研究者データベース

清水 健太(シミズ ケンタ)
医学研究院 病理系部門 微生物学免疫学分野
助教

基本情報

所属

  • 医学研究院 病理系部門 微生物学免疫学分野

職名

  • 助教

学位

  • 博士(獣医学)(岐阜大学)

科研費研究者番号

  • 20466840

J-Global ID

研究キーワード

  • 免疫学   微生物学   獣医学   実験動物学   人獣共通感染症   ウイルス学   

研究分野

  • ライフサイエンス / 実験動物学
  • ライフサイエンス / ウイルス学
  • ライフサイエンス / 獣医学

職歴

  • 2008年10月 〜 北海道大学大学院医学研究科助教
  • 2008年04月 - 2008年09月 同特任研究員
  • 2007年04月 - 2008年03月 東京大学医科学研究所産学官連携研究員

学歴

  •         - 2007年   岐阜大学   連合獣医学研究科   獣医学
  •         - 2007年   岐阜大学
  •         - 2003年   岐阜大学   農学部   獣医学科
  •         - 2003年   岐阜大学

所属学協会

  • 日本バイオセーフティ学会   日本実験動物学会   日本実験動物医学会   日本ウイルス学会   日本獣医学会   Japanese Association for Laboratory Animal Medicine   The Japanese Society for Virology   The Japanese Society of Veterinary Science   

研究活動情報

論文

  • Exposure to hantavirus is a risk factor associated with kidney diseases in Sri Lanka: A cross sectional study.
    Sarathkumara YD, Gamage CD, Lokupathirage SMW, Muthusinghe DS, Nanayakkara N, Gunarathne L, Shimizu K, Tsuda Y, Arikawa J, Yoshimatsu K
    Viruses 11 700  2019年08月 [査読有り][通常論文]
  • Serological evidence of Thailand orthohantavirus or antigenically related virus infection among rodents in a chronic kidney disease of unknown etiology endemic area, Girandurukotte, Sri Lanka.
    Lokupathirage SMW, Muthusinghe DS, Shimizu K, Nishigami K, Noda K, Tsuda Y, Sarathkumara YD, Gunawardana S, Arikawa J, Gamage CD, Yoshimatsu K
    Vecror Borne Zoonotic Dis. 2019年 [査読有り][通常論文]
  • Thailand orthohantavirus infection in patients with chronic kidney disease of unknown aetiology in Sri Lanka.
    Yoshimatsu K, Gamage CD, Sarathkumara YD, Kulendiran T, Muthusinghe DS, Nanayakkara N, Gunarathne L, Shimizu K, Tsuda Y, Arikawa J
    Archives of virology 164 1 267 - 271 2019年01月 [査読有り][通常論文]
  • Kenta Shimizu, Kumiko Yoshimatsu, Midori Taruishi, Yoshimi Tsuda, Jiro Arikawa
    Archives of Virology 163 6 1577 - 1584 2018年06月01日 [査読有り][通常論文]
     
    Hemorrhagic fever with renal syndrome (HFRS) is caused by hantavirus infection. Although host immunity is thought to be involved in the pathogenesis of HFRS, the mechanism remains to be elucidated. A mouse model of HFRS, which showed renal hemorrhage similar to that seen in patients, has been developed previously. In this study, we aimed to clarify whether CD4+ and CD8+ T cells are involved in the development of renal hemorrhage in the mouse model. At 2 days before virus inoculation, CD4+ or CD8+ T cells in 6-week-old BALB/c mice were depleted by administration of antibodies. The CD4+ T cell-depleted mice developed signs of disease such as transient weight loss, ruffled fur and renal hemorrhage as in non-depleted mice. In contrast, the CD8+ T cell-depleted mice showed no signs of disease. After determination of CTL epitopes on the viral glycoprotein in BALB/c mice, the quantity of virus-specific CTLs was analyzed using an MHC tetramer. The quantity of virus-specific CTLs markedly increased in spleens and kidneys of virus-infected mice. However, the quantity in high-pathogenic clone-infected mice was comparable to that in low-pathogenic clone-infected mice. We previously reported that the high-pathogenic clone propagated more efficiently than the low-pathogenic clone in kidneys of mice during the course of infection. Therefore, there is a possibility that the balance between quantities of the target and effector is important for disease outcome. In conclusion, this study showed that CD8+ T cells are involved in the development of renal hemorrhage in a mouse model of HFRS.
  • Tapiwa LUNDU, Yoshimi TSUDA, Ryo ITO, Kenta SHIMIZU, Shintaro KOBAYASHI, Kentaro YOSHII, Kumiko YOSHIMATSU, Jiro ARIKAWA, Hiroaki KARIWA
    Biomedical Research 39 1 27 - 38 2018年 [査読有り][通常論文]
  • Kenta Shimizu, Rie Isozumi, Kazutoshi Takami, Isao Kimata, Kanae Shiokawa, Kumiko Yoshimatsu, Yoshimi Tsuda, Sanae Nishio, Jiro Arikawa
    JOURNAL OF VETERINARY MEDICAL SCIENCE 79 7 1261 - 1263 2017年07月 [査読有り][通常論文]
     
    We examined 33 rodents captured in an urban area of Osaka City, Japan for IgG antibodies against Seoul virus, severe fever with thrombocytopenia syndrome virus, hepatitis E virus, Leptospira interrogans, Yersinia pestis, spotted fever, typhus and scrub typhus group rickettsiae. We found that 3 (9.1%) and 1 (3.0%) of the 33 rodents had antibodies against L. interrogans and spotted fever group rickettsiae, respectively. DNAs of leptospires were detected from 2 of the 3 seropositive rodents, but DNA of rickettsia was not detected. Phylogenetic analysis and multiple locus sequence typing revealed that the 2 leptospires were L. interrogans belonging to a novel sequence type. There is a potential risk for acquiring rodent-borne zoonotic pathogens even in cities in developed countries.
  • Sanae Nishio, Yoshimi Tsuda, Ryo Ito, Kenta Shimizu, Kumiko Yoshimatsu, Jiro Arikawa
    JAPANESE JOURNAL OF INFECTIOUS DISEASES 70 4 388 - 393 2017年07月 [査読有り][通常論文]
     
    The first clinical case of the YG1 strain of the severe fever with thrombocytopenia syndrome virus (SFTSV) has been isolated in Japan. We found that only some of the cells underwent low pH-dependent cell fusion, although all of the cells were confirmed to have been infected with the virus. This suggested that the YG1 strain consists of a heterogeneous mixture of related viruses. Here, we established 3 subclones (termed E3, A4, and B7) from the YG1 strain, using the limiting dilution method with the pH-dependent cell fusion activity. Subclone E3 showed weak fusion activity and cytopathic effects (CPE) in Vero E6 cells. The amino acid sequence of E3 was identical to the published sequence for the YG1 strain, and it likely comprises a subpopulation of the YG1 strain. Subclone A4 displayed strong fusion activity under acidic conditions. In contrast, subclone B7 showed strong fusion activity and CPE under neutral and acidic conditions. Two amino acid differences shared between B7 and A4 were found in the envelope glycoproteins. In addition, an amino acid variant of the RNA-dependent RNA polymerase was found only in B7. These subclones will be valuable tools to elucidate cell fusion mechanisms of SFTSV and the relationship between viral proteins and their functions.
  • Kumiko Yoshimatsu, Satoru Arai, Kenta Shimizu, Yoshimi Tsuda, Bazartseren Boldgiv, Bazartseren Boldbaatar, Erdenebaatar Sergelen, Dagvatseren Ariunzaya, Orsoo Enkhmandal, Sukhbaatar Tuvshintugs, Shigeru Morikawa, Jiro Arikawa
    JAPANESE JOURNAL OF VETERINARY RESEARCH 65 1 39 - 44 2017年02月 [査読有り][通常論文]
     
    Seroepizootiological surveys among wild rodents were carried out on the east side of Lake Khovsgol in Mongolia in 2010 and 2011. A total of 76 voles belonging to the genera Myodes and Microtus were captured. Most of the voles that were seropositive to Tula virus antigen were Middendorf's voles (Microtus middendorffii (6/31)). Two of the 18 Myodes voles were also seropositive to Tula virus antigen. On the other hand, only one vole was seropositive to Puumala virus antigen. The results suggest that Tula virus was maintained in Middendorf's vole. This is the first report of detection of anti-Tula virus antibody in the central part of the Eurasia continent.
  • Kenta Shimizu, Takaaki Koma, Kumiko Yoshimatsu, Yoshimi Tsuda, Yuji Isegawa, Jiro Arikawa
    VIROLOGY JOURNAL 14 1 13  2017年01月 [査読有り][通常論文]
     
