間違った知識とは?
  今内覚; 安富一郎; 田島誉士; 村田史郎; 大橋和彦, Dairy Jpn, 56, 14, 38, 40, 01 Nov. 2011
  Japanese
• 牛白血病の感染予防対策 牛白血病に対する風評被害と間違った知識とは？
  今内 覚; 安富一郎; 田島誉士; 村田史郎; 大橋和彦, Dairy Japan, 11, 57, 58, Nov. 2011, [Invited]
  Japanese, Introduction scientific journal
• 現場で活かす酪農技術 牛白血病の感染予防対策 2 今からできる予防策とは?
  今内覚; 安富一郎; 田島誉士; 村田史郎; 大橋和彦, Dairy Jpn, 56, 12, 34, 36, 01 Oct. 2011
  Japanese
• 現場で活かす酪農技術 牛白血病の感染予防対策 1 牛白血病はなぜ増えたのか?
  今内覚; 安富一郎; 田島誉士; 村田史郎; 大橋和彦, Dairy Jpn, 56, 11, 53, 55, 01 Sep. 2011
  Japanese
• 牛白血病の感染予防対策 —牛白血病はなぜ増えたのか？—
  今内 覚; 安富一郎; 田島誉士; 村田史郎; 大橋和彦, Dairy Japan, 9, 37, 39, Sep. 2011, [Invited]
  Japanese, Introduction scientific journal
• シュルツェマダニ(Ixodes persulcatus)唾液腺由来免疫抑制因子の同定および発現解析
  丹羽彩乃; 今内覚; GITHAKA Naftary; 伊東拓也; 川端寛樹; 村田史郎; 大橋和彦, 日本獣医学会学術集会講演要旨集, 152nd, 207, 31 Aug. 2011
  Japanese
• マレック病ウイルス強毒株由来Meqおよび弱毒株由来L‐Meqによるbcl‐2遺伝子発現調節の比較
  村田史郎; 伊勢崎政美; 松山あゆ美; 高崎紗蘭; 今内覚; 大橋和彦, 日本獣医学会学術集会講演要旨集, 152nd, 241, 31 Aug. 2011
  Japanese
• ウシ免疫抑制受容体Tim‐3の同定と牛白血病ウイルス感染牛における発現解析
  岡川朋弘; 今内覚; 池渕良洋; 鈴木紗織; 寸田祐嗣; 村田史郎; 大橋和彦, 日本獣医学会学術集会講演要旨集, 152nd, 224, 31 Aug. 2011
  Japanese
• 牛白血病ウイルス感染牛における制御性T細胞の動態解析
  鈴木紗織; 今内覚; 池渕良洋; 岡川朋弘; 寸田祐嗣; 村田史郎; 大橋和彦, 日本獣医学会学術集会講演要旨集, 152nd, 224, 31 Aug. 2011
  Japanese
• トリパノソーマ原虫感染において制御性樹状細胞は炎症反応を抑制して病態の改善に関与する
  目堅博久; 今内覚; 井上昇; 村田史郎; 大橋和彦, 日本獣医学会学術集会講演要旨集, 152nd, 205, 31 Aug. 2011
  Japanese
• Theileria parva原虫感染牛属のサイトカイン発現比較解析
  今内覚; 岡川朋弘; NAFTALY Githaka; 目堅博久; 鈴木紗織; EDWARD Kariuki; ROBERT Skilton; 石塚真由美; 村田史郎; 大橋和彦, 日本獣医学会学術集会講演要旨集, 152nd, 204, 31 Aug. 2011
  Japanese
• シュルツェマダニ(Ixodes persulcatus)におけるライム病ボレリア伝播関連因子の同定.
  今内覚; 村瀬優介; 伊東拓也; 高野愛; 川端寛樹; 村田史郎; 大橋和彦, 衛生動物, 62, 2, 133, 15 Jun. 2011
  Japanese
• バイオセキュリティを見直す 4)感染牛を把握し,伝播リスクを抑える ウイルスを原因とする牛白血病の防疫
  今内覚; 安富一郎; 田島誉士; 村田史郎; 大橋和彦, Dairyman, 61, 6, 30, 31, 01 Jun. 2011
  Japanese
• ウイルスを原因とする牛白血病の防疫（感染牛を把握し、伝播リスクを抑える）
  今内 覚; 安富一郎; 田島誉士; 村田史郎; 大橋和彦, DAIRYMAN, 61, 30, 31, Jun. 2011, [Invited]
  Japanese, Introduction scientific journal
• 牛の難治性疾病に対する多機能型ワクチン戦略
  今内覚; 池渕良洋; 寸田祐嗣; 小沼操; 村田史郎; 大橋和彦, 日本獣医学会学術集会講演要旨集, 151st, 120, 01 Mar. 2011
  Japanese
• 牛末梢血よりTheileria orientalisとBabesia ovataとの混合感染有核細胞培養系の樹立
  児玉道; 荻原喜久美; 納屋裕子; 久保正法; 榊原伸一; 村田史郎; 今内覚; 大橋和彦, 日本獣医学会学術集会講演要旨集, 151st, 217, 01 Mar. 2011
  Japanese
• ベクターコントロールによる東海岸熱新規防除法の探索
  山田慎二; 今内覚; 今村彩貴; SIMMUNZA Martin; NANBOTA Andrew; 小沼操; 村田史郎; 大橋和彦, 日本獣医学会学術集会講演要旨集, 151st, 79, 01 Mar. 2011
  Japanese
• 牛白血病の感染防御対策
  今内 覚; 田島誉士; 小沼 操; 村田史郎; 大橋和彦, 北海道しゃくなげ会会報, 50, 10, 12, Feb. 2011, [Invited]
  Japanese, Introduction scientific journal
• 国内の牛白血病の現状と今後の対策について
  今内 覚; 田島 誉士; 村田 史郎; 大橋 和彦, 牛臨床寄生虫研究会誌, 2, 2, 3, Feb. 2011, [Invited]
  Japanese, Introduction scientific journal
• 獣医学における保全医学の展開―生物多様性と野生動物感染症―マガンなどの野生水禽の疾病抵抗性とマレック病ウイルスの分布
  大橋和彦; 松原綾子; 村田史郎; 今内覚, 獣医畜産新報, 64, 1074, 21, 26, 01 Jan. 2011
  文永堂出版, Japanese
• 牛白血病の正体と予防を考察する
  今内 覚; 安富一郎; 田島誉士; 村田史郎; 大橋和彦, HOLSTEIN MAGAZINE, 505, 4, 7, Jan. 2011, [Invited]
  Japanese, Introduction scientific journal
• 増加傾向にある牛白血病の現状と対策~診療現場からの声に対して~
  今内覚; 田島誉士; 小沼操; 村田史郎; 大橋和彦, 産業動物臨床医学雑誌, 1, 2, 110, 114, 05 Nov. 2010
  日本家畜臨床学会, Japanese
• 難治性慢性疾病における免疫疲弊化機序の解明と新規治療法への応用―獣医医療への応用にむけて―
  池渕良洋; 今内覚; 寸田祐輔; 小沼操; 村田史郎; 大橋和彦, 日生研たより, 56, 6, 77, 81, 01 Nov. 2010
  Japanese
