山岸 潤也 (ヤマギシ ジユンヤ)
| 人獣共通感染症国際共同研究所 国際協力・教育部門 | 教授 |
| 総合イノベ-ション創発機構ワクチン研究開発拠点 | 教授 |
| Ｏｎｅ Ｈｅａｌｔｈリサーチセンター | 教授 |
Last Updated :2025/11/06
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担当教育組織
- 博士課程, 国際感染症学院
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論文
- A high prevalence of dogs seropositive to Leishmania in Zambia.
Herman M Chambaro, Kyoko Hayashida, Lavel C Moonga, Misheck Shawa, Walter Muleya, Joseph Chizimu, David Squarre, Tatsuki Sugi, Junya Yamagishi, Shohei Ogata, Masahiro Kajihara, Hirofumi Sawa, Chizu Sanjoba, Enala T Mwase, Roma Chilengi, Gilbert H Munsaka, Kelvin L Sarenje, Namwiinga R Mulunda, Mable M Mutengo, Boniface Namangala, Yasuyuki Goto
Parasitology international, 108, 103081, 103081, 2025年10月, ［国際誌］
英語, 研究論文（学術雑誌）, Domestic dogs are the main reservoir of Leishmania infantum, a causative agent of visceral leishmaniasis in humans. Although the disease is widespread in the world, the burden of visceral or any other disease form of leishmaniasis is poorly documented in Zambia, largely due to lack of surveillance. Recently, three cases of autochthonous canine leishmaniasis (CanL) were reported in Zambia following decades of presumed disease absence. This finding intimated probable disease emergence, raising the need for the identification of infection foci. Thus, in this study, we conducted the first mass serological survey for Leishmania infections in domestic dogs from two densely populated urban areas of Zambia in July 2022. In some of the study sites, seropositivity was up to ∼17 %, suggesting probable presence of Leishmania transmission hot spots. Moreover, on follow-up surveys of seropositive dogs, presence of antileishmanial antibodies was a risk factor for dog survival (relative risk = 7.9; odds ratio = 42.5). Our findings implies that Leishmania infection can be a health issue in domestic dogs in Zambia. Considering the risk of zoonotic transmission, the need for improved disease diagnosis and surveillance in both humans, dogs and sand fly vectors is highlighted in Zambia. - Emergence of livestock-associated MRSA clonal complex 398 in Thailand's swine industry.
Pawarut Narongpun, Pattrarat Chanchaithong, Wimonrat Tanomsridachchai, Narong Nuanmuang, Jeewan Thapa, Junya Yamagishi, Orasa Suthienkul, Pimlapas Leekitcharoenphon, Frank M Aarestrup, Chie Nakajima, Yasuhiko Suzuki
The Journal of antimicrobial chemotherapy, 2025年08月26日, ［国際誌］
英語, 研究論文（学術雑誌）, BACKGROUND: Livestock-associated MRSA (LA-MRSA) clonal complex (CC) 398 is primarily found among pigs across Europe. However, CC398 has also been sporadically identified in several Asian countries, including Thailand. This study aimed to examine the drivers contributing to the emergence of LA-MRSA CC398 in Thailand's swine industry. METHODS: Whole-genome sequencing data of 18 LA-MRSA CC398 isolates from pigs, retail pork and swine workers in Thailand were analysed. Additionally, a total of 1197 qualified genomes from various sources in 24 countries were retrieved from Pathogenwatch. Subsequently, two SNP-based phylogenetic trees were reconstructed using maximum likelihood (n = 1215) and Bayesian inference (n = 168). RESULTS: Our findings revealed that Staphylococcus aureus isolates with the CC398-SCCmec Vc (5C2&5)-t034 genotype were closely related to European isolates, including those from Denmark. In contrast, S. aureus isolates belonging to the CC398-composite SCCmec-t034 formed a phylogenetically distinct cluster. CONCLUSIONS: This study reports that the CC398-SCCmec Vc (5C2&5)-t034 genotype identified in Thai pigs may have originated from Danish swine populations, possibly introduced into Thailand through international livestock movement. The CC398-composite SCCmec-t034 genotype was also detected in domestic pigs; however, it appears to have evolved independently from the CC398-SCCmec Vc (5C2&5)-t034 genotype. Bayesian phylogenetic analysis estimates that the common ancestor of the composite genotype emerged earlier than that of the CC398-SCCmec Vc (5C2&5)-t034 genotype, indicating a separate evolutionary origin. - Vertical transmission of Leishmania donovani with placental degeneration in the pregnant mouse model of visceral leishmaniasis.
Haruka Mizobuchi, Junya Yamagishi, Chizu Sanjoba, Yasuyuki Goto
PLoS neglected tropical diseases, 19, 6, e0012650, 2025年06月, ［国際誌］
英語, 研究論文（学術雑誌）, Visceral leishmaniasis (VL) is a zoonotic disease caused by infection of macrophages by Leishmania donovani or L. infantum, and exhibits symptoms such as fever, anemia, and hepatosplenomegaly. VL during pregnancy has been reported to have negative effects such as miscarriage and vertical infection, but the mechanism is not clear. Here, we aimed to establish a pregnant VL mouse model and elucidate its immunopathology. Female BALB/c mice mated 6 months after L. donovani infection showed reduced pregnancy rates. The fetus was removed by caesarean section on the 18th day of pregnancy, and Leishmania parasite DNA was detected from fetal spleens and livers. As a result, the PCR positive rate was 68.9% (71/103 fetus), and vertical transmission was suspected in 66.7% of infected mothers (12/18 dams). Immunohistochemistry in the fetal livers detected cells positive for the Leishmania antigen, kinetoplastid membrane protein 11 (KMP11). In addition, pathological analysis of the VL placenta revealed trophoblast cell atrophy and vasodilation accompanied by CD3+ cell infiltration in the infected group. On the other hand, few KMP11+ cells were observed in the placenta of the infected group. Furthermore, RNA-Seq analysis revealed that IFN signal activation and cellular immune suppression were induced in the placenta of the infected group. These results suggest that VL in pregnancy induces suppression of placental cellular immunity through IFN and collapse of the placental barrier through trophoblast degeneration, leading to vertical transmission. Because few infected macrophages were observed in the placenta, it is possible that free Leishmania parasites in the blood contribute to transmission across the placenta. - Development of a semicomprehensive detection method for paramyxoviruses and its validation using Indonesian bats.
Dela Ria Nesti, Kyoko Hayashida, Tatsuki Sugi, Wayan T Artama, Hery Wijayanto, Naoko Kawai, Junya Yamagishi
Scientific reports, 15, 1, 19154, 19154, 2025年05月31日, ［国際誌］
英語, 研究論文（学術雑誌）, An outbreak of zoonotic diseases is one of the worldwide threats. Bats were reported as important reservoir hosts for many emerging zoonotic diseases. To mitigate the risk, understanding bat virome and their distribution is indispensable. Universal detection methods that can simultaneously identify multiple viruses are some of the most promising approaches. Here, we developed a semicomprehensive detection method integrating group-wide RT-PCR for paramyxoviruses and multiplex next-generation sequencing. The RT-PCR consists of three sets of degenerative primers covering viruses from Paramyxoviridae, including Pneumoviridae, which have now been reclassified into a distinct family. Index nucleotides were added to the primers to enable cost-effective multiplex sequencing, and the length of index was optimized to increase sensitivity. The method was applied to tracheal and rectal swabs from 135 bats captured in Indonesia. A conventional RT-PCR test validated the NGS results. Collectively, seven sequences of novel paramyxovirus-like similar to Pararubulavirus, Orthorubulavirus, and Henipavirus were successfully identified from seven bat samples. Furthermore, sequences between the two different target locations detected by NGS in the virus genomes were verified by RT-PCR. The similarity of the obtained sequences to the known paramyxoviruses sequences was relatively low, ranging from 70.88 to 82.44%. It suggests that the obtained sequences from novel viruses and the zoonotic risk of those novel viruses remain unknown. This cost-affordable, semi-comprehensive, pan-paramyxovirus test can be applied to other samples for viral genome surveillance, and the same strategy can be implemented to other pathogens for zoonosis control. - Facet joint arthritis as the presenting symptom for culture-negative Aggregatibacter aphrophilus native valve endocarditis in a patient without known cardiac disease: a case report.
Yurika Okuyama, Koki Kikuchi, Samuel David Stephenson, Naritomo Nishioka, Takahiro Doi, Junya Yamagishi, Satoshi Yuda
BMC infectious diseases, 25, 1, 682, 682, 2025年05月08日, ［国際誌］
英語, 研究論文（学術雑誌）, BACKGROUND: Aggregatibacter aphrophilus (A. aphrophilus) is a rare cause of infective endocarditis (IE), but is a recognized cause of culture-negative IE. The risk of developing IE is increased in patients with valvular disease or prosthetic valves. To our knowledge, A. aphrophilus has never previously been reported to cause lumbar facet joint arthritis in combination with IE. CASE PRESENTATION: We present the first case where facet joint arthritis was the presenting symptom for culture-negative A. aphrophilus native valve IE in a patient with no prior cardiac disease. A 58-year-old Japanese male without known cardiac disease, presented with high fever, chills, and lower back pain. Initial laboratory evaluation showed leukocytosis and transaminitis. Transthoracic echocardiography revealed an aortic valve vegetation with moderate aortic regurgitation. Magnetic resonance imaging (MRI) showed high-intensity areas in the right iliopsoas muscle and L4/L5 facet joint, indicative of fluid accumulation and disseminated lesions. Multiple sets of blood cultures showed no bacterial growth. Broad-range polymerase chain reaction (br-PCR) for 16S ribosomal RNA on both blood and hepatocytes (due to the patient's acute liver damage) also failed to identify the causative organism. The patient developed heart failure, and transesophageal echocardiography showed severe aortic regurgitation and an aneurysm at the noncoronary cusp of the aortic valve with perforation. He underwent aortic valve replacement and his symptoms were promptly improved. Although cultures from the excised valve were negative, br-PCR on the valve tissue eventually confirmed the presence of A. aphrophilus. CONCLUSION: This is the first reported case of culture-negative native valve IE caused by A. aphrophilus presenting with facet joint arthritis in a patient without known cardiac disease. Our case emphasizes the importance of considering IE in patients with fever and unexplained musculoskeletal pain, even without known cardiac disease. When conventional diagnostic tests are inconclusive, br-PCR on excised valve tissue is indispensable. Further improvements in non-invasive diagnostic methods are needed to facilitate early diagnosis and treatment. - ves1α genes expression is the major determinant of Babesia bovis-infected erythrocytes cytoadhesion to endothelial cells.
Hassan Hakimi, Junya Yamagishi, Miako Sakaguchi, Atefeh Fathi, Jae Seung Lee, Guilherme G Verocai, Shin-Ichiro Kawazu, Masahito Asada
PLoS pathogens, 21, 4, e1012583, 2025年04月, ［国際誌］
英語, 研究論文（学術雑誌）, Babesia bovis causes the most pathogenic form of babesiosis in cattle, resulting in high mortality in naive adults. This parasite invades red blood cells (RBCs) within the bovine hosts where they multiply and produce clinical disease. Babesia bovis exports numerous proteins into invaded RBCs changing its properties. Thus, the infected RBCs (iRBCs) are capable to cytoadhere in the microvasculature of internal organs and brain, leading to respiratory distress, neurologic signs, and mortality. Variant Erythrocyte Surface Antigen 1 (VESA1) is one of those exported proteins by B. bovis which represents a major virulence factor due to its central role in immune evasion by antigenic variation and intravascular parasite sequestration. VESA1 is a heterodimer protein encoded by ves1α and ves1β multigene family and localized on the ridges, the focal point for cytoadhesion. To gain further insights into the molecular mechanisms of cytoadhesion of B. bovis, we panned the parasites with bovine brain microvasculature endothelial cells, which resulted in obtaining several clones with different cytoadherence abilities. The transcriptome analysis of 2 high and 2 low cytoadherent clones revealed that ves1α sequences were diversified, likely resulting from genomic recombination. On the other hand, ves1β sequences were almost identical among these 4 clones. Insertion and expression of ves1α of a clone with high binding into ef-1α locus of a low binding clone increased cytoadherence confirming the role of ves1α suggested by our transcriptome data. Whole genome sequencing of cytoadherent clones revealed active locus of ves1 on chromosome 2. These results suggest that VESA1a proteins encoded by ves1α genes determine the cytoadherence strength of B. bovis and they are in the active site for recombination. - Unification of symbiotic bacteria during larva-to-adult transition in Culicoides circumscriptus (Diptera: Ceratopogonidae).
Ryo Ozuru, Junya Yamagishi, Ayumi Takeuchi, Yusuke Date, Takatoshi Fujii, Chihiro Sugimoto, Chie Nakajima, Yasuhiko Suzuki, Kaoru Aoki, Jun Fujii, Takashi Matsuba
FEMS microbiology letters, 372, 2025年01月10日, ［国際誌］
英語, 研究論文（学術雑誌）, Blood-sucking midges such as Leptoconops and Culicoides are of medical importance due to their role in causing skin irritation and potentially transmitting pathogens. Investigating their bacterial communities, including possible endosymbionts, may help clarify ecological adaptations and interactions with hosts. Leptoconops nipponensis Tokunaga (Lnt) and Culicoides circumscriptus (Cc), blood-sucking midges, cause severe itching and inflammation in humans. Cc was collected from a small sample of an outbreak swarm of Lnt in the peninsula area of Yonago City, Tottori Prefecture, Japan. This study compared the bacterial flora of Lnt and Cc, revealing distinct bacterial diversity shifts in these insect species between life stages. We analyzed the bacterial communities of adult and larval females of Cc and Lnt using MiSeq sequencing of the V3-V4 hypervariable region of the 16S rRNA gene. Notably, alpha diversity in Cc adults was significantly reduced to 1.5 (n = 43), indicating that Cc adults were dominated by a single bacterial genus, compared to 14.9 in Cc larvae (n = 19). BLAST (Basic Local Alignment Search Tool) analysis identified this dominant genus in adult Cc as Rickettsia (Candidatus Tisiphisa), which is known for transovarial transmission in arthropod vectors. In contrast, the bacterial diversity of Lnt showed no significant difference between adults (18.1, n = 32) and larvae (n = 15). These findings suggest that the dominance of Rickettsia in Cc (Candidatus Tisiphisa) adults is linked to their emergence, potentially reflecting differences in reproductive biology and ecological adaptations between these two insect species. Further research is needed to elucidate the functional role of Rickettsia in the life cycle and physiology of Cc. - Critical role of Babesia bovis spherical body protein 3 in ridge formation on infected red blood cells.
Atefeh Fathi, Hassan Hakimi, Miako Sakaguchi, Junya Yamagishi, Shin-Ichiro Kawazu, Masahito Asada
PLoS pathogens, 20, 11, e1012294, 2024年11月, ［国際誌］
英語, 研究論文（学術雑誌）, Babesia bovis, an apicomplexan intraerythrocytic protozoan parasite, causes serious economic loss to cattle industries around the world. Infection with this parasite leads to accumulation of infected red blood cells (iRBCs) in the brain microvasculature that results in severe clinical complications known as cerebral babesiosis. Throughout its growth within iRBCs, the parasite exports various proteins to the iRBCs that lead to the formation of protrusions known as "ridges" on the surface of iRBCs, which serve as sites for cytoadhesion to endothelial cells. Spherical body proteins (SBPs; proteins secreted from spherical bodies, which are organelles specific to Piroplasmida) are exported into iRBCs, and four proteins (SBP1-4) have been reported to date. In this study, we elucidated the function of SBP3 using an inducible gene knockdown (KD) system. Localization of SBP3 was assessed by immunofluorescence assay, and only partial colocalization was detected between SBP3 and SBP4 inside the iRBCs. In contrast, colocalization was observed with VESA-1, which is a major parasite ligand responsible for the cytoadhesion. Immunoelectron microscopy confirmed localization of SBP3 at the ridges. SBP3 KD was performed using the glmS system, and effective KD was confirmed by Western blotting, immunofluorescence assay, and RNA-seq analysis. The SBP3 KD parasites showed severe growth defect suggesting its importance for parasite survival in the iRBCs. VESA-1 on the surface of iRBCs was scarcely detected in SBP3 KD parasites, whereas SBP4 was still detected in the iRBCs. Moreover, abolition of ridges on the iRBCs and reduction of iRBCs cytoadhesion to the bovine brain endothelial cells were observed in SBP3 KD parasites. Immunoprecipitation followed by mass spectrometry analysis detected the host Band 3 multiprotein complex, suggesting an association of SBP3 with iRBC cytoskeleton proteins. Taken together, this study revealed the vital role of SBP3 in ridge formation and its significance in the pathogenesis of cerebral babesiosis. - Genome sequencing of canine distemper virus isolates from unvaccinated dogs in Mongolia.
Ariunbold Munkhtsetseg, Enkhbaatar Batmagnai, Myagmarsuren Odonchimeg, Gombodash Ganbat, Yondonjamts Enkhmandakh, Gantulga Ariunbold, Tsedenbal Dolgorsuren, Raadan Odbileg, Purevtseren Dulam, Bumduuren Tuvshintulga, Chihiro Sugimoto, Yoshihiro Sakoda, Junya Yamagishi, Dashzevge Erdenechimeg
Veterinary journal (London, England : 1997), 308, 106231, 106231, 2024年08月28日, ［国際誌］
英語, 研究論文（学術雑誌）, Canine distemper virus (CDV) triggers a severe, often fatal disease in dogs and wildlife known as canine distemper (CD). Prior research has noted significant genetic diversity and recombination among CDV isolates from different geographical regions, potentially contributing to vaccine failures. Despite this, no genetic characterization of Mongolian CDVs has been conducted. This study, isolated CDVs from three unvaccinated dogs: two 10-month-old mixed-breeds and an 18-month-old Samoyed. All exhibited CD symptoms and subsequently died. Virus isolation was conducted using Vero/dog SLAM cells, with genome sequencing performed via nanopore technology. The mixed-breed dogs were infected with non-recombinant CDV isolates, forming a sister clade to the Asia-1 lineage prevalent in Asia. The Samoyed was infected with a non-recombinant CDV isolate, classifying as Asia-4 lineage sporadically reported in some Asian countries. This sequencing data offers foundational information on genetic diversity, aiding CD control measure development and benefiting future Eurasia and Asian studies. - Correction: Narongpun et al. Whole-Genome Investigation of Zoonotic Transmission of Livestock-Associated Methicillin-Resistant Staphylococcus aureus Clonal Complex 398 Isolated from Pigs and Humans in Thailand. Antibiotics 2023, 12, 1745.
Pawarut Narongpun, Pattrarat Chanchaithong, Junya Yamagishi, Jeewan Thapa, Chie Nakajima, Yasuhiko Suzuki
Antibiotics (Basel, Switzerland), 13, 8, 2024年07月23日, ［国際誌］
英語, In the published publication [...]. - Inflammatory CD11b+ Macrophages Produce BAFF in Spleen of Mice Infected with Leishmania donovani
Kazuki Nagai, Wataru Fujii, Junya Yamagishi, Chizu Sanjoba, Yasuyuki Goto
Pathogens, 13, 3, 2024年03月06日, ［査読有り］, ［国際誌］
英語, 研究論文（学術雑誌）, Visceral leishmaniasis (VL) is an infectious disease caused by parasitic protozoa of the genus Leishmania and manifests clinical symptoms such as splenomegaly, hepatomegaly, anemia, and fever. It has previously been shown that B-cell-activating factor (BAFF) is involved in splenomegaly during VL. Although BAFF is known to be expressed by a variety of cells, the mechanism of elevated BAFF expression in VL is not clear. In this study, we aimed to identify BAFF-producing cells in the spleens of mice infected with Leishmania donovani. Splenocytes of L. donovani-infected mice showed elevated BAFF expression compared to that of naive mice. In the infected spleen, the number of both CD11b+ and F4/80+ cells increased, and the major BAFF-producing cells were CD11b+ cells, which did not serve as host cells of Leishmania. Immunohistochemical/immunofluorescent staining of spleens of infected mice revealed that the increased CD11b+ cells were primarily MRP14+ mononuclear cells. Together, these results suggest the increased BAFF expression in the spleen of L. donovani-infected mice involves a recruitment of inflammatory macrophages distinct from host macrophages for the parasites. - Extraction of the CDRH3 sequence of the mouse antibody repertoire selected upon influenza virus infection by subtraction of the background antibody repertoire
Masashi Shingai, Sayaka Iida, Naoko Kawai, Mamiko Kawahara, Toshiki Sekiya, Marumi Ohno, Naoki Nomura, Chimuka Handabile, Tomomi Kawakita, Ryosuke Omori, Junya Yamagishi, Kaori Sano, Akira Ainai, Tadaki Suzuki, Kazuo Ohnishi, Kimihito Ito, Hiroshi Kida
Journal of Virology, 98, 3, e0199523, American Society for Microbiology, 2024年02月07日, ［国際誌］
英語, 研究論文（学術雑誌）, As specific interactions between antigens and cell-surface antibodies trigger the proliferation of B-cell clones, the frequency of each antibody sequence in the samples reflects the size of each clonal population. Nevertheless, it is extremely difficult to extract antigen-specific antibody sequences from the comprehensive bulk antibody sequences obtained from blood samples due to repertoire bias influenced by exposure to dietary antigens and other infectious agents. This issue can be addressed by subtracting the background noise from the post-immunization or post-infection repertoire data. In the present study, we propose a method to quantify repertoire data from comprehensive repertoire data. This method allowed subtraction of the background repertoire, resulting in more accurate extraction of expanded antibody repertoires upon influenza virus infection. This accurate extraction of antigen- or infection-specific repertoire information is a useful tool for vaccine evaluation. - Upregulation of ATP6V0D2 benefits intracellular survival of Leishmania donovani in erythrocytes-engulfing macrophages
Jing Hong, Budhaditya Mukherjee, Chizu Sanjoba, Junya Yamagishi, Yasuyuki Goto
Frontiers in Cellular and Infection Microbiology, 14, 1332381, 1332381, Frontiers Media SA, 2024年01月31日, ［国際誌］
英語, 研究論文（学術雑誌）, Visceral leishmaniasis (VL) is the most severe type of leishmaniasis which is caused by infection of Leishmania donovani complex. In the BALB/c mouse model of VL, multinucleated giant cells (MGCs) with heavy parasite infection consist of the largest population of hemophagocytes in the spleen of L. donovani-infected mice, indicating that MGCs provide the parasites a circumstance beneficial for their survival. Although ATP6V0D2 is a demonstrated factor inducing the formation of hemophagocytic MGCs during L. donovani infection, functions of this protein in shaping the infection outcome in macrophages remain unclear. Here we evaluated the influence of upregulated ATP6V0D2 on intracellular survival of the parasites. L. donovani infection-induced hemophagocytosis of normal erythrocytes by macrophages was suppressed by RNAi-based knockdown of Atp6v0d2. The knockdown of Atp6v0d2 did not improve the survival of amastigotes within macrophages when the cells were cultured in the absence of erythrocytes. On the other hand, reduced intracellular survival of amastigotes in macrophages by the knockdown was observed when macrophages were supplemented with antibody-opsonized erythrocytes before infection. There, increase in cytosolic labile iron pool was observed in the L. donovani-infected knocked-down macrophages. It suggests that ATP6V0D2 plays roles not only in upregulation of hemophagocytosis but also in iron trafficking within L. donovani-infected macrophages. Superior access to iron in macrophages may be how the upregulated expression of the molecule brings benefit to Leishmania for their intracellular survival in the presence of erythrocytes. - Correction: Transcriptional profiling of Toll-like receptor 2-deficient primary murine brain cells during Toxoplasma gondii infection.
Kousuke Umeda, Sachi Tanaka, Fumiaki Ihara, Junya Yamagishi, Yutaka Suzuki, Yoshifumi Nishikawa
PloS one, 19, 5, e0303453, 2024年, ［国際誌］
英語, [This corrects the article DOI: 10.1371/journal.pone.0187703.]. - Combination therapy with oral antiviral and anti-inflammatory drugs improves the efficacy of delayed treatment in a COVID-19 hamster model.
Michihito Sasaki, Tatsuki Sugi, Shun Iida, Yuichiro Hirata, Shinji Kusakabe, Kei Konishi, Yukari Itakura, Koshiro Tabata, Mai Kishimoto, Hiroko Kobayashi, Takuma Ariizumi, Kittiya Intaruck, Haruaki Nobori, Shinsuke Toba, Akihiko Sato, Keita Matsuno, Junya Yamagishi, Tadaki Suzuki, William W Hall, Yasuko Orba, Hirofumi Sawa
EBioMedicine, 99, 104950, 104950, 2023年12月29日, ［国際誌］
英語, 研究論文（学術雑誌）, BACKGROUND: Pulmonary infection with SARS-CoV-2 stimulates host immune responses and can also result in the progression of dysregulated and critical inflammation. Throughout the pandemic, the management and treatment of COVID-19 has been continuously updated with a range of antiviral drugs and immunomodulators. Monotherapy with oral antivirals has proven to be effective in the treatment of COVID-19. However, treatment should be initiated in the early stages of infection to ensure beneficial therapeutic outcomes, and there is still room for further consideration on therapeutic strategies using antivirals. METHODS: We studied the therapeutic effects of monotherapy with the oral antiviral ensitrelvir or the anti-inflammatory corticosteroid methylprednisolone and combination therapy with ensitrelvir and methylprednisolone in a delayed dosing model of hamsters infected with SARS-CoV-2. FINDINGS: Combination therapy with ensitrelvir and methylprednisolone improved respiratory conditions and reduced the development of pneumonia in hamsters even when the treatment was started after 2 days post-infection. The combination therapy led to a differential histological and transcriptomic pattern in comparison to either of the monotherapies, with reduced lung damage and down-regulation of expression of genes involved in the inflammatory response. Furthermore, we found that the combination treatment is effective in case of infection with either the highly pathogenic delta or circulating omicron variants. INTERPRETATION: Our results demonstrate the advantage of combination therapy with antiviral and corticosteroid drugs in COVID-19 treatment from the perspective of lung pathology and host inflammatory responses. FUNDING: Funding bodies are described in the Acknowledgments section. - Whole-Genome Investigation of Zoonotic Transmission of Livestock-Associated Methicillin-Resistant Staphylococcus aureus Clonal Complex 398 Isolated from Pigs and Humans in Thailand.
