Papers - YAMADA Kentaro
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Development of an oncolytic mammalian orthoreovirus expressing the near-infrared fluorescent protein iRFP720. Reviewed International journal
Katsuhiro Ogawa, Kentaro Yamada, Tsuyoshi Etoh, Masahiro Kitagawa, Yoshinori Shirasaka, Kazuko Noguchi, Takeshi Kobayashi, Akira Nishizono, Masafumi Inomata
Journal of virological methods 308 114574 - 114574 2022.10
Authorship：Corresponding author Language：English Publishing type：Research paper (scientific journal)
Fluorescence-guided surgery (FGS) is a useful method for removing invasive tumor tissues. For this, near-infrared (NIR) fluorescence probes are suitable for visualizing cancer cells due to their low autofluorescence, and an oncolytic mammalian orthoreovirus (MRV) expressing an NIR fluorescent protein is expected to be a novel tool for FGS. In this study, we identified the optimal insertion site of the NIR fluorescent protein gene iRFP720 (915 nt) in the MRV genome. We constructed genome plasmids for the L1, M1, and S2 segments, where a gene cassette comprising iRFP720 and T2A self-cleaving peptide was inserted in the 5' or 3' region of each segment. Through virus recovery, the recombinant MRV with the gene cassette at the M1 segment's 3' end, T3D-L(M1/3'iRFP720), was capable of replication and passaging with bright NIR fluorescence. However, the replication of T3D-L(M1/3'iRFP720) was approximately 1,000-fold lower than that of the wild-type virus. T3D-L(M1/3'iRFP720) production improved due to the transfection of a fusion-associated small transmembrane protein gene of fusogenic reovirus. Further, fluorescence signals were detected in T3D-L(M1/3'iRFP720)-infected human gastric and pancreatic cancer cells. Thus, the M1 segment's 3' end tolerates the expression of the long iRFP720 gene, which may propel the development of recombinant MRV vectors for FGS.
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Kojima I, Onomoto K, Zuo W, Ozawa M, Okuya K, Naitou K, Izumi F, Okajima M, Fujiwara T, Ito N, Yoneyama M, Yamada K, Nishizono A, Sugiyama M, Fujita T, Masatani T
Journal of virology 96 ( 18 ) e0081022 2022.9
Language：English Publishing type：Research paper (scientific journal) Publisher：Journal of Virology
Stress granules (SGs) are dynamic structures that store cytosolic messenger ribonucleoproteins. SGs have recently been shown to serve as a platform for activating antiviral innate immunity; however, several pathogenic viruses suppress SG formation to evade innate immunity. In this study, we investigated the relationship between rabies virus (RABV) virulence and SG formation, using viral strains with different levels of virulence. We found that the virulent Nishigahara strain did not induce SG formation, but its avirulent offshoot, the Ni-CE strain, strongly induced SG formation. Furthermore, we demonstrated that the amino acid at position 95 in the RABV matrix protein (M95), a pathogenic determinant for the Nishigahara strain, plays a key role in inhibiting SG formation, followed by protein kinase R (PKR)-dependent phosphorylation of the a subunit of eukaryotic initiation factor 2a (eIF2a). M95 was also implicated in the accumulation of RIG-I, a viral RNA sensor protein, in SGs and in the subsequent acceleration of interferon induction. Taken together, our findings strongly suggest that M95-related inhibition of SG formation contributes to the pathogenesis of RABV by allowing the virus to evade the innate immune responses of the host.