    Background: Hemorrhagic fever with renal syndrome (HFRS) caused by hantavirus infection is characterized by fever, renal dysfunction and hemorrhage. An animal model mimicking symptoms of HFRS remains to be established. In this study, we evaluated the pathogenicity of an HFRS patient-derived Hantaan virus (HTNV) in adult mice. Methods: Five clones of HTNV strain KHF 83-61 BL (KHFV) that was derived from blood of an HFRS patient were obtained by plaque cloning. The pathogenicity of the virus clones was evaluated by using 6-week-old female BALB/c mice. Sequence analysis of the viral genome was performed by conventional methods. Results: All of the mice intravenously inoculated with KHFV clone (cl)-1, -2, -3 and -5 showed signs of disease such as transient body weight loss, ruffled fur, reduced activity and remarkably prominent hemorrhage in the renal medulla at 6 to 9 days post-inoculation (dpi) and then recovered. In contrast, mice intravenously inoculated with KHFV cl-4 did not show any signs of disease. We selected KHFV cl-5 and cl-4 as representative of high-pathogenic and low-pathogenic clones, respectively. Quantities of viral RNA in kidneys of KHFV cl-5-infected mice were larger than those in KHFV cl-4-infected mice at any time point examined (3, 6, 9 and 12 dpi). The quantities of viral RNA of KHFV cl-5 and cl-4 peaked at 3 dpi, which was before the onset of disease. Sequence analysis revealed that the amino acid at position 417 in the glycoprotein Gn was the sole difference in viral proteins between KHFV cl-5 and cl-4. The result suggests that amino acid at position 417 in Gn is related to the difference in pathogenicity between KHFV cl-5 and cl-4. When the inoculum of KHFV cl-5 was pretreated with a neutralizing antibody against HTNV strain 76-118, which belongs to the same serotype as KHFV clones, mice did not show any signs of disease, confirming that the disease was caused by KHFV infection. Conclusion: We found that an HFRS patient-derived HTNV caused renal hemorrhage in adult mice. We anticipate that this infection model will be a valuable tool for understanding the pathogenesis of HFRS.
  • Yoshimi Tsuda, Manabu Igarashi, Ryo Ito, Sanae Nishio, Kenta Shimizu, Kumiko Yoshimatsu, Jiro Arikawa
    BIOMEDICAL RESEARCH-TOKYO 38 2 89 - 97 2017年 [査読有り][通常論文]
     
    Severe fever with thrombocytopenia syndrome virus (SFTSV) is a novel phlebovirus responsible for causing an emerging zoonotic disease. We previously established subclones from SFTSV strain YG1 based on differences in low-pH-dependent cell fusion activities and found two amino acid substitutions, Y328H and R624W, in the envelope glycoprotein (GP) of high fusion subclones. In this study, we show that transiently expressed GP with the R624W mutation, but not the Y328H mutation, induced cell fusion under acidic conditions. GP possessing either tryptophan, serine, glycine or aspartic acid at position 624 induced cell fusion, whereas GP possessing basic amino acids such as arginine or lysine did not induce cell fusion. These results indicated that the amino acid at position 624 has an important role for inducing low-pH-dependent cell fusion.
  • Satomu Obana, Kenta Shimizu, Kumiko Yoshimatsu, Futoshi Hasebe, Kozue Hotta, Rie Isozumi, Hoa Thuy Nguyen, Mai Quynh Le, Tetsu Yamashiro, Yoshimi Tsuda, Jiro Arikawa
    JOURNAL OF VETERINARY MEDICAL SCIENCE 79 1 76 - 81 2017年01月 [査読有り][通常論文]
     
    There is concern about the zoonotic potential of rodent-borne hepatitis E virus, designated as HEV-C1. However, epizootiological information about HEV-C1 is limited. To address this issue, serum samples from 443 small mammals captured at 5 sites in Hanoi, Vietnam, were examined for anti-HEV-Cl IgG antibodies. In addition, livers of seropositive animals were examined for viral RNA. Anti-HEV-C1 antibodies were detected in 57 (12.9%) of the 443 serum samples. Seropositive animals were found in all of the sites (4.7% to 22.2%). Anti-HEV-C1 antibodies were detected from 48 (12.3%) of 389 Rattus norvegicus and 9 (19.6%) of 46 R. tanezumi, but were not detected from 8 Suncus murinus. Viral RNAs were detected from 13 (22.8%) of the 57 seropositive rodents. The detectioh rate of viral RNA in seropositive R. tanezumi (66.7%, 6/9) was significantly higher than that in seropositive R. norvegicus (14.6%, 7/48). The results suggest that R. tanezumi is more susceptible than R. norvegicus to HEV-Cl infection. Phylogenetic analysis revealed that Vietnamese strains were divided into 3 clusters in genetic group 2 of HEV-C1. Multiple clusters of viruses were detected at several sites without species specificity, suggesting that 3 clusters of HEV-C1 co-circulate in Hanoi, Vietnam.
  • Kenta Shimizu, Sugihiro Hamaguchi, Cuong Chi Ngo, Tian-Cheng Li, Shuji Ando, Kumiko Yoshimatsu, Shumpei P. Yasuda, Takaaki Koma, Rie Isozumi, Yoshimi Tsuda, Hiromi Fujita, Thuy Thanh Pham, Mai Quynh Le, Anh Duc Dang, Tuan Quang Nguyen, Lay-Myint Yoshida, Koya Ariyoshi, Jiro Arikawa
    JOURNAL OF VETERINARY MEDICAL SCIENCE 78 11 1677 - 1681 2016年11月 [査読有り][通常論文]
     
    Zoonotic potential of a rat-derived hepatitis E virus (HEV), designated as HEV-Cl, remains unknown. To evaluate the risk for HEV-Cl infection in humans, paired sera of 208 hospitalized febrile patients collected from 2001 to 2003 in Hanoi, Vietnam, were examined for IgG antibodies to HEV-Cl and genotype 1 HEV (HEV-1), which is common in humans. IgG antibodies to virus-like particles (VLPs) of HEV-Cl and/or HEV-1 were detected from 99 of the 208 convalescent sera in enzyme-linked immunosorbent assay (ELISA). IgG antibody titers to HEV-Cl antigen in 3 of the 99 sera were more than 8-fold higher than those to HEV-1 antigen. IgM antibodies to HEV-Cl antigen were detected in acute sera from 2 of the 3 patients in ELISA and Western blotting. However, no HEV genome was detected. Clinical information was available for 1 of the 2 patients. Hepatic enzymes, aspartate aminotransferase and alanine aminotransferase, were mildly elevated (156 IU/l and 68 IU/l, respectively), and hepatomegaly was detected by ultrasonography. The patient recovered from the illness after 17 days. These results indicated that HEV-Cl or its variants infect humans in Vietnam and may cause acute febrile illness with mild liver dysfunction.
  • Aaron Brice, Donna R. Whelan, Naoto Ito, Kenta Shimizu, Linda Wiltzer-Bach, Camden Y. Lo, Danielle Blondel, David A. Jans, Toby D. M. Bell, Gregory W. Moseley
    SCIENTIFIC REPORTS 6 33493  2016年09月 [査読有り][通常論文]
     