• フィリピン共和国で分離したTrypanosoma evansiの病原性比較解析
  目堅博久; 今内覚; MINGALA Claro; DARGANTES Alan; 井上昇; 小沼操; 村田史郎; 大橋和彦, 日本獣医学会学術集会講演要旨集, 150th, 191, 01 Sep. 2010
  Japanese
• シュルツェマダニ(Ixodes persulcatus)由来Salp15様因子の性状解析
  村瀬優介; 今内覚; 伊藤拓也; 高野愛; 川端寛樹; 小沼操; 村田史郎; 大橋和彦, 日本獣医学会学術集会講演要旨集, 150th, 193, 01 Sep. 2010
  Japanese
• アフリカスイギュウ(Syncerus caffer)のTh1,Th2および炎症性サイトカイン遺伝子の同定
  岡川朋弘; 鈴木紗織; 今内覚; GITHAKA Naftali; KARIUKI Edward; GAKUYA Francis; ESTHER Kanduma; 石塚真由美; 村田史郎; 大橋和彦, 日本獣医学会学術集会講演要旨集, 150th, 212, 01 Sep. 2010
  Japanese
• マレック病ウイルス感染鶏における免疫抑制受容体PD‐1とそのリガンドPD‐L1の発現解析
  河東あゆ美; 村田史郎; 加納里佳; 今内覚; 大橋和彦, 日本獣医学会学術集会講演要旨集, 150th, 214, 01 Sep. 2010
  Japanese
• 牛白血病ウイルス感染牛における免疫抑制因子PD‐1/PD‐L1の発現解析と機能解析
  池渕良洋; 今内覚; 白井達哉; 寸田祐嗣; 小沼操; 村田史郎; 大橋和彦, 日本獣医学会学術集会講演要旨集, 150th, 214, 01 Sep. 2010
  Japanese
• シュルツェマダニ(Ixodes persulcatus)由来Salp16の宿主免疫抑機構解析
  今内覚; 肥田野新; 村瀬優介; 伊東拓也; 川端寛樹; 樋口豪起; 小沼操; 村田史郎; 大橋和彦, 日本獣医学会学術集会講演要旨集, 150th, 193, 01 Sep. 2010
  Japanese
• ウシ免疫抑制因子LAG‐3の同定と牛白血病ウイルス感染牛における発現解析
  白井達哉; 今内覚; 池渕良洋; 寸田祐嗣; 小沼操; 村田史郎; 大橋和彦, 日本獣医学会学術集会講演要旨集, 150th, 214, 01 Sep. 2010
  Japanese
• ワクモDermanyssus gallinaeに対する新規防除法開発のためのトランスクリプトーム解析
  伊勢崎政美; 村田史郎; 今内覚; 大橋和彦, 日本獣医学会学術集会講演要旨集, 150th, 193, 01 Sep. 2010
  Japanese
• マガンおよび鶏におけるinterferon‐γ遺伝子プロモーター活性の比較解析
  松原綾子; 村田史郎; 加納里佳; 橋口知幸; 伊勢崎政美; 河東あゆ美; 小沼操; 今内覚; 大橋和彦, 日本獣医学会学術集会講演要旨集, 150th, 215, 01 Sep. 2010
  Japanese
• メガファームにおける取り組み紹介―牛白血病ウイルス感染伝播阻止トライアルの実例―
  今内覚; 安富一郎; 田島誉士; 村田史郎; 大橋和彦, 北海道獣医師会雑誌, 54, 8, 378, 10 Aug. 2010
  Japanese
• 難治性慢性疾病における免疫疲弊化機序の解明と新規治療法への応用〜獣医医療への応用に向けて〜
  池渕良洋; 今内 覚; 寸田祐嗣; 小沼 操; 村田史郎; 大橋和彦, NIBS Letter, 56, 6, 7, 11, Jun. 2010, [Invited]
  Japanese, Introduction scientific journal
• The detection and characterization of the meq variants of Marek's diseases virus
  Ohashi K; Murata S; Okada T; Konnai S, Animal Viruses, 16, 117, 132, Jan. 2010, [Peer-reviewed], [Invited]
  Japanese, Introduction scientific journal
• マガンなどの野生水禽の疾病抵抗性とマレック病ウイルスの分布
  大橋和彦; 松原綾子; 村田史郎; 今内覚, 日本獣医学会学術集会講演要旨集, 149th, 198, 2010
  Japanese
• マレック病ウイルスmeq遺伝子の多型に伴うMeqタンパクの転写活性化能の変化
  橋口知幸; 村田史郎; 岡田宰; 加納里佳; 今内覚; 小沼操; 大橋和彦, 日本獣医学会学術集会講演要旨集, 146th, 189, 05 Sep. 2008
  Japanese
• 下大静脈内進展と大胸筋内転移を伴った骨盤内平滑筋肉腫の1例
  耕崎志乃; 村田和子; 大西剛直; 福本光孝; 小川恭弘; 前田博教; 笹栗史郎; 松本学; 弘井誠, Abstracts. Annual Symposium. Japanese Society for the Advancement of Women’s Imaging, 9th (Web), 2008
• 弱毒および強毒株マレック病ウイルス感染鶏における免疫応答の比較
  加納里佳; 橋口知幸; 岡田宰; 村田史郎; 今内覚; 小沼操; 大橋和彦, 日本ウイルス学会北海道支部夏季シンポジウム講演抄録, 42nd, 14, 2008
  Japanese
• 帯広近郊,ロシアならびにカムチャッカにおける野生水禽類のToxoplasma gondii抗体保有状況
  村尾岳洋; 加納里佳; 村田史郎; 今内覚; 大橋和彦; 小沼操; 前田龍一郎; 小俣吉孝, 日本野生動物医学会大会・講演要旨集, 13th, 88, 06 Sep. 2007
  Japanese
• マレック病ウイルス弱毒株由来L‐Meqと強毒株由来Meqの機能における比較解析
  村田史郎; 岡田宰; 加納里佳; 小沼操; 今内覚; 大橋和彦, 日本獣医学会学術集会講演要旨集, 144th, 83, 27 Aug. 2007
  Japanese
• 弱毒および強毒マレック病ウイルス感染鶏における免疫応答の比較
  加納里佳; 村田史郎; 岡田宰; 今内覚; 小沼操; 大橋和彦, 日本獣医学会学術集会講演要旨集, 144th, 77, 27 Aug. 2007
  Japanese
• 帯広近郊,ロシアならびにカムチャッカにおける野生カモ類のトキソプラズマ抗体保有状況
  村尾岳洋; 加納里佳; 村田史郎; 今内覚; 大橋和彦; 小沼操; 前田龍一郎; 小俣吉孝, 日本獣医学会学術集会講演要旨集, 144th, 65, 27 Aug. 2007
  Japanese
• マレック病ウイルス発癌遺伝子meqのスプライス産物Δmeqの発現・性状解析
  岡田宰; 村田史郎; 加納里佳; 高木道浩; 今内覚; 小沼操; 大橋和彦, 日本獣医学会学術集会講演要旨集, 144th, 83, 27 Aug. 2007
  Japanese
• マレック病由来腫瘍細胞株におけるMeqバリアント(変異体)の機能解析