Pawarut Narongpun, Pattrarat Chanchaithong, Junya Yamagishi, Jeewan Thapa, Chie Nakajima, Yasuhiko Suzuki
Antibiotics (Basel, Switzerland), 12, 12, 2023年12月16日, ［国際誌］
英語, 研究論文（学術雑誌）, Livestock-associated methicillin-resistant Staphylococcus aureus (LA-MRSA) has been widespread globally in pigs and humans for decades. Nasal colonization of LA-MRSA is regarded as an occupational hazard to people who are regularly involved in livestock production. Our previous study suggested pig-to-human transmission caused by LA-MRSA clonal complex (CC) 398, using traditional molecular typing methods. Instead, this study aimed to investigate the zoonotic transmission of LA-MRSA CC398 using whole genome sequencing (WGS) technologies. A total of 63 LA-MRSA isolates were identified and characterized in Thailand. Further, the 16 representatives of LA-MRSA CC9 and CC398, including porcine and worker isolates, were subjected to WGS on the Illumina Miseq platform. Core-genome single nucleotide polymorphism (SNP)-based analyses verify the zoonotic transmission caused by LA-MRSA CC398 in two farms. WGS-based characterization suggests the emergence of a novel staphylococcal cassette chromosome (SCC) mec type, consisting of multiple cassette chromosome recombinase (ccr) gene complexes via genetic recombination. Additionally, the WGS analyses revealed putative multi-resistant plasmids and several cross-resistance genes, conferring resistance against drugs of last resort used in humans such as quinupristin/dalfopristin and linezolid. Significantly, LA-MRSA isolates, in this study, harbored multiple virulence genes that may become a serious threat to an immunosuppressive population, particularly for persons who are in close contact with LA-MRSA carriers. - Surveys of eleven species of wild and zoo birds and feeding experiments in white-tailed eagles reveal differences in the composition of the avian gut microbiome based on dietary habits between and within species.
Kohei Ogasawara, Naoki Yamada, Shouta Mm Nakayama, Yukiko Watanabe, Keisuke Saito, Akane Chiba, Yoshitaka Uchida, Kaoru Ueda, Yasunori Takenaka, Kentaro Kazama, Mami Kazama, Junya Yamagishi, Hazuki Mizukawa, Yoshinori Ikenaka, Mayumi Ishizuka
The Journal of veterinary medical science, 85, 12, 1355, 1365, 2023年10月31日, ［国内誌］
英語, 研究論文（学術雑誌）, The composition of the gut microbiome varies due to dietary habits. We investigated influences of diet on the composition of the gut microbiome using the feces of 11 avian species, which consumed grain-, fish- and meat-based diets. We analyzed gut microbiome diversity and composition by next-generation sequencing (NGS) of 16S ribosomal RNA. The grain-diet group had higher gut microbiome diversity than the meat- and fish-diet group. The ratio of Bacteroidetes and Firmicutes phyla was higher in the grain-diet group than in the meat- and fish-diet groups. The grain-diet group had a higher ratio of Veillonellaceae than the meat-diet group and a higher ratio of Eubacteriaceae than the fish-diet habit group. To clarify the influence of diet within the same species, white-tailed eagles (Haliaeetus albicilla, n=6) were divided into two groups, and given only deer meat or fish for approximately one month. The composition of the gut microbiome of individuals in both groups were analyzed by NGS. There were indications of fluctuation in the levels of some bacteria (Lactobacillus, Coriobacteriales, etc.) in each diet group. Moreover, one individual for each group which switched each diet in last week changed to each feature of composition of bacterial flora. The above results show that the composition of the gut microbiome differ depending on diet, even within the same species. - Bovine Piroplasma Populations in the Philippines Characterized Using Targeted Amplicon Deep Sequencing.
Eloiza May Galon, Adrian Miki Macalanda, Tatsuki Sugi, Kyoko Hayashida, Naoko Kawai, Taishi Kidaka, Rochelle Haidee Ybañez, Paul Franck Adjou Moumouni, Aaron Edmond Ringo, Hang Li, Shengwei Ji, Junya Yamagishi, Adrian Ybañez, Xuenan Xuan
Microorganisms, 11, 10, 2023年10月18日, ［国際誌］
英語, 研究論文（学術雑誌）, Molecular assays and capillary electrophoresis sequencing have been used to identify parasites in livestock. The low sample capacity, which increases labor and processing time, is one drawback. Targeted amplicon sequencing (Ampliseq) uses the fast and large sample capacity platform to identify parasites in the target host, overcoming this limitation. DNA was extracted from 162 whole blood samples collected from cattle in three provinces in the Philippines. Using Illumina's Miseq platform, the V4 hypervariable region of the piroplasma 18S rRNA gene was amplified and sequenced. The AMPtk pipeline was used to obtain distinct amplicon sequence variants (ASVs) and the NCBI BLAST non-redundant database was used to assign taxonomy. In total, 95 (58.64%) samples were positive for piroplasma. Using the AMPTk pipeline, 2179 ASVs were obtained. A total of 79 distinct ASVs were obtained after clustering and filtering, which belonged to genera Babesia (n = 58), Theileria (n = 17), Hepatozoon (n = 2), and Sarcocystis (n = 2). The ASV top hits were composed of 10 species: Babesia bovis, B. bigemina, Theileria orientalis, Babesia sp., Hepatozoon canis, Sarcocystis cruzi, T. annulata, T. equi, T. mutans, and Theileria sp. Thung Song. The results generated in this study demonstrated the applicability of Ampliseq in detecting piroplasmid parasites infecting cattle in the Philippines. - Prevalence and Genomic Characterization of Rotavirus A from Domestic Pigs in Zambia: Evidence for Possible Porcine–Human Interspecies Transmission
Joseph Ndebe, Hayato Harima, Herman Moses Chambaro, Michihito Sasaki, Junya Yamagishi, Annie Kalonda, Misheck Shawa, Yongjin Qiu, Masahiro Kajihara, Ayato Takada, Hirofumi Sawa, Ngonda Saasa, Edgar Simulundu
Pathogens, 12, 10, 1199, 1199, MDPI AG, 2023年09月27日, ［査読有り］, ［国際誌］
英語, 研究論文（学術雑誌）, Rotavirus is a major cause of diarrhea globally in animals and young children under 5 years old. Here, molecular detection and genetic characterization of porcine rotavirus in smallholder and commercial pig farms in the Lusaka Province of Zambia were conducted. Screening of 148 stool samples by RT-PCR targeting the VP6 gene revealed a prevalence of 22.9% (34/148). Further testing of VP6-positive samples with VP7-specific primers produced 12 positives, which were then Sanger-sequenced. BLASTn of the VP7 positives showed sequence similarity to porcine and human rotavirus strains with identities ranging from 87.5% to 97.1%. By next-generation sequencing, the full-length genetic constellation of the representative strains RVA/pig-wt/ZMB/LSK0137 and RVA/pig-wt/ZMB/LSK0147 were determined. Genotyping of these strains revealed a known Wa-like genetic backbone, and their genetic constellations were G4-P[6]-I5-R1-C1-M1-A8-N1-T1-E1-H1 and G9-P[13]-I5-R1-C1-M1-A8-N1-T1-E1-H1, respectively. Phylogenetic analysis revealed that these two viruses might have their ancestral origin from pigs, though some of their gene segments were related to human strains. The study shows evidence of reassortment and possible interspecies transmission between pigs and humans in Zambia. Therefore, the “One Health” surveillance approach for rotavirus A in animals and humans is recommended to inform the design of effective control measures. - Chromosome-level genome assembly of Babesia caballi reveals diversity of multigene families among Babesia species.
Akihiro Ochi, Taishi Kidaka, Hassan Hakimi, Masahito Asada, Junya Yamagishi
BMC genomics, 24, 1, 483, 483, 2023年08月24日, ［国際誌］
英語, 研究論文（学術雑誌）, BACKGROUND: Babesia caballi is an intraerythrocytic parasite from the phylum Apicomplexa, capable of infecting equids and causing equine piroplasmosis. However, since there is limited genome information available on B. caballi, molecular mechanisms involved in host specificity and pathogenicity of this species have not been fully elucidated yet. RESULTS: Genomic DNA from a B. caballi subclone was purified and sequenced using both Illumina and Nanopore technologies. The resulting assembled sequence consisted of nine contigs with a size of 12.9 Mbp, rendering a total of 5,910 protein-coding genes. The phylogenetic tree of Apicomplexan species was reconstructed using 263 orthologous genes. We identified 481 ves1-like genes and named "ves1c". In contrast, expansion of the major facilitator superfamily (mfs) observed in closely related B. bigemina and B. ovata species was not found in B. caballi. A set of repetitive units containing an open reading frame with a size of 297 bp was also identified. CONCLUSIONS: We present a chromosome-level genome assembly of B. caballi. Our genomic data may contribute to estimating gene expansion events involving multigene families and exploring the evolution of species from this genus. - Draft genome sequence data of Haemaphysalis longicornis Oita strain
Rika Umemiya-Shirafuji, Xuenan Xuan, Kozo Fujisaki, Junya Yamagishi
Data in Brief, 49, 109352, 109352, 2023年08月, ［国際誌］
英語, 研究論文（学術雑誌）, Haemaphysalis longicornis Neumann, 1901 is one of the most well-known hard ticks because of its medical and veterinary importance. Haemaphysalis longicornis transmit a wide range of pathogens among vertebrates, affecting humans and animals in Asia and Oceania. In Japan, the tick species is a major pest of cattle because it can spread a protozoan parasite Theileria orientalis, which causes theileriosis and produces economic losses to the livestock industry (Yokoyama et al. 2012 [1]). Apart from bovine theileriosis, H. longicornis is a vector of bovine babesiosis caused by Babesia ovata, canine babesiosis caused by Babesia gibsoni, and rickettsiosis and viral diseases in humans. Its habitats are mainly Japan, Australia, New Zealand, New Caledonia, the Fiji Islands, Korea, China, and Russia (Oliver et al. 1973 [2]). In the United States, heavy H. longicornis infestations on cattle and white-tailed deer were reported in 2019, making it now one of the tick species to be an increasing threat to livestock animals and humans globally. Ticks reproduce offspring after mating with female and male ticks, however, interestingly, there are two races of H. longicornis: bisexual (diploid) and parthenogenetic (triploid) races [2]. Parthenogenetic H. longicornis is distributed throughout Japan, while the northern limit of the bisexual race is believed to be Fukushima Prefecture on Honshu Island (Fujita et al. 2013 and Kitaoka et al. 1961 [3,4]). This tick species is also considered to be of great scientific importance, and the parthenogenetic race collected in Okayama prefecture has been reared since 1961, while the bisexual race collected in Oita prefecture has been reared since 2008 under laboratory conditions in Japan (Boldbaatar et al. 2010 and Fujisaki et al. 1976 [5,6]). Namely, the "Okayama strain" and "Oita strain" of H. longicornis have been maintained for more than six decades and 15 years, respectively, stably under laboratory conditions. To obtain reference data of bisexual H. longicornis, we sequenced unfed females with haploid genomes using Illumina and MinION Q20 kit then obtained a draft genome consisting of 2.48 Gbp. The number of the contig was 98,529 and N50 was 46.5 Kb. Genome information derived from our laboratory colony of bisexual H. longicornis ticks would provide fundamental insight into understanding how different reproductive lineages occur within the same species of the tick. - 一酪農家におけるクリプトスポリジウム症の清浄化への試み
高橋 和瑛, 松尾 智英, 山岸 潤也, 芝原 友幸, 笹井 和美, 松林 誠
家畜診療, 70, 7, 407, 416, (公社)全国農業共済協会, 2023年07月
日本語 - Surveillance, Isolation, and Genetic Characterization of Bat Herpesviruses in Zambia.
Hayato Harima, Yongjin Qiu, Junya Yamagishi, Masahiro Kajihara, Katendi Changula, Kosuke Okuya, Mao Isono, Tomoyuki Yamaguchi, Hirohito Ogawa, Naganori Nao, Michihito Sasaki, Edgar Simulundu, Aaron S Mweene, Hirofumi Sawa, Kanako Ishihara, Bernard M Hang'ombe, Ayato Takada
Viruses, 15, 6, 2023年06月13日, ［国際誌］
英語, 研究論文（学術雑誌）, Bats are of significant interest as reservoirs for various zoonotic viruses with high diversity. During the past two decades, many herpesviruses have been identified in various bats worldwide by genetic approaches, whereas there have been few reports on the isolation of infectious herpesviruses. Herein, we report the prevalence of herpesvirus infection of bats captured in Zambia and genetic characterization of novel gammaherpesviruses isolated from striped leaf-nosed bats (Macronycteris vittatus). By our PCR screening, herpesvirus DNA polymerase (DPOL) genes were detected in 29.2% (7/24) of Egyptian fruit bats (Rousettus aegyptiacus), 78.1% (82/105) of Macronycteris vittatus, and one Sundevall's roundleaf bat (Hipposideros caffer) in Zambia. Phylogenetic analyses of the detected partial DPOL genes revealed that the Zambian bat herpesviruses were divided into seven betaherpesvirus groups and five gammaherpesvirus groups. Two infectious strains of a novel gammaherpesvirus, tentatively named Macronycteris gammaherpesvirus 1 (MaGHV1), were successfully isolated from Macronycteris vittatus bats, and their complete genomes were sequenced. The genome of MaGHV1 encoded 79 open reading frames, and phylogenic analyses of the DNA polymerase and glycoprotein B demonstrated that MaGHV1 formed an independent lineage sharing a common origin with other bat-derived gammaherpesviruses. Our findings provide new information regarding the genetic diversity of herpesviruses maintained in African bats. - Identification of three members of the multidomain adhesion CCp family in Babesia gibsoni.
Hang Li, Shengwei Ji, Eloiza May Galon, Iqra Zafar, Zhuowei Ma, Thom Do, Moaz M Amer, Yihong Ma, Junya Yamagishi, Mingming Liu, Xuenan Xuan
Acta tropica, 241, 106890, 106890, 2023年05月, ［国際誌］
英語, 研究論文（学術雑誌）, Babesia gibsoni is an intraerythrocytic apicomplexan parasite transmitted by Haemaphysalis longicornis and causes canine babesiosis. Within the tick, the Babesia parasite undergoes sexual conjugation and the sporogony process of its life cycle. To control B. gibsoni infection, prompt and effective treatment of acute infections and curing chronic carriers are urgently needed. Gene disruption of Plasmodium CCps resulted in blocking the transition of sporozoites from the mosquito midgut to the salivary glands, showing that these proteins are potential targets for the development of a transmission-blocking vaccine. In this study, we described the identification and characterization of three members of the CCp family in B. gibsoni, named CCp1, CCp2, and CCp3. The B. gibsoni sexual stages were induced in vitro by exposing parasites to xanthurenic acid (XA), dithiothreitol (DTT), and tris (2-carboxyethyl) phosphine (TCEP) at serial concentrations. Among them, 100 µM XA-exposed and cultured at 27 °C without CO2B. gibsoni presented diverse morphologies, including parasites with long projections, gradually increased free merozoites, and aggregated and round forms, indicative of sexual stage induction. Then, the expression of CCp proteins of induced parasites was confirmed by real-time reverse transcription PCR, immunofluorescence, and western blot. The results showed that BgCCp genes were highly significantly increased at 24 h post-sexual stage induction (p < 0.01). The induced parasites were recognized by anti-CCp mouse antisera and anti-CCp 1, 2, and 3 antibodies weakly reacted with sexual stage proteins of expected molecular weights of 179.4, 169.8, and 140.0 KDa, respectively. Our observations on morphological changes and confirmation of sexual stage protein expression will advance elemental biological research and lay the foundation for the development of transmission-blocking vaccines against canine babesiosis. - タイにおける豚からヒトへのLA-MRSAの人獣共通感染の全ゲノム解析(Whole Genome Analysis of Zoonotic Transmission of LA-MRSA from Pigs to Humans in Thailand)
Narongpun Pawarut, チャンチャイトン・パッタラット, 山岸 潤也, 中島 千絵, 鈴木 定彦
日本細菌学雑誌, 78, 1, 109, 109, 日本細菌学会, 2023年02月
英語 - Current status and molecular epidemiology of rabies virus from different hosts and regions in Malawi.
Henson Kainga, Elisha Chatanga, Marvin Collen Phonera, John Pilate Kothowa, Precious Dzimbiri, Gladson Kamwendo, Malala Mulavu, Cynthia Sipho Khumalo, Katendi Changula, Herman Chambaro, Hayato Harima, Masahiro Kajihara, Kholiwe Mkandawire, Patrick Chikungwa, Julius Chulu, Gilson Njunga, Simbarashe Chitanga, Benjamin Mubemba, Michihito Sasaki, Yasuko Orba, Yongjin Qiu, Junya Yamagishi, Edgar Simulundu, Ayato Takada, Boniface Namangala, Hirofumi Sawa, Walter Muleya
Archives of virology, 168, 2, 61, 61, 2023年01月12日, ［国際誌］
英語, 研究論文（学術雑誌）, Although rabies is endemic in Malawi, there have been no studies in which rabies virus was systematically investigated and characterized in multiple animal hosts in that country. In order to provide molecular epidemiological data on rabies virus in Malawi, 683 suspected rabies case reports from 2008 to 2021 were examined, and 46 (dog = 40, cow = 5, and cat = 1) viable rabies-positive brain samples archived at the Central Veterinary Laboratory (CVL), Lilongwe, Malawi, were analyzed genetically. The results showed an increase in the submission of brain samples from 2008 to 2010, with the highest number of submissions observed in 2020. Of the 683 case reports analyzed for the period under review, 38.1% (260/683) (CI: 34.44 - 41.84) were confirmed by direct fluorescent antibody test. Among the confirmed cases, 65.4% (170/260) (CI: 59.23 - 71.09) were canine rabies. Further, phylogenetic analysis revealed that sequences from different animal hosts clustered together within the Africa 1b lineage, suggesting that the strains circulating in livestock are similar to those in domestic dogs. This finding supports the hypothesis that canine rabies is spilling over to livestock and emphasizes the need for further studies to provide data for effective control of rabies in Malawi. - Whole genome sequence and diversity in multigene families of Babesia ovis.
Junya Yamagishi, Onur Ceylan, Xuenan Xuan, Ferda Sevinc
Frontiers in cellular and infection microbiology, 13, 1194608, 1194608, 2023年, ［国際誌］
英語, 研究論文（学術雑誌）, Ovine babesiosis, caused by Babesia ovis, is an acute, lethal, and endemic disease worldwide and causes a huge economic loss to animal industry. Pathogen genome sequences can be utilized for selecting diagnostic markers, drug targets, and antigens for vaccine development; however, those for B. ovis have not been available so far. In this study, we obtained a draft genome sequence for B. ovis isolated from an infected sheep in Turkey. The genome size was 7.81 Mbp with 3,419 protein-coding genes. It consisted of 41 contigs, and the N50 was 526 Kbp. There were 259 orthologs identified among eight Babesia spp., Plasmodium falciparum, and Toxoplasma gondii. A phylogeny was estimated on the basis of the orthologs, which showed B. ovis to be closest to B. bovis. There were 43 ves genes identified using hmm model as well. They formed a discriminating cluster to other ves multigene family of Babesia spp. but showed certain similarities to those of B. bovis, B. caballi, and Babesia sp. Xinjiang, which is consistent with the phylogeny. Comparative genomics among B. ovis and B. bovis elucidated uniquely evolved genes in these species, which may account for the adaptation. - 一酪農場におけるクリプトスポリジウム症の清浄化への試み
高橋 和瑛, 松尾 智英, 山岸 潤也, 芝原 友幸, 笹井 和美, 松林 誠
家畜診療, 70, 1, 47, 48, (公社)全国農業共済協会, 2023年01月
日本語 - Field-deployable multiplex detection method of SARS-CoV-2 and influenza virus using loop-mediated isothermal amplification and DNA chromatography.
Kyoko Hayashida, Alejandro Garcia, Lavel Chinyama Moonga, Tatsuki Sugi, Kodera Takuya, Mitsuo Kawase, Fumihiro Kodama, Atsushi Nagasaka, Nobuhisa Ishiguro, Ayato Takada, Masahiro Kajihara, Naganori Nao, Masashi Shingai, Hiroshi Kida, Yasuhiko Suzuki, William W Hall, Hirofumi Sawa, Junya Yamagishi
PloS one, 18, 5, e0285861, 2023年, ［国際誌］
英語, 研究論文（学術雑誌）, A novel multiplex loop-mediated isothermal amplification (LAMP) method combined with DNA chromatography was developed for the simultaneous detection of three important respiratory disease-causing viruses: severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), influenza A virus, and influenza B virus. Amplification was performed at a constant temperature, and a positive result was confirmed by a visible colored band. An in-house drying protocol with trehalose was used to prepare the dried format multiplex LAMP test. Using this dried multiplex LAMP test, the analytical sensitivity was determined to be 100 copies for each viral target and 100-1000 copies for the simultaneous detection of mixed targets. The multiplex LAMP system was validated using clinical COVID-19 specimens and compared with the real-time qRT-PCR method as a reference test. The determined sensitivity of the multiplex LAMP system for SARS-CoV-2 was 71% (95% CI: 0.62-0.79) for cycle threshold (Ct) ≤ 35 samples and 61% (95% CI: 0.53-0.69) for Ct ≤40 samples. The specificity was 99% (95%CI: 0.92-1.00) for Ct ≤35 samples and 100% (95%CI: 0.92-1.00) for the Ct ≤40 samples. The developed simple, rapid, low-cost, and laboratory-free multiplex LAMP system for the two major important respiratory viral diseases, COVID-19 and influenza, is a promising field-deployable diagnosis tool for the possible future 'twindemic, ' especially in resource-limited settings. - Cloning, Expression and Evaluation of Thioredoxin Peroxidase-1 Antigen for the Serological Diagnosis of Schistosoma mekongi Human Infection.
Atcharaphan Wanlop, Jose Ma M Angeles, Adrian Miki C Macalanda, Masashi Kirinoki, Yuma Ohari, Aya Yajima, Junya Yamagishi, Kevin Austin L Ona, Shin-Ichiro Kawazu
Diagnostics (Basel, Switzerland), 12, 12, 2022年12月07日, ［国際誌］
英語, 研究論文（学術雑誌）, Schistosoma mekongi, a blood fluke that causes Asian zoonotic schistosomiasis, is distributed in communities along the Mekong River in Cambodia and Lao People's Democratic Republic. Decades of employing numerous control measures including mass drug administration using praziquantel have resulted in a decline in the prevalence of schistosomiasis mekongi. This, however, led to a decrease in sensitivity of Kato-Katz stool microscopy considered as the gold standard in diagnosis. In order to develop a serological assay with high sensitivity and specificity which can replace Kato-Katz, recombinant S. mekongi thioredoxin peroxidase-1 protein (rSmekTPx-1) was expressed and produced. Diagnostic performance of the rSmekTPx-1 antigen through ELISA for detecting human schistosomiasis was compared with that of recombinant protein of S. japonicum TPx-1 (rSjTPx-1) using serum samples collected from endemic foci in Cambodia. The sensitivity and specificity of rSmekTPx-1 in ELISA were 89.3% and 93.3%, respectively, while those of rSjTPx-1 were 71.4% and 66.7%, respectively. In addition, a higher Kappa value of 0.82 calculated between rSmekTPx-1 antigen ELISA and Kato-Katz confirmed better agreement than between rSjTPx-1 antigen ELISA and Kato-Katz (Kappa value 0.38). These results suggest that ELISA with rSmekTPx-1 antigen can be a potential diagnostic method for detecting active human S. mekongi infection. - Leishmania infection-induced multinucleated giant cell formation via upregulation of ATP6V0D2 expression
Jing Hong, Chizu Sanjoba, Wataru Fujii, Junya Yamagishi, Yasuyuki Goto
Frontiers in Cellular and Infection Microbiology, 12, Frontiers Media SA, 2022年09月23日
研究論文（学術雑誌）, Leishmaniasis is caused by infection with protozoan parasites of the genus Leishmania. In both clinical and experimental visceral leishmaniasis, macrophage multinucleation is observed in parasitized tissues. However, the feature and the mechanism of macrophage multinucleation remained unclear. Here, we report that infection of Leishmania donovani, a causative agent of visceral leishmaniasis, induces multinucleation of bone marrow-derived macrophages (BMDMs) in vitro. When these infection-induced multinucleated macrophages were compared with cytokine-induced multinucleated giant cells, the former had higher phagocytic activity on red blood cells but no apparent changes on phagocytosis of latex beads. BMDMs infected with L. donovani had increased expression of ATP6V0D2, one of the components of V-ATPase, which was also upregulated in the spleen of infected mice. Infection-induced ATP6V0D2 localized in a cytoplasmic compartment, which did not overlap with the mitochondria, endoplasmic reticulum, or lysosomes. When ATP6V0D2 expression was recombinantly induced in BMDMs, the formation of multinucleated macrophages was induced as seen in the infected macrophages. Taken together, L. donovani infection induces multinucleation of macrophages via ATP6V0D2 upregulation leading to a unique metamorphosis of the macrophages toward hemophagocytes. - Single-cell analytical technologies: uncovering the mechanisms behind variations in immune responses.
Yukie Kashima, Patrick Reteng, Yasuhiko Haga, Junya Yamagishi, Yutaka Suzuki
The FEBS journal, 2022年09月09日, ［国際誌］
英語, 研究論文（学術雑誌）, The immune landscape varies among individuals. It determines the immune response and results in surprisingly diverse symptoms, even in response to similar external stimuli. However, the detailed mechanisms underlying such diverse immune responses have remained mostly elusive. The utilization of recently developed single-cell multimodal analysis platforms has started to answer this question. Emerging studies have elucidated several molecular networks that may explain diversity with respect to age or other factors. An elaborate interplay between inherent physical conditions and environmental conditions has been demonstrated. Furthermore, the importance of modifications by the epigenome resulting in transcriptome variation among individuals is gradually being revealed. Accordingly, epigenomes and transcriptomes are direct indicators of the medical history and dynamic interactions with environmental factors. COVID-19 has recently become one of the most remarkable examples of the necessity of in-depth analyses of diverse responses with respect to various factors to improve treatment in severe cases and to prevent viral transmission from asymptomatic carriers. In fact, determining why some patients develop serious symptoms is still a pressing issue. Here, we review the current "state of the art" in single-cell analytical technologies and their broad applications to healthy individuals and representative diseases, including COVID-19. - SICA-mediated cytoadhesion of Plasmodium knowlesi-infected red blood cells to human umbilical vein endothelial cells.