DOI： 10.1128/jvi.00810-22
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Distribution of Severe Fever with Thrombocytopenia Syndrome Virus and Antiviral Antibodies in Wild and Domestic Animals in Oita Prefecture, Japan. Reviewed International journal
Takehiro Hashimoto, Takaaki Yahiro, Kentaro Yamada, Kazunori Kimitsuki, Minami W Okuyama, Akiko Honda, Miki Kato, Hiroshi Narimatsu, Kazufumi Hiramatsu, Akira Nishizono
The American journal of tropical medicine and hygiene 106 ( 5 ) 1547 - 1551 2022.2
Language：English Publishing type：Research paper (scientific journal)
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The Capsid (ORF2) Protein of Hepatitis E Virus in Feces Is C-Terminally Truncated. Reviewed International journal
Takashi Nishiyama, Koji Umezawa, Kentaro Yamada, Masaharu Takahashi, Satoshi Kunita, Mulyanto, Isao Kii, Hiroaki Okamoto
Pathogens (Basel, Switzerland) 11 ( 1 ) 2022.1
Language：English Publishing type：Research paper (scientific journal)
The hepatitis E virus (HEV) is a causative agent of hepatitis E. HEV virions in circulating blood and culture media are quasi-enveloped, while those in feces are nonenveloped. The capsid (ORF2) protein associated with an enveloped HEV virion is reported to comprise the translation product of leucine 14/methionine 16 to 660 (C-terminal end). However, the nature of the ORF2 protein associated with fecal HEV remains unclear. In the present study, we compared the molecular size of the ORF2 protein among fecal HEV, cell-culture-generated HEV (HEVcc), and detergent-treated protease-digested HEVcc. The ORF2 proteins associated with fecal HEV were C-terminally truncated and showed the same size as those of the detergent-treated protease-digested HEVcc virions (60 kDa), in contrast to those of the HEVcc (68 kDa). The structure prediction of the ORF2 protein (in line with previous studies) demonstrated that the C-terminal region (54 amino acids) of an ORF2 protein is in flux, suggesting that proteases target this region. The nonenveloped nondigested HEV structure prediction indicates that the C-terminal region of the ORF2 protein moves to the surface of the virion and is unnecessary for HEV infection. Our findings clarify the maturation of nonenveloped HEV and will be useful for studies on the HEV lifecycle.
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Comprehensive exploration of chemical space using trisubstituted carboranes. Reviewed International coauthorship International journal
Yasunobu Asawa, Saki Hatsuzawa, Atsushi Yoshimori, Kentaro Yamada, Akira Katoh, Hiroyuki Kouji, Hiroyuki Nakamura
Scientific reports 11 ( 1 ) 24101 - 24101 2021.12
Language：English Publishing type：Research paper (scientific journal)
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Lateral flow devices for samples collected by straw sampling method for postmortem canine rabies diagnosis. Reviewed International journal
Milagros R Mananggit, Daria L Manalo, Nobuo Saito, Kazunori Kimitsuki, Alyssa Marie G Garcia, Patricia Mae T Lacanilao, Joely T Ongtangco, Cornhlo R Velasco, Maria Victoria A Del Rosario, Maria Glofezita O Lagayan, Kentaro Yamada, Chun-Ho Park, Satoshi Inoue, Motoi Suzuki, Mariko Saito-Obata, Yasuhiko Kamiya, Catalino S Demetria, Beatriz P Quiambao, Akira Nishizono
PLoS neglected tropical diseases 15 ( 12 ) e0009891 2021.12
Language：English Publishing type：Research paper (scientific journal)
The direct fluorescent antibody test (dFAT) using brain sample after opening the skull is the standard rabies diagnostic test in animal rabies. However, it is not feasible in many resource-limited settings. Lateral flow devices (LFD) combined with a simple sampling methodology is quicker, simpler, and less hazardous than the standard test and can be a useful tool. We conducted a prospective on-site study to evaluate the diagnostic accuracy of the LFD with the straw sampling method compared with that of the dFAT with the skull opening procedure for post-mortem canine rabies diagnosis. We collected 97 rabies-suspected animals between December 1, 2020 and March 31, 2021. Among the 97 samples, 53 and 50 cases were positive tests for dFAT and LFD, respectively. The sensitivity and specificity of LFD with straw sampling method were 94.3% (95% confidence interval [CI], 84.3-98.8%) and 100% (95% CI, 92.0-100%), respectively. The performance of LFD by the straw sampling method showed relatively high sensitivity and 100% specificity compared with that of dFAT performed on samples collected after opening the skull. This methodology can be beneficial and is a strong tool to overcome limited animal surveillance in remote areas. However, because of our limited sample size, more data using fresh samples on-site and the optimizations are urgently needed for the further implementation in endemic areas.