  岡田宰; 高木道浩; 村田史郎; 加納里佳; 林裕子; 今内覚; 小沼操; 大橋和彦, 日本獣医学会学術集会講演要旨集, 142nd, 80, 31 Aug. 2006
  Japanese
• マレック病ウイルスの強毒化とmeq遺伝子の多型との関連
  村田史郎; 岡田宰; 加納里佳; 林裕子; 今内覚; 大橋和彦; 小沼操, 日本獣医学会学術集会講演要旨集, 142nd, 80, 31 Aug. 2006
  Japanese
• 鶏およびマガン由来マレック病ウイルスの遺伝子解析とその比較
  村田史郎; 山本裕己; 岡田宰; 浅川満彦; 長雄一; 池内俊雄; 今内覚; 大橋和彦; 小沼操, 日本獣医学会学術集会講演要旨集, 140th, 108, 31 Aug. 2005
  Japanese
• 見えてきたウイルス発癌 マレック病ウイルスによる潜伏感染・腫よう化におけるL‐meq遺伝子の役割
  大橋和彦; CHANG K‐S; LEE S‐I; 村田史郎; 高木道浩; 小沼操, 獣医畜産新報, 996, 594, 595, 01 Jul. 2004
  Japanese
• マレック病ウイルスによる潜伏感染・腫よう化におけるL‐meq遺伝子の役割
  大橋和彦; CHANG K‐S; LEE S‐I; 村田史郎; 高木道浩; 小沼操, 日本獣医学会学術集会講演要旨集, 137th, 86, 01 Mar. 2004
  Japanese
• マレック病ウイルス感染鶏の羽材料からのmeq遺伝子の検出
  村田史郎; CHANG K‐S; LEE S‐I; 大橋和彦; 小沼操, 日本獣医学会学術集会講演要旨集, 136th, 135, 10 Sep. 2003
  Japanese
• 北海道宮島沼における腫よう病変の認められた野生マガンからのマレック病ウイルスmeq遺伝子の検出
  張しゅ; 大橋和彦; 李成一; 村田史郎; 浅川満彦; 小沼操, 日本獣医学会学術集会講演要旨集, 134th, 134, 20 Aug. 2002
  Japanese
• 当院における血液浄化法の検討(CHDFを中心に)
  山口均; 村田厚夫; 三島史郎; 山口芳裕; 四蔵勇一; 高橋由典; 須田健二; 松田博青; 島崎修次, 日本救急医学会雑誌, 11, 10, 2000
• 急性すい炎はabdominal burnか? Cytocaine moduleを用いた検討
  山本明彦; 村田厚夫; 田中秀治; 山口芳裕; 菊地充; 三島史郎; 後藤英明; 小川道雄; 島崎修次, 日本救急医学会雑誌, 11, 10, 2000

• Infectious Disease of Animals, 5th Edition
  迫田, 義博; 秋庭, 正人; 末吉, 益雄; 高野, 友美; 長井, 誠; 芳賀, 猛; 村瀬, 敏之, 各論, 家きんおよび鳥類, 3.鶏白血病; 4.マレック病; 6.鶏伝染性喉頭気管炎; 11.鶏アデノウイルス感染症; 17.細網内皮症ウイルス感染症; 18.鶏腎炎ウイルス感染症; 19. ウイルス性胃腸炎
  近代出版, Mar. 2025, 9784874023037, xviii, 323p, 図版32p, Japanese, [Contributor]
• 感染症の生態学
  長雄一; 大橋和彦; 村田史郎, 第20章 鳥インフルエンザ
  共立出版, Mar. 2016, [Joint work]
• Ducks: Habitat, Behavior, and Diseases
  Asakawa, M; Nakade, T; Murata, S; Ohashi, K; Osa, Y; Taniyama, H, Recent viral disease of Japanese Anatid with a fatal case of Marek’s disease in an endangered species, white-fronted goose (Anser albifrons).
  Nova Science Publishers, 2013, [Joint work]

• Effect of Amino Acid Polymorphisms in the Basic Region of the Meq Oncoprotein of Marek’s Disease Virus on its Transactivation Activity and Virulence
  Jumpei Sato; Aoi Kurokawa; Yoshinosuke Motai; Shunsuke Yamagami; Shwe Win; Fumiya Horio; Hikaru Saeki; Naoya Maekawa; Tomohiro Okagawa; Benedikt Kaufer; Nikolaus Osterrieder; Mark Parcells; Satoru Konnai; Kazuhiko Ohashi; Shiro Murata
  48th Annual International Herpesvirus Workshop, 27 Jun. 2025, Oral presentation
  26 Jun. 2025 - 30 Jun. 2025
• Effects of Polymorphisms in Proline-Rich Repeats of the Meq Oncoprotein of Marek’s Disease Virus on Virulence
  Shunsuke Yamagami; Jumpei Sato; Yoshinosuke Motai; Shwe Win; Fumiya Horio; Naoya Maekawa; Tomohiro Okagawa; Benedikt Kaufer; Nikolaus Osterrieder; Mark Parcells; Satoru Konnai; Kazuhiko Ohashi; Shiro Murata
  48th Annual International Herpesvirus Workshop, 26 Jun. 2025, Oral presentation
  26 Jun. 2025 - 30 Jun. 2025
• Effect of Polymorphisms in the Meq Oncogene of Marek’s Disease Virus Distributed in Japan on the Pathogenicity
  Yoshinosuke Motai; Shiro Murata; Jumpei Sato; Naoya Maekawa; Tomohiro Okagawa; Benedikt B. Kaufer; Nikolaus Osterrieder; Mark S. Parcells; Satoru Konnai; Kazuhiko Ohashi
  THE 14TH INTERNATIONAL SYMPOSIUM ON MAREK’S DISEASE AND AVIAN HERPESVIRUSES, 13 Jul. 2024, Oral presentation
  12 Jul. 2024 - 14 Jul. 2024
• Amino-acid insertion in the Meq protein of Marek’s disease virus accelerates tumorigenesis.