Huai Chuang, Miako Sakaguchi, Amuza Byaruhanga Lucky, Junya Yamagishi, Yuko Katakai, Satoru Kawai, Osamu Kaneko
Scientific reports, 12, 1, 14942, 14942, 2022年09月02日, ［国際誌］
英語, 研究論文（学術雑誌）, Zoonotic malaria due to Plasmodium knowlesi infection in Southeast Asia is sometimes life-threatening. Post-mortem examination of human knowlesi malaria cases showed sequestration of P. knowlesi-infected red blood cells (iRBCs) in blood vessels, which has been proposed to be linked to disease severity. This sequestration is likely mediated by the cytoadhesion of parasite-iRBCs to vascular endothelial cells; however, the responsible parasite ligands remain undetermined. This study selected P. knowlesi lines with increased iRBC cytoadhesion activity by repeated panning against human umbilical vein endothelial cells (HUVECs). Transcriptome analysis revealed that the transcript level of one gene, encoding a Schizont Infected Cell Agglutination (SICA) protein, herein termed SICA-HUVEC, was more than 100-fold increased after the panning. Transcripts of other P. knowlesi proteins were also significantly increased, such as PIR proteins exported to the iRBC cytosol, suggesting their potential role in increasing cytoadhesion activity. Transgenic P. knowlesi parasites expressing Myc-fused SICA-HUVEC increased cytoadhesion activity following infection of monkey as well as human RBCs, confirming that SICA-HUVEC conveys activity to bind to HUVECs. - Advances in understanding red blood cell modifications by Babesia.
Hassan Hakimi, Junya Yamagishi, Shin-Ichiro Kawazu, Masahito Asada
PLoS pathogens, 18, 9, e1010770, 2022年09月, ［国際誌］
英語, 研究論文（学術雑誌）, Babesia are tick-borne protozoan parasites that can infect livestock, pets, wildlife animals, and humans. In the mammalian host, they invade and multiply within red blood cells (RBCs). To support their development as obligate intracellular parasites, Babesia export numerous proteins to modify the RBC during invasion and development. Such exported proteins are likely important for parasite survival and pathogenicity and thus represent candidate drug or vaccine targets. The availability of complete genome sequences and the establishment of transfection systems for several Babesia species have aided the identification and functional characterization of exported proteins. Here, we review exported Babesia proteins; discuss their functions in the context of immune evasion, cytoadhesion, and nutrient uptake; and highlight possible future topics for research and application in this field. - Behavior and toxic effects of Pb in a waterfowl model with oral exposure to Pb shots: Investigating Pb exposure in wild birds.
Hiroshi Sato, Chihiro Ishii, Shouta M M Nakayama, Takahiro Ichise, Keisuke Saito, Yukiko Watanabe, Kohei Ogasawara, Ryota Torimoto, Atsushi Kobayashi, Takashi Kimura, Yukiko Nakamura, Junya Yamagishi, Yoshinori Ikenaka, Mayumi Ishizuka
Environmental pollution (Barking, Essex : 1987), 308, 119580, 119580, 2022年09月01日, ［国際誌］
英語, 研究論文（学術雑誌）, Among wild birds, lead (Pb) exposure caused by ingestion of ammunition is a worldwide problem. We aimed to reveal the behavior and toxic effect of Pb caused by ingesting Pb shots in waterfowl. Four male, eight-week old Muscovy ducks (Cairina moschata) were given three Pb shots (approximately 240 mg in total) orally and then fed for 29 days after exposure, simulating a low-dose Pb exposure in wild waterfowl. During the breeding period, blood samples were collected 10 times, and fecal samples every day. Additionally, 22 fresh tissue and 6 bone samples were obtained from each duck through the dissection. Although there were no gross abnormalities, the maximum blood Pb concentration of each duck ranged from 0.6 to 3.7 mg/L, reaching a threshold concentration indicative of clinical symptoms (>0.5 mg/L). δ-aminolevulinic acid dehydratase declined one day after exposure and remained low throughout the feeding period. Hematocrit also tended to decrease, indicating signs of anemia. The highest Pb accumulation was observed in the bones, followed by the kidneys, intestinal tracts, and liver. High Pb accumulation in the bones, which are known to have a long Pb half-life, suggested that Pb would remain in the body and possibly affect bird health beyond 28 days after exposure. Gene expression analysis showed a significant increase in the expression of the toll-like receptor-3 gene, which is involved in virus discrimination in the liver, suggesting a disruption of the immune system. Microbiota analyses showed a correlation between the blood Pb concentration and the abundances of Lachnospiraceae and Ruminococcaceae, suggesting that Pb affects lipid metabolism. These results provide fundamental data on Pb exposure in wild birds and a new perspective on the damage such exposure causes. - Circular Whole-Transcriptome Amplification (cWTA) and mNGS Screening Enhanced by a Group Testing Algorithm (mEGA) Enable High-Throughput and Comprehensive Virus Identification.
Patrick Reteng, Linh Nguyen Thuy, Mizanur Rahman, Ana Maria Bispo de Filippis, Kyoko Hayashida, Tatsuki Sugi, Gabriel Gonzalez, William W Hall, Lan Anh Nguyen Thi, Junya Yamagishi
mSphere, e0033222, 2022年08月25日, ［国際誌］
英語, 研究論文（学術雑誌）, Metagenomic next-generation sequencing (mNGS) offers a hypothesis-free approach for pathogen detection, but its applicability in clinical diagnosis, in addition to other factors, remains limited due to complicated library construction. The present study describes a PCR-free isothermal workflow for mNGS targeting RNA, based on a multiple displacement amplification, termed circular whole-transcriptome amplification (cWTA), as the template is circularized before amplification. The cWTA approach was validated with clinical samples and nanopore sequencing. Reads homologous to dengue virus 2 and chikungunya virus were detected in clinical samples from Bangladesh and Brazil, respectively. In addition, the practicality of a high-throughput detection system that combines mNGS and a group testing algorithm termed mNGS screening enhanced by a group testing algorithm (mEGA) was established. This approach enabled significant library size reduction while permitting trackability between samples and diagnostic results. Serum samples of patients with undifferentiated febrile illnesses from Vietnam (n = 43) were also amplified with cWTA, divided into 11 pools, processed for library construction, and sequenced. Dengue virus 2, hepatitis B virus, and parvovirus B19 were successfully detected without prior knowledge of their existence. Collectively, cWTA with the nanopore platform opens the possibility of hypothesis-free on-site comprehensive pathogen diagnosis, while mEGA contributes to the scaling up of sample throughput. IMPORTANCE Given the breadth of pathogens that cause infections, a single approach that can detect a wide range of pathogens is ideal but is impractical due to the available tests being highly specific to a certain pathogen. Recent developments in sequencing technology have introduced mNGS as an alternative that provides detection of a wide-range of pathogens by detecting the presence of their nucleic acids in the sample. However, sequencing library preparation is still a bottleneck, as it is complicated, costly, and time-consuming. In our studies, alternative approaches to optimize library construction for mNGS were developed. This included isothermal nucleic acid amplification and expansion of sample throughput with a group testing algorithm. These methods can improve the utilization of mNGS as a diagnostic tool and can serve as a high-throughput screening system aiding infectious disease surveillance. - Genomic Surveillance of SARS-CoV-2 in the Southern Province of Zambia: Detection and Characterization of Alpha, Beta, Delta, and Omicron Variants of Concern.
Ben Katowa, Annie Kalonda, Benjamin Mubemba, Japhet Matoba, Doreen Mainza Shempela, Jay Sikalima, Boniface Kabungo, Katendi Changula, Simbarashe Chitanga, Mpanga Kasonde, Otridah Kapona, Nathan Kapata, Kunda Musonda, Mwaka Monze, John Tembo, Matthew Bates, Alimuddin Zumla, Catherine G Sutcliffe, Masahiro Kajihara, Junya Yamagishi, Ayato Takada, Hirofumi Sawa, Roma Chilengi, Victor Mukonka, Walter Muleya, Edgar Simulundu
Viruses, 14, 9, 2022年08月24日, ［国際誌］
英語, 研究論文（学術雑誌）, Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) variants of concern (VOCs) have significantly impacted the global epidemiology of the pandemic. From December 2020 to April 2022, we conducted genomic surveillance of SARS-CoV-2 in the Southern Province of Zambia, a region that shares international borders with Botswana, Namibia, and Zimbabwe and is a major tourist destination. Genetic analysis of 40 SARS-CoV-2 whole genomes revealed the circulation of Alpha (B.1.1.7), Beta (B.1.351), Delta (AY.116), and multiple Omicron subvariants with the BA.1 subvariant being predominant. Whereas Beta, Delta, and Omicron variants were associated with the second, third, and fourth pandemic waves, respectively, the Alpha variant was not associated with any wave in the country. Phylogenetic analysis showed evidence of local transmission and possible multiple introductions of SARS-CoV-2 VOCs in Zambia from different European and African countries. Across the 40 genomes analysed, a total of 292 mutations were observed, including 182 missense mutations, 66 synonymous mutations, 23 deletions, 9 insertions, 1 stop codon, and 11 mutations in the non-coding region. This study stresses the need for the continued monitoring of SARS-CoV-2 circulation in Zambia, particularly in strategically positioned regions such as the Southern Province which could be at increased risk of introduction of novel VOCs. - Phylogenetic analyses of the mitochondrial, plastid, and nuclear genes of Babesia sp. Mymensingh and its naming as Babesia naoakii n. sp.
Thillaiampalam Sivakumar, Bumduuren Tuvshintulga, Davaajav Otgonsuren, Enkhbaatar Batmagnai, Believe Ahedor, Hemal Kothalawala, Singarayar Caniciyas Vimalakumar, Seekkuge Susil Priyantha Silva, Junya Yamagishi, Naoaki Yokoyama
Parasites & vectors, 15, 1, 299, 299, 2022年08月24日, ［国際誌］
英語, 研究論文（学術雑誌）, BACKGROUND: The recently discovered Babesia sp. Mymensingh, which causes clinical bovine babesiosis, has a wide geographical distribution. We investigated the phylogenetic position of Babesia sp. Mymensingh using its mitochondrial, plastid, and nuclear genes. Based on morphological and molecular data, Babesia sp. Mymensingh is a unique species and we named it as Babesia naoakii n. sp. METHODS: A blood DNA sample from a Babesia sp. Mymensingh-infected cow was subjected to genome sequencing to obtain the sequences of mitochondrial, plastid, and nuclear genes. Six phylogenetic trees were then constructed with (1) concatenated amino acid sequences of cytochrome oxidase subunit I, cytochrome oxidase subunit III, and cytochrome b genes of the mitochondrial genome; (2) 16S rRNA of the plastid genome; (3) nucleotide sequences of the elongation factor Tu gene of the plastid genome; (4) ITS1-5.8S rRNA-ITS2; (5) concatenated nucleotide sequences of 89 nuclear genes; and (6) concatenated amino acid sequences translated from the 89 nuclear genes. RESULTS: In all six phylogenetic trees, B. naoakii n. sp. formed a sister clade to the common ancestor of Babesia bigemina and B. ovata. The concatenated nuclear genes of B. naoakii n. sp. and their translated amino acid sequences shared lower identity scores with the sequences from B. bigemina (82.7% and 84.7%, respectively) and B. ovata (83.5% and 85.5%, respectively) compared with the identity scores shared between the B. bigemina and B. ovata sequences (86.3% and 87.9%, respectively). CONCLUSIONS: Our study showed that B. naoakii n. sp. occupies a unique phylogenetic position distinct from existing Babesia species. Our findings, together with morphological differences, identify B. naoakii n. sp. as a distinct parasite species. - Inactivated whole influenza virus particle vaccines induce neutralizing antibodies with an increase in immunoglobulin gene subclones of B-lymphocytes in cynomolgus macaques.
Masanori Shiohara, Saori Suzuki, Shintaro Shichinohe, Hirohito Ishigaki, Misako Nakayama, Naoki Nomura, Masashi Shingai, Toshiki Sekiya, Marumi Ohno, Sayaka Iida, Naoko Kawai, Mamiko Kawahara, Junya Yamagishi, Kimihito Ito, Ryotarou Mitsumata, Tomio Ikeda, Kenji Motokawa, Tomoyoshi Sobue, Hiroshi Kida, Kazumasa Ogasawara, Yasushi Itoh
Vaccine, 40, 30, 4026, 4037, 2022年06月26日, ［査読有り］, ［国際誌］
英語, 研究論文（学術雑誌）, The All-Japan Influenza Vaccine Study Group has been developing a more effective vaccine than the current split vaccines for seasonal influenza virus infection. In the present study, the efficacy of formalin- and/or β-propiolactone-inactivated whole virus particle vaccines for seasonal influenza was compared to that of the current ether-treated split vaccines in a nonhuman primate model. The monovalent whole virus particle vaccines or split vaccines of influenza A virus (H1N1) and influenza B virus (Victoria lineage) were injected subcutaneously into naïve cynomolgus macaques twice. The whole virus particle vaccines induced higher titers of neutralizing antibodies against H1N1 influenza A virus and influenza B virus in the plasma of macaques than did the split vaccines. At challenge with H1N1 influenza A virus or influenza B virus, the virus titers in nasal swabs and the increases in body temperatures were lower in the macaques immunized with the whole virus particle vaccine than in those immunized with the split vaccine. Repertoire analyses of immunoglobulin heavy chain genes demonstrated that the number of B-lymphocyte subclones was increased in macaques after the 1st vaccination with the whole virus particle vaccine, but not with the split vaccine, indicating that the whole virus particle vaccine induced the activation of vaccine antigen-specific B-lymphocytes more vigorously than did the split vaccine at priming. Thus, the present findings suggest that the superior antibody induction ability of the whole virus particle vaccine as compared to the split vaccine is attributable to its stimulatory properties on the subclonal differentiation of antigen-specific B-lymphocytes. - Autochthonous Leishmania infantum in Dogs, Zambia, 2021.
David Squarre, Herman M Chambaro, Kyoko Hayashida, Lavel C Moonga, Yongjin Qiu, Yasuyuki Goto, Elizabeth Oparaocha, Chisoni Mumba, Walter Muleya, Patricia Bwalya, Joseph Chizimu, Mwelwa Chembensofu, Edgar Simulundu, Wizaso Mwasinga, Nelly Banda, Racheal Mwenda, Junya Yamagishi, King S Nalubamba, Fredrick Banda, Musso Munyeme, Hirofumi Sawa, Paul Fandamu
Emerging infectious diseases, 28, 4, 888, 890, 2022年04月, ［国際誌］
英語, 研究論文（学術雑誌）, Leishmaniases are neglected tropical diseases of humans and animals. We detected Leishmania infantum in 3 mixed-breed dogs in Zambia that had no travel history outside the country. Our findings suggest presence of and probable emergence of leishmaniasis in Zambia, indicating the need for physicians and veterinarians to consider the disease during diagnosis. - Global research alliance in infectious disease: a collaborative effort to combat infectious diseases through dissemination of portable sequencing.
Lucky R Runtuwene, Nuankanya Sathirapongsasuti, Raweewan Srisawat, Narumon Komalamisra, Josef S B Tuda, Arthur E Mongan, Gabriel O Aboge, Victoria Shabardina, Wojciech Makalowski, Dela Ria Nesti, Wayan T Artama, Lan Anh Nguyen-Thi, Kiew-Lian Wan, Byoung-Kuk Na, William Hall, Arnab Pain, Yuki Eshita, Ryuichiro Maeda, Junya Yamagishi, Yutaka Suzuki
BMC research notes, 15, 1, 44, 44, 2022年02月12日, ［国際誌］
英語, 研究論文（学術雑誌）, OBJECTIVE: To disseminate the portable sequencer MinION in developing countries for the main purpose of battling infectious diseases, we found a consortium called Global Research Alliance in Infectious Diseases (GRAID). By holding and inviting researchers both from developed and developing countries, we aim to train the participants with MinION's operations and foster a collaboration in infectious diseases researches. As a real-life example in which resources are limited, we describe here a result from a training course, a metagenomics analysis from two blood samples collected from a routine cattle surveillance in Kulan Progo District, Yogyakarta Province, Indonesia in 2019. RESULTS: One of the samples was successfully sequenced with enough sequencing yield for further analysis. After depleting the reads mapped to host DNA, the remaining reads were shown to map to Theileria orientalis using BLAST and OneCodex. Although the reads were also mapped to Clostridium botulinum, those were found to be artifacts derived from the cow genome. An effort to construct a consensus sequence was successful using a reference-based approach with Pomoxis. Hence, we concluded that the asymptomatic cow might be infected with T. orientalis and showed the usefulness of sequencing technology, specifically the MinION platform, in a developing country. - TSS-seq of Toxoplasma gondii sporozoites revealed a novel motif in stage-specific promoters.
Taishi Kidaka, Tatsuki Sugi, Kyoko Hayashida, Yutaka Suzuki, Xuenan Xuan, Jitender P Dubey, Junya Yamagishi
Infection, genetics and evolution : journal of molecular epidemiology and evolutionary genetics in infectious diseases, 98, 105213, 105213, 2022年01月15日, ［国際誌］
英語, 研究論文（学術雑誌）, Toxoplasma gondii is one of the most common zoonotic protozoan parasites. It has three major infectious stages: rapidly multiplying tachyzoites (Tz), slowly replicating bradyzoites (Bz) and a resting/free-living stage, sporozoites (Sz). The regulatory mechanisms governing stage-specific gene expression are not fully understood. Few transcriptional start sites (TSS) are known for Sz. In this study, we obtained TSS of Sz using an oligo-capping method and RNA-seq analysis. We identified 1,043,503 TSS in the Sz transcriptome. These defined 38,973 TSS clusters, of which, 11,925 were expressed in Sz and 1535 TSS differentially expressed in Sz. Based on these data, we defined promoter regions and novel sporozoite stage-specific motifs using MEME. TGTANNTACA was distributed around -55 to -75 regions from each TSS. Interestingly, the same motif was reported in another apicomplexan, Plasmodium berghei, as a cis-element of female-specific gametocyte genes, implying the presence of common regulatory machinery. Further comparative analysis should better define the distribution and function of these elements in other members of this important parasitic phylum. - COVID-19 Whole-Genome Resequencing with Redundant Tiling PCR and Subtract-Based Amplicon Normalization Successfully Characterized SARS-CoV-2 Variants in Clinical Specimens.
Tatsuki Sugi, Mizanur Rahman, Rummana Rahim, Abu Hasan, Naoko Kawai, Kyoko Hayashida, Junya Yamagishi
Interdisciplinary perspectives on infectious diseases, 2022, 2109641, 2109641, 2022年, ［国際誌］
英語, 研究論文（学術雑誌）, With an increasing number of severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) sequences gathered worldwide, we recognize that deletion mutants and nucleotide substitutions that may affect whole-genome sequencing are accumulating. Here, we propose an additional strategy for tiling PCR for whole-genome resequencing, which can make the pipeline robust for mutations at the primer annealing site by a redundant amplicon scheme. We further demonstrated that subtracting overrepresented amplicons from the multiplex PCR products reduced the bias of the next-generation sequencing (NGS) library, resulting in decreasing required sequencing reads per sample. We applied this sequencing strategy to clinical specimens collected in Bangladesh. More than 80% out of the 304 samples were successfully sequenced. Less than 5% were ambiguous nucleotides, and several known variants were detected. With the additional strategies presented here, we believe that whole-genome resequencing of SARS-CoV-2 from clinical samples can be optimized. - Single Cell Transcriptomes of In Vitro Bradyzoite Infected Cells Reveals Toxoplasma gondii Stage Dependent Host Cell Alterations.
Tatsuki Sugi, Tadakimi Tomita, Taishi Kidaka, Naoko Kawai, Kyoko Hayashida, Louis M Weiss, Junya Yamagishi
Frontiers in cellular and infection microbiology, 12, 848693, 848693, 2022年, ［国際誌］
英語, 研究論文（学術雑誌）, Toxoplasma gondii bradyzoites establish chronic infections within their host cells. Recent studies have demonstrated that several parasite effector proteins are translocated to host cells during the bradyzoite stage of chronic infection. To understand the interaction between host cells and bradyzoites at the transcriptomic landscape level, we utilized single-cell RNA-sequencing (scRNA-Seq) to characterize the bradyzoite-induced host cell response. Distinct gene expression profiles were observed in infected host, cells with low parasite mapped reads, and mock (non-exposed) control cells. Gene set enrichment analysis showed that c-Myc and NF-κB signaling and energy metabolic pathways were upregulated by infection. Type I and II interferon response pathways were upregulated in cells with low parasite mapped reads compared to the non-exposed host control cells, and this upregulation effect was reversed in infected cells. Differences were observed in the host cells depending on the differentiation status of the parasites, as determined by BAG1 and SAG1 expression. NF-κB, inflammatory response pathways, and IFN-γ response pathways were downregulated in host cells containing T. gondii BAG1+/SAG1-, whereas this downregulation effect was reversed in case of T. gondii BAG1-/SAG1+. We also identified two distinct host cell subsets that contained T. gondii BAG1+/SAG1-, one of which displayed distinct transcriptomes with upregulated c-Myc expression. Overall, these data clearly demonstrate that host cell transcriptional alteration by bradyzoite infection is different from that of tachyzoite infection, indicating fine-tuning of the host immune response. - SARS-CoV-2 Genome Sequences, Including One with an ORF7a Deletion, Obtained from Patients in Bangladesh.
Mizanur Rahman, Rummana Rahim, Abu Hasan, Naoko Kawai, Lavel Chinyama Moonga, Tatsuki Sugi, Kyoko Hayashida, Junya Yamagishi
Microbiology resource announcements, 10, 49, e0076421, 2021年12月09日, ［国際誌］
英語, 研究論文（学術雑誌）, Genomic sequences from a complete SARS-CoV-2 open reading frame (ORF) were obtained from 24 patients diagnosed in May 2020 in Dhaka, Bangladesh. All sequences belonged to clade 20A or 20B, and none were variants of concern. Interestingly, one sequence showed a 161-nucleotide deletion in ORF7a. - Structural Requirements in the Hemagglutinin Cleavage Site-Coding RNA Region for the Generation of Highly Pathogenic Avian Influenza Virus.
Yurie Kida, Kosuke Okuya, Takeshi Saito, Junya Yamagishi, Aiko Ohnuma, Takanari Hattori, Hiroko Miyamoto, Rashid Manzoor, Reiko Yoshida, Naganori Nao, Masahiro Kajihara, Tokiko Watanabe, Ayato Takada
Pathogens (Basel, Switzerland), 10, 12, 2021年12月09日, ［国際誌］
英語, 研究論文（学術雑誌）, Highly pathogenic avian influenza viruses (HPAIVs) with H5 and H7 hemagglutinin (HA) subtypes are derived from their low pathogenic counterparts following the acquisition of multiple basic amino acids in their HA cleavage site. It has been suggested that consecutive adenine residues and a stem-loop structure in the viral RNA region that encodes the cleavage site are essential for the acquisition of the polybasic cleavage site. By using a reporter assay to detect non-templated nucleotide insertions, we found that insertions more frequently occurred in the RNA region (29 nucleotide-length) encoding the cleavage site of an H5 HA gene that was predicted to have a stem-loop structure containing consecutive adenines than in a mutated corresponding RNA region that had a disrupted loop structure with fewer adenines. In virus particles generated by using reverse genetics, nucleotide insertions that created additional codons for basic amino acids were found in the RNA region encoding the cleavage site of an H5 HA gene but not in the mutated RNA region. We confirmed the presence of virus clones with the ability to replicate without trypsin in a plaque assay and to cause lethal infection in chicks. These results demonstrate that the stem-loop structure containing consecutive adenines in HA genes is a key molecular determinant for the emergence of H5 HPAIVs. - Evidence of Borrelia theileri in Wild and Domestic Animals in the Kafue Ecosystem of Zambia.
Yongjin Qiu, David Squarre, Yukiko Nakamura, Alice C C Lau, Lavel Chinyama Moonga, Naoko Kawai, Aiko Ohnuma, Kyoko Hayashida, Ryo Nakao, Junya Yamagishi, Hirofumi Sawa, Boniface Namangala, Hiroki Kawabata
Microorganisms, 9, 11, 2021年11月22日, ［国際誌］
英語, 研究論文（学術雑誌）, Members of the genus Borrelia are arthropod-borne spirochetes that are human and animal pathogens. Vertebrate hosts, including wild animals, are pivotal to the circulation and maintenance of Borrelia spirochetes. However, information on Borrelia spirochetes in vertebrate hosts in Zambia is limited. Thus, we aimed to investigate the presence of Borrelia spirochetes in wild animals and cattle in Zambia. A total of 140 wild animals of four species and 488 cattle DNA samples from /near the Kafue National Park were collected for real-time PCR screening, followed by characterization using three different genes with positive samples. Five impalas and 20 cattle tested positive using real-time PCR, and sequence analysis revealed that the detected Borrelia were identified to be Borrelia theileri, a causative agent of bovine borreliosis. This is the first evidence of Borrelia theileri in African wildlife and cattle in Zambia. Our results suggest that clinical differentiation between bovine borreliosis and other bovine diseases endemic in Zambia is required for better treatment and control measures. As this study only included wild and domestic animals in the Kafue ecosystem, further investigations in other areas and with more wildlife and livestock species are needed to clarify a comprehensive epidemiological status of Borrelia theileri in Zambia. - Hepatomegaly Associated with Non-Obstructive Sinusoidal Dilation in Experimental Visceral Leishmaniasis.
Kota Maeda, Sonya Sadoughi, Ayako Morimoto, Kazuyuki Uchida, James K Chambers, Chizu Sanjoba, Junya Yamagishi, Yasuyuki Goto
Pathogens (Basel, Switzerland), 10, 11, 2021年10月20日, ［国際誌］
英語, 研究論文（学術雑誌）, Visceral leishmaniasis (VL) is the most severe form of leishmaniasis caused by protozoan parasites of the genus Leishmania. Hepatomegaly is one of the most frequent clinical manifestations of VL, whereas immunopathology of the symptom has not been well investigated. Using our chronic model of experimental VL, we examined the influence of Leishmania donovani infection on the liver by clinical, histological, and biochemical analyses. The infected mice showed increased liver weight 24 weeks post-infection. Although an increase in serum ALT and inflammatory cell accumulation were observed in the livers of infected mice, no apparent parenchymal necrosis or fibrosis was observed. Tissue water content analyses demonstrated that increased liver weight was predominantly due to an increase in water weight. Together with the finding of hepatic sinusoidal dilation, these results suggested that edema associated with sinusoidal dilation causes hepatomegaly in L. donovani infection. Immunostaining of platelets and erythrocytes showed no thrombus formation or damage to the sinusoidal endothelium in the liver of infected mice. Taken together, these results suggest that hepatomegaly during experimental VL is caused by non-obstructive sinusoidal dilation. - Rickettsia lusitaniae in Ornithodoros Porcinus Ticks, Zambia.