    Although microtubules (MTs) are known to have important roles in intracellular transport of many viruses, a number of reports suggest that specific viral MT-associated proteins (MAPs) target MTs to subvert distinct MT-dependent cellular processes. The precise functional importance of these interactions and their roles in pathogenesis, however, remain largely unresolved. To assess the association with disease of the rabies virus (RABV) MAP, P3, we quantitatively compared the phenotypes of P3 from a pathogenic RABV strain, Nishigahara (Ni) and a non-pathogenic Ni-derivative strain, Ni-CE. Using confocal/live-cell imaging and dSTORM super-resolution microscopy to quantify protein interactions with the MT network and with individual MT filaments, we found that the interaction by Ni-CE-P3 is significantly impaired compared with Ni-P3. This correlated with an impaired capacity to effect association of the transcription factor STAT1 with MTs and to antagonize interferon (IFN)/STAT1-dependent antiviral signaling. Importantly, we identified a single mutation in Ni-CE-P3 that is sufficient to inhibit MT-association and IFN-antagonist function of Ni-P3, and showed that this mutation alone attenuates the pathogenicity of RABV. These data provide evidence that the viral protein-MT interface has important roles in pathogenesis, suggesting that this interface could provide targets for vaccine/antiviral drug development.
  • Kanae Shiokawa, Chandika D. Gamage, Nobuo Koizumi, Yoshihiro Sakoda, Kenta Shimizu, Yoshimi Tsuda, Kumiko Yoshimatsu, Jiro Arikawa
    JOURNAL OF VETERINARY MEDICAL SCIENCE 78 2 221 - 230 2016年02月 [査読有り][通常論文]
     
    The applicability of the recombinant LipL32 for serodiagnosis of leptospiral infection in field rodents was assessed in this study. An immunodominant region of LipL32 was determined by monoclonal antibodies, and then, truncated LipL32 (tLipL32) was designed to contain the region (87-188th amino acid). The tLipL32 was compared between two recombinant expression hosts Escherichia coli and Pichia pastoris in ELISA. With field rat sera, tLipL32 expressed by P. pastoris (tLipL32p) had high antigenicity without background reactions, while tLipL32 expressed by E. coli (tLipL32e) showed high background reactions, which were reduced by pre-adsorption of sera with E. coli. To evaluate tLipL32-ELISA, field rat sera were tentatively divided into a Leptospira infection positive (12 sera) and a negative group (12 sera) based on the results from flaB gene PCR of kidney samples and WB with whole Leptospira cell. Consequently, the sensitivity of tLipL32p-ELISA for field rat sera was 83%. A similar result was obtained from tLipL32e-ELISA with adsorbed sera, (92%). However, sensitivity of tLipL32e-ELISA using sera without an adsorption treatment was 50%. Regardless of the expression host, tLipL32-ELISA had 100% specificity and sensitivity in experimentally infected laboratory rats. These results suggest that recombinant LipL32 expressed by P pastoris is more applicable for serodiagnosis in field rats due to a lack of background reaction.
  • Kozue Hotta, Hang T. T. Pham, Huong T. Hoang, Tu C. Trang, Thuy N. Vu, Trang T. H. Ung, Kenta Shimizu, Jiro Arikawa, Akio Yamada, Hoa T. Nguyen, Hang L. K. Nguyen, Mai T. Q. Le, Daisuke Hayasaka
    VECTOR-BORNE AND ZOONOTIC DISEASES 16 2 96 - 102 2016年02月 [査読有り][通常論文]
     
    Rodents are important reservoirs of many human pathogens transmitted via arthropod vectors. Arthropod-borne bacteria belonging to the family Rickettsiaceae cause acute febrile diseases in humans worldwide, but the real burdens of rickettsial diseases appear to be underestimated in Hanoi, Vietnam, because differential diagnosis on the basis of clinical signs and symptoms is confounded by the presence of other tropical infectious diseases with similar signs and symptoms. To know the prevalence of bacteria of the family Rickettsiaceae among small mammals in Hanoi, 519 animals thriving in the public places were captured and examined for the presence of bacterial sequences using duplex PCR. Nucleotide sequences specific for Orientia tsutsugamushi were detected in seven samples (1.3%). Out of seven animals, two were captured in a market, whereas five were in hospitals. None of the captured small mammals tested positive for the genus Rickettsia. The nucleotide sequence analysis of the genes encoding the 47-kDa high-temperature requirement A (47-kDa HtrA) and 56-kDa type-specific antigen (TSA) showed that these seven isolates were indistinguishable from each other. O. tsutsugamushi isolated in this study was closely related phylogenetically to the Gilliam strain, which was originally isolated at the border of Assam and Burma, rather than to those isolated in the central to southern part of Vietnam. It should be emphasized that Vietnamese hospitals were heavily infested by small rodents and some of them harbored O. tsutsugamushi. Strict hygienic control should be implemented to mitigate the potential risk posed by O. tsutsugamushi in hospital settings.
  • Sugihiro Hamaguchi, Ngo Chi Cuong, Doan Thu Tra, Yen Hai Doan, Kenta Shimizu, Nguyen Quang Tuan, Lay-Myint Yoshida, Le Quynh Mai, Dang Duc-Anh, Shuji Ando, Jiro Arikawa, Christopher M. Parry, Koya Ariyoshi, Pham Thanh Thuy
    AMERICAN JOURNAL OF TROPICAL MEDICINE AND HYGIENE 92 5 972 - 978 2015年05月 [査読有り][通常論文]
     
    A descriptive study on rickettsiosis was conducted at the largest referral hospital in Hanoi, Vietnam, to identify epidemiological and clinical characteristics of specific rickettsiosis. Between March 2001 and February 2003, we enrolled 579 patients with acute undifferentiated fever (AUF), excluding patients with malaria, dengue fever, and typhoid fever, and serologically tested for Orientia tsutsugamushi and Rickettsia typhi. Of the patients, 237 (40.9%) and 193 (33.3%) had scrub and murine typhus, respectively, and 149 (25.7%) had neither of them (non scrub and murine typhus [non-ST/MT]). The proportion of murine typhus was highest among patients living in Hanoi whereas that of scrub typhus was highest in national or regional border areas. The presence of an eschar, dyspnea, hypotension, and lymphadenopathy was significantly associated with a diagnosis of scrub typhus (OR = 46.56, 10.90, 9.01, and 7.92, respectively). Patients with murine typhus were less likely to have these findings but more likely to have myalgia, rash, and relative bradycardia (OR = 1.60, 1.56, and 1.45, respectively). Scrub typhus and murine typhus were shown to be common causes of AUF in northern Vietnam although the occurrence of spotted fever group rickettsiae was not determined. Clinical and epidemiological information may help local clinicians make clinical diagnosis of specific rickettsioses in a resource-limited setting.
  • Shumpei P. Yasuda, Chandika D. Gamage, Nobuo Koizumi, Sanae Nishio, Rie Isozumi, Kenta Shimizu, Takaaki Koma, Takako Amada, Hitoshi Suzuki, Kumiko Yoshimatsu, Jiro Arikawa
    Genes and Genetic Systems 89 2 71 - 80 2014年09月01日 [査読有り][通常論文]
     
    We examined genetic variation in black rats (the Rattus rattus complex) from Kandy District, Sri Lanka using mitochondrial cytochrome b (cytb, 1140 bp) and nuclear melanocortin 1 receptor (Mc1r, 954 bp) gene sequences together with database sequences. We confirmed the existence of two divergent mitochondrial lineages in Sri Lankan black rats, with genetic distance of 2.2% and estimated divergence time of 0.3 million years ago. Because one lineage is unique to the island and the other is closely related to R. rattus populations on the Indian subcontinent, two migration events of R. rattus from the subcontinent are inferred, one ancient and one recent. Mc1r analyses revealed 12 haplotypes among the Sri Lankan black rats. A median-joining network together with other availablesequences separated the 12 haplotypes into two groups, one unique to the island and the other related to previously reported R. rattus sequences. Notably, most individuals possessed various combinations of both haplotype groups which had no association with the cytb clades. These results imply that old and new R. rattuslineages are now intermingled as a result of hybridization in Sri Lanka. Specimensof the lesser bandicoot rat (Bandicota bengalensis) collected from Sri Lanka (n = 24) were shown to have no genetic variability in the cytb sequence. Our results indicate that the two most abundant groups of commensal rats in Sri Lanka, black rats and lesser bandicoot rats, are the product of contrasting evolutionary histories on different timescales.
  • Takaaki Koma, Kumiko Yoshimatsu, Noriyo Nagata, Yuko Sato, Kenta Shimizu, Shumpei P. Yasuda, Takako Amada, Sanae Nishio, Hideki Hasegawa, Jiro Arikawa
    JOURNAL OF VIROLOGY 88 13 7178 - 7188 2014年07月 [査読有り][通常論文]
     