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Background and descriptive features of rabies-suspected animals in Central Luzon, Philippines. Reviewed International coauthorship
Milagros R Mananggit, Kazunori Kimitsuki, Nobuo Saito, Alyssa Marie G Garcia, Patricia Mae T Lacanilao, Joely T Ongtangco, Cornelio R Velasco, Maria Victoria D Rosario, Maria Glofezita O Lagayan, Kentaro Yamada, Chun-Ho Park, Satoshi Inoue, Motoi Suzuki, Mariko Saito-Obata, Yasuhiko Kamiya, Daria L Manalo, Catalino S Demetria, Beatriz P Quiambao, Akira Nishizono
Tropical medicine and health 49 ( 1 ) 59 - 59 2021.12
Language：English Publishing type：Research paper (scientific journal)
BACKGROUND: The Philippines is one of the major endemic countries for canine rabies in Southeast Asia. However, detailed description and analysis of laboratory-confirmed animal rabies are limited. Highly accurate surveillance requires a thorough understanding of the target area-specific problems and obstacles. Therefore, we aim to describe and analyze the rabies suspect animals in Central Luzon, Philippines, to clarify the characteristics of management and clinical signs by conducting interviews with the owners. METHODS: We prospectively collected information on the rabies suspect animals submitted to the Regional animal laboratory in Central Luzon through passive laboratory-based rabies surveillance between 1st April 2019 and 30th September 2020. We performed active interviews directly or telephonically with the owner. The direct fluorescent antibody test was performed on the hippocampus, brain stem, and cerebellum for laboratory confirmation. Descriptive statistics were used to characterize the number of rabies cases according to management methods and characteristics of suspected animals during the observation period. Clinical symptoms of suspected rabid animals were analyzed by univariate logistic regression analysis. RESULTS: There were 292 sample submissions during the study period. Of these, 160 were positive for dFAT. Samples of pet animals (85.3%) provided by owners or their acquaintances (59.2%) accounted for the majority of laboratory confirmed cases. Case mapping showed that more rabies-suspected cases were sent from areas near the regional laboratory than from those far from the laboratory, despite the incidence of rabies being high in these areas. The management and clinical symptoms of 227 animal cases showed that most owners were managing their animals at home and were allowing them to roam outside (69.6%) and be unvaccinated (78.9%). Rabid animals were more likely to manifest aimless running, restlessness, and agitation. CONCLUSIONS: Our study provided some features of animals with laboratory-confirmed rabies in Central Luzon. However, most of the samples were submitted from areas near the rabies diagnosis laboratory, and the number of samples submitted from remote areas was low. To improve the surveillance capacity, it is necessary to increase sample submissions from remote areas.
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Validation of serum apolipoprotein A1 in rabies virus-infected mice as a biomarker for the preclinical diagnosis of rabies. Reviewed International journal
Kentaro Yamada, Koji Kuribayashi, Naotaka Inomata, Kazuko Noguchi, Kazunori Kimitsuki, Catalino S Demetria, Nobuo Saito, Satoshi Inoue, Chun-Ho Park, Ryo Kaimori, Motoi Suzuki, Mariko Saito-Obata, Yasuhiko Kamiya, Daria L Manalo, Beatriz P Quiambao, Akira Nishizono
Microbiology and immunology 65 ( 10 ) 438 - 448 2021.10
Language：English Publishing type：Research paper (scientific journal)
Rabies is a type of acute fetal encephalitis caused by rabies virus (RABV). While it becomes incurable after symptom onset, it can be prevented by post-exposure prophylaxis (PEP) during the long incubation period. While preclinical diagnosis aids the appropriate PEP administration, it is mostly nonfeasible owing to the absence of viremia or a specific antibody response during the incubation period. Here, an attempt was made to identify a serum biomarker for the preclinical diagnosis of rabies. Using the serum from a mouse inoculated intramuscularly (i.m.) with 5 × 105 focus-forming units (FFU) of recombinant RABV expressing red firefly luciferase (1088/RFLuc) immediately before symptom onset, two-dimensional differential gel electrophoresis was conducted, followed by mass spectrometry, and it was confirmed that apolipoprotein A1 (ApoA1) was up-regulated. ELISA showed that the serum ApoA1 and specific antibody levels increased during the incubation period and on the day of symptom onset. Since a lower infectious dose can be used to induce the unstable and long incubation period generally observed in natural infection, the ApoA1 level in mice inoculated i.m. with 103 FFU of 1088/RFLuc was examined by monitoring viral dynamics using in vivo imaging. The serum ApoA1 and specific antibody levels were up-regulated in 50% and 58.3% of mice exhibiting robust RABV replication, respectively, but not in mice exhibiting weak RABV replication. In addition, it was reported that ApoA1 was found to be a biomarker for neuronal damage. Additional biomarker candidates will be needed for the effective preclinical diagnosis of rabies.