Simbarashe Chitanga, Herman M Chambaro, Lavel C Moonga, Kyoko Hayashida, Junya Yamagishi, Walter Muleya, Katendi Changula, Benjamin Mubemba, Manyando Simbotwe, David Squarre, Paul Fandamu, King S Nalubamba, Yongjin Qiu, Sawa Hirofumi, Edgar Simulundu
Pathogens (Basel, Switzerland), 10, 10, 2021年10月12日, ［国際誌］
英語, 研究論文（学術雑誌）, Rickettsial pathogens are amongst the emerging and re-emerging vector-borne zoonoses of public health importance. Though traditionally considered to be transmitted by ixodid ticks, the role of argasid ticks as vectors of these pathogens is increasingly being recognized. While bat-feeding (Ornithodoros faini) and chicken-feeding (Argas walkerae) argasid ticks have been shown to harbor Rickettsia pathogens in Zambia, there are currently no reports of Rickettsia infection in southern Africa from warthog-feeding (Phacochoerus africanus) soft ticks, particularly Ornithodoros moubata and Ornithodoros porcinus. Our study sought to expand on the existing knowledge on the role of soft ticks in the epidemiology of Rickettsia species through screening for Rickettsia pathogens in warthog burrow-dwelling soft ticks from two national parks in Zambia. The tick species from which Rickettsia were detected in this study were identified as Ornithodoros porcinus, and an overall minimal Rickettsia infection rate of 19.8% (32/162) was observed. All of the sequenced Rickettsia were identified as Rickettsia lusitaniae based on nucleotide sequence similarity and phylogenetic analysis of the citrate synthase (gltA) and 17kDa common antigen (htrA) genes. Utilizing all of the gltA (n = 10) and htrA (n = 12) nucleotide sequences obtained in this study, BLAST analysis showed 100% nucleotide similarity to Rickettsia lusitaniae. Phylogenetic analysis revealed that all of the Zambian gltA and htrA gene sequences could be grouped with those of Rickettsia lusitaniae obtained in various parts of the world. Our data suggest that Rickettsia lusitaniae has a wider geographic and vector range, enhancing to our understanding of Rickettsia lusitaniae epidemiology in sub-Saharan Africa. - A targeted approach with nanopore sequencing for the universal detection and identification of flaviviruses.
Patrick Reteng, Linh Nguyen Thuy, Tam Tran Thi Minh, Maria Angélica Monteiro de Mello Mares-Guia, Maria Celeste Torres, Ana Maria Bispo de Filippis, Yasuko Orba, Shintaro Kobayashi, Kyoko Hayashida, Hirofumi Sawa, William W Hall, Lan Anh Nguyen Thi, Junya Yamagishi
Scientific reports, 11, 1, 19031, 19031, 2021年09月24日, ［国際誌］
英語, 研究論文（学術雑誌）, Nucleic acid test (NAT), most typically quantitative PCR, is one of the standard methods for species specific flavivirus diagnosis. Semi-comprehensive NATs such as pan-flavivirus PCR which covers genus Flavivirus are also available; however, further specification by sequencing is required for species level differentiation. In this study, a semi-comprehensive detection system that allows species differentiation of flaviviruses was developed by integration of the pan-flavivirus PCR and Nanopore sequencing. In addition, a multiplexing method was established by adding index sequences through the PCR with a streamlined bioinformatics pipeline. This enables defining cut-off values for observed read counts. In the laboratory setting, this approach allowed the detection of up to nine different flaviviruses. Using clinical samples collected in Vietnam and Brazil, seven different flaviviruses were also detected. When compared to a commercial NAT, the sensitivity and specificity of our system were 66.7% and 95.4%, respectively. Conversely, when compared to our system, the sensitivity and specificity of the commercial NAT were 57.1% and 96.9%, respectively. In addition, Nanopore sequencing detected more positive samples (n = 8) compared to the commercial NAT (n = 6). Collectively, our study has established a semi-comprehensive sequencing-based diagnostic system for the detection of flaviviruses at extremely affordable costs, considerable sensitivity, and only requires simple experimental methods. - Molecular Detection and Characterization of Rickettsia asembonensis in Human Blood, Zambia.
Lavel C Moonga, Kyoko Hayashida, Namwiinga R Mulunda, Yukiko Nakamura, James Chipeta, Hawela B Moonga, Boniface Namangala, Chihiro Sugimoto, Zephaniah Mtonga, Mable Mutengo, Junya Yamagishi
Emerging infectious diseases, 27, 8, 2237, 2239, 2021年08月, ［査読有り］, ［国際誌］
英語, 研究論文（学術雑誌）, Rickettsia asembonensis is a flea-related Rickettsia with unknown pathogenicity to humans. We detected R. asembonensis DNA in 2 of 1,153 human blood samples in Zambia. Our findings suggest the possibility of R. asembonensis infection in humans despite its unknown pathogenicity. - DNA methylation landscape of 16 canine somatic tissues by methylation-sensitive restriction enzyme-based next generation sequencing.
Jumpei Yamazaki, Yuki Matsumoto, Jaroslav Jelinek, Teita Ishizaki, Shingo Maeda, Kei Watanabe, Genki Ishihara, Junya Yamagishi, Mitsuyoshi Takiguchi
Scientific reports, 11, 1, 10005, 10005, 2021年05月11日, ［査読有り］, ［国際誌］
英語, 研究論文（学術雑誌）, DNA methylation plays important functions in gene expression regulation that is involved in individual development and various diseases. DNA methylation has been well studied in human and model organisms, but only limited data exist in companion animals like dog. Using methylation-sensitive restriction enzyme-based next generation sequencing (Canine DREAM), we obtained canine DNA methylation maps of 16 somatic tissues from two dogs. In total, we evaluated 130,861 CpG sites. The majority of CpG sites were either highly methylated (> 70%, 52.5-64.6% of all CpG sites analyzed) or unmethylated (< 30%, 22.5-28.0% of all CpG sites analyzed) which are methylation patterns similar to other species. The overall methylation status of CpG sites across the 32 methylomes were remarkably similar. However, the tissue types were clearly defined by principle component analysis and hierarchical clustering analysis with DNA methylome. We found 6416 CpG sites located closely at promoter region of genes and inverse correlation between DNA methylation and gene expression of these genes. Our study provides basic dataset for DNA methylation profiles in dogs. - Detection of B.1.351 SARS-CoV-2 Variant Strain - Zambia, December 2020.
Mulenga Mwenda, Ngonda Saasa, Nyambe Sinyange, George Busby, Peter J Chipimo, Jason Hendry, Otridah Kapona, Samuel Yingst, Jonas Z Hines, Peter Minchella, Edgar Simulundu, Katendi Changula, King Shimumbo Nalubamba, Hirofumi Sawa, Masahiro Kajihara, Junya Yamagishi, Muzala Kapin'a, Nathan Kapata, Sombo Fwoloshi, Paul Zulu, Lloyd B Mulenga, Simon Agolory, Victor Mukonka, Daniel J Bridges
MMWR. Morbidity and mortality weekly report, 70, 8, 280, 282, 2021年02月26日, ［査読有り］, ［国際誌］
英語, 研究論文（学術雑誌）, The first laboratory-confirmed cases of coronavirus disease 2019 (COVID-19), the illness caused by SARS-CoV-2, in Zambia were detected in March 2020 (1). Beginning in July, the number of confirmed cases began to increase rapidly, first peaking during July-August, and then declining in September and October (Figure). After 3 months of relatively low case counts, COVID-19 cases began rapidly rising throughout the country in mid-December. On December 18, 2020, South Africa published the genome of a SARS-CoV-2 variant strain with several mutations that affect the spike protein (2). The variant included a mutation (N501Y) associated with increased transmissibility.†,§ SARS-CoV-2 lineages with this mutation have rapidly expanded geographically.¶,** The variant strain (PANGO [Phylogenetic Assignment of Named Global Outbreak] lineage B.1.351††) was first detected in the Eastern Cape Province of South Africa from specimens collected in early August, spread within South Africa, and appears to have displaced the majority of other SARS-CoV-2 lineages circulating in that country (2). As of January 10, 2021, eight countries had reported cases with the B.1.351 variant. In Zambia, the average number of daily confirmed COVID-19 cases increased 16-fold, from 44 cases during December 1-10 to 700 during January 1-10, after detection of the B.1.351 variant in specimens collected during December 16-23. Zambia is a southern African country that shares substantial commerce and tourism linkages with South Africa, which might have contributed to the transmission of the B.1.351 variant between the two countries. - Genetic Diversity of African Trypanosomes in Tsetse Flies and Cattle From the Kafue Ecosystem.
Yukiko Nakamura, Kyoko Hayashida, Victoire Delesalle, Yongjin Qiu, Ryosuke Omori, Martin Simuunza, Chihiro Sugimoto, Boniface Namangala, Junya Yamagishi
Frontiers in veterinary science, 8, 599815, 599815, 2021年, ［国際誌］
英語, 研究論文（学術雑誌）, We clarified the genetic diversity of Trypanosoma spp. within the Kafue ecosystem, using PCR targeting the internal transcribed spacer 1 and the cathepsin L-like cysteine protease (CatL) sequences. The overall prevalence of Trypanosoma spp. in cattle and tsetse flies was 12.65 and 26.85%, respectively. Cattle positive for Trypanosoma vivax had a significantly lower packed cell volume, suggesting that T. vivax is the dominant Trypanosoma spp. causing anemia in this area. Among the 12 operational taxonomic units (OTUs) of T. vivax CatL sequences detected, one was from a known T. vivax lineage, two OTUs were from known T. vivax-like lineages, and nine OTUs were considered novel T. vivax-like lineages. These findings support previous reports that indicated the extensive diversity of T. vivax-like lineages. The findings also indicate that combining CatL PCR with next generation sequencing is useful in assessing Trypanosoma spp. diversity, especially for T. vivax and T. vivax-like lineages. In addition, the 5.42% prevalence of Trypanosoma brucei rhodesiense found in cattle raises concern in the community and requires careful monitoring of human African trypanosomiasis. - Investigation of the piroplasm diversity circulating in wildlife and cattle of the greater Kafue ecosystem, Zambia.
David Squarre, Yukiko Nakamura, Kyoko Hayashida, Naoko Kawai, Herman Chambaro, Boniface Namangala, Chihiro Sugimoto, Junya Yamagishi
Parasites & vectors, 13, 1, 599, 599, 2020年11月30日, ［国際誌］
英語, 研究論文（学術雑誌）, BACKGROUND: Piroplasms are vector-borne intracellular hemoprotozoan parasites that infect wildlife and livestock. Wildlife species are reservoir hosts to a diversity of piroplasms and play an important role in the circulation, maintenance and evolution of these parasites. The potential for likely spillover of both pathogenic and non-pathogenic piroplasm parasites from wildlife to livestock is underlined when a common ecological niche is shared in the presence of a competent vector. METHOD: To investigate piroplasm diversity in wildlife and the cattle population of the greater Kafue ecosystem, we utilized PCR to amplify the 18S rRNA V4 hyper-variable region and meta-barcoding strategy using the Illumina MiSeq sequencing platform and amplicon sequence variant (ASV)-based bioinformatics pipeline to generate high-resolution data that discriminate sequences down to a single nucleotide difference. RESULTS: A parasite community of 45 ASVs corresponding to 23 species consisting of 4 genera of Babesia, Theileria, Hepatozoon and Colpodella, were identified in wildlife and the cattle population from the study area. Theileria species were detected in buffalo, impala, hartebeest, sable antelope, sitatunga, wild dog and cattle. In contrast, Babesia species were only observed in cattle and wild dog. Our results demonstrate possible spillover of these hemoprotozoan parasites from wildlife, especially buffalo, to the cattle population in the wildlife-livestock interface. CONCLUSION: We demonstrated that the deep amplicon sequencing of the 18S rRNA V4 hyper-variable region for wildlife was informative. Our results illustrated the diversity of piroplasma and the specificity of their hosts. They led us to speculate a possible ecological cycle including transmission from wildlife to domestic animals in the greater Kafue ecosystem. Thus, this approach may contribute to the establishment of appropriate disease control strategies in wildlife-livestock interface areas. - Development of a Multiplex Loop-Mediated Isothermal Amplification (LAMP) Method for Simultaneous Detection of Spotted Fever Group Rickettsiae and Malaria Parasites by Dipstick DNA Chromatography.
Lavel Chinyama Moonga, Kyoko Hayashida, Naoko Kawai, Ryo Nakao, Chihiro Sugimoto, Boniface Namangala, Junya Yamagishi
Diagnostics (Basel, Switzerland), 10, 11, 2020年11月02日, ［国際誌］
英語, 研究論文（学術雑誌）, Spotted fever group (SFG) rickettsiae causes febrile illness in humans worldwide. Since SFG rickettsiosis's clinical presentation is nonspecific, it is frequently misdiagnosed as other febrile diseases, especially malaria, and complicates proper treatment. Aiming at rapid, simple, and simultaneous detection of SFG Rickettsia spp. and Plasmodium spp., we developed a novel multiple pathogen detection system by combining a loop-mediated isothermal amplification (LAMP) method and dipstick DNA chromatography technology. Two primer sets detecting SFG Rickettsia spp. and Plasmodium spp. were mixed, and amplified products were visualized by hybridizing to dipstick DNA chromatography. The multiplex LAMP with dipstick DNA chromatography distinguished amplified Rickettsia and Plasmodium targeted genes simultaneously. The determined sensitivity using synthetic nucleotides was 1000 copies per reaction for mixed Rickettsia and Plasmodium genes. When genomic DNA from in vitro cultured organisms was used, the sensitivity was 100 and 10 genome equivalents per reaction for Rickettsia monacensis and Plasmodium falciparum, respectively. Although further improvement will be required for more sensitive detection, our developed simultaneous diagnosis technique will contribute to the differential diagnosis of undifferentiated febrile illness caused by either SFG Rickettsia spp. or Plasmodium spp. in resource-limited endemic areas. Importantly, this scheme is potentially versatile for the simultaneous detection of diverse infectious diseases. - The Lethal(2)-Essential-for-Life [L(2)EFL] Gene Family Modulates Dengue Virus Infection in Aedes aegypti.
Lucky R Runtuwene, Shuichi Kawashima, Victor D Pijoh, Josef S B Tuda, Kyoko Hayashida, Junya Yamagishi, Chihiro Sugimoto, Shoko Nishiyama, Michihito Sasaki, Yasuko Orba, Hirofumi Sawa, Tomohiko Takasaki, Anthony A James, Takashi Kobayashi, Yuki Eshita
International journal of molecular sciences, 21, 20, 2020年10月12日, ［国際誌］
英語, 研究論文（学術雑誌）, Efforts to determine the mosquito genes that affect dengue virus replication have identified a number of candidates that positively or negatively modify amplification in the invertebrate host. We used deep sequencing to compare the differential transcript abundances in Aedes aegypti 14 days post dengue infection to those of uninfected A. aegypti. The gene lethal(2)-essential-for-life [l(2)efl], which encodes a member of the heat shock 20 protein (HSP20) family, was upregulated following dengue virus type 2 (DENV-2) infection in vivo. The transcripts of this gene did not exhibit differential accumulation in mosquitoes exposed to insecticides or pollutants. The induction and overexpression of l(2)efl gene products using poly(I:C) resulted in decreased DENV-2 replication in the cell line. In contrast, the RNAi-mediated suppression of l(2)efl gene products resulted in enhanced DENV-2 replication, but this enhancement occurred only if multiple l(2)efl genes were suppressed. l(2)efl homologs induce the phosphorylation of eukaryotic initiation factor 2α (eIF2α) in the fruit fly Drosophila melanogaster, and we confirmed this finding in the cell line. However, the mechanism by which l(2)efl phosphorylates eIF2α remains unclear. We conclude that l(2)efl encodes a potential anti-dengue protein in the vector mosquito. - Novel Babesia bovis exported proteins that modify properties of infected red blood cells.
Hassan Hakimi, Thomas J Templeton, Miako Sakaguchi, Junya Yamagishi, Shinya Miyazaki, Kazuhide Yahata, Takayuki Uchihashi, Shin-Ichiro Kawazu, Osamu Kaneko, Masahito Asada
PLoS pathogens, 16, 10, e1008917, 2020年10月, ［国際誌］
英語, 研究論文（学術雑誌）, Babesia bovis causes a pathogenic form of babesiosis in cattle. Following invasion of red blood cells (RBCs) the parasite extensively modifies host cell structural and mechanical properties via the export of numerous proteins. Despite their crucial role in virulence and pathogenesis, such proteins have not been comprehensively characterized in B. bovis. Here we describe the surface biotinylation of infected RBCs (iRBCs), followed by proteomic analysis. We describe a multigene family (mtm) that encodes predicted multi-transmembrane integral membrane proteins which are exported and expressed on the surface of iRBCs. One mtm gene was downregulated in blasticidin-S (BS) resistant parasites, suggesting an association with BS uptake. Induced knockdown of a novel exported protein encoded by BBOV_III004280, named VESA export-associated protein (BbVEAP), resulted in a decreased growth rate, reduced RBC surface ridge numbers, mis-localized VESA1, and abrogated cytoadhesion to endothelial cells, suggesting that BbVEAP is a novel virulence factor for B. bovis. - Development of a bio-inkjet printed LAMP test kit for detecting human African trypanosomiasis.
Kyoko Hayashida, Peter Nambala, Nick Van Reet, Philippe Büscher, Naoko Kawai, Mable Mwale Mutengo, Janelisa Musaya, Boniface Namangala, Chihiro Sugimoto, Junya Yamagishi
PLoS neglected tropical diseases, 14, 10, e0008753, 2020年10月, ［国際誌］
英語, 研究論文（学術雑誌）, Human African trypanosomiasis (HAT) is one of the neglected tropical diseases in sub-Saharan Africa. Early diagnosis and treatment prior to disease progression are crucial for the survival of HAT patients. We had previously established a loop-mediated isothermal amplification (LAMP) method for HAT diagnosis in which the reagents were dried for field-use purposes. In this study, we used a semi-automated process to produce the test tubes using a bio-inkjet printer to achieve an accurate production. The performance of the inkjet printer-produced dried LAMP test (CZC-LAMP) was found to be stable after storage for up to 180 days at 30 °C. The diagnostic accuracy of CZC-LAMP HAT was evaluated using DNA samples that were extracted from 116 Trypanosoma brucei gambiense patients and 66 T. b. rhodesiense patients. The sensitivity was 72% for T. b. gambiense (95%CI: 63%-80%) and 80% for T. b. rhodesiense (95%CI: 69%-89%). The specificity determined using DNA from 116 endemic control DNA samples was 95% (95%CI: 89%-98%). The performance of the CZC-LAMP HAT and CZC-LAMP rHAT were also evaluated using 14 crude blood lysate samples obtained from T. b. rhodesiense patients and endemic control samples collected from Rumphi District in Malawi. The sensitivity and specificity were both 100% (95%CI: 77%-100%). As the developed CZC-LAMP test does not require a cold chain or a sophisticated laboratory, it holds promise for use as a routine simple molecular tool for point-of-care HAT diagnosis in endemic areas. - Diversity of trypanosomes in wildlife of the Kafue ecosystem, Zambia.
David Squarre, Kyoko Hayashida, Alex Gaithuma, Herman Chambaro, Naoko Kawai, Ladslav Moonga, Boniface Namangala, Chihiro Sugimoto, Junya Yamagishi
International journal for parasitology. Parasites and wildlife, 12, 34, 41, 2020年08月, ［国際誌］
英語, 研究論文（学術雑誌）, The Kafue ecosystem is a vast conservation protected area comprising the Kafue National Park (KNP) and the Game Management Areas (GMA) that act as a buffer around the national park. The KNP has been neglected as a potential foci for rhodesiense sleeping sickness despite the widespread presence of the tsetse vector and abundant wildlife reservoirs. The aim of this study was to generate information on circulating trypanosomes and their eminent threat/risk to public health and livestock production of a steadily growing human and livestock population surrounding the park. We detected various trypanosomes circulating in different mammalian wildlife species in KNP in Zambia by applying a high throughput ITS1-polymerase chain reaction (PCR)/nanopore sequencing method in combination with serum resistant associated-PCR/Sanger sequencing method. The prevalence rates of trypanosomes in hartebeest, sable antelope, buffalo, warthog, impala and lechwe were 6.4%, 37.2%, 13.2%, 11.8%, 2.8% and 11.1%, respectively. A total of six trypanosomes species or subspecies were detected in the wildlife examined, including Trypanosoma brucei brucei, T. godfreyi, T. congolense, T. simiae and T. theileri. Importantly we detected human infective T. b. rhodesiense in buffalo and sable antelope with a prevalence of 9.4% and 12.5%, respectively. In addition, T. b. rhodesiense was found in the only vervet monkey analyzed. The study thus reaffirmed that the Kafue ecosystem is a genuine neglected and re-emerging foci for human African trypanosomiasis. This is the first assessment of the trypanosome diversity circulating in free-ranging wildlife of the KNP. - Morphological and molecular identification of Eimeria spp. in breeding chicken farms of Japan.
Makoto Matsubayashi, Tomoyuki Shibahara, Tomohide Matsuo, Toshimitsu Hatabu, Junya Yamagishi, Kazumi Sasai, Takashi Isobe
The Journal of veterinary medical science, 82, 5, 516, 519, 2020年05月30日, ［査読有り］, ［国内誌］
英語, 研究論文（学術雑誌）, There have been no reports of the prevalence of Eimeria spp. in poultry breeding farms in Japan unlike those of broiler farms. From 2017 to 2018, we examined the prevalence of Eimeria spp. on breeding farms in Japan by oocyst morphology and PCR analyses. A total of 143 samples was collected from 37 breeding farms in 21 prefectures of Japan. We detected oocysts of seven species at 34 of 37 breeding farms by PCR, and we identified E. brunetti at 51.5% of farms found to be positive for Eimeria. The differences in the identification of Eimeria spp. between the morphology and PCR assay methods of oocysts were pronounced for E. maxima and E. necatrix. We confirmed that molecular tools were more suitable for accurately estimating prevalence of Eimeria spp., and these findings suggest that E. brunetti could be widespread in Japan. - Draft Genome Sequence of Leishmania tarentolae Parrot Tar II, Obtained by Single-Molecule Real-Time Sequencing.
Yasuyuki Goto, Akihiro Kuroki, Kengo Suzuki, Junya Yamagishi
Microbiology resource announcements, 9, 21, 2020年05月21日, ［国際誌］
英語, 研究論文（学術雑誌）, Leishmania tarentolae is a protozoan parasite of lizards and is nonpathogenic to mammals. Genome information for the nonpathogenic species will facilitate an understanding of the pathology caused by species pathogenic to mammals. Here, we report resequencing of the L. tarentolae genome with a single-molecule real-time (SMRT) sequencer to provide a more complete genome assembly. - Longitudinal plasma amino acid profiling with maternal genomic background throughout human pregnancy
Matsuyuki Shirota, Daisuke Saigusa, Riu Yamashita, Yasutake Kato, Mitsuyo Matsumoto, Junya Yamagishi, Noriko Ishida, Kazuki Kumada, Yuji Oe, Hisaaki Kudo, Junji Yokozawa, Yoko Kuroki, Ikuko Motoike, Fumiki Katsuoka, Masao Nagasaki, Seizo Koshiba, Keiko Nakayama, Osamu Tanabe, Jun Yasuda, Shigeo Kure, Kengo Kinoshita, Hirohito Metoki, Shinichi Kuriyama, Nobuo Yaegashi, Masayuki Yamamoto, Junichi Sugawara
Medical Mass Spectrometry, 4, 1, 36, 49, 2020年04月16日, ［査読有り］, ［国際誌］
英語, 研究論文（学術雑誌） - Blood meal sources and bacterial microbiome diversity in wild-caught tsetse flies.
Alex Gaithuma, Junya Yamagishi, Kyoko Hayashida, Naoko Kawai, Boniface Namangala, Chihiro Sugimoto
Scientific reports, 10, 1, 5005, 5005, 2020年03月19日, ［国際誌］
英語, 研究論文（学術雑誌）, Tsetse flies are the vectors of African trypanosomiasis affecting 36 sub-Saharan countries. Both wild and domestic animals play a crucial role in maintaining the disease-causing parasites (trypanosomes). Thus, the identification of animal reservoirs of trypanosomes is vital for the effective control of African trypanosomiasis. Additionally, the biotic and abiotic factors that drive gut microbiome diversity in tsetse flies are primarily unresolved, especially under natural, field conditions. In this study, we present a comprehensive DNA metabarcoding approach for individual tsetse fly analysis in the identification of mammalian blood meal sources and fly bacterial microbiome composition. We analyzed samples from two endemic foci, Kafue, Zambia collected in June 2017, and Hurungwe, Zimbabwe sampled in April 2014 (pilot study) and detected DNA of various mammals including humans, wild animals, domestic animals and small mammals (rat and bat). The bacterial diversity was relatively similar in flies with different mammalian species DNA, trypanosome infected and uninfected flies, and female and male flies. This study is the first report on bat DNA detection in wild tsetse flies. This study reveals that small mammals such as bats and rats are among the opportunistic blood meal sources for tsetse flies in the wild, and the implication on tsetse biology and ecology needs to be studied. - Genetic and Biological Diversity of Porcine Sapeloviruses Prevailing in Zambia.