    Hantavirus infections are characterized by vascular hyperpermeability and neutrophilia. However, the pathogenesis of this disease is poorly understood. Here, we demonstrate for the first time that pulmonary vascular permeability is increased by Hantaan virus infection and results in the development of pulmonary edema in C. B-17 severe combined immunodeficiency ( SCID) mice lacking functional T cells and B cells. Increases in neutrophils in the lung and blood were observed when pulmonary edema began to be observed in the infected SCID mice. The occurrence of pulmonary edema was inhibited by neutrophil depletion. Moreover, the pulmonary vascular permeability was also significantly suppressed by neutrophil depletion in the infected mice. Taken together, the results suggest that neutrophils play an important role in pulmonary vascular hyperpermeability and the occurrence of pulmonary edema after hantavirus infection in SCID mice.
  • Takako Amada, Kumiko Yoshimatsu, Takaaki Koma, Kenta Shimizu, Chandika D. Gamage, Kanae Shiokawa, Sanae Nishio, Clas Ahlm, Jiro Arikawa
    VIROLOGY JOURNAL 11 1 87  2014年05月 [査読有り][通常論文]
     
    Background: Hantaviruses are causative agents of hemorrhagic fever with renal syndrome (HFRS) and nephropathia epidemica (NE) in the Old World and hantavirus pulmonary syndrome (HPS) in the New World. There is a need for time-saving diagnostic methods. In the present study, recombinant N antigens were used as antigens in an immunochromatography strip (ICG) test to detect specific IgG antibodies. Methods: The N-terminal 103 amino acids (aa) of Hantaan virus (HTNV), Puumala virus (PUUV) and Andes virus (ANDV) nucleocapsid (N) protein were expressed in E. coli as representative antigens of three groups (HFRS, NE and HPS-causing viruses) of hantavirus. Five different types of ICG test strips, one antigen line on one strip for each of the three selected hantaviruses (HTNV, PUUV and ANDV), three antigen lines on one strip and a mixed antigen line on one strip, were developed and sensitivities were compared. Results: A total of 87 convalescent-phase patient sera, including sera from 35 HFRS patients, 36 NE patients and 16 HPS patients, and 25 sera from healthy seronegative people as negative controls were used to evaluate the ICG test. Sensitivities of the three-line strip and mixed-line strip were similar to those of the single antigen strip (97.2 to 100%). On the other hand, all of the ICG test strips showed high specificities to healthy donors. Conclusion: These results indicated that the ICG test with the three representative antigens is an effective serodiagnostic tool for screening and typing of hantavirus infection in humans.
  • Kenta Shimizu, Kumiko Yoshimatsu, Takaaki Koma, Shumpei P. Yasuda, Jiro Arikawa
    Virus Research 178 2 349 - 356 2013年12月26日 [査読有り][通常論文]
     
    To understand the role of nucleocapsid protein (NP) of hantaviruses in viral assembly, the effect of NP on intracellular traffic of viral glycoproteins Gn and Gc was investigated. Double staining of viral and host proteins in Hantaan virus (HTNV)-infected Vero E6 cells showed that Gn and Gc were localized to cis-Golgi, in which virus particles are thought to be formed. When HTNV Gn and Gc were expressed by a plasmid encoding glycoprotein precursor (GPC), which is posttranslationaly cleaved into Gn and Gc, Gn was localized to cis-Golgi, whereas Gc showed diffuse distribution in the cytoplasm in 32.9% of Gc-positive cells. The ratio of the diffused Gc-positive cells was significantly decreased to 15.0% by co-expression of HTNV NP. Co-expression of HTNV GPC with NPs of other hantaviruses, such as Seoul virus, Puumala virus and Sin Nombre virus, also reduced the ratios of diffused Gc-positive cells to 13.5%, 25.2%, and 11.6%, respectively. Among amino- and carboxyl-terminally truncated HTNV NPs, NP75-429, NP116-429, NP1-333, NP1-233, and NP1-155 possessed activity to reduce the ratio of diffused Gc-positive cells, while NP155-429 and NP1-116 did not. NP30-429 has partial activity. These results indicate that amino acid region 116-155 of NP is important for the activity, although amino acid region 1-30 is partially related. Truncation of the HTNV Gc cytoplasmic tail caused an increase in diffused Gc-positive cells. In addition, the effect of coexpression of HTNV NP was weakened. These results suggest that HTNV NP has a role to promote Golgi localization of Gc through a mechanism possibly mediated by the Gc cytoplasmic tail. © 2013 Elsevier B.V.
  • Takako Arnada, Kumiko Yoshimatsu, Shumpei P. Yasuda, Kenta Shimizu, Takaaki Koma, Nobuhito Hayashimoto, Chandika D. Gamage, Sanae Nishio, Akira Takakura, Jiro Arikawa
    JOURNAL OF VIROLOGICAL METHODS 193 1 42 - 49 2013年10月 [査読有り][通常論文]
     
    Hantavirus is a causative agent of rodent-borne viral zoonoses, hemorrhagic fever with renal syndrome (HFRS) and hantavirus pulmonary syndrome. Seoul virus (SEOV) is a causative agent of urban and laboratory rat-associated HFRS worldwide. Surveillance of rodents has been done mainly by serological detection of hantavirus-specific antibodies by enzyme linked immunosorbent assay (ELISA) and immunofluorescent antibody assay (IFA). An immunochromatographic (ICG) test was developed with the N-terminal 103 amino acids of nucleocapsid protein of Hantaan virus expressed by Escherichia coli as an antigen to detect IgG antibody specific to hantavirus in sera from Rattus sp. animals. Antibody-detecting sensitivity of the ICG test was the same as that of ELISA and about 100-times higher than that of IFA. Overall sensitivities and specificities of the ICG test in comparison to ELISA and IFA for sera from 192 urban rats and 123 laboratory rats were 99.3% and 100%, respectively. Diluted whole blood samples without separation could be used for the ICG test. The ICG test enabled detection of antibodies to SEOV, Hantaan, Dobrava/Belgrade, and Thailand viruses, which are causative agents of HFRS throughout Eurasia. The ICG test is a rapid, simple and safe method for diagnosis of SEOV infection in rats. (c) 2013 Elsevier B.V. All rights reserved.
  • T. Koma, K. Yoshimatsu, S. P. Yasuda, T. Li, T. Amada, K. Shimizu, R. Isozumi, L. T.Q. Mai, N. T. Hoa, V. Nguyen, T. Yamashiro, F. Hasebe, J. Arikawa
    Epidemiology and Infection 141 9 1876 - 1884 2013年09月 [査読有り][通常論文]
     
    To examine the prevalence of human pathogens carried by rats in urban areas in Hanoi and Hai Phong, Vietnam, we live-trapped 100 rats in January 2011 and screened them for a panel of bacteria and viruses. Antibodies against Leptospira interrogans (22·0%), Seoul virus (14·0%) and rat hepatitis E virus (23·0%) were detected in rats, but antibodies against Yersinia pestis were not detected. Antibodies against L. interrogans and Seoul virus were found only in adult rats. In contrast, antibodies to rat hepatitis E virus were also found in juvenile and sub-adult rats, indicating that the transmission mode of rat hepatitis E virus is different from that of L. interrogans and Seoul virus. Moreover, phylogenetic analyses of the S and M segments of Seoul viruses found in Rattus norvegicus showed that Seoul viruses from Hai Phong and Hanoi formed different clades. Human exposure to these pathogens has become a significant public health concern. Copyright © Cambridge University Press 2012.
  • Ima-Nurisa Ibrahim, Kenta Shimizu, Kumiko Yoshimatsu, Andre Yunianto, Ervi Salwati, Shumpei P. Yasuda, Takaaki Koma, Rika Endo, Jiro Arikawa
    JOURNAL OF VETERINARY MEDICAL SCIENCE 75 8 1003 - 1008 2013年08月 [査読有り][通常論文]
     