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Elisabet Tangkonda, Meiko Kubo, Satoshi Sekiguchi, Taisuke Shinki, Satomi Sasaki, Kentaro Yamada, Takako Taniguchi, Torrung Vetchapitak, Naoaki Misawa
The Journal of veterinary medical science 83 ( 8 ) 1306 - 1314 2021.8
Language：English Publishing type：Research paper (scientific journal) Publisher：JAPANESE SOCIETY OF VETERINARY SCIENCE
Workers in poultry abattoirs may be frequently exposed to Campylobacter jejuni, which is a leading cause of bacterial food poisoning in Japan. The present study was conducted to measure the titers of IgG and IgA antibodies against C. jejuni among 104 female workers in a chicken processing plant in Miyazaki prefecture, Japan. Information regarding habitual ingestion of raw chicken meat and potential occupational risk factors was collected using a questionnaire. Acid extracts of four C. jejuni strains representing the genotypes most dominant in Miyazaki were used as antigens. The levels of both immunoglobulins measured by ELISA were not correlated with ingestion of edible raw chicken meat, the amount consumed in one sitting, or its frequency. Although age was correlated with antibody levels, the length of employment was not. Furthermore, the IgG and IgA levels in workers at the evisceration step were significantly higher than those at other locations in the plant. To identify the bacterial proteins recognized by the workers' IgG and IgA antibodies, Western blotting followed by LC/MS was conducted. Flagellin was identified as the common protein recognized in the sera of workers for whom ELISA demonstrated both the highest and lowest antibody levels. We concluded that the titers of IgG and IgA against C. jejuni in workers at the processing plant had been increased by occupational exposure to Campylobacter, regardless of raw chicken meat ingestion.
DOI： 10.1292/jvms.21-0244
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Synthesis of Three-Dimensional (Di)Azatricyclododecene Scaffold and Its Application to Peptidomimetics. Reviewed International journal
Kohei Umedera, Taiki Morita, Atsushi Yoshimori, Kentaro Yamada, Akira Katoh, Hiroyuki Kouji, Hiroyuki Nakamura
Chemistry (Weinheim an der Bergstrasse, Germany) 27 ( 46 ) 11888 - 11894 2021.8
Language：English Publishing type：Research paper (scientific journal)
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Evaluation of the diagnostic accuracy of lateral flow devices as a tool to diagnose rabies in post-mortem animals. Reviewed International coauthorship International journal
Kazunori Kimitsuki, Nobuo Saito, Kentaro Yamada, Chun-Ho Park, Satoshi Inoue, Motoi Suzuki, Mariko Saito-Obata, Yasuhiko Kamiya, Daria L Manalo, Catalino S Demetria, Milagros R Mananggit, Beatriz P Quiambao, Akira Nishizono
PLoS neglected tropical diseases 14 ( 11 ) e0008844 2020.11
Language：English Publishing type：Research paper (scientific journal)
Implementation of lateral flow devices (LFDs) for rabies antigen detection is expected to improve surveillance through the efficient detection of rabid animals in resource-limited settings; however, the use of LFDs for diagnosis remains controversial because some commercially available kits show low sensitivity. Therefore, we compared the diagnostic efficacy of three LFDs (ADTEC, Bionote, and Elabscience kits) paralleled with the direct fluorescent antibody test (dFAT) using fresh samples and investigated the diagnostic accuracies. To do so, we evaluated rabies-suspected samples submitted to the Regional Animal Disease Diagnostic Laboratory III, Philippines. Furthermore, we conducted real-time RT-PCR and sequencing to measure the accuracy of field laboratory diagnosis. The total number of animals submitted during this study period was 184 cases, including negative control samples. Of these, 53.9% (84 cases) were positive in the dFAT. Dogs were the most common rabies-suspected animal (n = 135). The sensitivities of the ADTEC and Bionote kits were 0.88 (74 cases) and 0.95 (80 cases), respectively. The specificity of both kits was 1.00 (100 cases). Furthermore, the sensitivity and specificity of the ADTEC kit after directly homogenizing the samples in assay buffer without dilution in phosphate-buffered saline (ADTEC kit DM) were 0.94 (79 cases) and 1.00 (100 cases), respectively. By contrast, there were no positive results using the Elabscience kit among all dFAT-positive samples. The sensitivity and specificity of LFDs make these tests highly feasible if properly used. Therefore, LFD tests can be used to strengthen the surveillance of rabies-infected animals in endemic and resource-limited settings.