Hayato Harima, Masahiro Kajihara, Edgar Simulundu, Eugene Bwalya, Yongjin Qiu, Mao Isono, Kosuke Okuya, Gabriel Gonzalez, Junya Yamagishi, Bernard M Hang'ombe, Hirofumi Sawa, Aaron S Mweene, Ayato Takada
Viruses, 12, 2, 2020年02月05日, ［査読有り］, ［国際誌］
英語, 研究論文（学術雑誌）, Porcine sapelovirus (PSV) has been detected worldwide in pig populations. Although PSV causes various symptoms such as encephalomyelitis, diarrhea, and pneumonia in pigs, the economic impact of PSV infection remains to be determined. However, information on the distribution and genetic diversity of PSV is quite limited, particularly in Africa. In this study, we investigated the prevalence of PSV infection in Zambia and characterized the isolated PSVs genetically and biologically. We screened 147 fecal samples collected in 2018 and found that the prevalences of PSV infection in suckling pigs and fattening pigs were high (36.2% and 94.0%, respectively). Phylogenetic analyses revealed that the Zambian PSVs were divided into three different lineages (Lineages 1-3) in the clade consisting of Chinese strains. The Zambian PSVs belonging to Lineages 2 and 3 replicated more efficiently than those belonging to Lineage 1 in Vero E6 and BHK cells. Bioinformatic analyses revealed that genetic recombination events had occurred and the recombination breakpoints were located in the L and 2A genes. Our results indicated that at least two biologically distinct PSVs could be circulating in the Zambian pig population and that genetic recombination played a role in the evolution of PSVs. - First molecular identification of trypanosoma evansi from cattle in Syria
Megasari Marsela, Kyoko Hayashida, Alaa Terkawi, Xuenan Xuan, Chihiro Sugimoto, Junya Yamagishi
Japanese Journal of Veterinary Research, 68, 2, 117, 127, Hokkaido University, 2020年
英語, 研究論文（学術雑誌） - Molecular identification of trypanosomes in cattle in Malawi using PCR methods and nanopore sequencing: epidemiological implications for the control of human and animal trypanosomiases.
Megasari Marsela, Kyoko Hayashida, Ryo Nakao, Elisha Chatanga, Alex Kiarie Gaithuma, Kawai Naoko, Janelisa Musaya, Chihiro Sugimoto, Junya Yamagishi
Parasite (Paris, France), 27, 46, 46, 2020年, ［国際誌］
英語, 研究論文（学術雑誌）, This study aimed to identify trypanosomes infecting cattle in Malawi in order to understand the importance of cattle in the transmission dynamics of Human African Trypanosomiasis (HAT) and Animal African Trypanosomosis (AAT). A total of 446 DNA samples from cattle blood from three regions of Malawi were screened for African trypanosomes by ITS1 PCR. The obtained amplicons were sequenced using a portable next-generation sequencer, MinION, for validation. Comparison of the results from ITS1 PCR and MinION sequencing showed that combining the two methods provided more accurate species identification than ITS1 PCR alone. Further PCR screening targeting the serum resistance-associated (SRA) gene was conducted to detect Trypanosoma brucei rhodesiense. Trypanosoma congolense was the most prevalent Trypanosoma sp., which was found in Nkhotakota (10.8%; 20 of 185), followed by Kasungu (2.5%; 5 of 199). Of note, the prevalence of T. b. rhodesiense detected by SRA PCR was high in Kasungu and Nkhotakota showing 9.5% (19 of 199) and 2.7% (5 of 185), respectively. We report the presence of animal African trypanosomes and T. b. rhodesiense from cattle at the human-livestock-wildlife interface for the first time in Malawi. Our results confirmed that animal trypanosomes are important causes of anemia in cattle and that cattle are potential reservoirs for human African trypanosomiasis in Malawi. - Legionella pneumophila Infection Rewires the Acanthamoeba castellanii Transcriptome, Highlighting a Class of Sirtuin Genes.
Pengfei Li, Dane Vassiliadis, Sze Ying Ong, Vicki Bennett-Wood, Chihiro Sugimoto, Junya Yamagishi, Elizabeth L Hartland, Shivani Pasricha
Frontiers in cellular and infection microbiology, 10, 428, 428, 2020年, ［国際誌］
英語, 研究論文（学術雑誌）, Legionella pneumophila is an environmental bacterium that has evolved to survive predation by soil and water amoebae such as Acanthamoeba castellanii, and this has inadvertently led to the ability of L. pneumophila to survive and replicate in human cells. L. pneumophila causes Legionnaire's Disease, with human exposure occurring via the inhalation of water aerosols containing both amoebae and the bacteria. These aerosols originate from aquatic biofilms found in artifical water sources, such as air-conditioning cooling towers and humidifiers. In these man-made environments, A. castellanii supports L. pneumophila intracellular replication, thereby promoting persistence and dissemination of the bacteria and providing protection from external stress. Despite this close evolutionary relationship, very little is known about how A. castellanii responds to L. pneumophila infection. In this study, we examined the global transcriptional response of A. castellanii to L. pneumophila infection. We compared A. castellanii infected with wild type L. pneumophila to A. castellanii infected with an isogenic ΔdotA mutant strain, which is unable to replicate intracellularly. We showed that A. castellanii underwent clear morphological and transcriptional rewiring over the course of L. pneumophila infection. Through improved annotation of the A. castellanii genome, we determined that these transcriptional changes primarily involved biological processes utilizing small GTPases, including cellular transport, signaling, metabolism and replication. In addition, a number of sirtuin-encoding genes in A. castellanii were found to be conserved and upregulated during L. pneumophila infection. Silencing of sirtuin gene, sir6f (ACA1_153540) resulted in the inhibition of A. castellanii cell proliferation during infection and reduced L. pneumophila replication. Overall our findings identified several biological pathways in amoebae that may support L. pneumophila replication and A. castellanii proliferation in environmental conditions. - Molecular characterization of Cryptosporidium spp. from patients with diarrhoea in Lusaka, Zambia.
Namwiinga Rozaria Mulunda, Kyoko Hayashida, Junya Yamagishi, Sandie Sianongo, Gilbert Munsaka, Chihiro Sugimoto, Mable Mwale Mutengo
Parasite (Paris, France), 27, 53, 53, 2020年, ［国際誌］
英語, 研究論文（学術雑誌）, Cryptosporidium is a major etiological agent of diarrhoeal diseases among children and immune-compromised individuals in sub-Saharan African countries. We conducted a study to determine the prevalence and genetic characteristics of Cryptosporidium spp. in stool samples from patients with diarrhoea who presented at the University Teaching Hospital in Lusaka, Zambia. Cryptosporidium species and subtypes from 71 microscopically confirmed cryptosporidiosis stool samples collected between 2017 and 2019 were determined by polymerase chain reaction followed by partial sequencing of the small subunit rRNA and 60-kDa glycoprotein (gp60) gene. Additionally, data for the period between 2014 and 2019 were reviewed and analysed for cryptosporidiosis seasonal and age distribution. Cryptosporidium was more prevalent in the rainy season. The highest number of cases was reported among the 1-4 year age group. By sequence analysis of the 71 positive isolates, Cryptosporidium hominis (n = 42; 59.2%), C. parvum (n = 27; 38%), C. felis (n = 1; 1.4%), and C. meleagridis (n = 1; 1.4%) were identified. Four C. hominis subtype families (Ia, Ib, Id, and Ie) and three C. parvum subtype families (IIc, IIe, and IIs) were identified. The most frequent subtypes were IeA11G3T3 (n = 20; 28.2%), IIcA5G3 (n = 12; 16.9%), IIeA12G1 (n = 11; 15.5%) and IaA30R3 (n = 10; 14.1%). The observed species/subtypes of C. hominis and C. parvum indicated that the infection was mainly transmitted through the anthroponotic route. The identification of C. felis and C. meleagridis suggests that an atypical zoonotic transmission cycle also exists. - Complete genome and bimodal genomic structure of the amoebal symbiont Neochlamydia strain S13 revealed by ultra-long reads obtained from MinION.
Yamagishi J, Hayashida K, Matsuo J, Okubo T, Kuroda M, Nagai H, Sekizuka T, Yamaguchi H, Sugimoto C
Journal of human genetics, 65, 1, 41, 48, Springer Science and Business Media LLC, 2019年11月, ［査読有り］
研究論文（学術雑誌） - Rapid sequencing-based diagnosis of infectious bacterial species from meningitis patients in Zambia.
Nakagawa S, Inoue S, Kryukov K, Yamagishi J, Ohno A, Hayashida K, Nakazwe R, Kalumbi M, Mwenya D, Asami N, Sugimoto C, Mutengo MM, Imanishi T
Clinical & Translational Immunology, 8, 11, e01087, 2019年11月, ［査読有り］, ［国際誌］
英語, 研究論文（学術雑誌）, Objectives: We have developed a portable system for the rapid determination of bacterial composition for the diagnosis of infectious diseases. Our system comprises of a nanopore technology-based sequencer, MinION, and two laptop computers. To examine the accuracy and time efficiency of our system, we provided a proof-of-concept for the detection of the causative bacteria of 11 meningitis patients in Zambia. Methods: We extracted DNA from cerebrospinal fluid samples of each patient and amplified the 16S rRNA gene regions. The sequencing library was prepared, and the sequenced reads were simultaneously processed for bacterial composition determination using the minimap2 software and the representative prokaryote genomes. Results: The sequencing results of four of the six culture-positive samples were consistent with those of conventional culture-based methods. The dominant bacterial species in each of these samples were identified from the sequencing data within only 3 min. Although the major bacterial species were also detected from the other two culture-positive samples and five culture-negative samples, their presence could not be confirmed. Moreover, as a whole, although the number of sequencing reads obtained within a short sequencing run was small, there was no change in the major bacterial species over time with prolonged sequencing. In addition, the processing time strongly correlated with the number of sequencing reads used for the analysis. Conclusion: Our results suggest that time-effective analysis could be achieved by determining the number of sequencing reads required for the rapid diagnosis of infectious bacterial species depending on the complexity of bacterial species in a sample. - Transcriptome analysis of the effect of C-C chemokine receptor 5 deficiency on cell response to Toxoplasma gondii in brain cells.
Kobayashi K, Umeda K, Ihara F, Tanaka S, Yamagishi J, Suzuki Y, Nishikawa Y
BMC genomics, 20, 1, 705, 2019年09月, ［査読有り］
研究論文（学術雑誌） - Complete Genome Sequence of Rickettsia asiatica Strain Maytaro1284, a Member of Spotted Fever Group Rickettsiae Isolated from an Ixodes ovatus Tick in Japan.
Thu MJ, Qiu Y, Yamagishi J, Kusakisako K, Ogata S, Moustafa MAM, Isoda N, Sugimoto C, Katakura K, Nonaka N, Nakao R
Microbiology resource announcements, 8, 37, American Society for Microbiology, 2019年09月, ［査読有り］
研究論文（学術雑誌）, This is the first report of the complete genome sequence of
Rickettsia asiatica
strain Maytaro1284, isolated from an
Ixodes ovatus
tick in Japan. The genome contains a 1,344,324-bp circular chromosome and one plasmid of 74,761 bp. There was no outer membrane protein A (
ompA
) gene encoded in the genome. - Prevalence of hemoprotozoan parasites in small ruminants along a human-livestock-wildlife interface in western Uganda
Keneth Iceland Kasozi, Monica Namayanja, Alex Kiarie Gaithuma, Michael Mahero, Enock Matovu, Junya Yamagishi, Chihiro Sugimoto, Ewan MacLeod
Veterinary Parasitology: Regional Studies and Reports, 17, Elsevier B.V., 2019年08月01日
英語, 研究論文（学術雑誌） - Transitions in morphological forms and rapid development of the asexual schizonts of Eimeria tenella through serial passaging in chicks.
Matsubayashi M, Yamaguchi H, Hatta T, Kawahara F, Hatabu T, Iseki H, Yamagishi J, Isobe T, Teramoto I, Kaneko A, Kita K, Tsuji N, Sasai K
Infection, genetics and evolution : journal of molecular epidemiology and evolutionary genetics in infectious diseases, 75, 103993, 2019年08月, ［査読有り］
研究論文（学術雑誌） - Initiated Babesia ovata Sexual Stages under In Vitro Conditions Were Recognized by Anti-CCp2 Antibodies, Showing Changes in the DNA Content by Imaging Flow Cytometry.
Nguyen TT, Dang-Trinh MA, Higuchi L, Mosqueda J, Hakimi H, Asada M, Yamagishi J, Umemiya-Shirafuji R, Kawazu SI
Pathogens (Basel, Switzerland), 8, 3, 2019年07月, ［査読有り］, ［国際誌］
英語, 研究論文（学術雑誌）, Sexual stage induction under in vitro conditions is useful for biological and molecular studies of Babesia parasites. Therefore, in the present study, we induced B. ovata tick stages using the chemical inducers: xanthurenic acid (XA), dithiothreitol (DTT) and tris (2-carboxyethyl) phosphine (TCEP) at 27 °C or 37 °C conditions. Cultures at low temperature (27 °C) or treated with XA/TCEP induced a large number of extra-erythrocytic merozoites, which transformed into round shape cells at 12-24 h post-induction (pi). However, typical forms of tick stages (aggregation forms and the spiky forms/ray bodies) were only observed in the cultures treated with 40 mM or 60 mM of DTT during 3-6 h pi. The induced cells were recognized by anti-CCp2 rabbit antisera. DNA content of the cell population treated with 40 mM of DTT was analyzed by imaging flow cytometry at 0, 12 and 48 h pi. The results indicated that the parasite population with diploid-like double DNA content increased at 48 h pi. Our observations on morphological and changes in the DNA content provide useful information for understanding the life cycle of B. ovata under in vitro conditions, which will facilitate further studies on basic biology and the development of transmission blocking vaccines against bovine babesiosis. - Genetic diversity and population structure of Glossina morsitans morsitans in the active foci of human African trypanosomiasis in Zambia and Malawi.
Nakamura Y, Yamagishi J, Hayashida K, Osada N, Chatanga E, Mweempwa C, Chilongo K, Chisi J, Musaya J, Inoue N, Namangala B, Sugimoto C
PLoS neglected tropical diseases, 13, 7, e0007568, 2019年07月, ［査読有り］ - Field diagnosis and genotyping of chikungunya virus using a dried reverse transcription loop-mediated isothermal amplification (LAMP) assay and MinION sequencing.
Hayashida K, Orba Y, Sequeira PC, Sugimoto C, Hall WW, Eshita Y, Suzuki Y, Runtuwene L, Brasil P, Calvet G, Rodrigues CDS, Santos CCD, Mares-Guia MAM, Yamagishi J, Filippis AMB, Sawa H
PLoS neglected tropical diseases, 13, 6, e0007480, 2019年06月, ［査読有り］, ［国際誌］
英語, 研究論文（学術雑誌）, Detection and sequencing of chikungunya virus (CHIKV) genome was performed using a combination of a modified reverse transcription loop-mediated isothermal amplification (RT-LAMP) method and a MinION sequencer. We developed the protocol for drying all the reagents for the RT-LAMP in a single reaction tube. Using this system, the CHIKV genome was effectively amplified under isothermal conditions, and used as a template for MinION sequencing with a laptop computer. Our in-house RT-LAMP method and MinION sequencing system were also validated with RNAs and serum samples from recent outbreaks of CHIKV patients in Brazil. The obtained sequence data confirmed the CHIKV outbreaks and identified the genotype. In summary, our established inexpensive on-site genome detection and sequencing system is applicable for both diagnosis of CHIKV infected patients and genotyping of the CHIKV virus in future outbreak in remote areas. - Molecular detection of Rickettsia felis in dogs, rodents and cat fleas in Zambia.
Moonga LC, Hayashida K, Nakao R, Lisulo M, Kaneko C, Nakamura I, Eshita Y, Mweene AS, Namangala B, Sugimoto C, Yamagishi J
Parasites & vectors, 12, 1, 168, 2019年04月, ［査読有り］ - Identification of Mouse and Human Antibody Repertoires by Next-Generation Sequencing.
Lin Sun, Naoko Kono, Hiroyuki Toh, Hanbing Xue, Kaori Sano, Tadaki Suzuki, Akira Ainai, Yasuko Orba, Junya Yamagishi, Hideki Hasegawa, Yoshimasa Takahashi, Shigeyuki Itamura, Kazuo Ohnishi
Journal of visualized experiments : JoVE, 145, 2019年03月15日, ［査読有り］, ［国際誌］
英語, 研究論文（学術雑誌）, The immense adaptability of antigen recognition by antibodies is the basis of the acquired immune system. Despite our understanding of the molecular mechanisms underlying the production of the vast repertoire of antibodies by the acquired immune systems, it has not yet been possible to arrive at a global view of a complete antibody repertoire. In particular, B cell repertoires have been regarded as a black box because of their astronomical number of antibody clones. However, next-generation sequencing technologies are enabling breakthroughs to increase our understanding of the B cell repertoire. In this report, we describe a simple and efficient method to visualize and analyze whole individual mouse and human antibody repertoires. From the immune organs, representatively from spleen in mice and peripheral blood mononuclear cells in humans, total RNA was prepared, reverse transcribed, and amplified using the 5'-RACE method. Using a universal forward primer and antisense primers for the antibody class-specific constant domains, antibody mRNAs were uniformly amplified in proportions reflecting their frequencies in the antibody populations. The amplicons were sequenced by next-generation sequencing (NGS), yielding more than 105 antibody sequences per immunological sample. We describe the protocols for antibody sequence analyses including V(D)J-gene-segment annotation, a bird's-eye view of the antibody repertoire, and our computational methods. - A single test approach for accurate and sensitive detection and taxonomic characterization of Trypanosomes by comprehensive analysis of internal transcribed spacer 1 amplicons.
Gaithuma AK, Yamagishi J, Martinelli A, Hayashida K, Kawai N, Marsela M, Sugimoto C
PLoS neglected tropical diseases, 13, 2, e0006842, 2019年02月, ［査読有り］ - Draft Genome Sequence of Trypanosoma equiperdum Strain IVM-t1.
Davaasuren B, Yamagishi J, Mizushima D, Narantsatsral S, Otgonsuren D, Myagmarsuren P, Battsetseg B, Battur B, Inoue N, Suganuma K
Microbiology resource announcements, 8, 9, 2019年02月, ［査読有り］, ［国際誌］
英語 - Hemophagocytosis induced by Leishmania donovani infection is beneficial to parasite survival within macrophages
Ayako Morimoto, Kazuyuki Uchida, James K. Chambers, Kai Sato, Jing Hong, Chizu Sanjoba, Yoshitsugu Matsumoto, Junya Yamagishi, Yasuyuki Goto
PLoS Neglected Tropical Diseases, 13, 11, 2019年
研究論文（学術雑誌） - Consensus and variations in cell line specificity among human metapneumovirus strains.
Nao N, Sato K, Yamagishi J, Tahara M, Nakatsu Y, Seki F, Katoh H, Ohnuma A, Shirogane Y, Hayashi M, Suzuki T, Kikuta H, Nishimura H, Takeda M
PloS one, 14, 4, e0215822, 2019年, ［査読有り］, ［国際誌］
英語, 研究論文（学術雑誌）, Human metapneumovirus (HMPV) has been a notable etiological agent of acute respiratory infection in humans, but it was not discovered until 2001, because HMPV replicates only in a limited number of cell lines and the cytopathic effect (CPE) is often mild. To promote the study of HMPV, several groups have generated green fluorescent protein (GFP)-expressing recombinant HMPV strains (HMPVGFP). However, the growing evidence has complicated the understanding of cell line specificity of HMPV, because it seems to vary notably among HMPV strains. In addition, unique A2b clade HMPV strains with a 180-nucleotide duplication in the G gene (HMPV A2b180nt-dup strains) have recently been detected. In this study, we re-evaluated and compared the cell line specificity of clinical isolates of HMPV strains, including the novel HMPV A2b180nt-dup strains, and six recombinant HMPVGFP strains, including the newly generated recombinant HMPV A2b180nt-dup strain, MG0256-EGFP. Our data demonstrate that VeroE6 and LLC-MK2 cells generally showed the highest infectivity with any clinical isolates and recombinant HMPVGFP strains. Other human-derived cell lines (BEAS-2B, A549, HEK293, MNT-1, and HeLa cells) showed certain levels of infectivity with HMPV, but these were significantly lower than those of VeroE6 and LLC-MK2 cells. Also, the infectivity in these suboptimal cell lines varied greatly among HMPV strains. The variations were not directly related to HMPV genotypes, cell lines used for isolation and propagation, specific genome mutations, or nucleotide duplications in the G gene. Thus, these variations in suboptimal cell lines are likely intrinsic to particular HMPV strains. - Novel Characteristics of Mitochondrial Electron Transport Chain from Eimeria tenella.
Matsubayashi M, Inaoka DK, Komatsuya K, Hatta T, Kawahara F, Sakamoto K, Hikosaka K, Yamagishi J, Sasai K, Shiba T, Harada S, Tsuji N, Kita K
Genes, 10, 1, 2019年01月, ［査読有り］, ［国際誌］
英語, 研究論文（学術雑誌）, Eimeria tenella is an intracellular apicomplexan parasite, which infects cecal epithelial cells from chickens and causes hemorrhagic diarrhea and eventual death. We have previously reported the comparative RNA sequence analysis of the E. tenella sporozoite stage between virulent and precocious strains and showed that the expression of several genes involved in mitochondrial electron transport chain (ETC), such as type II NADH dehydrogenase (NDH-2), complex II (succinate:quinone oxidoreductase), malate:quinone oxidoreductase (MQO), and glycerol-3-phosphate dehydrogenase (G3PDH), were upregulated in virulent strain. To study E. tenella mitochondrial ETC in detail, we developed a reproducible method for preparation of mitochondria-rich fraction from sporozoites, which maintained high specific activities of dehydrogenases, such as NDH-2 followed by G3PDH, MQO, complex II, and dihydroorotate dehydrogenase (DHODH). Of particular importance, we showed that E. tenella sporozoite mitochondria possess an intrinsic ability to perform fumarate respiration (via complex II) in addition to the classical oxygen respiration (via complexes III and IV). Further analysis by high-resolution clear native electrophoresis, activity staining, and nano-liquid chromatography tandem-mass spectrometry (nano-LC-MS/MS) provided evidence of a mitochondrial complex II-III-IV supercomplex. Our analysis suggests that complex II from E. tenella has biochemical features distinct to known orthologues and is a potential target for the development of new anticoccidian drugs. - Establishment of a mouse-tick infection model for Theileria orientalis and analysis of its transcriptome.
Kyoko Hayashida, Rika Umemiya-Shirafuji, Thillaiampalam Sivakumar, Junya Yamagishi, Yutaka Suzuki, Chihiro Sugimoto, Naoaki Yokoyama
International journal for parasitology, 48, 12, 915, 924, 2018年10月, ［査読有り］, ［国際誌］
英語, 研究論文（学術雑誌）, Oriental theileriosis caused by Theileria orientalis is an economically significant disease in cattle farming. The lack of laboratory animal models and in vitro culture systems is a major obstacle in the drive to better understand the biology of this parasite. Notably, research on the sporozoite stage of T. orientalis has rarely been undertaken, although such investigations are of paramount importance for vaccine development based on blocking sporozoite invasion of its host animals. In the present study, we established a mouse-tick infection model for propagating T. orientalis in mice and for producing the sporozoite stage in tick salivary glands. Splenectomized severe combined immunodeficient mice transfused with bovine erythrocytes were infected with T. orientalis. The larval ticks of Haemaphysalis longicornis were then fed on the T. orientalis-infected mice. The piroplasm and sporozoite stages were microscopically observed in the mouse blood and nymphal salivary glands, respectively. The transcriptomics data generated from the piroplasm and sporozoite stages revealed a stage-specific expression pattern for the parasite genes. The mouse-tick infection model and the transcriptomics data it has provided will contribute to a better understanding of T. orientalis biology and will also provide much needed information for the design of effective control measures targeting oriental theileriosis. - Targeting of RNA Polymerase II by a nuclear Legionella pneumophila Dot/Icm effector SnpL.
Schuelein R, Spencer H, Dagley LF, Li PF, Luo L, Stow JL, Abraham G, Naderer T, Gomez-Valero L, Buchrieser C, Sugimoto C, Yamagishi J, Webb AI, Pasricha S, Hartland EL
Cellular microbiology, 20, 9, e12852, 2018年09月, ［査読有り］ - Eimeria tenella 弱毒株の作出と比較ゲノム解析による弱毒化分子機構の解明
松林 誠, 川原 史也, 山岸 潤也, 八田 岳士, 畑生 俊光, 山口 浩貴, 寺本 勲, 金子 明, 磯部 尚, 北 潔, 辻 尚利, 笹井 和美
日本獣医学会学術集会講演要旨集, 161回, 316, 316, (公社)日本獣医学会, 2018年08月
日本語 - ザンビア、マラウイにおけるツェツェバエの集団遺伝学解析 ツェツェバエ媒介性アフリカトリパノソーマ症対策への応用
中村 有紀子, 林田 京子, 山岸 潤也, 杉本 千尋
日本獣医学会学術集会講演要旨集, 161回, 320, 320, (公社)日本獣医学会, 2018年08月
日本語 - Genetic analysis of Babesia isolates from cattle with clinical babesiosis in Sri Lanka.
Sivakumar T, Tuvshintulga B, Zhyldyz A, Kothalawala H, Yapa PR, Kanagaratnam R, Vimalakumar SC, Abeysekera TS, Weerasingha AS, Yamagishi J, Igarashi I, Silva SSP, Yokoyama N
Journal of clinical microbiology, 56, 11, 2018年08月, ［査読有り］, ［国際誌］
英語, 研究論文（学術雑誌）, Bovine babesiosis is a serious threat to the cattle industry. We prepared blood DNA samples from 13 cattle with clinical babesiosis from the Badulla (n = 8), Jaffna (n = 3), and Kilinochchi (n = 2) districts in Sri Lanka. These DNA samples tested positive in PCR assays specific for Babesiabovis (n = 9), Babesia bigemina (n = 9), and Babesiaovata (n = 1). Twelve cattle were positive for B. bovis and/or B. bigemina One cow was negative for the tested Babesia species but was positive for Babesia on microscopic examination; the phylogenetic positions of 18S rRNA and cytochrome oxidase subunit III gene sequences suggested that the cow was infected with Babesia sp. Mymensingh, which was recently reported from a healthy cow in Bangladesh. We then developed a novel Babesia sp. Mymensingh-specific PCR assay and obtained positive results for one other sample. Analysis of gene sequences from the cow with positive B. ovata-specific PCR results demonstrated that the animal was infected not with B. ovata but with Babesia sp. Hue-1, which was recently reported from asymptomatic cattle in Vietnam. The virulence of Babesia sp. Hue-1 is unclear, as the cow was coinfected with B. bovis and B. bigemina However, Babesia sp. Mymensingh probably causes severe clinical babesiosis, as it was the sole Babesia species detected in a clinical case. The present study revealed the presence of two bovine Babesia species not previously reported in Sri Lanka, plus the first case of severe bovine babesiosis caused by a Babesia species other than B. bovis, B. bigemina, and Babesiadivergens. - Development and validation of direct dry loop mediated isothermal amplification for diagnosis of Trypanosoma evansi.