    Hemorrhagic fever with renal syndrome (HFRS) is a rodent-borne zoonotic disease caused by hantavirus infection. Many HFRS cases have been reported in East Asia and North Europe, while the situation in Southeast Asia remains unclear. In this study, the prevalence of hantavirus infection in rodents and humans in Thousand Islands regency, which is close to the port of Jakarta, one of the largest historic ports in Indonesia, was investigated. A total of 170 rodents were captured in 2005, and 27 (15.9%) of the rodents were antibody-positive against Hantaan virus antigen in an immunofluorescence assay (IFA) and Western blotting. Despite the high prevalence in rodents, human sera collected from 31 patients with fever of -unknown origin and 20 healthy volunteers in the islands in 2009 did not show positive reaction to the antigen in IFA. To identify the virus in rodents genetically, a total of 59 rodents were captured in 2009. Sera from the rodents were screened for antibody by ELISA, and lung tissues were subjected to RT-PCR. 20 (33.9%) of the 59 rodents were antibody-positive, and 3 of those 20 rodents were positive for S and M genome segments of hantaviruses. Genetic analysis showed that the viruses belonged to Seoul virus and formed a cluster with those in Vietnam and Singapore. These results suggest that a unique group of Seoul viruses has spread widely in Southeast Asia.
  • Mathias Schlegel, Erdenesaikhan Tegshduuren, Kumiko Yoshimatsu, Rasa Petraityte, Kestutis Sasnauskas, Baerbel Hammerschmidt, Robert Friedrich, Marc Mertens, Martin H. Groschup, Satoru Arai, Rika Endo, Kenta Shimizu, Takaaki Koma, Shumpei Yasuda, Chiaki Ishihara, Rainer G. Ulrich, Jiro Arikawa, Bernd Koellner
    ARCHIVES OF VIROLOGY 157 11 2179 - 2187 2012年11月 [査読有り][通常論文]
     
    We developed serological tools for the detection of hantavirus-specific antibodies and hantavirus antigens in shrews. The work was focussed to generate Thottapalayam virus (TPMV)-specific monoclonal antibodies (mAbs) and anti-shrew immunoglobulin G (IgG) antibodies. The mAbs against TPMV nucleocapsid (N) protein were produced after immunization of BALB/c mice with recombinant TPMV N proteins expressed in Escherichia coli, baculovirus and Saccharomyces cerevisiae-mediated expression systems. In total, six TPMV N-protein-specific mAbs were generated that showed a characteristic fluorescent pattern in indirect immunofluorescence assay (IFA) using TPMV-infected Vero cells. Out of the six mAbs tested, five showed no cross-reaction to rodent-associated hantaviruses (Hantaan, Seoul, Puumala, Tula, Dobrava-Belgrade and Sin Nombre viruses) in IFA and enzyme-linked immunosorbent assay (ELISA), although one mAb reacted to Sin Nombre virus in IFA. None of the mAbs cross-reacted with an amino-terminal segment of the shrew-borne Asama virus N protein. Anti-shrew-IgG sera were prepared after immunization of rabbits and BALB/c-mice with protein-G-purified shrew IgG. TPMV-N-protein-specific sera were raised by immunisation of Asian house shrews (Suncus murinus) with purified yeast-expressed TPMV N protein. Using these tools, an indirect ELISA was developed to detect TPMV-N-protein-specific antibodies in the sera of shrews. Using an established serological assay, high TPMV N protein specific antibody titres were measured in the sera of TPMV-N-protein-immunized and experimentally TPMV-infected shrews, whereas no cross-reactivity to other hantavirus N proteins was found. Therefore, the generated mAbs and the established ELISA system represent useful serological tools to detect TPMV, TPMV-related virus antigens or hantavirus-specific antibodies in hantavirus-infected shrews.
  • Takaaki Koma, Kumiko Yoshimatsu, Midori Taruishi, Daisuke Miyashita, Rika Endo, Kenta Shimizu, Shumpei P. Yasuda, Takako Amada, Takahiro Seto, Ryo Murata, Haruka Yoshida, Hiroaki Kariwa, Ikuo Takashima, Jiro Arikawa
    JOURNAL OF VIROLOGICAL METHODS 185 1 74 - 81 2012年10月 [査読有り][通常論文]
     
    New World hantaviruses were divided into five groups based on the amino acid sequence variability of the internal variable region (around 230-302 amino acids) of hantavirus nucleocapsid protein (NP). Sin Nombre virus (SNV), Andes virus, Black Creek Canal virus (BCCV). Carrizal virus (CARV) and Cano Delgadito virus belong to groups 1, 2, 3,4 and 5, respectively. Patient and rodent sera were serotyped successfully by an enzyme-linked immunosorbent assay (ELISA) with recombinant truncated NP lacking 99 N-terminal amino acids (trNP100) of SNV, CARV and BCCV. The trNP100 of BCCV showed lower reactivity to heterologous sera. In contrast, whole recombinant NP antigens detected both homologous and heterologous antibodies equally. The results together with results of a previous study suggest that trNP100 can distinguish infections among viruses in groups 1, 2, 3 and 4 of New World hantaviruses. The serotyping ELISA with trNP100 is useful for epidemiological surveillance in humans and rodents. (C) 2012 Elsevier BM. All rights reserved.
  • Vu Dinh Luan, Kumiko Yoshimatsu, Rika Endo, Midori Taruishi, Vo Thi Huong, Dang Tuan Dat, Pham Cong Tien, Kenta Shimzu, Takaaki Koma, Shumpei P. Yasuda, Le Nhi, Vu Thi Que Huong, Jiro Arikawa
    JOURNAL OF VETERINARY MEDICAL SCIENCE 74 9 1155 - 1162 2012年09月 [査読有り][通常論文]
     
    To investigate the distribution of hantaviruses among animals in Southern and Central Highland area of Vietnam, a total of 1311 serum samples were obtained from rats and Asian house shrews (Suncus murinus) captured at 11 locations between 2006 and 2009. A total of 1066 serum samples from rats were examined for IgG antibodies against Hantaan virus, and there were 30 antibody-positive serum samples from rats that had been captured mainly in a port area and urban area in Ho Chi Minh City (HCMC) (2.8%). All of the antibody-positive rats were Rattus norvegicus, and they had Seoul virus (SEOV) genome in their lungs. SEOV sequences detected from rats captured in Southern Vietnam belonged to the same lineage as those from rats captured at Haiphong Port and a market area in Hanoi City. SEOV strain CSG5 was isolated from a rat captured at Saigon Harbor. Strain CSG5 showed a cross-neutralization pattern almost the same as that of a representative strain of SEOV. A total of 245 Asian house shrews were captured in the Central Highland area and near HCMC. Sera were examined for IgG antibodies against Thottapalayam virus (TPMV), and 32 (13.1%) of the antibody-positive shrews were mainly from the Central Highland area and showed a neutralizing antibody against TPMV. These results indicated that SEOV is distributed among R. norvegicus inhabiting harbor and urban areas of Southern Vietnam and that TPMV or an antigenically related virus is distributed among Asian house shrews in Central Highland area.
  • Rie Isozumi, Kumiko Yoshimatsu, Tetsu Yamashiro, Futoshi Hasebe, Binh Minh Nguyen, Tuan Cuong Ngo, Shumpei P. Yasuda, Takaaki Koma, Kenta Shimizu, Jiro Arikawa
    EMERGING INFECTIOUS DISEASES 18 8 1383 - 1385 2012年08月 [査読有り][通常論文]
  • Ngonda Saasa, Haruka Yoshida, Kenta Shimizu, Cornelio Sanchez-Hernandez, Maria de Lourdes Romero-Almaraz, Takaaki Koma, Takahiro Sanada, Takahiro Seto, Kentaro Yoshii, Celso Ramos, Kumiko Yoshimatsu, Jiro Arikawa, Ikuo Takashima, Hiroaki Kariwa
    VIROLOGY 428 1 48 - 57 2012年06月 [査読有り][通常論文]
     
    The hantavirus nucleocapsid (N) protein is an important immunogen that stimulates a strong and cross-reactive immune response in humans and rodents. A large proportion of the response to N protein has been found to target its N-terminus. However, the exact nature of this bias towards the N-terminus is not yet fully understood. We characterized six monoclonal antibodies (mAbs) against the N protein of Montano virus (MTNV), a Mexican hantavirus. Five of these mAbs recognized eight American hantaviruses and six European and Asian hantaviruses, but not the Soricomorpha-borne Thottapalayam hantavirus. The N protein-reactive binding regions of the five mAbs were mapped to discontinuous epitopes within the N-terminal 13-51 amino acid residues, while a single serotype-specific mAb was mapped to residues 1-25 and 49-75. Our findings suggest that discontinuous epitopes at the N-terminus are conserved, at least in rodent-borne hantaviruses, and that they contribute considerably to N protein cross-reactivity. (C) 2012 Elsevier Inc. All rights reserved.
  • Shumpei P. Yasuda, Kumiko Yoshimatsu, Takaaki Koma, Kenta Shimizu, Rika Endo, Rie Isozumi, Jiro Arikawa
    JOURNAL OF VETERINARY MEDICAL SCIENCE 74 2 215 - 219 2012年02月 [査読有り][通常論文]
     