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Genetic and Phenotypic Characterization of a Rabies Virus Strain Isolated from a Dog in Tokyo, Japan in the 1940s. Reviewed International journal
Tatsuki Takahashi, Maho Inukai, Michihito Sasaki, Madlin Potratz, Supasiri Jarusombuti, Yuji Fujii, Shoko Nishiyama, Stefan Finke, Kentaro Yamada, Hiroki Sakai, Hirofumi Sawa, Akira Nishizono, Makoto Sugiyama, Naoto Ito
Viruses 12 ( 9 ) 2020.9
Language：English Publishing type：Research paper (scientific journal)
The rabies virus strain Komatsugawa (Koma), which was isolated from a dog in Tokyo in the 1940s before eradication of rabies in Japan in 1957, is known as the only existent Japanese field strain (street strain). Although this strain potentially provides a useful model to study rabies pathogenesis, little is known about its genetic and phenotypic properties. Notably, this strain underwent serial passages in rodents after isolation, indicating the possibility that it may have lost biological characteristics as a street strain. In this study, to evaluate the utility of the Koma strain for studying rabies pathogenesis, we examined the genetic properties and in vitro and in vivo phenotypes. Genome-wide genetic analyses showed that, consistent with previous findings from partial sequence analyses, the Koma strain is closely related to a Russian street strain within the Arctic-related phylogenetic clade. Phenotypic examinations in vitro revealed that the Koma strain and the representative street strains are less neurotropic than the laboratory strains. Examination by using a mouse model demonstrated that the Koma strain and the street strains are more neuroinvasive than the laboratory strains. These findings indicate that the Koma strain retains phenotypes similar to those of street strains, and is therefore useful for studying rabies pathogenesis.
DOI： 10.3390/v12090914
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狂犬病発症前診断用バイオマーカー候補の探索と評価
西園 晃, 齊藤 信夫, 鈴木 基, 山田 健太郎
感染症学雑誌 94 ( 臨増 ) 329 - 329 2020.3
Language：Japanese Publishing type：Research paper (scientific journal) Publisher：(一社)日本感染症学会
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フィリピンにおける狂犬病迅速診断キットの有用性評価中間解析(ヒト・動物) Reviewed International coauthorship
齊藤 信夫, 鈴木 基, 西園 晃, 君付 和範, 山田 健太郎, 齊藤 麻理子[小畑], 朴 天鎬, 井上 智, Quiambao Beatriz
感染症学雑誌 94 ( 臨増 ) 322 - 322 2020.3
Language：Japanese Publishing type：Research paper (scientific journal) Publisher：(一社)日本感染症学会
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狂犬病発症前診断用バイオマーカー候補の探索と評価
西園 晃, 齊藤 信夫, 鈴木 基, 山田 健太郎
感染症学雑誌 94 ( 臨増 ) 329 - 329 2020.3
Language：Japanese Publishing type：Research paper (scientific journal) Publisher：(一社)日本感染症学会
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Reevaluation of the efficacy of favipiravir against rabies virus using in vivo imaging analysis.