Salim B, Hayashida K, Mossaad E, Nakao R, Yamagishi J, Sugimoto C
Veterinary parasitology, 260, 53, 57, Elsevier BV, 2018年08月, ［査読有り］
研究論文（学術雑誌） - An innovative diagnostic technology for the codon mutation C580Y in kelch13 of Plasmodium falciparum with MinION nanopore sequencer.
Kazuo Imai, Norihito Tarumoto, Lucky Ronald Runtuwene, Jun Sakai, Kyoko Hayashida, Yuki Eshita, Ryuichiro Maeda, Josef Tuda, Hideaki Ohno, Takashi Murakami, Shigefumi Maesaki, Yutaka Suzuki, Junya Yamagishi, Takuya Maeda
Malaria journal, 17, 1, 217, 217, 2018年05月29日, ［査読有り］, ［国際誌］
英語, 研究論文（学術雑誌）, BACKGROUND: The recent spread of artemisinin (ART)-resistant Plasmodium falciparum represents an emerging global threat to public health. In Southeast Asia, the C580Y mutation of kelch13 (k13) is the dominant mutation of ART-resistant P. falciparum. Therefore, a simple method for the detection of C580Y mutation is urgently needed to enable widespread routine surveillance in the field. The aim of this study is to develop a new diagnostic procedure for the C580Y mutation using loop-mediated isothermal amplification (LAMP) combined with the MinION nanopore sequencer. RESULTS: A LAMP assay for the k13 gene of P. falciparum to detect the C580Y mutation was successfully developed. The detection limit of this procedure was 10 copies of the reference plasmid harboring the k13 gene within 60 min. Thereafter, amplicon sequencing of the LAMP products using the MinION nanopore sequencer was performed to clarify the nucleotide sequences of the gene. The C580Y mutation was identified based on the sequence data collected from MinION reads 30 min after the start of sequencing. Further, clinical evaluation of the LAMP assay in 34 human blood samples collected from patients with P. falciparum malaria in Indonesia revealed a positive detection rate of 100%. All LAMP amplicons of up to 12 specimens were simultaneously sequenced using MinION. The results of sequencing were consistent with those of the conventional PCR and Sanger sequencing protocol. All procedures from DNA extraction to variant calling were completed within 3 h. The C580Y mutation was not found among these 34 P. falciparum isolates in Indonesia. CONCLUSIONS: An innovative method combining LAMP and MinION will enable simple, rapid, and high-sensitivity detection of the C580Y mutation of P. falciparum, even in resource-limited situations in developing countries. - Nanopore sequencing of drug-resistance-associated genes in malaria parasites, Plasmodium falciparum.
Lucky R Runtuwene, Josef S B Tuda, Arthur E Mongan, Wojciech Makalowski, Martin C Frith, Mallika Imwong, Suttipat Srisutham, Lan Anh Nguyen Thi, Nghia Nguyen Tuan, Yuki Eshita, Ryuichiro Maeda, Junya Yamagishi, Yutaka Suzuki
Scientific reports, 8, 1, 8286, 8286, 2018年05月29日, ［査読有り］, ［国際誌］
英語, 研究論文（学術雑誌）, Here, we report the application of a portable sequencer, MinION, for genotyping the malaria parasite Plasmodium falciparum. In the present study, an amplicon mixture of nine representative genes causing resistance to anti-malaria drugs is diagnosed. First, we developed the procedure for four laboratory strains (3D7, Dd2, 7G8, and K1), and then applied the developed procedure to ten clinical samples. We sequenced and re-sequenced the samples using the obsolete flow cell R7.3 and the most recent flow cell R9.4. Although the average base-call accuracy of the MinION sequencer was 74.3%, performing >50 reads at a given position improves the accuracy of the SNP call, yielding a precision and recall rate of 0.92 and 0.8, respectively, with flow cell R7.3. These numbers increased significantly with flow cell R9.4, in which the precision and recall are 1 and 0.97, respectively. Based on the SNP information, the drug resistance status in ten clinical samples was inferred. We also analyzed K13 gene mutations from 54 additional clinical samples as a proof of concept. We found that a novel amino-acid changing variation is dominant in this area. In addition, we performed a small population-based analysis using 3 and 5 cases (K13) and 10 and 5 cases (PfCRT) from Thailand and Vietnam, respectively. We identified distinct genotypes from the respective regions. This approach will change the standard methodology for the sequencing diagnosis of malaria parasites, especially in developing countries. - Establishment of a stable transfection system for genetic manipulation of Babesia gibsoni
Mingming Liu, Paul Franck Adjou Moumouni, Masahito Asada, Hassan Hakimi, Tatsunori Masatani, Patrick Vudriko, Seung-Hun Lee, Shin-Ichiro Kawazu, Junya Yamagishi, Xuenan Xuan
Parasites and Vectors, 11, 1, 260, BioMed Central Ltd., 2018年04月23日, ［査読有り］
英語, 研究論文（学術雑誌） - Comparative genomics and proteomic analyses between lethal and nonlethal strains of Plasmodium berghei
Mamoru Niikura, Shin Ichi Inoue, Toshiyuki Fukutomi, Junya Yamagishi, Hiroko Asahi, Fumie Kobayashi
Experimental Parasitology, 185, 1, 9, 2018年02月, ［査読有り］
研究論文（学術雑誌） - 【どこでも 誰でも より長く ナノポアシークエンサーが研究の常識を変える!】フィールドでの熱帯病原性微生物タイピング
山岸 潤也, Runtuwene Lucky, 林田 京子, 鈴木 穣
実験医学, 36, 1, 27, 31, (株)羊土社, 2018年01月
日本語 - Transcriptional profiling of Toll-like receptor 2-deficient primary murine brain cells during Toxoplasma gondii infection
Kousuke Umeda, Sachi Tanaka, Fumiaki Ihara, Junya Yamagishi, Yutaka Suzuki, Yoshifumi Nishikawa
PLOS ONE, 12, 11, e0187703, 2017年11月, ［査読有り］
英語, 研究論文（学術雑誌） - Identification and characterization of interchangeable cross-species functional promoters between Babesia gibsoni and Babesia bovis.
Liu M, Adjou Moumouni PF, Cao S, Asada M, Wang G, Gao Y, Guo H, Li J, Vudriko P, Efstratiou A, Ringo AE, Lee SH, Hakimi H, Masatani T, Sunaga F, Kawazu SI, Yamagishi J, Jia L, Inoue N, Xuan X
Ticks and tick-borne diseases, 2017年11月, ［査読有り］ - Whole-genome assembly of Babesia ovata and comparative genomics between closely related pathogens
Junya Yamagishi, Masahito Asada, Hassan Hakimi, Takeshi Q. Tanaka, Chihiro Sugimoto, Shin-ichiro Kawazu
BMC GENOMICS, 18, 1, 832, 2017年10月, ［査読有り］
英語, 研究論文（学術雑誌） - Transient transfection of intraerythrocytic Babesia gibsoni using elongation factor-1 alpha promoter
Mingming Liu, Masahito Asada, Shinuo Cao, Paul Franck Adjou Moumouni, Patrick Vudriko, Artemis Efstratiou, Hassan Hakimi, Tatsunori Masatani, Fujiko Sunaga, Shin-ichiro Kawazu, Junya Yamagishi, Xuenan Xuan
MOLECULAR AND BIOCHEMICAL PARASITOLOGY, 216, 56, 59, 2017年09月, ［査読有り］
英語, 研究論文（学術雑誌） - A novel diagnostic method for malaria using loop-mediated isothermal amplification (LAMP) and MinION™ nanopore sequencer.
Imai K, Tarumoto N, Misawa K, Runtuwene LR, Sakai J, Hayashida K, Eshita Y, Maeda R, Tuda J, Murakami T, Maesaki S, Suzuki Y, Yamagishi J, Maeda T
BMC infectious diseases, 17, 1, 621, 2017年09月, ［査読有り］
英語, 研究論文（学術雑誌） - Eimeria tenella弱毒株の作出および病理組織学的比較解析
山口 浩貴, 松林 誠, 川原 史也, 八田 岳士, 畑生 俊光, 山岸 潤也, 磯部 尚, 寺本 勲, 金子 明, 北 潔, 辻 尚利, 笹井 和美
日本獣医学会学術集会講演要旨集, 160回, 336, 336, (公社)日本獣医学会, 2017年08月
日本語 - PEGylation of a TLR2-agonist-based vaccine delivery system improves antigen trafficking and the magnitude of ensuing antibody and CD8(+) T cell responses
Toshiki Sekiya, Junya Yamagishi, John Henry V. Gray, Paul G. Whitney, Axel Martinelli, Weiguang Zeng, Chinn Yi Wong, Chihiro Sugimoto, David C. Jackson, Brendon Y. Chua
BIOMATERIALS, 137, 61, 72, 2017年08月, ［査読有り］
英語, 研究論文（学術雑誌） - Use of the Oxford Nanopore MinION sequencer for MLST genotyping of vancomycin-resistant enterococci.
N Tarumoto, J Sakai, K Sujino, T Yamaguchi, M Ohta, J Yamagishi, L R Runtuwene, T Murakami, Y Suzuki, T Maeda, S Maesaki
The Journal of hospital infection, 96, 3, 296, 298, 2017年07月, ［査読有り］, ［国際誌］
英語 - Use of the Oxford Nanopore MinION sequencer for MLST genotyping of vancomycin-resistant enterococci.
Tarumoto N, Sakai J, Sujino K, Yamaguchi T, Ohta M, Yamagishi J, Runtuwene LR, Murakami T, Suzuki Y, Maeda T, Maesaki S
J Hosp Infect, 96, 3, 296, 298, 2017年06月, ［査読有り］
研究論文（学術雑誌） - Serotyping dengue virus with isothermal amplification and a portable sequencer
Junya Yamagishi, Lucky R. Runtuwene, Kyoko Hayashida, Arthur E. Mongan, Lan Anh Nguyen Thi, Linh Nguyen Thuy, Cam Nguyen Nhat, Kriengsak Limkittikul, Chukiat Sirivichayakul, Nuankanya Sathirapongsasuti, Martin Frith, Wojciech Makalowski, Yuki Eshita, Sumio Sugano, Yutaka Suzuki
SCIENTIFIC REPORTS, 7, 1, 3510, 2017年06月, ［査読有り］
英語, 研究論文（学術雑誌） - A TLR3-Specific Adjuvant Relieves Innate Resistance to PD-L1 Blockade without Cytokine Toxicity in Tumor Vaccine Immunotherapy
Yohei Takeda, Keisuke Kataoka, Junya Yamagishi, Seishi Ogawa, Tsukasa Seya, Misako Matsumoto
CELL REPORTS, 19, 9, 1874, 1887, 2017年05月, ［査読有り］
英語, 研究論文（学術雑誌） - Genetic polymorphisms in Plasmodium falciparum chloroquine resistance genes, pfcrt and pfmdr1, in North Sulawesi, Indonesia
Patrick Reteng, Visia Vrisca, Inka Sukarno, Ilham Habib Djarkoni, Jane Angela Kalangi, George Eduardo Jacobs, Lucky Ronald Runtuwene, Yuki Eshita, Ryuichiro Maeda, Yutaka Suzuki, Arthur Elia Mongan, Sarah Maria Warouw, Junya Yamagishi, Josef Tuda
BMC Research Notes, 10, 1, 147, BioMed Central Ltd., 2017年04月04日, ［査読有り］
英語, 研究論文（学術雑誌） - Genetic Predisposition To Acquire a Polybasic Cleavage Site for Highly Pathogenic Avian Influenza Virus Hemagglutinin
Naganori Nao, Junya Yamagishi, Hiroko Miyamoto, Manabu Igarashi, Rashid Manzoor, Aiko Ohnuma, Yoshimi Tsuda, Wakako Furuyama, Asako Shigeno, Masahiro Kajihara, Noriko Kishida, Reiko Yoshida, Ayato Takada
MBIO, 8, 1, 2017年01月, ［査読有り］
英語, 研究論文（学術雑誌） - MinIONを利用したLAMP法による革新的マラリア診断系の作成
今井 一男, 三沢 和央, 樽本 憲人, 前崎 繁文, 前田 卓哉, 鈴木 穰, 山岸 潤也, 林田 京子
日本臨床微生物学雑誌, 27, Suppl., 255, 255, (一社)日本臨床微生物学会, 2016年12月
日本語 - Identification and functional analysis of a novel mitochondria-localized 2-Cys peroxiredoxin, BbTPx-2, from Babesia bovis
Tatsunori Masatani, Masahito Asada, Hassan Hakimi, Kei Hayashi, Junya Yamagishi, Shin-ichiro Kawazu, Xuenan Xuan
PARASITOLOGY RESEARCH, 115, 8, 3139, 3145, 2016年08月, ［査読有り］
英語, 研究論文（学術雑誌） - Mycophenolic Acid and Its Derivatives as Potential Chemotherapeutic Agents Targeting Inosine Monophosphate Dehydrogenase in Trypanosoma congolense
Keisuke Suganuma, Albertus Eka Yudistira Sarwono, Shinya Mitsuhashi, Marcin Jakalski, Tadashi Okada, Molefe Nthatisi, Junya Yamagishi, Makoto Ubukata, Noboru Inoue
ANTIMICROBIAL AGENTS AND CHEMOTHERAPY, 60, 7, 4391, 4393, 2016年07月, ［査読有り］
英語, 研究論文（学術雑誌） - Transcriptional profiles of virulent and precocious strains of Eimeria tenella at sporozoite stage; novel biological insight into attenuated asexual development
Makoto Matsubayashi, Fumiya Kawahara, Takeshi Hatta, Junya Yamagishi, Takeharu Miyoshi, Anisuzzaman, Kazumi Sasai, Takashi Isobe, Kiyoshi Kita, Naotoshi Tsuji
INFECTION GENETICS AND EVOLUTION, 40, 54, 62, 2016年06月, ［査読有り］
英語, 研究論文（学術雑誌） - Characterization of an epimastigote -stage-specific hemoglobin receptor of Trypanosoma congolense
Shino Yamasaki, Keisuke Suganuma, Junya Yamagishi, Masahito Asada, Naoaki Yokoyama, Shin-ichiro Kawazu, Noboru Inoue
PARASITES & VECTORS, 9, 1, 299, 2016年05月, ［査読有り］
英語, 研究論文（学術雑誌） - Comparison of Boiling and Robotics Automation Method in DNA Extraction for Metagenomic Sequencing of Human Oral Microbes
Junya Yamagishi, Yukuto Sato, Natsuko Shinozaki, Bin Ye, Akito Tsuboi, Masao Nagasaki, Riu Yamashita
PLOS ONE, 11, 4, e0154389, 2016年04月, ［査読有り］
英語, 研究論文（学術雑誌） - Establishment of transient and stable transfection systems for Babesia ovata
Hassan Hakimi, Junya Yamagishi, Yuto Kegawa, Osamu Kaneko, Shin-ichiro Kawazu, Masahito Asada
PARASITES & VECTORS, 9, 171, 2016年03月, ［査読有り］
英語, 研究論文（学術雑誌） - Characterization of Toxoplasma gondii glyoxalase 1 and evaluation of inhibitory effects of curcumin on the enzyme and parasite cultures
Youn-Kyoung Goo, Junya Yamagishi, Akio Ueno, Mohamad Alaa Terkawi, Gabriel Oluga Aboge, Dongmi Kwak, Yeonchul Hong, Dong-Il Chung, Makoto Igarashi, Yoshifumi Nishikawa, Xuenan Xuan
PARASITES & VECTORS, 8, 654, 2015年12月, ［査読有り］
英語, 研究論文（学術雑誌） - Eimeria tenellaのミトコンドリア呼吸鎖酵素活性の測定と阻害剤による虫体殺滅効果
松林 誠, 稲岡 ダニエル健, 小松谷 啓介, 八田 岳士, 三好 猛晴, 磯部 尚, 川原 史也, 山岸 潤也, 彦坂 健児, 佐藤 暖, 志波 智生, 原田 繁春, 北 潔, 辻 尚利
日本獣医学会学術集会講演要旨集, 158回, 305, 305, (公社)日本獣医学会, 2015年08月
日本語 - Inter-Individual Differences in the Oral Bacteriome Are Greater than Intra-Day Fluctuations in Individuals
Yukuto Sato, Junya Yamagishi, Riu Yamashita, Natsuko Shinozaki, Bin Ye, Takuji Yamada, Masayuki Yamamoto, Masao Nagasaki, Akito Tsuboi
PLOS ONE, 10, 6, e0131607, 2015年06月, ［査読有り］
英語, 研究論文（学術雑誌） - DB-AT: a 2015 update to the Full-parasites database brings a multitude of new transcriptomic data for apicomplexan parasites
Marcin Jakalski, Hiroyuki Wakaguri, Tabea G. Kischka, Yoshifumi Nishikawa, Shin-ichiro Kawazu, Makoto Matsubayashi, Fumiya Kawahara, Naotoshi Tsuji, Shinuo Cao, Fujiko Sunaga, Xuenan Xuan, Kazuhiro Okubo, Ikuo Igarashi, Josef Tuda, Arthur E. Mongan, Yuki Eshita, Ryuichiro Maeda, Wojciech Makalowski, Yutaka Suzuki, Junya Yamagishi
NUCLEIC ACIDS RESEARCH, 43, D1, D631, D636, 2015年01月, ［査読有り］
英語, 研究論文（学術雑誌） - Transcriptome and Histopathological Changes in Mouse Brain Infected with Neospora caninum
Maki Nishimura, Sachi Tanaka, Fumiaki Ihara, Yoshikage Muroi, Junya Yamagishi, Hidefumi Furuoka, Yutaka Suzuki, Yoshifumi Nishikawa
SCIENTIFIC REPORTS, 5, 7936, 2015年01月, ［査読有り］
英語, 研究論文（学術雑誌） - Cloning and characterization of aspartic protease 3 of Toxoplasma gondii.
Kamyingkird K, Goo YK, Cao S, Adjou Moumouni PF, Aboge GO, Yamagishi J, Terkawi MA, Masatani T, Yu L, Nishikawa Y, Xuan X
Journal of Protozoology Research, 24, 1, 18, 25, National Research Center for Protozoan Diseases, 2014年12月
英語, 研究論文（学術雑誌） - Defining the roles of the baculovirus regulatory proteins IE0 and IE1 in genome replication and early gene transactivation
Nadia Sokal, Yingchao Nie, Leslie G. Willis, Junya Yamagishi, Gary W. Blissard, Mark R. Rheault, David A. Theilmann
VIROLOGY, 468, 160, 171, 2014年11月, ［査読有り］
英語, 研究論文（学術雑誌） - Interactive transcriptome analysis of malaria patients and infecting Plasmodium falciparum
Junya Yamagishi, Anna Natori, Mohammed E. M. Tolba, Arthur E. Mongan, Chihiro Sugimoto, Toshiaki Katayama, Shuichi Kawashima, Wojciech Makalowski, Ryuichiro Maeda, Yuki Eshita, Josef Tuda, Yutaka Suzuki
GENOME RESEARCH, 24, 9, 1433, 1444, 2014年09月, ［査読有り］
英語, 研究論文（学術雑誌） - Eimeria tenellaスポロゾイトのミトコンドリア精製法の確立と呼吸鎖酵素活性
松林 誠, 稲岡 ダニエル健, 小松谷 啓介, 八田 岳士, 三好 猛晴, 磯部 尚, 川原 史也, 山岸 潤也, 彦坂 健児, 佐藤 暖, 志波 智生, 原田 繁春, 北 潔, 辻 尚利
日本獣医学会学術集会講演要旨集, 157回, 355, 355, (公社)日本獣医学会, 2014年08月
日本語 - The Babesia bovis gene and promoter model: an update from full-length EST analysis
Junya Yamagishi, Hiroyuki Wakaguri, Naoaki Yokoyama, Riu Yamashita, Yutaka Suzuki, Xuenan Xuan, Ikuo Igarashi
BMC GENOMICS, 15, 678, 2014年08月, ［査読有り］
英語, 研究論文（学術雑誌） - Transcriptome Analysis of Mouse Brain Infected with Toxoplasma gondii
Sachi Tanaka, Maki Nishimura, Fumiaki Ihara, Junya Yamagishi, Yutaka Suzuki, Yoshifumi Nishikawa
INFECTION AND IMMUNITY, 81, 10, 3609, 3619, 2013年10月, ［査読有り］
英語, 研究論文（学術雑誌） - Actin polymerization mediated by Babesia gibsoni aldolase is required for parasite invasion
Youn-Kyoung Goo, Akio Ueno, Mohamad Alaa Terkawi, G. Oluga Aboge, Yamagishi Junya, Makoto Igarashi, Jung-Yeon Kim, Yeon-Chul Hong, Dong-Il Chung, Yoshifumi Nishikawa, Xuenan Xuan
EXPERIMENTAL PARASITOLOGY, 135, 1, 42, 49, 2013年09月, ［査読有り］
英語, 研究論文（学術雑誌） - Adenosine-uridine-rich element is one of the required cis-elements for epimastigote form stage-specific gene expression of the congolense epimastigote specific protein
Keisuke Suganuma, Kennedy Miyoro Mochabo, Hassan Hakimi, Shino Yamasaki, Junya Yamagishi, Masahito Asada, Shin-ichiro Kawazu, Noboru Inoue
MOLECULAR AND BIOCHEMICAL PARASITOLOGY, 191, 1, 36, 43, 2013年09月, ［査読有り］
英語, 研究論文（学術雑誌） - TgGRA23, a novel Toxoplasma gondii dense granule protein associated with the parasitophorous vacuole membrane and intravacuolar network
Tatsunori Masatani, Tomohide Matsuo, Tetsuya Tanaka, Mohamad Alaa Terkawi, Eung-Goo Lee, Youn-Kyoung Goo, Gabriel Oluga Aboge, Junya Yamagishi, Kei Hayashi, Kyohko Kameyama, Shinuo Cao, Yoshifumi Nishikawa, Xuenan Xuan
PARASITOLOGY INTERNATIONAL, 62, 4, 372, 379, 2013年08月, ［査読有り］
英語, 研究論文（学術雑誌） - Epidemiological survey of Babesia bovis and Babesia bigemina infections of cattle in Philippines
Longzheng Yu, Mohmad Alaa Terkawi, Mary Jane Cruz-Flores, Florencia G. Claveria, Gabriel Oluga Aboge, Junya Yamagishi, Youn-Kyoung Goo, Shinuo Cao, Tatsunori Masatani, Yoshifumi Nishikawa, Xuenan Xuan
Journal of Veterinary Medical Science, 75, 7, 995, 998, 2013年, ［査読有り］
英語 - Protective effect of a prime-boost strategy with plasmid DNA followed by recombinant adenovirus expressing TgAMA1 as vaccines against Toxoplasma gondii infection in mice
Longzheng Yu, Junya Yamagishi, Shoufa Zhang, Chunmei Jin, Gabriel Oluga Aboge, Houshuang Zhang, Guohong Zhang, Tetsuya Tanaka, Kozo Fujisaki, Yoshifumi Nishikawa, Xuenan Xuan
PARASITOLOGY INTERNATIONAL, 61, 3, 481, 486, 2012年09月, ［査読有り］
英語, 研究論文（学術雑誌） - Molecular detection and identification of Babesia bovis and Babesia bigemina in cattle in northern Thailand
Shinuo Cao, Gabriel Oluga Aboge, Mohamad Alaa Terkawi, Longzheng Yu, Ketsarin Kamyingkird, Yuzi Luo, Yan Li, Youn-Kyoung Goo, Junya Yamagishi, Yoshifumi Nishikawa, Naoaki Yokoyama, Hiroshi Suzuki, Ikuo Igarashi, Ryuichiro Maeda, Tawin Inpankaew, Sathaporn Jittapalapong, Xuenan Xuan
PARASITOLOGY RESEARCH, 111, 3, 1259, 1266, 2012年09月, ［査読有り］
英語, 研究論文（学術雑誌） - Four promising antigens, BgP32, BgP45, BgP47, and BgP50, for serodiagnosis of Babesia gibsoni infection were classified as B. gibsoni merozoite surface protein family
Youn-Kyoung Goo, Gabriel Oluga Aboge, M. Alaa Terkawi, Honglin Jia, Junya Yamagishi, Fujiko Sunaga, Kazuhiko Namikawa, Se-Yeoun Cha, Hyung-Kwan Jang, Suk Kim, Yoshifumi Nishikawa, Xuenan Xuan
PARASITOLOGY INTERNATIONAL, 61, 2, 364, 368, 2012年06月, ［査読有り］
英語, 研究論文（学術雑誌） - Characterization of Toxoplasma gondii 5’ UTR with encyclopedic TSS information
Yamagishi J, Watanabe J, Goo YK, Masatani T, Suzuki Y, Xuan X
The Journal of Parasitology, 98, 2, 445, 447, American Society of Parasitologists, 2012年02月, ［査読有り］
英語, 研究論文（学術雑誌）, The 5′ UTR is widely involved in gene expression via post-transcriptional regulation. However, a detailed profile of the 5′ UTR for Toxoplasma gondii has not yet been demonstrated. To investigate the issue, we compared the predicted open reading frames (ORFs) and transcription start sites (TSSs) of T. gondii obtained by TSS-seq, a method that enables analysis of encyclopedic TSSs with next-generation sequencers. As a result, it was demonstrated that the mode length of the 5′ UTR is between 120 and 140 nucleotides (nts) when a subset of genes with predicted signal peptides was examined. However, when genes without the signal peptide were examined, the length was extended to approximately 600 nts. Because additional information on the predicted signal peptide generates increased reliability to the 5′ end estimation of each ORF, we believe that the former value was more reliable as a representative of the 5′ UTR length of T. gondii. The discrepancy suggests that current predictions of the 5′ end of the ORF were less accurate and considerably more discordant with the natural status. The 5′ untranslated region (5′ UTR) is defined as that between the 5′ end of the transcripts and just in front of a start codon of an ORF. Therefore, the 5′ UTR does not contain any information for a protein sequence; however, it is involved in the control of protein expression via the modulation of translational efficiency (Kozak, 1991b; Hughes, 2006).http://www.journalofparasitology.org/doi/abs/10.1645/GE-2864.1 - Macrophages Are Critical for Cross-Protective Immunity Conferred by Babesia microti against Babesia rodhaini Infection in Mice
Yan Li, Mohamad Alaa Terkawi, Yoshifumi Nishikawa, Gabriel Oluga Aboge, Yuzi Luo, Hideo Ooka, Youn-Kyoung Goo, Longzheng Yu, Shinuo Cao, Yongfeng Sun, Junya Yamagishi, Tatsunori Masatani, Naoaki Yokoyama, Ikuo Igarashi, Xuenan Xuan
INFECTION AND IMMUNITY, 80, 1, 311, 320, 2012年01月, ［査読有り］
英語, 研究論文（学術雑誌） - Epidemiological Survey of Theileria Parasite Infection of Cattle in Northeast China by Allele-Specific PCR
Longzheng Yu, Shoufa Zhang, Wanfeng Liang, Chunmei Jin, Lijun Ji, Yuzi Luo, Yan Li, Shinuo Cao, Junya Yamagishi, Yoshifumi Nishikawa, Suguru Kawano, Kozo Fujisaki, Xuenan Xuan
JOURNAL OF VETERINARY MEDICAL SCIENCE, 73, 11, 1509, 1512, 2011年11月, ［査読有り］
英語, 研究論文（学術雑誌） - Identification and characterization of a novel secreted antigen 1 of Babesia microti and evaluation of its potential use in enzyme-linked immunosorbent assay and immunochromatographic test
Yuzi Luo, Honglin Jia, M. Alaa Terkawi, Youn-Kyoung Goo, Suguru Kawano, Hideo Ooka, Yan Li, Longzheng Yu, Shinuo Cao, Junya Yamagishi, Kozo Fujisaki, Yoshifumi Nishikawa, Atsuko Saito-Ito, Ikuo Igarashi, Xuenan Xuan
PARASITOLOGY INTERNATIONAL, 60, 2, 119, 125, 2011年06月, ［査読有り］
英語, 研究論文（学術雑誌） - Construction and analysis of full-length cDNA library of Cryptosporidium parvum
Junya Yamagishi, Hiroyuki Wakaguri, Sumio Sugano, Suguru Kawano, Kozo Fujisaki, Chihiro Sugimoto, Junichi Watanabe, Yutaka Suzuki, Isao Kimata, Xuenan Xuan
PARASITOLOGY INTERNATIONAL, 60, 2, 199, 202, 2011年06月, ［査読有り］
英語, 研究論文（学術雑誌） - Babesia microti: Molecular and antigenic characterizations of a novel 94-kDa protein (BmP94)
Hideo Ooka, Mohamad Alaa Terkawi, Youn-Kyoung Goo, Yuzi Luo, Yan Li, Junya Yamagishi, Yoshifumi Nishikawa, Ikuo Igarashi, Xuenan Xuan
EXPERIMENTAL PARASITOLOGY, 127, 1, 287, 293, 2011年01月, ［査読有り］
英語, 研究論文（学術雑誌） - Full-parasites: database of full-length cDNAs of apicomplexa parasites, 2010 update
Josef Tuda, Arthur E. Mongan, Mohammed E. M. Tolba, Mihoko Imada, Junya Yamagishi, Xuenan Xuan, Hiroyuki Wakaguri, Sumio Sugano, Chihiro Sugimoto, Yutaka Suzuki
NUCLEIC ACIDS RESEARCH, 39, Database issue, D625, D631, 2011年01月, ［査読有り］
英語, 研究論文（学術雑誌） - Construction of Neospora caninum stably expressing TgSAG1 and evaluation of its protective effects against Toxoplasma gondii infection in mice
Guohong Zhang, Xiaohong Huang, Damdinsuren Boldbaatar, Banzragch Battur, Badgar Battsetseg, Houshuang Zhang, Longzheng Yu, Yan Li, Yuzi Luo, Shinuo Cao, Youn-Kyong Goo, Junya Yamagishi, Jinlin Zhou, Shoufa Zhang, Hiroshi Suzuki, Ikuo Igarashi, Takeshi Mikami, Yoshifumi Nishikawa, Xuenan Xuan
VACCINE, 28, 45, 7243, 7247, 2010年10月, ［査読有り］
英語, 研究論文（学術雑誌） - High-Resolution Characterization of Toxoplasma gondii Transcriptome with a Massive Parallel Sequencing Method
Junya Yamagishi, Hiroyuki Wakaguri, Akio Ueno, Youn-Kyoung Goo, Mohammed Tolba, Makoto Igarashi, Yoshifumi Nishikawa, Chihiro Sugimoto, Sumio Sugano, Yutaka Suzuki, Junichi Watanabe, Xuenan Xuan
DNA RESEARCH, 17, 4, 233, 243, 2010年08月, ［査読有り］
英語, 研究論文（学術雑誌） - Serodiagnosis of ovine toxoplasmosis in Mongolia by an enzyme-linked immunosorbent assay with recombinant Toxoplasma gondii matrix antigen 1
Buyannemekh Tumurjav, Mohamad Alaa Terkawi, Houshuang Zhang, Guohong Zhang, Honglin Jia, Youn-Kyoung Goo, Junya Yamagishi, Yoshifumi Nishikawa, Ikuo Igarashi, Chihiro Sugimoto, Xuenan Xuan
JAPANESE JOURNAL OF VETERINARY RESEARCH, 58, 2, 111, 119, 2010年08月, ［査読有り］
英語, 研究論文（学術雑誌） - Neospora caninum: Application of apical membrane antigen 1 encapsulated in the oligomannose-coated liposomes for reduction of offspring mortality from infection in BALB/c mice
Houshuang Zhang, Yoshifumi Nishikawa, Junya Yamagishi, Jinlin Zhou, Yuzuru Ikehara, Naoya Kojima, Naoaki Yokoyama, Xuenan Xuan
EXPERIMENTAL PARASITOLOGY, 125, 2, 130, 136, 2010年06月, ［査読有り］
英語, 研究論文（学術雑誌） - Characterization of a leucine aminopeptidase from Toxoplasma gondii
Honglin Jia, Yoshifumi Nishikawa, Yuzi Luo, Junya Yamagishi, Chihiro Sugimoto, Xuenan Xuan
MOLECULAR AND BIOCHEMICAL PARASITOLOGY, 170, 1, 1, 6, 2010年03月, ［査読有り］
英語, 研究論文（学術雑誌） - Molecular characterization and expression of a 47-kDa merozoite surface protein of Babesia gibsoni for serodiagnosis by enzyme-linked immunosorbent assay.