    Truncated recombinant nucleocapsid proteins (trNs) that lack N-terminally located cross-reactive epitopes of four Murinae rodent-associated hantaviruses, Seoul virus (SEOV). Thailand virus, Hantaan virus (HTNV) and Dobrava-Belgrade virus, were produced by using a baculovirus expression system. ELISA with the trNs as antigens enabled serotyping of immune sera from rats experimentally inoculated with the corresponding hantaviruses with cut-off OD values of 60% of those of whole N of HTNV. The trN-based ELISA could serotype 12 out of 13 sera obtained from wild rodents (Ratios norvegicus) naturally infected with SEOV using the 60% cut-off value. These results indicate that screening with whole N followed by serotyping with trNs using a cut-off OD value of 60% of that of whole N is a useful method for serological surveillance of Murinae-associated hantavirus infection among rodents.
  • Naoto Ito, Tetsuo Mita, Kenta Shimizu, Yuki Ito, Tatsunori Masatani, Keisuke Nakagawa, Satoko Yamaoka, Masako Abe, Kota Okadera, Nobuyuki Minamoto, Makoto Sugiyama
    JOURNAL OF VETERINARY MEDICAL SCIENCE 73 10 1363 - 1366 2011年10月 [査読有り][通常論文]
     
    We previously reported that rabies virus strain CE(NiM), but not the parental Ni-CE strain, killed mice after intracerebral inoculation. CE(NiM) and Ni-CE are genetically identical except for two amino acids at positions 29 and 95 in the M protein. In this study, to identify which residue determines the pathogenicity, we examined pathogenicities of two Ni-CE mutants, CE(NiM29) and CE(NiM95), which were established by replacement of an amino acid residue at position 29 or 95 in the Ni-CE M protein with the corresponding residue CE(NiM), respectively. We found that CE(NiM95), but not CE(NiM29), killed mice, indicating that the amino acid at position 95 in the M protein is the pathogenic determinant.
  • Kenta Shimizu, Chengjun Li, Yukiko Muramoto, Shinya Yamada, Jiro Arikawa, Hualan Chen, Yoshihiro Kawaoka
    JOURNAL OF GENERAL VIROLOGY 92 7 1645 - 1649 2011年07月 [査読有り][通常論文]
     
    Since their emergence in 1996 in southern China, highly pathogenic H5N1 avian influenza viruses have spread widely and continue to circulate in some countries. Genetic reassortment has created multiple H5N1 virus lineages, some of which are dominant in nature. However, the mechanism by which certain H5N1 influenza virus lineages (or genotypes) become dominant in avian species remains unknown. Here, we used competitive inoculation and genetic analysis of the resultant viruses to show that the nucleoprotein (NP) and matrix protein (M) segments of Fujian-like viruses (clade 2.3.4), which became predominant in southern China in mid-2006, are responsible for viral dominance in embryonated eggs. We further found that specific residues in the NP and M proteins play key roles in conferring this viral dominance; specifically, a glutamic acid at position 66 in M2 was conserved among the Fujian-like viruses. These results suggest roles for these viral proteins in influenza virus dominance.
  • Tatsunori Masatani, Naoto Ito, Kenta Shimizu, Yuki Ito, Keisuke Nakagawa, Masako Abe, Satoko Yamaoka, Makoto Sugiyama
    VIRUS RESEARCH 155 1 168 - 174 2011年01月 [査読有り][通常論文]
     
    We previously reported that nucleoprotein (N) is related to the different pathogenicities of the virulent rabies virus strain Nishigahara (Ni) and avirulent strain Ni-CE and also that Ni N, but not Ni-CE N, functions to evade retinoic acid-inducible gene I (RIG-I)-mediated innate immunity. There are three amino acid differences between Ni and Ni-CE N (at positions 273, 394 and 395), indicating that one of these mutations or a combination of mutations is important for the pathogenicity and evasion of innate immunity. We generated Ni-CE mutants in which the amino acids in Ni-CE N were replaced with those of Ni in all combinations. Among the mutants, CE(NiN273/394) with mutations at positions 273 and 394 evaded activation of RIG-I-mediated signaling most efficiently and also showed the highest pathogenicity. This correlation reinforces the relation between evasion of host RIG-I-mediated innate immunity and pathogenicity of rabies virus. (C) 2010 Elsevier B.V. All rights reserved.
  • Erdenesaikhan Tegshduuren, Kumiko Yoshimatsu, Midori Taruishi, Rika Endo, Kenta Shimizu, Takaaki Koma, Shumpei P. Yasuda, Hiroaki Kariwa, Jiro Arikawa, Chiaki Ishihara
    COMPARATIVE IMMUNOLOGY MICROBIOLOGY AND INFECTIOUS DISEASES 33 6 E67 - E73 2010年12月 [査読有り][通常論文]
     
    Tula virus (TULV) and Puumala virus (PUUV) are hantaviruses carried by the bank vole (Myodes glareolus) and European common vole (Microtus arvalis), respectively. PUUV is a causative agent of hemorrhagic fever with renal syndrome (HFRS), while TULV is thought to be apathogenic to humans. The N-terminal regions of the N proteins from TULV and PUUV were expressed and applied as enzyme-linked immunosorbent assay (ELISA) antigens. Colonized Japanese grass voles (Microtus montebelli) and BALB/c mice were used for experimental inoculation of the vole-borne hantaviruses TULV and PUUV. Voles and mice showed significant antibody production toward both viruses, but these antisera showed little cross-reactivity between TULV and PUUV in the immunofluorescence antibody assay and ELISA. In contrast, sera from patients with HFRS caused by PUUV exhibited high cross-reactivity against the TULV antigen, and sera from a natural rodent reservoir showed moderate cross-reactivity against the heterologous antigen, indicating that the antigenic cross-reactivity between TULV and PUUV differs in sera from rodents and humans. (C) 2010 Elsevier Ltd. All rights reserved.
  • Naoto Ito, Gregory W. Moseley, Danielle Blondel, Kenta Shimizu, Caitlin L. Rowe, Yuki Ito, Tatsunori Masatani, Keisuke Nakagawa, David A. Jans, Makoto Sugiyama
    JOURNAL OF VIROLOGY 84 13 6699 - 6710 2010年07月 [査読有り][通常論文]
     
    The fixed rabies virus (RV) strain Nishigahara kills adult mice after intracerebral inoculation, whereas the chicken embryo fibroblast cell-adapted strain Ni-CE causes nonlethal infection in adult mice. We previously reported that the chimeric CE(NiP) strain, which has the phosphoprotein (P protein) gene from the Nishigahara strain in the genetic background of the Ni-CE strain, causes lethal infection in adult mice, indicating that the P gene is responsible for the different pathogenicities of the Nishigahara and Ni-CE strains. Previous studies demonstrated that RV P protein binds to the interferon (IFN)-activated transcription factor STAT1 and blocks IFN signaling by preventing its translocation to the nucleus. In this study, we examine the molecular mechanism by which RV P protein determines viral pathogenicity by comparing the IFN antagonist activities of the Nishigahara and Ni-CE P proteins. The results, obtained from both RV-infected cells and cells transfected to express P protein only, show that Ni-CE P protein is significantly impaired for its capacity to block IFN-activated STAT1 nuclear translocation and, consequently, inhibits IFN signaling less efficiently than Nishigahara P protein. Further, it was demonstrated that a defect in the nuclear export of Ni-CE P protein correlates with a defect in its ability to cause the mislocalization of STAT1. These data provide the first evidence that the capacity of the RV P protein to inhibit STAT1 nuclear translocation and IFN signaling correlates with the viral pathogenicity.
  • Takaaki Koma, Kumiko Yoshimatsu, Noemi Pini, David Safronetz, Midori Taruishi, Silvana Levis, Rika Endo, Kenta Shimizu, Shumpei P. Yasuda, Hideki Ebihara, Heinz Feldmann, Delia Enria, Jiro Arikawa
    JOURNAL OF CLINICAL MICROBIOLOGY 48 5 1635 - 1642 2010年05月 [査読有り][通常論文]
     