  Jumpei Sato; Shiro Murata; Yoshinosuke Motai; Shwe Yee Win; Hikari Seo; Naoya Maekawa; Tomohiro Okagawa; Satoru Konnai; Kazuhiko Ohashi
  THE 14TH INTERNATIONAL SYMPOSIUM ON MAREK’S DISEASE AND AVIAN HERPESVIRUSES, 13 Jul. 2024, Oral presentation
  12 Jul. 2024 - 14 Jul. 2024
• In vitro evaluation of cysteine protease and ferritin 2 as vaccine antigens with broad efficacy across avian mites, poultry red mites, tropical fowl mites, and northern fowl mites
  Shwe Yee Win; Shiro Murata; Sotaro Fujisawa; Hikari Seo; Jumpei Sato; Yoshinosuke Motai; Takumi Sato; Eiji Oishi; Akira Taneno; Lat Lat Htun; Saw Bawm; Tomohiro Okagawa; Naoya Maekawa; Satoru Konnai; Kazuhiko Ohashi
  13th International Veterinary Immunology Symposium, 18 Nov. 2023, Oral presentation
  17 Nov. 2023 - 21 Nov. 2023
• Effects of an Insertion and a Deletion in the Marek’s Disease Virus Oncogene Meq on Transactivation Activity and Virulence
  Jumpei Sato; Shiro Murata; Zhiyuan Yang; Benedikt B. Kaufer; Sotaro Fujisawa; Hikari Seo; Naoya Maekawa; Tomohiro Okagawa; Satoru Konnai; Nikolaus Osterrieder; Mark S. Parcells; Kazuhiko Ohashi
  46th annual international herpesvirus workshop, 25 Jul. 2022, English, Oral presentation
  25 Jul. 2022 - 29 Jul. 2022
• Transcriptome analysis in blood-fed and starved poultry red mites, Dermanyssus gallinae
  Fujisawa, S; Murata, S; Takehara, M; Isezaki, M; Ogawa, R; Uno, Y; Taneno, A; Okagawa, T; Maekawa, N; Konnai, S; Ohashi, K
  The 4th International Symposium on Parasite Infections in Poultry, Vieena, 28 Jun. 2019, English, Poster presentation
  [International presentation]
• Cysteine protease and ferritin 2 as vaccine antigens to control poultry red mites, Dermanyssus gallinae
  S. Murata; M. Isezaki; A. Taniguchi; S. Fujisawa; A. Taneno; E. Sakai; Y. Uno; R. Ogawa; O. Ichii; T. Ito; M. Takehara; A. Morita; N. Maekawa; T. Okagawa; S. Konnai; K. Ohashi
  The 4th International Symposium on Parasite Infections in Poultry, Vienna, 28 Jun. 2019, English, Oral presentation
  [International presentation]
• マレック病に関する最近の知見について ―日本に分布するマレック病ウイルスの性状
  村田史郎; 今内覚; 大橋和彦
  鶏病研究会佐賀県支部鶏病技術研修会, 06 Jun. 2019, Japanese, Invited oral presentation
  [Invited], [Domestic Conference]
• 抗ワクモワクチン開発に向けた試み
  村田 史郎
  第２８４回鶏病事例検討会, 20 Sep. 2018, Japanese, Invited oral presentation
  [Invited], [Domestic Conference]
• Molecular detection of Marek’s disease virus in poultry farms in Myanmar
  Shiro Murata; Masaki Takehara; Ken Katakura; Myint Myint Hmoon; Shwe Yee Win; Masayoshi Isezaki; Satoru Konnai; Kazuhiko Ohashi
  The 12th International Symposium on Marek's Disease and Avian Herpesviruses, 29 Jul. 2018, English, Poster presentation
  [International presentation]
• Enhanced immune responses after vaccination with a gallid alphaherpesvirus 2 (Marek’s disease virus, MDV) expressing PD-1
  Shiro Murata; Benedikt B Kaufer; Nikolaus Osterrieder; Masayoshi Isezaki; Satoru Konnai; Kazuhiko Ohashi
  42nd International Herpesvirus Workshop, 29 Jul. 2017, English, Poster presentation
  [International presentation]
• The Role of Meq Mutations on Marek’s Disease Virus Pathogenesis
  Nirajan Bhandari; Phaedra Tavlarides-Hontz; Upendra Katneni; Benedikt B. Kaufer; Shiro Murata; Mark S. Parcells
  The 11th International Symposium on Marek's Disease and Avian herpesviruses, 07 Jul. 2016, English, Oral presentation
  [International presentation]
• Sequence analysis of a Marek’s disease virus strain isolated in Japan
  Shiro Murata; Yuka Machida; Masayoshi Isezaki; Satoru Konnai; Kazuhiko Ohashi
  The 11th International Symposium on Marek's Disease and Avian herpesviruses, 07 Jul. 2016, English, Poster presentation
  [International presentation]
• Expression analysis of programmed death ligand 2 in tumors caused by the avian oncovirus Marek’s disease virus
  Shiro Murata; Ayumi Matsuyama-Kato; Masayoshi Isezaki; Satoru Konnai; Kazuhiko Ohashi
  The 39th Annual International Herpesvirus Workshop, 19 Jul. 2014, English, Poster presentation
  [International presentation]
• Characterization of Meq proteins from field isolates of Marek’s disease virus in Japan
  Shiro Murata; Tomoyuki Hashiguchi; Yuko Hayashi; Yuki Yamamoto; Ayumi Kato; Sarah Takasaki; Masayoshi Isezaki; Satoru Konnai; Kazuhiko Ohashi
  The 9th International Symposium on Marek’s disease and Avian Herpesviruses, 28 Jun. 2012, English, Oral presentation
  [International presentation]
• Characterization of immunoinhibitory factors PD-1/PD-L1 in chickens infected with Marek’s disease virus
  Ayumi Matsuyama-Kato; Shiro Murata; Masayoshi Isezaki; Rika Kano; Sarah Takasaki; Osamu Ichii; Satoru Konnai; Kazuhiko Ohashi
  The 9th International Symposium on Marek’s disease and Avian Herpesviruses, 26 Jun. 2012, English, Oral presentation
  [International presentation]
• Amino acid substitutions or insertion in the Meq proteins could affect their transactivation and transformation abilities
  Shiro Murata; Tomoyuki Hashiguchi; Tsukasa Okada; Rika Kano; Misao Onuma; Satoru Konnai; Kazuhiko Ohashi