Aboge, G. O, Batbaatar, V, Goo, Y-K, Yamagishi, J, Nishikawa, Y, Sunaga, F, Namikawa, K, Igarashi, I, Fujisaki, K, Suzuki, H, Xuan, X
J. Protozool. Res., 20, 59, 69, 2010年, ［査読有り］ - Babesia gibsoni: Identification, expression, localization, and serological characterization of a Babesia gibsoni 22-kDa protein
Youn-Kyoung Goo, Honglin Jia, Mohamad Alaa Terkawi, Gabriel Oluga Aboge, Junya Yamagishi, Yoshifumi Nishikawa, Suk Kim, Hyung-Kwan Jang, Kozo Fujisaki, Xuenan Xuan
EXPERIMENTAL PARASITOLOGY, 123, 3, 273, 276, 2009年11月, ［査読有り］
英語, 研究論文（学術雑誌） - Molecular characterizations of three distinct Babesia gibsoni rhoptry-associated protein-1s (RAP-1s).
Terkawi MA, Amornthep A, Ooka H, Aboge G, Jia H, Goo YK, Nelson B, Yamagishi J, Nishikawa Y, Igarashi I, Kawazu SI, Fujisaki K, Xuan X
Parasitology, 136, 10, 1147, 1160, 10, 2009年09月, ［査読有り］, ［国際誌］
英語, 研究論文（学術雑誌）, Three cDNAs encoding rhoptry-associated protein 1 (RAP-1) homologues were found in the Babesia gibsoni EST database. Based on similarities to BgRAP-1a, which was identified previously by serological screening of a cDNA merozoite library, the two new genes were designated BgRAP-1b (33.7%) and BgRAP-1c (57%). Mice antiserum raised against each recombinant protein reacted specifically with B. gibsoni parasites as determined by Western blotting, which showed native molecular sizes of the BgRAP-1a (51 kDa), BgRAP-1b (53 kDa) and BgRAP-1c (47 kDa) consistent with predictable molecular weights. Immunofluoresence using these antibodies revealed localization of all BgRAP-1s within the matrix of merozoites; however, BgRAP-1a appeared to diverge from the other two when it was found secreted into the cytoplasm of infected erythrocytes. Apical localization of all 3 BgRAP-1s during the extracellular stage of the parasite combined with their ability to bind a canine erythrocyte membrane fraction was suggestive of a role for these proteins in erythrocyte attachment. Lastly, the ability of these recombinant proteins to be used as diagnostic reagents was tested by ELISA and the sensitivities of BgRAP-1a and BgRAP-1c were found increased through N-terminal truncation. Taken together, our data suggest divergent roles for the 3 BgRAP-1s in the merozoite stage of B. gibsoni. - Characterization of a leucine aminopeptidase of Babesia gibsoni.
Jia H, Terkawi MA, Aboge GO, Goo YK, Luo Y, Li Y, Yamagishi J, Nishikawa Y, Igarashi I, Sugimoto C, Fujisaki K, Xuan X
Parasitology, 136, 9, 945, 952, 9, 2009年08月, ［査読有り］, ［国際誌］
英語, 研究論文（学術雑誌）, Peptidases of parasitic protozoa are currently under intense investigation in order to identify novel virulence factors, drug targets, and vaccine candidates, except in Babesia. Leucine aminopeptidases in protozoa, such as Plasmodium and Leishmania, have been identified to be involved in free amino acid regulation. We report here the molecular and enzymatic characterization, as well as the localization of a leucine aminopeptidase, a member of the M17 cytosolic aminopeptidase family, from B. gibsoni (BgLAP). A functional recombinant BgLAP (rBgLAP) expressed in Escherichia coli efficiently hydrolysed synthetic substrates for aminopeptidase, a leucine substrate. Enzyme activity of the rBgLAP was found to be optimum at pH 8.0 and at 37 degrees C. The substrate profile was slightly different from its homologue in P. falciprum. The activity was also strongly dependent on metal divalent cations, and was inhibited by bestatin, which is a specific inhibitor for metalloprotease. These results indicated that BgLAP played an important role in free amino acid regulation. - Molecular and immunological characterization of Babesia gibsoni and Babesia microti heat shock protein-70.
Terkawi MA, Aboge G, Jia H, Goo YK, Ooka H, Yamagishi J, Nishikawa Y, Yokoyama N, Igarashi I, Kawazu SI, Fujisaki K, Xuan X
Parasite immunology, 31, 6, 328, 340, 6, 2009年06月, ［査読有り］, ［国際誌］
英語, 研究論文（学術雑誌）, Serological immunoscreening was used to identify a gene encoding heat shock protein-70 from Babesia gibsoni (BgHSP-70) that showed high homology with HSP-70s from other apicomplexan parasites. This gene corresponded to a full-length cDNA containing an open reading frame of 1968 bp predicted to result in a 70-kDa mature protein consisting of 656 amino acids. Analysis of the expression levels of BgHSP-70 indicated elevated transcription from cultured parasites incubated at 40 degrees C for 1 h, but not at 30 degrees C. Interestingly, antiserum raised against recombinant BgHSP-70 protein reacted specifically not only with a 70-kDa protein of B. gibsoni but also with a corresponding native protein of B. microti (BmHSP-70), indicating the high degree of conservation of this protein. The BmHSP-70 gene was then isolated and characterized and the immunoprotective properties of recombinant BgHSP-70 (rBgHSP-70) and rBmHSP-70 were compared in vitro and in vivo. Both proteins had potent mitogenic effects on murine and canine mononuclear cells as evidenced by high proliferative responses and IFN-gamma production after stimulation. Immunization regimes in BALB/c and C57BL/6 mice using rBgHSP-70 and rBmHSP-70 elicited high antibody levels, with concurrent significant reductions in peripheral parasitaemias. Taken together, these results emphasize the potential of HSP-70s as a molecular adjuvant vaccine. - Toxoplasma gondii: Sensitive and rapid detection of infection by loop-mediated isothermal amplification (LAMP) method
Houshuang Zhang, Oriel M. M. Thekisoe, Gabriel O. Aboge, Hisako Kyan, Junya Yamagishi, Noboru Inoue, Yoshifumi Nishikawa, Satoshi Zakimi, Xuenan Xuan
EXPERIMENTAL PARASITOLOGY, 122, 1, 47, 50, 2009年05月, ［査読有り］
英語, 研究論文（学術雑誌） - Characterization of the Babesia gibsoni 12-kDa protein as a potential antigen for the serodiagnosis
Youn-Kyoung Goo, Honglin Jia, G. Oluga Aboge, M. Alaa Terkawi, Eung-Goo Lee, Junya Yamagishi, Yoshifumi Nishikawa, Hyung-Kwan Jang, Fujiko Sunaga, Kazuhiko Namikawa, Kozo Fujisaki, Xuenan Xuan
PARASITOLOGY INTERNATIONAL, 58, 1, 55, 60, 2009年03月, ［査読有り］
英語, 研究論文（学術雑誌） - The AcMNPV pp31 gene is not essential for productive AcMNPV replication or late gene transcription but appears to increase levels of most viral transcripts
Junya Yamagishi, Erik D. Burnett, Steven H. Harwood, Gary W. Blissard
VIROLOGY, 365, 1, 34, 47, 2007年08月, ［査読有り］
英語, 研究論文（学術雑誌） - Expression of Autographa californica multiple nucleopolyhedrovirus genes in mammalian cells and Upregulation of the host beta-actin gene
R Fujita, T Matsuyama, J Yamagishi, K Sahara, S Asano, H Bando
JOURNAL OF VIROLOGY, 80, 5, 2390, 2395, 2006年03月, ［査読有り］
英語, 研究論文（学術雑誌） - The use of a recombinant baculovirus expressing a chitinase from the hard tick Haemaphysalis longicornis and its potential application as a bioacaricide for tick control
SP Assenga, M You, CH Shy, J Yamagishi, T Sakaguchi, JL Zhou, MK Kibe, XN Xuan, K Fujisaki
PARASITOLOGY RESEARCH, 98, 2, 111, 118, 2006年01月, ［査読有り］
英語, 研究論文（学術雑誌） - [Baculovirus vector--gene transfer into mammalian cells].
Tani H, Abe T, Limn CK, Mochizuki R, Yamagishi J, Kitagawa Y, Watanabe R, Moriishi K, Matsuura Y
Uirusu, 53, 2, 185, 193, 2003年12月, ［査読有り］ - DNA microarrays of baculovirus genomes: differential expression of viral genes in two susceptible insect cell lines
J Yamagishi, R Isobe, T Takebuchi, H Bando
ARCHIVES OF VIROLOGY, 148, 3, 587, 597, 2003年03月, ［査読有り］
英語, 研究論文（学術雑誌） - Genome organization and mRNA structure of Periplaneta fuliginosa densovirus imply alternative splicing involvement in viral gene expression
J Yamagishi, Y Hu, J Zheng, H Bando
ARCHIVES OF VIROLOGY, 144, 11, 2111, 2124, 1999年, ［査読有り］
英語, 研究論文（学術雑誌）
その他活動・業績
- Ampliseqを用いたフィリピンにおけるウシピロプラズマ集団の特性化【JST・京大機械翻訳】
MAY Galon Eloiza, MAY Galon Eloiza, MIKI Macalanda Adrian, ADRIAN Ybanez, 山岸潤也, 玄学南, 日本獣医学会学術集会講演要旨集, 166th, 2023年 - ザンビアのコウモリが保有する多様なヘルペスウイルスの遺伝学的性状の解明
播磨勇人, 邱永晋, 山岸潤也, 梶原将大, CHANGULA Katendi, 佐々木道仁, MWEENE Aaron S, 澤洋文, 石原加奈子, HANG’OMBE Bernard M, 高田礼人, 日本ウイルス学会学術集会プログラム・予稿集(Web), 70th, 2023年 - SARS-CoV-2感染ハムスターを用いた経口抗ウイルス薬と抗炎症薬の併用治療効果の解析
佐々木道仁, 杉達紀, 飯田俊, 平田雄一郎, 日下部伸治, 日下部伸治, 小西慧, 小西慧, 板倉友香里, 板倉友香里, 田畑耕史郎, 田畑耕史郎, 岸本麻衣, 岸本麻衣, 小林広子, 有泉拓馬, KITTIYA Intaruck, 登治謙, 鳥羽晋輔, 鳥羽晋輔, 佐藤彰彦, 佐藤彰彦, 佐藤彰彦, 松野啓太, 松野啓太, 山岸潤也, 山岸潤也, 鈴木忠樹, 大場靖子, 大場靖子, 澤洋文, 澤洋文, 澤洋文, 日本ウイルス学会学術集会プログラム・予稿集(Web), 70th, 2023年 - Babesia caballiの全ゲノム配列解析
山岸潤也, 越智章仁, 木高大志, HAKIMI Hassan, 麻田正仁, 日本寄生虫学会大会プログラム・抄録集, 92nd, 2023年 - サルマラリア原虫Plasmodium knowlesi寄生赤血球におけるシントン・マリガン斑点と関連分子
坂口美亜子, LUCKY Amuza Byaruhanga, LUCKY Amuza Byaruhanga, ASARE Kwame Kumi, ASARE Kwame Kumi, 莊淮, 莊淮, 莊淮, 麻田正仁, 麻田正仁, 宮崎真也, 山岸潤也, 片貝祐子, 川合覚, SONG Chihong, 村田和義, TEMPLETON Thomas J, 矢幡一英, 金子修, 金子修, 日本寄生虫学会大会プログラム・抄録集, 91st, 2022年 - ヒト静脈内皮細胞に付着するPlasmodium knowlesi感染赤血球の分子基盤
CHUANG Huai, AMUZA Lucky Byaruhanga, YAMAGISHI Junya, KATAKAI Yuko, KAWAI Satoru, KANEKO Osamu, SAKAGUCHI Miako, 日本熱帯医学会大会プログラム抄録集, 62nd (CD-ROM), 2021年 - ヒト静脈内皮細胞付着に対するPlasmodium knowlesi感染赤血球の分子基礎
莊淮, 莊淮, BYARUHANGA Lucky Amuza, BYARUHANGA Lucky Amuza, 山岸潤也, 片貝裕子, 河合覚, 金子修, 金子修, 坂口美亜子, 日本寄生虫学会大会プログラム・抄録集, 89th, 2020年 - Haemaphysalis longicornisが保有するCoxiella様共生細菌のビタミンB群およびその補因子の代謝に関わる遺伝子の探索
草木迫浩大, 山本佳代子, 今里裕平, 山岸潤也, 白藤(梅宮)梨可, 玄学南, 野中成晃, 中尾亮, 日本寄生虫学会大会プログラム・抄録集, 89th, 2020年 - ウシはマラウイでの眠り病の潜在的動物リザーバーである
MARSELA Megasari, 林田京子, 中尾亮, CHATANGA Elisha, GAITHUMA Alex Kiari, 河合直子, MUSAYA Janelisa, 杉本千尋, 山岸潤也, 日本寄生虫学会大会プログラム・抄録集, 89th, 2020年 - East/central/south African genotype in a chikungunya outbreak, Dhaka, Bangladesh, 2017
Mizanur Rahman, Junya Yamagishi, Rummana Rahim, Abu Hasan, Abu Sobhan, Emerging Infectious Diseases, 25, 2, 370, 372, 2019年02月01日
Centers for Disease Control and Prevention (CDC), 英語, 速報，短報，研究ノート等（学術雑誌） - Corrigendum: Interactive transcriptome analysis of malaria patients and infecting Plasmodium falciparum.
Junya Yamagishi, Anna Natori, Mohammed E M Tolba, Arthur E Mongan, Chihiro Sugimoto, Toshiaki Katayama, Shuichi Kawashima, Wojciech Makalowski, Ryuichiro Maeda, Yuki Eshita, Josef Tuda, Yutaka Suzuki, Genome research, 28, 8, 1253, 1253, 2018年08月, ［国際誌］
英語 - 日本産ヒトスジシマカのジカウイルス媒介能
江下優樹, 江下優樹, 江下優樹, 江下優樹, 和田雄治, 林田京子, 山岸潤也, 杉本千尋, 佐々木道仁, 大場靖子, 澤洋文, 飛弾野真也, 神山長慶, 小林隆志, 高崎智彦, 加藤文博, 田島茂, 倉根一郎, LIM Chang Kweng, 衛生動物, 69, Supplement, 51, 2018年03月30日
日本語 - Investigating the roles of IFN gamma and IFN gamma-stimulated GTPases during Legionella pneumophila replication in alveolar macrophages and monocyte-derived cells
Chao Yang, Shivani Pasricha, Sze Ying Ong, Andrew Stephen Brown, Junya Yamagishi, Chihiro Sugimoto, Sammy Bedoui, Ian R. van Driel, Elizabeth L. Hartland, CYTOKINE, 100, 149, 149, 2017年12月
英語, 研究発表ペーパー・要旨（国際会議） - Theileria orientalisのマウスとマダニを用いた感染実験系の確立と遺伝子発現解析
林田京子, 林田京子, 白藤梨可, SIVAKUMAR Thillaiampalam, 山岸潤也, 鈴木穣, 杉本千尋, 横山直明, 日本寄生虫学会大会プログラム・抄録集, 86th, 89, 2017年
日本語 - Babesia ovataのゲノム・トランスクリプトーム解析
山岸潤也, 麻田正仁, HASSAN Hakimi, 田中健, 田中健, 杉本千尋, 河津信一郎, 日本獣医学会学術集会講演要旨集, 159th, 334, 334, 2016年08月30日
(公社)日本獣医学会, 日本語 - 2種類のインビトロ培養条件下における大型ピロプラズマ原虫Babesia ovataの比較トランスクリプトーム解析
田中健, 山岸潤也, HASSAN Hakimi, 麻田正仁, 河津信一郎, 日本獣医学会学術集会講演要旨集, 158th, 304, 304, 2015年08月30日
(公社)日本獣医学会, 日本語 - Babesia ovataの短時間かつ安定的な遺伝子導入系の確立(Establishment of transient and stable transfection system for Babesia ovata)
Hakimi Hassan, 山岸 潤也, 外川 裕人, 田中 健, 金子 修, 河津 信一郎, 麻田 正仁, 日本獣医学会学術集会講演要旨集, 158回, 304, 304, 2015年08月
(公社)日本獣医学会, 英語 - 1‐Cys型ペルオキシレドキシン遺伝子欠損Plasmodium bergheiの赤内型RNA‐seq解析
薄井美帆, 増田(菅沼)裕乃, 元岡大祐, 中村昇太, 山岸潤也, 福本晋也, 井上昇, 堀井俊宏, 河津信一郎, 日本寄生虫学会大会プログラム・抄録集, 84th, 79, 2015年02月
日本語 - ヒト口腔細菌叢の日内変動性
佐藤行人, 山岸潤也, 山下理宇, 篠崎夏子, 長崎正朗, 坪井明人, 日本ゲノム微生物学会年会要旨集, 9th, 2015年 - 口腔細菌叢メタゲノム解析におけるDNA抽出方法の検討
山岸潤也, 佐藤行人, 山下理宇, 篠崎夏子, 長崎正朗, 坪井明人, 日本ゲノム微生物学会年会要旨集, 9th, 2015年 - メタゲノム解析における歯垢試料の影響の評価指標について
篠崎夏子, 篠崎夏子, 山岸潤也, 佐藤行人, 長崎正朗, 坪井明人, 山下理宇, 日本ゲノム微生物学会年会要旨集, 9th, 2015年 - 歯垢を中心とした口腔内細菌叢解析の基盤整備
山下理宇, 佐藤行人, 山岸潤也, 篠崎夏子, 長崎正朗, 坪井明人, 日本ゲノム微生物学会年会要旨集, 9th, 2015年 - 8.一過性的にデングウイルス血症をマウスに起こして、定量的感染蚊を大量に作る新規の方法(一般講演(1),都市有害生物管理学会第35回大会講演要旨)
江下 優樹, Runtuwene Lucky R, 牧野 芳大, 野口 香緒里, 川上 絵理, 福田 昌子, 小林 隆志, 小西 英二, 山中 敦史, Srisawat Raweewan, Komalamisra Narumon, 成田 弘成, 牛島 廣治, Mon-gan Arthur E, 今田 美穂子, 山岸 潤也, 鈴木 穣, 中井 謙太, 前田 龍一郎, 杉本 千尋, 倉根 一郎, 高崎 智彦, 都市有害生物管理, 4, 2, 120, 121, 2014年12月20日
都市有害生物管理学会, 日本語 - Eimeria tenellaスポロゾイトのミトコンドリア精製法の確立と呼吸鎖酵素活性
稲岡 ダニエル健, 松林 誠, 小松谷 啓介, 八田 岳士, 三好 猛晴, 磯部 尚, 川原 史也, 山岸 潤也, 彦坂 健児, 佐藤 暖, 志波 智生, 原田 繁春, 辻 尚利, 北 潔, 日本生化学会大会プログラム・講演要旨集, 87回, [4T09p, 15], 2014年10月
(公社)日本生化学会, 日本語 - 歯垢検体の保存温度および保存期間による細菌叢への影響
篠崎夏子, 山岸潤也, 佐藤行人, 山下理宇, 山田拓司, 長崎正朗, 坪井明人, 日本ゲノム微生物学会年会要旨集, 8th, 66, 2014年
日本語 - Babesia gibsoniのドラフトゲノム解析
山岸 潤也, 麻田 正仁, 正谷 達謄, 于 龍政, 曹 世諾, 須永 藤子, 玄 学南, 日本獣医学会学術集会講演要旨集, 156回, 238, 238, 2013年08月
(公社)日本獣医学会, 日本語 - Validation of 40S ribosomal protein 17S as internal control for qRT-PCR of dengue-infected Aedes aegypti.