Kentaro Yamada, Kazuko Noguchi, Kazunori Kimitsuki, Ryo Kaimori, Nobuo Saito, Takashi Komeno, Nozomi Nakajima, Yousuke Furuta, Akira Nishizono
Antiviral research 172 104641 - 104641 2019.12
Language：Japanese Publishing type：Research paper (scientific journal)
Rabies virus (RABV) is a highly neurotropic virus and the causative agent of rabies, an encephalitis with an almost 100% case-fatality rate that remains incurable after the onset of symptoms. Favipiravir (T-705), a broad-spectrum antiviral drug against RNA viruses, has been shown to be effective against RABV in vitro but ineffective in vivo. We hypothesized that favipiravir is effective in infected mice when RABV replicates in the peripheral tissues/nerves but not after virus neuroinvasion. We attempted to clarify this point in this study using in vivo bioluminescence imaging. We generated a recombinant RABV from the field isolate 1088, which expressed red firefly luciferase (1088/RFLuc). This allowed semiquantitative detection and monitoring of primary replication at the inoculation site and viral spread in the central nervous system (CNS) in the same mice. Bioluminescence imaging revealed that favipiravir (300 mg/kg/day) treatment commencing 1 h after intramuscular inoculation of RABV efficiently suppressed viral replication at the inoculation site and the subsequent replication in the CNS. However, virus replication in the CNS was not inhibited when the treatment began 2 days after inoculation. We also found that higher doses (600 or 900 mg/kg/day) of favipiravir could suppress viral replication in the CNS even when administration started 2 days after inoculation. These results support our hypothesis and suggest that a highly effective drug-delivery system into the CNS and/or the enhancement of favipiravir conversion to its active form are required to improve favipiravir treatment of rabies. Furthermore, the bioluminescence imaging system established in this study will facilitate the development of treatment for symptomatic rabies.
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Near-infrared fluorescent protein iRFP720 is optimal for &ITin vivo&IT fluorescence imaging of rabies virus infection Reviewed
Minori Isomura, Kentaro Yamada, Kazuko Noguchi, Akira Nishizono
JOURNAL OF GENERAL VIROLOGY 98 ( 11 ) 2689 - 2698 2017.11
Authorship：Corresponding author Language：English Publishing type：Research paper (scientific journal) Publisher：MICROBIOLOGY SOC
In vivo imaging is a noninvasive method that enables real-time monitoring of viral infection dynamics in a small animal, which allows a better understanding of viral pathogenesis. In vivo bioluminescence imaging of virus infection is widely used but, despite its advantage over bioluminescence that no substrate administration is required, fluorescence imaging is not used because of severe autofluorescence. Recently, several far-red and near-infrared (NIR) fluorescent proteins (FPs) have been developed and shown to be useful for whole-body fluorescence imaging. Here, we report comparative testing of far-red and NIR FPs in the imaging of rabies virus (RABV) infection. Using the highly neuroinvasive 1088 strain, we generated recombinant RABV that expressed FPs such as Katushka2S, E2-Crimson, iRFP670 or iRFP720. After intracerebral inoculation to nude mice, the 1088 strain expressing iRFP720, the most red-shifted FP, was detected the earliest with the highest signal-to-noise ratio using a filter set for >700 nm, in which the background signal level was very low. Furthermore, we could also track viral dissemination from the spinal cord to the brain in nude mice after intramuscular inoculation of iRFP720-expressing 1088 into the hind limb. Hence, we conclude that the NIR FP iRFP720 used with a filter set for >700 nm is useful for in vivo fluorescence imaging not only for RABV infection but also for other virus infections. Our findings will also be useful for developing dual-optical imaging of virus-host interaction dynamics using bioluminescence reporter mice for inflammation imaging.
DOI： 10.1099/jgv.0.000950
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A Comparative Study of the RAPINA and the Virus-Neutralizing Test (RFFIT) for the Estimation of Antirabies-Neutralizing Antibody Levels in Dog Samples Reviewed International coauthorship
D. L. Manalo, K. Yamada, I. Watanabe, M. E. G. Miranda, S. M. D. Lapiz, E. Tapdasan, W. Petspophonsakul, S. Inoue, P. Khawplod, A. Nishizono
ZOONOSES AND PUBLIC HEALTH 64 ( 5 ) 355 - 362 2017.8
Language：Japanese Publishing type：Research paper (scientific journal) Publisher：WILEY
The mass vaccination of dogs against rabies is a highly rational strategy for interrupting the natural transmission of urban rabies. According to the World Organization for Animal Health (OIE) and the World Health Organization (WHO), the immunization of at least 70% of the total dog population minimizes the risk of endemic rabies. Knowledge of the virus-neutralizing antibody (VNA) level against the rabies virus (RABV) is required to evaluate protective immunity and vaccine coverage of dogs in the field. The rapid focus fluorescent inhibition test (RFFIT) and the fluorescent antibody virus neutralization (FAVN) test are recommended by OIE and WHO to determine the VNA levels in serum. However, these tests are cell culture based and require the use of live viruses and specialized equipment. The rapid neutralizing antibody test (RAPINA) is a novel, immunochromatographic test that uses inactivated virus to estimate the VNA level qualitatively. It is a simple, rapid and inexpensive, although indirect, assay for the detection of VNA levels. The RAPINA has shown good positive and negative predictive values and a high concordance with the RFFIT results. In this study, we compared the performance of the two tests for evaluating the vaccination status of dogs in the Philippines, Thailand and Japan. A total of 1135 dog sera were analysed by the RAPINA and compared to the VNA levels determined by the RFFIT. The overall positive and negative predictive values of the RAPINA were 96.2-99.3% and 84.5-94.8%, respectively, with a concordance (kappa) of 0.946-0.97 among the three countries. The RAPINA results were highly homologous and reproducible among different laboratories. These results suggest that this test is appropriate to survey vaccination coverage in countries with limited resources.