  International Union of Microbiological Societies 2011 Congress, 11 Sep. 2011, English, Oral presentation
  [International presentation]
• Regulation of the interferon gamma expression by an oncoprotein of Marek’s disease virus in wild waterfowls and chickens
  Ayako Matsubara; Shiro Murata; Rika Kano; Tomoyuki Hashiguchi; Masayoshi Isezaki; Ayumi Kato; Misao Onuma; Satoru Konnai; Kazuhiko Ohashi
  The 9th International Veterinary Immunology Symposium, 16 Aug. 2010, English, Oral presentation
  [International presentation]
• Analysis of transcriptional activities by the Meq proteins with highly virulent Marek’s disease virus strains
  Shiro MURATA; Tsukasa OKADA; Rika KANO; Yuko HAYASHI; Satoru KONNAI; Misao ONUMA; Kazuhiko OHASHI
  4th International Workshop of the Molecular Pathogenesis of Marek's disease Virus, 05 Aug. 2006, English, Oral presentation
  [International presentation]

• 免疫学特論, 2024年, 博士後期課程, 獣医学院
• 免疫学特論, 2024年, 博士後期課程, 国際感染症学院
• 伝染病学実習, 2024年, 学士課程, 獣医学部
• 獣医疫学演習, 2024年, 学士課程, 獣医学部
• 家畜育種学, 2024年, 学士課程, 獣医学部
• 動物衛生学実習, 2024年, 学士課程, 獣医学部
• アドバンスト演習, 2024年, 学士課程, 獣医学部
• アドバンスト演習, 2024年, 学士課程, 獣医学部

• JAPANESE SOCIETY OF VETERINARY SCIENCE
• 日本ウイルス学会

• 家禽の飼養衛生管理向上を目指したワクモによる宿主免疫応答調節機構の解明
  科学研究費助成事業
  15 Nov. 2024 - 31 Mar. 2027
  村田 史郎
  日本学術振興会, 特別研究員奨励費, 北海道大学, 24KF0168
• Development of cocktail vaccines against poultry red mites using the recombinant turkey herpesvirus
  Grants-in-Aid for Scientific Research
  01 Apr. 2023 - 31 Mar. 2027
  村田 史郎; 今内 覚; 大橋 和彦
  鳥類の外部寄生虫であるワクモによる吸血被害は、世界中の養鶏場で深刻な問題を引き起こしており、最近では薬剤に耐性を示すワクモの出現がその防除をさらに困難にさせている。そのため薬剤に変わる新たな防除法の開発研究が行われているが、その中でもワクチンによる防除法開発が注目されている。本ワクチンは、ワクモの生理機能に重要な分子を抗原として免疫し、血液に含まれる抗体を吸血したワクモに致死的影響を与えることを作用機序とする。しかし、単一の抗原による効果には限界があり、効果増強のためには複数抗原を用いたカクテルワクチンの開発が求められる。また実用化に際しては、従来のワクチンプログラムに抗ワクモワクチンを追加することになり、飼養者の負担が増えることになる。そこで本研究では、家禽の監視伝染病であるマレック病の予防に使用される七面鳥ヘルペスウイルス（HVT）をベクターとして用い、ウイルスゲノムに複数の抗原遺伝子を挿入することで、組換えHVT抗ワクモカクテルワクチンを開発することを目的とした。本年度は、ウイルスベクターとして用いるHVTの感染性クローンの作製に必要なベクター作製と、カクテルワクチンの作製に用いるワクチン抗原の評価を行なった。感染性クローン作製にはpBeloBac11ベクターを用い、ウイルスゲノム中のUS2遺伝子と相同組換えにより置換させることで行う。そのため、HVTのUS2遺伝子の両端の領域をpBeloBac11にクローニングした。さらに薬剤による組換えHVTの選抜のため、ecogpt遺伝子の発現カセットをクローニングした。カクテルワクチンの作製に用いるワクチン抗原として、複数の抗原候補の効果をin vitroにて評価した。その際には、ワクモと形態が類似するその他の家禽の吸血性ダニへの効果も期待される、ユニバーサルワクチンとしての応用の可能性を持つ抗原を選抜した。
  Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (B), Hokkaido University, 23K27063
• Development of cocktail vaccines against poultry red mites using the recombinant turkey herpesvirus
  Grants-in-Aid for Scientific Research
  01 Apr. 2023 - 31 Mar. 2027
  村田 史郎; 今内 覚; 大橋 和彦
  Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (B), Hokkaido University, 23H02370
• The roles of polymorphism in Meq on the pathogenicity of Marek's disease virus and their application for the vaccine development
  Grants-in-Aid for Scientific Research
  01 Apr. 2023 - 31 Mar. 2026
  大橋 和彦; 村田 史郎; 今内 覚
  マレック病ウイルス(MDV)の病原性進化の分子機構を明らかにして新規防除法を確立するために、MDVの病原性に最も重要なMeqについて、挿入や欠損を含むMeqアイソフォームを導入した組換えMDVを作出して、MDVの病原性発現機構を明らかにすることを目的として実施した。
  これまでMeqにおける挿入配列が病原性を増強することを明らかにしたが、その詳細について転写活性化能やmeq遺伝子をL-meq遺伝子に置換した組換えMDV（rMDV_L-Meq）を用いた感染実験等で検討した。L-MeqはMeqに比べ高い転写活性化能を示し、rMDV_Meq感染鶏群と比較して、rMDV_L-Meq感染鶏群ではがん関連因子の発現や末梢血におけるCD4+T細胞や制御性T細胞の増加が早期に認められた。一方で腫瘍病変部での発現遺伝子には両群に有意な差は認められなかった。以上よりMeqの挿入配列は、転写活性化能を増強させ早期に病態を形成させることが示唆された。
  当研究室では、野外株で欠損配列を持つmeq遺伝子（S-meq遺伝子）の同定・性状解析を報告したが、2022年に過去に報告例のない欠損配列を持つmeq遺伝子が同定されたため、その性状について解析を行なった。検出されたS-meq遺伝子よりも小さく（VS-meq遺伝子）、meq遺伝子と比べて転写活性化調節領域内に192bpの欠損が認められた。またMDV強毒株であるRB-1B株のMeqの配列と比べて、欠損以外に2箇所のアミノ酸配列の相違が認められた。RB-1B MeqとVS-Meqの転写活性化能に差は認められなかったが、欠損以外のアミノ酸配列をそろえたところ、VS-Meqの転写活性化能はRB-1B Meqより有意に高く、VS-Meqの欠損配列が転写活性化能を増加させることが示された。今後、組換えMDVを用いた感染実験により病原性への影響を調査する必要がある。
  Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (B), Hokkaido University, 23K27068
• The roles of polymorphism in Meq on the pathogenicity of Marek's disease virus and their application for the vaccine development
  Grants-in-Aid for Scientific Research
  01 Apr. 2023 - 31 Mar. 2026
  大橋 和彦; 村田 史郎; 今内 覚
  Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (B), Hokkaido University, 23H02375
• Development of evolving therapeutic technologies to address unmet medical needs for intractable diseases in animals
  Grants-in-Aid for Scientific Research
  01 Apr. 2022 - 31 Mar. 2025
  今内 覚; 村田 史郎; 大橋 和彦
  アンメット・メディカルニーズとは、未だ有効な治療方法や薬剤がない医療へのニーズを示す。獣医療においてもアンメット・メディカルニーズは難治性疾病を中心に存在する。そこでこれまで開発を進めてきた動物用免疫チェックポイント阻害薬を基盤とした副作用が小さく、より高い奏効率を発揮する発展的な新規治療・予防技術の構築を行い将来的な社会実装・実用化に向けた臨床知見を収集する。牛の慢性感染症では、免疫チェックポイント分子PD-1/PD-L1がT細胞の疲弊を引き起こし、病気の進行に関与している。我々は、ウシのPD-1/PD-L1の結合を特異的に阻害するモノクローナル抗体（mAbs）を確立したが、従来の抗PD-1mAbsは抗原との結合親和性が低いため、治療薬としては適さない。そこで今年度は、高い結合親和性を示す抗ウシPD-1ウサギmAbおよびキメラ抗体を樹立した。抗ウシPD-1ウサギmAbは、従来の抗PD-1 mAbと比較してウシのPD-1に強く結合し、PD-1/PD-L1相互作用に対して顕著な阻害活性を示した。また、キメラ抗体はIFN-γ産生の増強などの免疫活性化を示すことも確認された。さらに、牛伝染性リンパ腫ウイルス感染牛にキメラ抗体を投与し、抗ウイルス効果を調べた結果、抗ウイルス効果が確認された。一方、抗PD-L1抗体による治療を行った肺転移のある口腔内悪性黒色腫のイヌについて、低分割放射線療法と抗PD-L1抗体療法の組み合わせが奏効に及ぼす影響を検討した。その結果、抗PD-L1抗体治療を開始する直前(8 週間以内)に放射線治療を受けていたイヌで肺転移病変に対する奏効率が高く、また免疫療法における生存期間も長くなる傾向が認められた。
  Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (B), Hokkaido University, 23K23768
• Development of evolving therapeutic technologies to address unmet medical needs for intractable diseases in animals
  Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B)
  01 Apr. 2022 - 31 Mar. 2025
  今内 覚; 村田 史郎; 大橋 和彦
  Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (B), Hokkaido University, 22H02503
• 家禽の吸血性外部寄生虫ワクモに由来する鶏免疫応答調節因子の解明
  2023 年度 旗影会研究助成
  Apr. 2023 - Mar. 2024
  村田史郎
  一般財団法人旗影会, Principal investigator
• Application of a viral vector to develop anti-poultry red mite vaccines targeting antigens expressed on the plasma membrane of midgut cells
  Grants-in-Aid for Scientific Research Grant-in-Aid for Challenging Research (Exploratory)
  30 Jul. 2020 - 31 Mar. 2023
  村田 史郎
  ワクモ（鳥類の外部寄生虫）による吸血被害は、世界中の養鶏場で深刻な問題となっている。最近では薬剤に耐性を示すワクモの出現により、現行の薬剤による防除では制御が困難になりつつある。薬剤に変わる新規防除法としてワクチンが注目されている。本ワクチンは、ワクモの生理機能に重要な分子を抗原として免疫し、血液に含まれる抗体を吸血したワクモに致死的影響を与えることを作用機序とする。そのため腸管の細胞膜上に発現するタンパク質は最も影響を受けやすいと考えられ、これまでに腸管細胞の細胞膜上に発現が予想される分子を複数同定してきた。しかしそれらの多くは、複数回膜貫通型の分子や、大きな細胞膜領域を持つ分子であり、組換えタンパク質の作製には困難が予想され、ワクチン抗原としての利用には工夫が必要とされる。そこで本研究では、ウイルスベクターを用いて、ワクチン抗原を組換えウイルス感染細胞に細胞膜タンパク質として発現させることで、ワクチン効果に有効な免疫応答を誘導すること目的とした。本年度は組換えウイルスゲノムに挿入するためのワクチン抗原の選抜を行った。以前に実施したRNA sequencingのデータより、細胞膜上に発現が予想される分子を抽出し、本研究の趣旨に合致する、組換えタンパク質としての発現に困難が予想される立体構造を持つ抗原を選抜した。複数の抗原候補の中において、細胞膜上に発現が予想されるadipocyte plasma membrane-associated protein (APMAP) は、一回膜貫通型の大きな細胞外領域を持つ構造であると予想された。また、APMAPは腸管に発現することを確認することができた。そのためAPMAPは腸管細胞の細胞膜上に発現が予想されることから、APMAPを組換えウイルスゲノムに挿入する抗原として用いることとした。
  Japan Society for the Promotion of Science, Grant-in-Aid for Challenging Research (Exploratory), Hokkaido University, 20K21357
• The search for the candidate molecule for the vaccine to control a wide range of ectoparasite species of chickens
  Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B)
  01 Apr. 2020 - 31 Mar. 2023
  大橋 和彦; 村田 史郎; 今内 覚
  ワクモやトリサシダニは、鳥類の代表的な外部寄生虫であり、鶏に貧血や削痩、産卵率低下など養鶏産業に大きな経済的被害をもたらしている。現在、その防除には主に駆虫剤が使用されているが、薬剤耐性個体の出現や駆虫剤残存による衛生環境の悪化などが問題となっており、抗ダニワクチンなど新規防除法の開発が必要となっている。そこで本研究では、ワクモとトリサシダニ及び近縁なミナミトリサシダニを同時に防除できるダニ種横断的な新規“ユニバーサル”ワクチンの開発を目指して、次世代シークエンサーを用いたRNA-SEQ解析や全ゲノム解析と相同性検索により、これらのダニ間で共通の非暴露型抗原の探索・同定を行い、吸血したダニ個体に致死的に作用して鶏舎内の寄生虫個体数を持続的に減少できるワクチンへの応用を検討することを目的とした。
  今年度は、最初にワクモにおいて、飢餓および吸血後のワクモより試料を調製して次世代シークエンサーを用いたRNA-SEQを行い、発現する遺伝子群の網羅的解析を行った。その結果、吸血後のワクモでは、細胞内の代謝やエネルギー合成に関わる遺伝子群の発現が上昇し、飢餓ワクモでは、脂肪酸酸化や糖質合成など関わる遺伝子群の発現が顕著であった。
  新規“ユニバーサル”ワクチン候補分子を同定する際には、候補分子の発育ステージ毎あるいは組織別での詳細な発現解析を行う必要があるが、ワクモなどでは相対的発現量に計測に必須のreference遺伝子の情報が乏しい。そこで適当な２つのreference遺伝子を同定して、定量的real-time PCR法を樹立した。
  Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (B), Hokkaido University, 20H03137
• Experimental study for practical application of cross-sectional cancer therapy in companion animals
  Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B)
  01 Apr. 2019 - 31 Mar. 2022
  KONNAI SATORU
  Our previous studies demonstrated programmed cell death-ligand 1 (PD-L1) overexpression in some canine cancers, and suggested the therapeutic potential of a canine chimeric anti-PD-L1 monoclonal antibody (mAb). Thus, canine PD-L1 expression was assessed in various cancer types and the safety and efficacy of therapeutic anti-PD-L1 mAb were explored in dogs with pulmonary metastatic oral malignant melanoma (OMM). PD-L1 expression was detected in most canine malignant cancers including OMM, and survival was significantly longer in the treatment group when compared to a historical control group. We show that PD-L1 is a promising target for cancer immunotherapy in dogs.
  Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (B), Hokkaido University, 19H03114
• Development of a method to control poultry red mites using recombinant Marek's disease virus that secretes a vaccine antigen
  Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B)
  01 Apr. 2018 - 31 Mar. 2021
  Murata Shiro
  Poultry red mites (PRMs, Dermanyssus gallinae) are haematophagous ectoparasites which causes serious economic losses to the poultry industry worldwide, and the development of novel methods to control PRMs is required. We have investigated the application of vaccination and have shown that immunization with vaccine antigens could contribute to the reduction in the number of PRMs. In addition, we investigate the application of virus vectors to develop easy-to-use vaccines for farmers. In this study, we designed the recombinant virus that secrets vaccine antigens to improve the efficiency of vaccines using the recombinant virus. To produce an antigen-secreting virus, we inserted the signal peptides of a virus-derived secretory protein to the virus genome. However, the secretion of antigens was not confirmed, although the expression was observed in infected cells. Further manipulation is required to generate effective vaccines using virus vectors for controlling PRMs.
  Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (B), Hokkaido University, 18H02332
• Analysis of the molecular mechanism for the increase in the virulence of Marek's disease virus
  Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B)
  01 Apr. 2017 - 31 Mar. 2020
  Ohashi Kazuhiko
  Marek’s disease virus (MDV), a causative agents of Marek’s disease (MD), which is characterized by the formation of malignant lymphoma, appears to increase its virulence in the fields. To elucidate the molecular mechanisms for the increase in its virulence, we isolated MDVs from vaccinated chickens which still developed clinical MD, and analyzed their virological properties. MDV recently isolated in Japan caused clinical MD in inoculated chickens, although less virulent to chickens compared to virulent MDV strains isolated in United States. In addition, it is suggested that virus-derived factors other than polymorphisms in viral oncogene reported in the United States would be involved in the increase in MDV virulence.
  Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (B), Hokkaido University, 17H03911
• Analysis of genetic polymorphism of poultry red mites in the world and search for candidate molecules of anti-mite vaccines
  Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B)
  01 Apr. 2016 - 31 Mar. 2019
  OHASHI Kazuhiko
  Poultry red mites (PRMs), Dermanyssus gallinae, are blood-sucking ectoparasites of chickens, which cause huge economic losses in the poultry industries worldwide. To establish a new effective control method for PRMs, we have searched for and identified candidate antigens which are not polymorphic among PRMs from different areas in the world to develop anti-PRM vaccines. Several antigens identified in this study showed anti-PRM effects determined by in vitro feeding assay using sera from immunized chickens.
  Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (B), Hokkaido University, 16H05804
• Reconstitution of a novel recombinant Marek's disease virus as a vector vaccine against poultry red mites
  Grants-in-Aid for Scientific Research Grant-in-Aid for Young Scientists (B)
  01 Apr. 2016 - 31 Mar. 2018
  Shiro Murata
  The poultry red mite (PRM), is one of the major hematophagous ectoparasites of poultry farming, and the decreased productivity due to blood-sucking by PRMs is a severe problem for the poultry industry. The use of acaricides often leads to the spread of acaricide-resistant PRMs. Therefore, alternative strategies are required to reduce economic losses in poultry farming. Here, we focus on preventive methods such as vaccination to control PRMs, and we have already identified several vaccine candidates. In order to prepare an easy-to-use anti-PRM vaccine, the recombinant Marek's disease (MD) virus (rMDV) expressing a vaccine antigen was reconstituted. Reconstituted rMDV showed no significant differences in the growth kinetics compared to the parental strain, and the vaccine antigen was expressed in the infected cells. Thus, the rMDV live vaccine expressing a vaccine antigen against PRMs may be applicable as a candidate dual vaccine that provides protection against both PRMs and MD.
  Japan Society for the Promotion of Science, Grant-in-Aid for Young Scientists (B), Hokkaido University, 16K18798
• Comprehensive analysis of molecules expressed on the midgut of poultry red mites for the development of new anti-mite vaccines
  Grants-in-Aid for Scientific Research Grant-in-Aid for Challenging Exploratory Research
  01 Apr. 2016 - 31 Mar. 2018
  OHASHI Kazuhiko
  For the development of new vaccines against poultry red mite, Dermanyssus gallinae, a search was made for novel vaccine candidates expressed on the cell membrane of the midgut of the mite. Three kinds of molecules, whose expressions are predicted on the cell membrane, were identified, and their expressions on the midgut were confirmed by laser capture microdissection and RT-PCR. In addition, cathepsin D-like molecule, catD2, which was already identified in the course of the comprehensive analysis of genes expressed in the red mites done in our laboratory, was shown to be expressed in protonymph, deutonymph and adult and expressed regardless engorgement or starvation. The recombinant catD2 protein was not recognized by the sera from chickens fed by the red mites, suggesting that catD2 is a concealed antigen. Further analyses, including the detailed functions, the expression sites in red mites and the evaluation as vaccine antigens, are required to develop effective vaccines.
  Japan Society for the Promotion of Science, Grant-in-Aid for Challenging Exploratory Research, Hokkaido University, 16K15043
• Empirical research for bovine leukemia control strategy
  Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (A)
  01 Apr. 2014 - 31 Mar. 2018
  KONNAI SATORU
  Bovine leukemia virus (BLV) is highly endemic in many countries including Japan. In this study, we evaluated the risk for BLV transmission among the infected animals to establish the guideline for effective eradication of EBL. We found that BLV-infected cattle with persistent lymphocytosis (PL) or high proviral load or EBL are considered major sources of both horizontal and vertical BLV transmission. Furthermore, maternal proviral load is closely correlated with the frequency of vertical transmission to calves. However, no direct evidence of intrauterine BLV infection has been confirmed in infected cattle. We confirmed intrauterine BLV infection in two pregnant dams with high viral load by cesarean delivery, and BLV was detected in the newborns. In the study, we presented a direct evidence of intrauterine BLV transmission in pregnant dam with a high proviral load. These results could aid the development of BLV control measures targeting viral load.
  Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (A), Hokkaido University, 26252051
• Studies on the molecular mechanisms for the increase in MDV virulence in the field
  Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B)
  01 Apr. 2014 - 31 Mar. 2017
  Kazuhiko Ohashi
  Marek’s disease virus (MDV), a causative agent of Marek’s disease (MD), which is characterized by the formation of malignant lymphomas, tends to increase its virulence in the fields. To clarify the molecular mechanism for the increase in MDV virulence, recent MDV strains isolated from vaccinated chickens which developed clinical MD were virologically and molecular biologically characterized. Several new diversities/mutations were detected in viral genes of these strains, and a plaque-purified MDV strain was shown to be pathogenic to chicks. However, phylogenetic analysis of the whole genome of the MDV strain and reference strains showed that domestic MDV may increase its virulence through unknown mechanisms which are different from those suggested previously in MDV strains isolated in the United States.
  Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (B), Hokkaido University, 26292147
• Pathologic analysis of malignant infectious diseases in foreign livestock and its applications for establishment of novel control strategy.
  Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (A)
  01 Apr. 2013 - 31 Mar. 2017
  Satoru KONNAI; Cruz L.; Abes N.; Mingala C.; Skilton R.; Bishop R.; Nene V.; Kanduma E.; Githaka N.; Gakuya F.; Kariuki E.; Simmunza M.; Logullo C.; Itabajara S.; Brown W.; Mulenga A.; Morrison I.; Connelley T.; Whittington R.; Silva K. de; Chultemdorj T.; Sharav T.; Odbileg R.