RUNTUWENE Lucky R, KAWASHIMA Shuichi, YAMAGISHI Junya, SUZUKI Yutaka, SUGANO Sumio, NAKAI Kenta, MAEDA Ryuichiro, SUGIMOTO Chihiro, TAKASAKI Tomohiko, KURANE Ichiro, KOBAYASHI Takashi, ESHITA Yuki, 衛生動物, 64, 2, 138, 2013年06月15日
英語 - Trypanosoma congolenseエピマスティゴート型原虫のヘモグロビン取込みに関する研究
山崎詩乃, 菅沼啓輔, 麻田正仁, NGASAMAN Ruttayaporn, 周末, 山岸潤也, 河津信一郎, 井上昇, 日本獣医学会学術集会講演要旨集, 155th, 216, 216, 2013年03月04日
(公社)日本獣医学会, 日本語 - Trypanosoma congolense EMFステージ特異的遺伝子の発現調節機構解析
菅沼啓輔, 山崎詩乃, 山岸潤也, 麻田正仁, 河津信一郎, 井上昇, 日本獣医学会学術集会講演要旨集, 155th, 118, 118, 2013年03月04日
(公社)日本獣医学会, 日本語 - congolense epimastigote specific protein(CESP)のEMF特異的発現は3'非翻訳領域によって調節されている
菅沼 啓輔, 山崎 詩乃, 山岸 潤也, 麻田 正仁, 河津 信一郎, 井上 昇, 獣医寄生虫学会誌, 11, 1, 49, 49, 2012年10月
日本獣医寄生虫学会, 日本語 - Trypanosoma congolenseハプトグロビン-ヘモグロビン複合体レセプター様タンパク質(TcHpHbR)に関する研究
山崎 詩乃, 菅沼 啓輔, 山岸 潤也, 麻田 正仁, 河津 信一郎, 井上 昇, 獣医寄生虫学会誌, 11, 1, 50, 50, 2012年10月
日本獣医寄生虫学会, 日本語 - congolense epimastigote specific protein(CESP)のEMF特異的発現は3’非翻訳領域によって調節されている
菅沼啓輔, 山崎詩乃, 山岸潤也, 麻田正仁, 河津信一郎, 井上昇, 日本獣医学会学術集会講演要旨集, 153rd, 224, 224, 2012年03月01日
(公社)日本獣医学会, 日本語 - Trypanosoma congolenseハプトグロビン‐ヘモグロビン複合体レセプター様タンパク質(TcHpHbR)に関する研究
山崎詩乃, 菅沼啓輔, 山岸潤也, 麻田正仁, 河津信一郎, 井上昇, 日本獣医学会学術集会講演要旨集, 153rd, 225, 225, 2012年03月01日
(公社)日本獣医学会, 日本語 - トキソプラズマのステージ間比較トランスクリプトーム解析
山岸潤也, 若栗浩幸, 河津信一郎, 鈴木穣, XUAN Xuenan, 日本寄生虫学会大会プログラム・抄録集, 81st, 87, 2012年02月
日本語 - アピコンプレクス門原虫のプロモーター構造種間比較
山岸潤也, 山下理宇, 鈴木穣, 五十嵐郁男, 河津信一郎, XUAN Xuenan, 日本分子生物学会年会プログラム・要旨集(Web), 35th, 1P-0029 (WEB ONLY), 2012年
日本語 - Babesia bovisメロゾイト滑走運動のバイオイメージング解析
麻田正仁, 山岸潤也, 後藤康之, 横山直明, 井上昇, 河津信一郎, 日本寄生虫学会東日本支部大会プログラム・講演要旨, 72nd, 19, 2012年
日本語 - ヒトと熱帯熱マラリア原虫のトランスクリプトーム相互作用データの収集と解析
名取杏奈, TUDA Josef, MONGAN Arthur, TOLBA Mohammed, TOLBA Mohammed, 今田美穂子, 山岸潤也, XUAN Xuenan, 若栗浩幸, 菅野純夫, 杉本千尋, 鈴木穣, 日本分子生物学会年会プログラム・要旨集(Web), 35th, WEB ONLY 4P-0211, 2012年
日本語 - Identification of an immunodominant Babesia gibsoni 47-kDa antigen
Goo Jia Y-K, Aboge G. O, Terkawi M. A, Yamagishi J, Nishikawa Y, Igarashi I, Xuan X, The Journal of protozoology research, 19, 2, 16, 21, 2009年12月
National Research Center for Protozoan Diseases, 英語 - Identification of a novel B. gibsoni 27-kDa protein as a serodiagnostic antigen
Terkawi M. A., Aboge G., Jia H., Goo Y-K., Ooka H., Yamagishi Junya, Nishikawa Yoshifumi, Kawazu Shin-ichiro, Fujisaki K., Xuan Xuenan, 山岸 潤也, 西川 義文, 河津 信一郎, 玄 学南, The Journal of Protozoology Research, 18, 2, 48, 56, 2008年12月
A novel gene encoding 27-kDa protein was identified by the screening of Babesia gibsoni cDNA library with acutely infected dog serum. The BgP27 is a single copy gene with a predicted open reading frame of 762 bp and 254 amino acids. The phylogenic analysis of the deduced amino acid of BgP27 demonstrated considerable identities with members of Plasmodium berghei circumsporozoite protein family that ranged between 18.4% and 22.8%. The BgP27 was expressed as a glutathione S-transferase fusion protein in Escherichia coli. The serum raised in mice against the recombinant protein specifically reacted with a 27-kDa protein in the extracts of B. gibsoni parasites. Confocal laser scanning microscopic observation showed high fluorescent reactivity with both extracellular and intracellular merozoite. Furthermore, recombinant BgP27 was used as an antigen in an enzyme-linked immunosorbent assay (ELISA). Thus, the kinetics of the anti-BgP27 antibody was detected in serial serum samples collected from a B. gibsoni-infected dog. IgG levels were high throughout the course of infection. In addition, the ELISA was able to differentiate between B. gibsoni-infected dog serum and B. canis subspecies-infected dog serum or normal dog serum. The diagnostic performance of BgP27-ELISA revealed the potential use of the antigen for detection of infection in dogs., National Research Center for Protozoan Diseases, 英語 - 【ウイルスベクター 最新の話題】 バキュロウイルスベクター 哺乳動物細胞への遺伝子導入
谷 英樹, 阿部 隆之, 林 昌宏, 望月 理加, 山岸 潤也, 北川 善紀, 渡辺 理恵, 宮本 大伸, 森石 恆司, 松浦 善治, ウイルス, 53, 2, 185, 193, 2003年12月
日本ウイルス学会, 日本語 - PfDNV遺伝子産物によるウイルスRNAスプライシングの抑制
山岸 潤也, 浅野 真一郎, 佐原 健, 飯塚 敏彦, 伴戸 久徳, 日本蠶絲學雜誌, 69, 4, 271, 276, 2000年08月31日
日本蠶絲學會, 日本語 - ＰｆＤＮＶ遺伝子産物によるウイルスＲＮＡスプライシングの抑制
山岸 潤也, 浅野 真一郎, 佐原 健, 飯塚 敏彦, 伴戸 久徳, 日本蚕糸学雑誌, 69, 4, 271, 276, 2000年
The Japanese Society of Sericultural Science, 英語
共同研究・競争的資金等の研究課題
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山岸 潤也, 麻田 正仁, 林田 京子, 杉 達紀
日本学術振興会, 基盤研究(B), 北海道大学, 24K02270 - アフリカトリパノソーマ原虫伝播動態理解のためのツェツェバエ微生物総体解析
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林田 京子, 山岸 潤也
日本学術振興会, 基盤研究(C), 北海道大学, 24K09240 - 内臓型リーシュマニア症の免疫病態
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2024年04月01日 - 2027年03月31日
後藤 康之, 山岸 潤也, 藤井 渉, 片岡 直行, 溝渕 悠代
日本学術振興会, 基盤研究(B), 東京大学, 24K02271 - 犬バベシア症における宿主防御免疫機構の解明と新型組換えワクチンの開発
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2022年04月01日 - 2025年03月31日
玄 学南, 麻田 正仁, 正谷 達謄, 山岸 潤也
日本学術振興会, 基盤研究(B), 帯広畜産大学, 22H02509 - 犬バベシア症における宿主防御免疫機構の解明と新型組換えワクチンの開発
科学研究費助成事業 基盤研究（B）
2024年04月 - 2025年03月
玄学南, 麻田正仁, 正谷達謄, 山岸潤也, 川瀬摂, 平田 晴之
日本学術振興会, 科学研究費助成事業 基盤研究（B）, 帯広畜産大学, 23K23774 - 内臓型リーシュマニア症の免疫病態
科学研究費助成事業
2021年04月01日 - 2024年03月31日
後藤 康之, 藤井 渉, 片岡 直行, 山岸 潤也
内臓型リーシュマニア症（VL）はリーシュマニア原虫の感染により引き起こされる人獣共通感染症である。本症の化学療法はさまざまな問題点を抱えており、宿主免疫を適切に調節することで効果を発揮する免疫療法は新たなVLの治療法として期待される。一方、発熱、貧血、肝脾腫といったVLの症状が免疫応答に起因することから、不適切な免疫刺激による逆効果も予想される。つまり、VLに対する効果的な免疫療法の確立にはその感染・発症機序を詳細にとらえる必要がある。本研究では、発生工学を駆使したマウスモデルと、ヒト患者やイヌ由来材料を用いた解析を有機的に組み合わせることで、VLにおける病態免疫を明らかにし、症状の改善を促す免疫療法の開発を目指している。
2021年度は特に貧血の原因となる脾臓での血球貪食に焦点を当てて研究を進めた。血球貪食が亢進する感染マウスの脾臓では多核化マクロファージ（MGC）の出現が顕著になる。そこで本年度はin vitroにて感染誘導性マクロファージ多核化現象の構築を行った。その結果、L. donovani感染によって引き起こされる多核化はGM-CSFなどのサイトカインによって誘導される多核化とは異なり、血球貪食能力を特に引き上げることが明らかとなった。感染マウスでの血球貪食が肝臓では見られず脾臓で見られることから、感染臓器トランスクリプトーム解析により脾臓のみで上昇する遺伝子の探索を行ったところ、その一つがin vitro誘導MGCでも顕著に上昇することが明らかとなった。プラスミドを用いてマクロファージにその遺伝子を過発現させたところ、感染なしでもマクロファージの多核化が誘導された。つまり、原虫感染による当該遺伝子発現の上昇は、血球貪食型のMGCを誘導することが示唆された。
日本学術振興会, 基盤研究(B), 東京大学, 21H02722 - バベシアのマダニ体内発育ステージ抗原の網羅的解析：伝搬阻止ワクチン開発の基盤整備
科学研究費助成事業
2019年04月01日 - 2022年03月31日
河津 信一郎, 白藤 梨可, 麻田 正仁, 山岸 潤也
本研究はバベシアのマダニ体内発育ステージに特異的な細胞表在性蛋白質を網羅的に検出し、その機能を解析するとともに伝搬阻止ワクチン（TBV）抗原としての性能を評価することを目的とする。
【材料と方法】Babesia ovata（三宅株）赤血球内発育ステージ（Blood stage）をin vitro誘導法（PubMed ID: 1549158）によりマダニ体内発育ステージ（Tick stage）へと誘導し、RNA-sequence法（RNA-seq.）を用いてTick stage分子の探索を行った。まず、誘導後何時間の原虫をRNA-seq.に用いるか検討した。次に、特徴的な形態が観察され、またRT-PCR法でTick stage遺伝子の発現が確認出来た分画をパーコール密度勾配遠心法で濃縮した。得られた原虫からRNAを抽出し、RNA-seq.に供した。次世代シークエンシング（NGS)でデータを取得し、同時に実施したBlood stage RNA-seq.データとの間で比較解析を行った。【結果】Tick stage誘導後の原虫は様々な形態が認められた。特に誘導後6時間では核を１つまたは２つ有し、短い突起を持つray bodyと呼ばれる特徴的な形態が確認された。また、Tick stageに発現すると予想された遺伝子群の発現が認められた。そこで、TBV標的に適した細胞外発育ステージに相当する、誘導後6時間のTick stage原虫をRNA-seq.に用いることにした。分画濃縮法にて、ウシ赤血球が除かれたTick stage原虫分画を精製し、RNA-seq.に供した。NGS RNA-seq.データをBlood stage/Tick stage間で比較解析したところ、Blood stageよりも Tick stageで発現量が2倍以上大きい遺伝子は413個あり、特にTick stageにおいて十分な転写産物発現量があると考えられる遺伝子は253個あった。【考察】Tick stage原虫では、DNAの複製に関連する遺伝子の発現上昇が認められ、このステージでの活発な細胞分裂（原虫増殖）が裏付けられた。
日本学術振興会, 基盤研究(B), 帯広畜産大学, 19H03120 - マダニ体内におけるバベシア原虫発育の分子基盤の解明と伝播阻止ワクチンの開発
科学研究費助成事業
2018年04月01日 - 2022年03月31日
玄 学南, 白藤 梨可, 正谷 達謄, 山岸 潤也
バベシア原虫はマダニによって媒介される病原体であり、種々の動物に重篤な貧血を引き起こし、家畜・ペット産業に重大な損害をもたらしている。本研究は、実験室レベルで全ライフサイクル解析が可能なバベシア原虫とフタトゲチマダ二を材料とし、マダニ体内におけるバベシア原虫発育の分子基盤解明と、それに基づくバベシア原虫伝播阻止ワクチンの開発を目指す。当該年度に得られた成果は、下記の通りである。１）in vitro培養系におけるB. gibsoniの有性生殖期虫体の誘導方法（キサンツレン酸の添加による誘導法）を確立した。２）有性生殖期マーカー分子とされるCCp1、CCp2、CCp3の発現が確認された。３）CCp1、CCp２、CCp3遺伝子をクローニングし、組換えタンパク質を作製した。４）組換えCCp1、CCp2、CCp3をマウスに免疫し、特異抗体を作製した。５）蛍光抗体法により、CCp1、CCp2、CCp3の特異抗体は、有性生殖期虫体と特異的反応を示すことを確認した。
日本学術振興会, 基盤研究(B), 帯広畜産大学, 18H02336 - サイトカイン発現住血原虫の開発研究
科学研究費助成事業
2018年06月29日 - 2021年03月31日
河津 信一郎, 麻田 正仁, 新川 武, 山岸 潤也
本研究では、ウシに不顕性感染する赤血球寄生性のバベシア原虫にインターフェロン等のサイトカインを発現させ、ウイルス・細菌・寄生虫といった病原体の種類に応じて、宿主動物の免疫系を抗原非特異的に自在に制御して、これら病原体による慢性消耗性感染症による被害を軽減する技術を開発することを目的とする。サイトカインを発現するバベシア原虫を作製する基盤技術を開発する目的で、原虫ゲノムをCRISPR/Cas9系で編集する実験系及びGlcN誘導性glmS リボザイムを応用したノックダウンの実験系を外国産のバベシア原虫（Babesia bovis：バベシア ボービス）で確立した。
日本学術振興会, 挑戦的研究(萌芽), 帯広畜産大学, 18K19258 - 内臓型リーシュマニア症の免疫病態
科学研究費助成事業
2018年04月01日 - 2021年03月31日
後藤 康之, 藤井 渉, 山岸 潤也
内臓型リーシュマニア症（VL）はリーシュマニア原虫の感染により引き起こされる人獣共通感染症である。本研究では、ヒトVL患者と同様に貧血や肝脾腫を引き起こすマウス感染モデルを用いて、これら病態に影響する免疫応答を明らかにすることを目的とした。貧血については、原虫の感染がマクロファージの抑制性因子SIRPαの発現を変化させ、血球貪食を誘導することが明らかとなった。この赤血球貪食は原虫数の増加に寄与することから、原虫が宿主分子を制御することで自身の生存に有利な環境を作り出していることが示唆される。また、この貧血や脾腫に宿主炎症性因子MRP14が関与することも明らかとなった。
日本学術振興会, 基盤研究(B), 東京大学, 18H02649 - Single cell応答に基づく原虫病研究の基盤構築と有用性の評価
科学研究費助成事業
2017年04月01日 - 2021年03月31日
山岸 潤也, 鈴木 穣
原虫は、そのサイズ、形態、偏性通性寄生性等において多様であり、近年利用が進んでいる１細胞トランスクリプトーム解析を行う際には、それに応じた技術基盤を確立する必要があった。そこで本研究では、上市されているscRNA-seqライブラリー構築を評価し、BD Rhapsodyが好適であることを見出した。さらに、トキソプラズマ原虫のタキゾイト→ブラディゾイト間のステージ変換をモデルにscRNA-seq解析を行い、トキソプラズマ原虫の感染により、IFNα経路が活性化されること、しかも、当該経路が活性化されるのは原虫感染細胞の周囲の非感染細胞であり、感染細胞の当該経路は逆に抑制されることを見出した。
日本学術振興会, 基盤研究(B), 北海道大学, 17H03912 - マラリア肝障害を引き起こす免疫応答の解明
科学研究費助成事業
2016年04月01日 - 2018年03月31日
後藤 康之, 藤井 渉, 山岸 潤也
肝障害はマラリアにおける代表的な症状であるが、その発症メカニズムは不明な点が多い。MRP14はミエロイド細胞に豊富に発現している蛋白質であり、様々な炎症性疾患との関わりが報告されている。そこで、本研究ではMRP14がマラリア肝障害に与える影響について明らかにするために、原虫感染マウスに対して組換えMRP14の投与を行った。結果、投与群では肝臓へのMRP14陽性細胞の浸潤が増強され、肝臓の壊死、それに伴う血中肝酵素の上昇が見られた。以上より、マラリアにおいてMRP14陽性細胞の浸潤にともなうMRP14の分泌が炎症反応の亢進による肝障害の悪化に寄与していることが明らかとなった。
日本学術振興会, 挑戦的萌芽研究, 東京大学, 16K15051 - ナノポアシークエンサーを用いた不明熱病原体ゲノムの同定
科学研究費助成事業
2015年04月01日 - 2018年03月31日
鈴木 穣, 山岸 潤也
本研究ではナノポアシークエンサーを用いて熱帯感染症不明熱病原体を同定する方法論開発を試みた。ランダムプライマーを用いた増幅系については実験ノイズが大きく安定したプロトコールを確立することができなかったが、代表的なウィルス性、細菌性、寄生虫性感染源であるデングウィルス、16S rRNAによる汎細菌性感染源、熱帯熱マラリア原虫については安定した検出系を構築することができた。また開発された実験系を用いて、インドネシアマナド地域で50症例の感染症患者から感染源を同定できた。そのゲノムについて薬剤耐性関連遺伝子の塩基多型を解析することが可能であり患者治療戦略の策定に際し重要な情報を与えると考えられた。
日本学術振興会, 挑戦的萌芽研究, 東京大学, 15K14417 - 感染症ゲノム遺伝子型迅速診断法の開発と臨床検体での評価
科学研究費助成事業
2015年04月01日 - 2018年03月31日
山岸 潤也, 鈴木 穣
本研究課題では、１）小型迅速次世代シーケンサーMinIONを、核酸等温増幅系である２）Loop-mediated Isothermal Amplification（LAMP）法、あるいは３）非特異的multiple displacement amplification (MDA)法と組み合わせることで、病原体ゲノム配列に基づいた感染症迅速診断系の開発を試みた。本系は、シーケンサーやサンプル処理に関わるシステム一式の持ち運びが容易であることを特徴とするため、研究室において基本的な系を確立した後、インドネシアをはじめとした種々の感染症の流行地に赴き、現場でも実施可能であることを確認した。
日本学術振興会, 基盤研究(B), 北海道大学, 15H05272 - インドネシアにおける人獣共通原虫病の疫学調査と社会実装可能な診断方法の開発
科学研究費助成事業
2014年04月01日 - 2017年03月31日
西川 義文, 関 まどか, 鈴木 穣, 山岸 潤也, 下田 直美, 梅田 剛佑
本研究ではインドネシアにおける人獣共通原虫病の疫学調査と社会実装可能な診断方法の開発を目的とした。インドネシアのスラウェシ島とジャワ島を調査地域とし、トキソプラズマやクリプトスポリジウムなどの原虫感染症の疫学調査により、当該地域における潜在的な感染リスクが明らかにされた。特にトキソプラズマのヒトの感染リスクは１０～２０歳で極めて高く、食肉や環境中からの感染が危惧された。実際、当該地域のウシやブタからはトキソプラズマ感染が確認された。また、疫学調査に利用できるクリプトスポリジウムの簡易迅速診断法を開発した。本研究期間中に共同研究を通じた人材育成を行うため、セミナー及び技術講習会を開催した。
日本学術振興会, 基盤研究(B), 帯広畜産大学, 26304037 - 細胞性免疫誘導型リバースワクチノロジーの確立
科学研究費助成事業
2012年04月01日 - 2014年03月31日
後藤 康之, 松本 芳嗣, 山岸 潤也
Leishmania major感染モデルを用いて、ワクチン抗原としての有用性に寄与するパラメータの同定を試みた。統計学的解析の結果、アミノ酸組成、発現量、エクソソームへの局在が有意なパラメータと同定され、これらを基にした数式によりプロテオーム中の大多数と既知のワクチン抗原を区別できた。さらに、新規に同定された蛋白は既知のワクチン抗原と同等の抗原性を示した。このように、in silicoアプローチで同定された新規タンパクが既知のワクチン抗原と同様の挙動を示したことは本手法がワクチン候補探索手法として有用であることを示唆している。
日本学術振興会, 挑戦的萌芽研究, 東京大学, 24658253 - 創薬へ向けた原虫トランスクリプトームの高解像度解析
科学研究費助成事業
2010年 - 2012年
山岸 潤也
RNA-seqやTSS-seqに代表される次世代型シークエンサーを用いた新しい技術によって種々の生物の遺伝子発現を網羅的に解析することが可能になっている。我々は当該技術を哺乳類一般を宿主とする原虫の1つであるトキソプラズマに適応し、その生活環を構成する代表的な3ステージの解析を行うことで、各ステージの遺伝子発現プロファイルを明らかにし、その制御に関わるDNAcis-elementを推定した。
日本学術振興会, 若手研究(B), 22780258 - トキソプラズマ原虫の発育ステージ変換機構の解明と新規次世代型ワクチンの開発
科学研究費助成事業
2010年 - 2010年
玄 学南, 山岸 潤也
本研究は、トキソプラズマ原虫の発育ステージ(急性感染期のタキゾイト→慢性感染期のブラディゾイト)変換の分子機構の解明と新規次世代型ワクチン開発を目指して実施する。本年度に実施した研究内容と得られた研究成果は以下の通りである。
1. 試験管内ステージ変換系を確立した。試験管内で人為的にタキゾイトからブラディゾイトへのステージ変換を促し、大量のブラディゾイト虫体を得ることができた。
2. ブラディゾイト虫体の完全長cDNAラブラリーとESTデータベースを作成した。ブラディゾイト虫体のRNAを抽出し、オリゴキャッピング法にて完全長cDNAライブラリーを構築した後、約10,000クローンの全塩基配列を解読し、ESTデータベースを作成した。
3. トランスクリプトーム解析を行った。以前当研究グループで構築したタキゾイトESTデータベースと2)で作成したブラディゾイトESTデータベースの比較により、タキゾイト或いはブラディゾイト単一ステージのみに特異的に発現する遺伝子を網羅的に探索した。その結果、タキゾイトとブラディゾイト特異遺伝子がそれぞれ424個と759個が同定された。そのうち転写制御因子と推定されるAP2遺伝子がそれぞれ4個と6個が含まれていた。また.ブラディゾイト特異的に発現するAP2はCCAGTGモチーフに結合することで、ステージ特異的な転写制御が行われている可能性が示唆された。
日本学術振興会, 基盤研究(A), 帯広畜産大学, 22248034 - 異種抗原発現用原虫ベクターの構築と次世代型組換えワクチン開発への応用
科学研究費助成事業
2009年 - 2010年
玄 学南, 山岸 潤也
本研究は異種抗原発現用新規原虫ベクターの構築と組換えワクチン開発への応用を目指して実施する。本年度に実施した研究内容と得られた研究成果は以下の通りである。
1. 過年度に異種抗原発現用ネオスポラ原虫ベクターを構築し、トキソプラズマ原虫ワクチン候補遺伝子TgSAG1の発現(Nc/TgSAG1)に成功した。
2. 今年度はまずNc/TgSAG1をBALB/cマウスに接種し、TgSAG1に対する特異抗体反応を誘導することを確認した。特異抗体のサブクラスを調べたところ、Th1型優勢免疫が誘導されていることが示唆された。また、Nc/TgSAG1を接種したマウスにおいてはIFN-γの産生がベクターのみを接種した対照群と比べ有意に高いことが示された。なお、IL-4の産生には対照群と比べ有意な変化がなかった。
3. 次に、Nc/TgSAG1にて免疫したマウスに致死量のトキソプラズマ原虫を接種したところ、約80%のマウスが生残した。これらの結果より、異種抗原発現用原虫ベクターは次世代型ワクチン開発に新しいツールを提供しうることが示唆された。
日本学術振興会, 挑戦的萌芽研究, 帯広畜産大学, 21658098 - 東南アジアにおけるダニ媒介性動物原虫感染症の流行実態の解明と予防対策の確立
科学研究費助成事業
2009年 - 2010年
玄 学南, 山岸 潤也
本研究は東南アジアにおけるダニ媒介性動物原虫感染症の流行実態の解明と予防対策の確立を目指して実施する。本年度に実施した研究内容と得られた研究成果は以下の通りである。
1. 昨年度に引き続きタイ国北部地域における牛のマダニ媒介性原虫感染症の流行実態を調べた。計200頭の牛の血液サンプルを採集し、全DNAを抽出した。ウシバベシア原虫特異PCR法を用いて、血液サンプル中の原虫DNAの検出を行ったところ、Babesia bovisとBabesia bigeminaの陽性率がそれぞれ12%(24/200)と21%(42/200)であった。この結果より、ウシバベシア症はタイ国北部地域広く流行しており、本症の制圧対策は当地域の牛の生産性向上に重要であることが示唆された。
2. フィリピンマニラ周辺地域における牛のマダニ媒介性原虫感染症の流行実態を調べた。計250頭の牛より血液サンプルを採集し、全DNAを抽出した。ウシバベシア原虫特異PCR法とウシタイレリア原虫特異PCR法を用いて予備実験を行ったところ、2種類のウシバベシア原虫(Babesia bovisとBabesia bigemina)と1種類のウシタイレリア原虫(Theileria orientalis)DNAが高率に検出された。これらの結果により、フィリピンマニラ周辺地域の牛にはマダニ媒介性バベシア原虫とタイレリア原虫感染症が高率に流行していることが示唆された。
日本学術振興会, 基盤研究(B), 帯広畜産大学, 21405036