DOI： 10.1111/zph.12313
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Pathological lesions in the central nervous system and peripheral tissues of ddY mice with street rabies virus (1088 strain) Reviewed
Kazunori Kimitsuki, Kentaro Yamada, Nozomi Shiwa, Satoshi Inoue, Akira Nishizono, Chun-Ho Park
JOURNAL OF VETERINARY MEDICAL SCIENCE 79 ( 6 ) 970 - 978 2017.6
Language：English Publishing type：Research paper (scientific journal) Publisher：JAPAN SOC VET SCI
Most studies on rabies virus pathogenesis in animal models have employed fixed rabies viruses, and the results of those employing street rabies viruses have been inconsistent. Therefore, to clarify the pathogenesis of street rabies virus (1088 strain) in mice, 10(6) focus forming units were inoculated into the right hindlimb of ddY mice (6 weeks, female). At 3 days postinoculation (DPI), mild inflammation was observed in the hindlimb muscle. At 5 DPI, ganglion cells in the right lumbosacral spinal dorsal root ganglia showed chromatolysis. Axonal degeneration and inflammatory cells increased with infection progress in the spinal dorsal horn and dorsal root ganglia. Right hindlimb paralysis was observed from 7 DPI, which progressed to quadriparalysis. However, no pathological changes were observed in the ventral horn and root fibers of the spinal cord. Viral antigen was first detected in the right hindlimb muscle at 3 DPI, followed by the right lumbosacral dorsal root ganglia, dorsal horn of spinal cord, left red nuclei, medulla oblongata and cerebral cortex (M1 area) at 5 DPI. These results suggested that the 1088 virus ascended the lumbosacral spinal cord via mainly afferent fibers at early stage of infection and moved to cerebral cortex (M1 area) using descending spinal tract. Additionally, we concluded that significant pathological changes in mice infected with 1088 strain occur in the sensory tract of the spinal cord; this selective susceptibility results in clinical features of the disease.
DOI： 10.1292/jvms.17-0028
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Increased pathogenicity of rabies virus due to modification of a non-coding region Reviewed International coauthorship
Phatthamon Virojanapirom, Kentaro Yamada, Pakamatz Khawplod, Akira Nishizono, Thiravat Hemachudha
ARCHIVES OF VIROLOGY 161 ( 11 ) 3255 - 3261 2016.11
Language：English Publishing type：Research paper (scientific journal) Publisher：SPRINGER WIEN
Sub-passaging of QS-05, a street rabies virus (RABV) isolate, in non-neuronal cells resulted in a virus with higher pathogenicity, QS-BHK-P7. Four full-length cDNA plasmids were constructed and the corresponding recombinant viruses were recovered: rQS-05, rQS-BHK-P7 and rQS05-2475G/rQS-BHK-P7-2475A (made by switching of intergenic P-M between these two backbones). rQS-BHK-P7-2475 A virus had eight instead of seven adenosines in its poly(A) sequence. Interestingly, mutant viruses with 6 or 8 adenosines infected more neuroblastoma cells than their parental ones. Mice that were infected intracerebrally and intramuscularly with rQS05-2475G and rQS-BHK-P7 exhibited highest mortality. However, mice infected with rQS-BHK-P7-2475AA had the shortest survival time. This study demonstrates that modifications in the non-coding region may play a role in determining the virulence of RABV.