    Sin Nombre virus (SNV), Andes virus (ANDV), and Laguna Negra virus (LANV) have been known as the dominant causative agents of hantavirus pulmonary syndrome (HPS). ANDV and LANV, with different patterns of pathogenicity, exist in a sympatric relationship. Moreover, there is documented evidence of person-to-person transmission of ANDV. Therefore, it is important in clinical medicine and epidemiology to know the serotype of a hantavirus causing infection. Truncated SNV, ANDV, and LANV recombinant nucleocapsid proteins (trNs) missing 99 N-terminal amino acids (trN100) were expressed using a baculovirus system, and their applicability for serotyping SNV, ANDV, and LANV infection by the use of enzyme-linked immunosorbent assays (ELISA) was examined. HPS patient sera and natural-reservoir rodent sera infected with SNV, ANDV, and LANV showed the highest optical density (OD) values for homologous trN100 antigens. Since even patient sera with lower IgM and IgG antibody titers were serotyped, the trN100s are therefore considered useful for serotyping with early-acute-phase sera. In contrast, assays testing whole recombinant nucleocapsid protein antigens of SNV, ANDV, and LANV expressed in Escherichia coli detected homologous and heterologous antibodies equally. These results indicated that a screening ELISA using an E. coli-expressed antigen followed by a serotyping ELISA using trN100s is useful for epidemiological surveillance in regions where two or more hantavirus species cocirculate.
  • Tatsunori Masatani, Naoto Ito, Kenta Shimizu, Yuki Ito, Keisuke Nakagawa, Yoshiharu Sawaki, Hiroyuki Koyama, Makoto Sugiyama
    JOURNAL OF VIROLOGY 84 8 4002 - 4012 2010年04月 [査読有り][通常論文]
     
    The rabies virus Ni-CE strain causes nonlethal infection in adult mice after intracerebral inoculation, whereas the parental Nishigahara (Ni) strain kills mice. We previously reported that the chimeric CE(NiN) strain with the N gene from the Ni strain in the genetic background of the Ni-CE strain kills adult mice, indicating that the N gene is related to the different pathogenicities of Ni and Ni-CE strains. In the present study, to obtain an insight into the mechanism by which the N gene determines viral pathogenicity, we compared the effects of Ni, Ni-CE, and CE(NiN) infections on host gene expressions using a human neuroblastoma cell line. Microarray analysis of these infected cells revealed that the expression levels of particular genes in Ni- and CE(NiN)-infected cells, including beta interferon (IFN-beta) and chemokine genes (i.e., CXCL10 and CCL5) were lower than those in Ni-CE-infected cells. We also demonstrated that Ni-CE infection activated the interferon regulatory factor 3 (IRF-3)-dependent IFN-beta promoter and induced IRF-3 nuclear translocation more efficiently than did Ni or CE(NiN) infection. Furthermore, we showed that Ni-CE infection, but not Ni or CE(NiN) infection, strongly activates the IRF-3 pathway through activation of RIG-I, which is known as a cellular sensor of virus infection. These findings indicate that the N protein of rabies virus (Ni strain) has a function to evade the activation of RIG-I. To our knowledge, this is the first report that the Mononegavirales N protein functions to evade induction of host IFN and chemokines.
  • Thua Thang Truong, Kumiko Yoshimatsu, Koichi Araki, Byoung-Hee Lee, Ichiro Nakamura, Rika Endo, Kenta Shimizu, Shumpei P. Yasuda, Takaaki Koma, Midori Taruishi, Megumi Okumura, Uyen Ninh Truong, Jiro Arikawa
    JOURNAL OF VETERINARY MEDICAL SCIENCE 71 10 1357 - 1363 2009年10月 [査読有り][通常論文]
     
    The distribution of anti-hantavirus antibodies in humans and rodents in northern Vietnam was examined. In total, 837 seruml samples from healthy humans (617) and patients with fever (220), living in six different areas were screened for IgG antibodies against Hantaan or Seoul virus (SEOV) by ELISA, IFA, and Western blot analysis. Antibody-positive sera were identified ill 7/617 (1.1%) healthy donors, 51150 port workers in the port of Hai Phong, and 2/185 residents of Ha Nam Province. In comparison, positive sera were detected in 5/220 (2.3%) fever patients ill the provinces of Ha Nam (1/58) and Thanh Hoa (4/146). Antibody-positive Rattus norvegicus were found in the provinces of Ha Nam (7/52) and Thanh Hoa (1/67), in Haibatrung District (7/43) in Hanoi, and in Hai Phong" Port (21/62), while antibody-positive R. rattus (2/17) were found in Hai Phong Port. Part of the Gc region from the viral genome was amplified by RT-PCR using lung tissue samples front R. norvegicus in Haibatrung (2/7) and Hai Phong Port (7/9), bill not from R. rattus (0/2). Viral sequences were located in the SEOV clade and formed a single lineage with Indonesian SEOV. suggesting that Vietnamese SEOV is part of a distinct lineage among Asian SEOVs.
  • Tetsuo Mita, Kenta Shimizu, Naoto Ito, Kentaro Yamada, Yuki Ito, Makoto Sugiyama, Nobuyuki Minamoto
    VIRUS RESEARCH 137 1 33 - 39 2008年10月 [査読有り][通常論文]
     
    The molecular mechanism involved in cytopathogenicity of rabies virus has not been fully elucidated yet. A fixed rabies virus Nishigahara strain does not induce clear cytopathic effect (CPE) in mouse neuroblastoma (NA) cells, whereas Ni-CE strain, which was established after 100 passages of Nishigahara strain in chicken embryo fibroblast cells, induces CIPE that is characterized by rounding, shrinkage and detachment of the cells. In this study, to identify which viral gene is associated with the CPE of Ni-CE Strain, we analyzed chimeric viruses between Nishigahara and Ni-CE strains generated by reverse genetics systems of both strains. We showed that the matrix gene of Ni-CE strain is responsible for the CPE in NA cells. It was also demonstrated by infection of Nishigahara and Ni-CE mutants with a single amino acid substitution in the matrix protein (M) that an amino acid at position 95 of M is a cytopathic determinant of the virus. We also demonstrated that the CPF is, at least partly, due to apoptosis. This is the first report of identification of an amino acid residue in a rabies virus protein that is important for the cytopathogenicity of the virus. (C) 2008 Elsevier B.V. All rights reserved.
  • Kenta Shimizu, Naoto Ito, Tetsuo Mita, Kentaro Yamada, Junji Hosokawa-Muto, Makoto Sugiyama, Nobuyuki Minamoto
    VIRUS RESEARCH 123 2 154 - 160 2007年02月 [査読有り][通常論文]
     
    Rabies virus Ni-CE strain causes nonlethal infection in adult mice after intracerebral inoculation, whereas the parental Nishigahara strain kills mice. In this study, to identify viral gene(s) related to the difference in pathogenicity between Ni-CE and Nishigahara strains, we generated chimeric viruses with respective genes of the virulent Nishigahara strain in the background of the avirudent Ni-CE genome. Since chimeric viruses, which had the N, P, or M genes of the Nishigahara strain, respectively, killed adult mice after intracerebral inoculation, it became evident that the N, P, and M genes are related to the difference in pathogenicity between Ni-CE and Nishigahara strains. Previously, we showed that the G gene is a major contributor to the difference in pathogenicity between another avirulent strain, RC-HL, and the parental Nishigahara strain. These results imply that the attenuation mechanism of the Ni-CE strain is different from that of the RC-HL strain, thus suggesting that rabies virus can be attenuated by diverse mechanisms. This is the first report of changes in viral genes other than the G gene of rabies virus causing the reversion of pathogenicity of an avirulent strain. (c) 2006 Elsevier B.V. All rights reserved.
  • J Hosokawa-Muto, N Ito, K Yamada, K Shimizu, M Sugiyama, N Minamoto
    MICROBIOLOGY AND IMMUNOLOGY 50 3 187 - 196 2006年 [査読有り][通常論文]
     