  Although there are many malignant infectious diseases in livestock involving immune abnormalities (impairments), the mechanisms underlying these diseases remain unknown. In this research, we analyzed the mechanism of immune abnormalities in the malignant infectious diseases associated with economic loss in livestock industry, such as bovine anaplasmosis, East Coast fever, Surra, bovine tuberculosis, Johne’s disease, bovine leukemia, Equine infectious anemia. Our analyses have shown that immunosuppressive factors, such as Programmed death 1 (PD-1) and Programmed death-ligand 1 (PD-L1), are involved in immune suppression seen in the malignant infectious diseases. Furthermore, in vitro blockade of the PD-1/PD-L1 pathway restored anti-pathogen-specific T-cell proliferation and cytokine production.
  Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (A), Hokkaido University, 25257415
• Development of a novel vaccine to control virulent strains of Marek's disease virus: enhancement of vaccine efficacy by modulating the host immunosuppression pathway
  Grants-in-Aid for Scientific Research Grant-in-Aid for Young Scientists (B)
  01 Apr. 2013 - 31 Mar. 2015
  MURATA Shiro
  Field isolates of Marek’s disease virus (MDV) have tended to increase in virulence, and therefore, the development of more effective vaccines would be desirable. To develop a novel vaccine, we focused on the PD1/PD-L1 pathway that contributes to the immune suppression, and reconstituted the recombinant MDVs that carry the soluble-PD1 gene to competitively inhibit the immunosuppression regulated by the endogenous PD1. The recombinant vaccine expressing soluble PD1 induced higher production of interferon γ in the immune cells from the vaccinated chickens, indicating that this vaccine potentially possesses an ability to induce more effective cell-mediated immunity in chickens.
  Japan Society for the Promotion of Science, Grant-in-Aid for Young Scientists (B), Hokkaido University, Principal investigator, Competitive research funding, 25850194
• Establishment of control measure for bovine leukemia based on risk analysis of virus transmission
  Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (A)
  18 Nov. 2011 - 31 Mar. 2015
  KONNAI SATORU; OHASHI Kazuhiko; MURATA Shiro
  Bovine leukaemia virus (BLV) causes enzootic bovine leukosis (EBL) in cattle and represents a serious problem in Japanese meat and dairy industries. In this study, BLV from newly infected cattle was characterized to determine the virus source in a herd and a vector control measure was used to prevent the spread of the infection. The env sequences from positive conversion cattle showed 100% nucleotide identity with the sequence from the PL cattle with high provirus, suggesting that animals may serve as a reservoir of BLV infection. In the farms, where many blood-feeding stable flies were observed, vector control performed using a combination of insect repellents was very efficient as no new cases of BLV infection were observed after the treatment. This study indicates that BLV-infected cattle with high virus is one of high risk factors for the transmission, and vector-borne transmission might have been the major transmission route in the farm.
  Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (A), Hokkaido University, 23248051
• Studies on the immunoinhibitory mechanisms of Marek's disease virus and its application for adjuvant
  Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B)
  01 Apr. 2011 - 31 Mar. 2014
  OHASHI KAZUHIKO; KONNAI Satoru; MURATA Shiro
  The programmed death 1 (PD-1) receptor and its ligands, programmed death ligand 1 (PD-L1) and 2 (PD-L2) are known to have an immunoinhibitory function which contribute to the pathogenesis of chronic infections and neoplastic diseases via the PD-1/PD-L pathway. In this study, it has been shown that chicken PD-1 and PD-L1/PD-L2 also have an immunoinhibitory function, such as inhibition of cytokine production. The expressions of PD-1 and PD-L2 were significantly higher in the spleens of infected chickens in the latent phase and in tumor lesions caused by Marek's disease virus (MDV). These results suggest that chicken PD-1/PD-L2 pathway is involved in the establishment of latency and tumor formation by MDV.
  Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (B), Hokkaido University, 23380176
• 秋山記念生命科学振興財団研究助成
  2010 - 2010
  村田史郎
  秋山記念生命科学振興財団, Principal investigator, Competitive research funding

• 新規ベクターおよびその利用
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  JP2019018899, 13 May 2019
  特許第7264353号
  202303011844880950
• 抗ワクモワクチン組成物及びその使用
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  特願2021-080691, 11 May 2021
  特開2022-174692, 24 Nov. 2022
  202203006614155970
• イヌTGF-β受容体II
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  特願2021-075425, 27 Apr. 2021
  特開2022-169389, 09 Nov. 2022
  202203018896478340
• PD-1/PD-L1を標的とする阻害薬とCOX-2阻害薬との併用
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  JP2018027041, 19 Jul. 2018
  WO2019-017420, 24 Jan. 2019
  特許第7134417号
  202203000928000549
• 抗LAG-3抗体
  Patent right, 今内 覚; 大橋 和彦; 村田 史郎; 岡川 朋弘; 西森 朝美; 前川 直也; 鈴木 定彦; 中島 千絵, 国立大学法人北海道大学, 扶桑薬品工業株式会社
  JP2017029057, 10 Aug. 2017
  WO2018-034227, 22 Feb. 2018
  特許第7054144号
  202203006529697836
• PD-L1検出用抗PD-L1抗体
  Patent right, 今内 覚; 大橋 和彦; 村田 史郎; 岡川 朋弘; 西森 朝美; 前川 直也; 高木 哲; 賀川 由美子; 鈴木 定彦; 中島 千絵, 国立大学法人北海道大学, 扶桑薬品工業株式会社
  JP2018011895, 23 Mar. 2018
  WO2018-181064, 04 Oct. 2018
  特許第7036388号
  202203010523660792
• 抗PD-1抗体
  Patent right, 今内 覚; 大橋 和彦; 村田 史郎; 岡川 朋弘; 西森 朝美; 前川 直也; 鈴木 定彦; 中島 千絵, 国立大学法人北海道大学, 扶桑薬品工業株式会社
  特願2018-534378, 10 Aug. 2017
  WO2018-034226, 22 Feb. 2018
  特許第6960635号
  202103001370378735
• 抗PD-L1抗体
  Patent right, 今内 覚; 大橋 和彦; 村田 史郎; 岡川 朋弘; 西森 朝美; 前川 直也; 鈴木 定彦; 中島 千絵, 国立大学法人北海道大学, 扶桑薬品工業株式会社
  特願2018-534377, 10 Aug. 2017
  WO2018-034225, 22 Feb. 2018
  特許第6960634号
  202103010624578216
• ワクモの防除方法及び該方法で利用されるワクモタンパク質
  Patent right, 大橋 和彦; 村田 史郎; 伊勢崎 政美; 今内 覚; 谷口 綾香; 北條 巧, 国立大学法人北海道大学
  特願2016-523581, 29 May 2015
  WO2015-182754, 03 Dec. 2015
  特許第6765088号
  202003006944716043
• 新規ベクターおよびその利用
  Patent right, 鈴木 定彦; 中川 美樹; 亀田 弥生; 今内 覚; 岡川 朋弘; 前川 直也; 後藤 伸也; 佐治木 大和; 大橋 和彦; 村田 史郎; 北原 譲; 山本 啓一, 国立大学法人北海道大学, 扶桑薬品工業株式会社
  JP2019018899, 13 May 2019
  WO2019-225372, 28 Nov. 2019
  202103019221572843