    A recombinant rabies virus carrying double glycoprotein (G) genes, R(NPMGGL) strain, was generated by a reverse genetics system utilizing cloned cDNA of the RC-HL strain, and the biological properties of the virus were compared to those of the recombinant RC-HL (rRC-HL) strain. The extents of virus growth in cultured cells and virulence for adult mice of the R(NPMGGL) strain were almost same as those of the rRC-HL strain, while G protein content of the purified R(NPMGGL) virion and G protein expression level in R(NPMGGL)-infected cells were 1.5-fold higher than those of the rRC-HL strain. As a result of serial passages of the R(NPMGGL) strain in cultured cells, the expression level of G protein in cultured cells infected with the passaged R(NPMGGL) strain was maintained and virus titers rose with adaptation to the cultured cells. Furthermore, analysis of neutralization titers in mice immunized with UV-inactivated virus suggested that the R(NPMGGL) strain had higher immunogenicity than that of the rRC-HL strain. The results suggest that the R(NPMGGL) strain carrying double G genes might be a useful candidate for development of a new inactivated rabies vaccine.
  • Kenta Shimizu, Naoto Ito, Makoto Sugiyama, Nobuyuki Minamoto
    MICROBIOLOGY AND IMMUNOLOGY 50 12 975 - 978 2006年 [査読有り][通常論文]
     
    The growth of a virulent strain of fixed rabies virus, Nishigahara, in mouse neuroblastoma NA cells treated with type I interferon (IFN) was compared with that of a derivative avirulent strain, Ni-CE. Nishigahara strain was slightly sensitive to IFN treatment but still grew more efficiently than did Ni-CE strain in IFN-treated NA cells. Furthermore, a virulent chimeric virus with the phosphoprotein gene from Nishigahara strain in the Ni-CE genome was less sensitive to IFN treatment than was Ni-CE strain, indicating that the IFN sensitivity is determined by the phosphoprotein gene of the virus.
  • N Ito, M Sugiyama, K Yamada, K Shimizu, M Takayama-Ito, J Hosokawa, N Minamoto
    MICROBIOLOGY AND IMMUNOLOGY 49 11 973 - 981 2005年 [査読有り][通常論文]
     
    Matrix (M) protein of rabies virus is known to play an important role in assembly and budding of the progeny virus. We generated an M gene-deficient rabies virus, RC-HL Delta M, using a reverse genetics system of rabies virus RC-HL strain to develop a novel type of vaccine. RC-HL Delta M infection was confined within a single cell in mouse neuroblastoma cells. This deficient virus failed to generate the progeny virus in the cells. In contrast, RC-HL Delta M propagated in BHK cells inductively expressing M protein. Suckling and adult mice inoculated intracerebrally with the parental RC-HL strain showed lethal infection and transient body weight loss, respectively, whereas both suckling and adult mice inoculated with RC-HL Delta M showed no symptoms. The neutralizing antibody against rabies virus was successfully induced by intramuscular immunization with 10(5) focus-forming units of RC-HL Delta M but not UV-inactivated RC-HL. Intranasal immunization with RC-HL Delta M resulted in almost the same antibody titer to rabies virus as that in the case of immunization with live RC-HL strain. These findings indicate that RC-HL Delta M is a candidate for a novel rabies vaccine that is safer and more effective than are current vaccines.

受賞

  • 2019年07月 北海道大学医学部 平成29年度 優秀科目賞 (最優秀賞)
     
    受賞者: 微生物学;免疫学実習;授業担当教員
  • 2017年09月 日本獣医学会 JVMS優秀論文賞 (2016)
     Serological evidence of infection with rodent-borne hepatitis E virus HEV-C1 or antigenically related virus in humans 
    受賞者: Shimizu K;Hamaguchi S;Ngo CC;Li TC;Ando S;Yoshimatsu K;Yasuda SP;Koma T;Isozumi R;Tsuda Y;Fujita H;Pham TT;Le MQ;Dang AD;Nguyen TQ;Yoshida LM;Ariyoshi K;Arikawa J

共同研究・競争的資金等の研究課題

  • 熱帯性慢性腎臓病に関連する新規ハンタウイルス、ランカウイルスの解析
    日本学術振興会:科学研究費助成事業 基盤研究(C) (一般)
    研究期間 : 2019年04月 -2022年03月 
    代表者 : 吉松組子, 研究分担者, 清水健太
  • スリランカに急増する原因不明慢性腎臓病 (CKDu) の原因究明
    平和中島財団:アジア地域重点学術研究助成
    研究期間 : 2019年04月 -2020年03月 
    代表者 : 吉松組子, 共同研究者, 清水健太
  • ハンタウイルスが誘導する免疫病原性発現機構
    日本学術振興会:科学研究費助成事業 基盤 (C)
    研究期間 : 2016年04月 -2020年03月 
    代表者 : 清水 健太
  • 動物モデルを用いたハンタウイルス感染症の治療法の開発
    日本学術振興会:科学研究費助成事業 若手 (B)
    研究期間 : 2014年04月 -2017年03月 
    代表者 : 清水 健太
  • 動物モデルを用いたハンタウイルス感染症の病態解明
    日本学術振興会:科学研究費助成事業 若手 (B)
    研究期間 : 2012年04月 -2015年03月 
    代表者 : 清水 健太
  • コウモリにおける節足動物媒介性ウイルスの保有状況の調査
    北海道大学医学研究科:国際交流基金 若手研究者の派遣・補助事業
    研究期間 : 2012年04月 -2013年03月 
    代表者 : 清水 健太
  • ラットE型肝炎ウイルスは人獣共通感染症の病原体か?
    長崎大学熱帯医学研究所:熱帯医学研究拠点一般共同研究
    研究期間 : 2012年04月 -2013年03月 
    代表者 : 清水 健太

教育活動情報

主要な担当授業

  • 医学総論
    開講年度 : 2020年
    課程区分 : 博士後期課程
    開講学部 : 医学研究科
  • 基本医学研究
    開講年度 : 2020年
    課程区分 : 修士課程
    開講学部 : 医学院
    キーワード : ハンタウイルス、E型肝炎ウイルス、レプトスピラ、病原性、感染症、人獣共通感染症、実験動物 hantavirus,hepatitis E virus, leptospira, pathogenicity, infectious disease, zoonoses, laboratory animal
  • 基本医学総論
    開講年度 : 2020年
    課程区分 : 修士課程
    開講学部 : 医学院
    キーワード : ハンタウイルス、E型肝炎ウイルス、レプトスピラ、病原性、感染症、人獣共通感染症、実験動物 viruses, hepatitis E virus, leptospira, pathogenicity, infectious disease, zoonoses, laboratory animal
  • 医学総論
    開講年度 : 2020年
    課程区分 : 博士後期課程
    開講学部 : 医学院
    キーワード : ウイルス、細菌、病原性、感染症、人獣共通感染症、実験動物 viruses, bacteria, pathogenicity, infection, zoonotic infection, laboratory animal
  • 基盤医学研究
    開講年度 : 2020年
    課程区分 : 博士後期課程
    開講学部 : 医学院
    キーワード : ハンタウイルス、E型肝炎ウイルス、レプトスピラ、病原性、感染症、人獣共通感染症、実験動物 viruses, hepatitis E virus, leptospira, pathogenicity, infectious disease, zoonoses, laboratory animal
  • 微生物・免疫学実習
    開講年度 : 2020年
    課程区分 : 学士課程
    開講学部 : 医学部
    キーワード : 細菌、ウイルス、無菌操作、グラム染色、抗酸菌染色、培養、細胞培養、定量、抗生物質、CPE、PCR、ELISA、IFA、蛍光抗体法、抗体、ブロッティング、炎症性サイトカイン、フローサイトメトリー、リンパ球、マクロファージ
  • 微生物学
    開講年度 : 2020年
    課程区分 : 学士課程
    開講学部 : 医学部
    キーワード : 細菌、ウイルス、グラム陽性菌、グラム陰性菌、抗酸性菌、感染症、ワクチン、プリオン、寄生虫、原虫、マイコプラズマ、クラミジア、スピロヘータ、真菌

大学運営

委員歴

  • 2014年11月 - 現在   Frontiers in Microbiology   Review Editor
  • 2018年02月 - 2018年09月   日本実験動物医学専門医協会   試験問題検討委員会副委員長
  • 2013年06月 - 2018年09月   日本実験動物医学専門医協会   試験問題検討委員会委員


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