研究者詳細 - 小原　恭子

論文 - 小原　恭子

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Kayesh MEH, Kohara M, Tsukiyama-Kohara K . Toll-like Receptor Response to Human Immunodeficiency Virus Type 1 or Co-Infection with Hepatitis B or C Virus: An Overview. . International journal of molecular sciences24 ( 11 ) 2023年6月査読国際誌

Mohammad Enamul Hoque Kayesh, Md Abul Hashem, Takahiro Sanada, Bouchra Kitab, Md Haroon Or Rashid, Lipi Akter, Sayeh Ezzikouri, Shuko Murakami, Shintaro Ogawa, Yasuhito Tanaka, Michinori Kohara and Kyoko Tsukiyama-Kohara . Characterization of innate immune response to hepatitis B virus genotype F acute infection in tree shrew (Tupaia belangeri) model . Frontiers in Virology2 926831 2022年8月査読国際共著国際誌

担当区分：責任著者記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Frontiers Media SA

Hepatitis B virus (HBV) infection is a global public health problem. The clinical outcomes of HBV infections are influenced by host as well as viral factors, including viral genotypes and subgenotypes. The interplay between HBV and host innate immunity remains unclear because of the lack of a suitable small animal model. Tree shrews (Tupaia belangeri) have been utilized as a useful animal model for hepatitis viruses such as hepatitis B and C viruses. In this study, we characterized acute infections by HBV genotype F (HBV-F) wild type (Wt) and mutant type (Mt) viruses in adult tree shrews. Serum alanine aminotransferase levels were measured before and post- infection 7 and 14 dpi. Both HBV-F-Wt and Mt were detected in the HBV-F-infected tree shrew serum and liver tissue at 7 and 14 dpi. We examined the intrahepatic expression patterns of Toll-like receptors (TLRs) (TLR1–9 mRNAs), cGAS, several transcription factors such as STAT1, STAT2, IRF7, HNF4, PD-L1, and cytokines, including IFN-β, IFN-γ, IL-6, and TNF-α in HBV-F Wt/Mt-infected tree shrews. When compared with uninfected animal group, significant suppression of TLR8 in HBV-F-Wt infected animals and significant suppression of PD-L1 in both HBV-F-Wt and Mt infected animals were observed. Thus, tree shrew can be a useful animal model to characterize HBV-F pathogenesis.

DOI： 10.3389/fviro.2022.926831

Hashem M.A., Kayesh M.E.H., Maetani F., Goto A., Nagata N., Kasori A., Imanishi T., Tsukiyama-Kohara K. . Subtype distribution and expression of the koala retrovirus in the Japanese zoo koala population . Infection, Genetics and Evolution102 105297 - 105297 2022年8月査読国際誌

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Infection, Genetics and Evolution

We investigated the proviral copies and RNA expression in koala retrovirus (KoRV)-infected koalas. To ascertain any variation in viral load by institution, age, sex, or body condition score, we quantified KoRV proviral DNA and RNA loads in captive koalas (n = 37) reared in Japanese zoos. All koalas were positive for KoRV genes (pol, LTRs, and env of KoRV-A) in genomic DNA (gDNA), and 91.89% were positive for the pol gene in RNA. In contrast, the distribution rates of KoRV-B, KoRV-C, KoRV-D, and KoRV-F env genes in gDNA were 94.59%, 27.03%, 67.57%, and 54.05%, respectively. A wide inter-individual variation and/or a significant inter-institutional difference in proviral DNA (p < 0.0001) and RNA (p < 0.001) amounts (copies/103 koala β-actin copies) were observed in Awaji Farm England Hill Zoo koalas, which were obtained from southern koala populations, suggesting exogenous incorporation of KoRV in these koalas. Significant (p < 0.05) age differences were noted in KoRV RNA load (p < 0.05) and median total RNA load (p < 0.001), with loads higher in younger koalas (joeys and juveniles). Thus, the current study provides the distribution of KoRV subtypes in Japanese zoo koala populations and identifies several additional risk factors (sex, age, and body condition) associated with KoRV expression.

DOI： 10.1016/j.meegid.2022.105297

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Reverse vaccinology-based prediction of a multi-epitope SARS-CoV-2 vaccine and its tailoring to new coronavirus variants. . 2022年5月査読国際共著

Kayesh MEH, Kohara M, Tsukiyama-Kohara K. . Toll-like Receptor Response to Hepatitis C Virus Infection: A Recent Overview. . Int J Mol Sci.23 ( 10 ) 5475 2022年5月査読国際共著

Kayesh M.E.H., Kohara M., Tsukiyama-Kohara K. . Toll-like Receptor Response to Hepatitis C Virus Infection: A Recent Overview . International Journal of Molecular Sciences23 ( 10 ) 2022年5月査読国際誌

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：International Journal of Molecular Sciences

Hepatitis C virus (HCV) infection remains a major global health burden, causing chronic hepatitis, cirrhosis, and hepatocellular carcinoma. Toll-like receptors (TLRs) are evolutionarily con-served pattern recognition receptors that detect pathogen-associated molecular patterns and acti-vate downstream signaling to induce proinflammatory cytokine and chemokine production. An in-creasing number of studies have suggested the importance of TLR responses in the outcome of HCV infection. However, the exact role of innate immune responses, including TLR response, in controlling chronic HCV infection remains to be established. A proper understanding of the TLR response in HCV infection is essential for devising new therapeutic approaches against HCV infection. In this review, we discuss the progress made in our understanding of the host innate immune response to HCV infection, with a particular focus on the TLR response. In addition, we discuss the mechanisms adopted by HCV to avoid immune surveillance mediated by TLRs.

DOI： 10.3390/ijms23105475

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Ezzemani W., Kettani A., Sappati S., Kondaka K., El Ossmani H., Tsukiyama-Kohara K., Altawalah H., Saile R., Kohara M., Benjelloun S., Ezzikouri S. . Reverse vaccinology-based prediction of a multi-epitope SARS-CoV-2 vaccine and its tailoring to new coronavirus variants . Journal of Biomolecular Structure and Dynamics 1 - 22 2022年5月査読国際誌

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Journal of Biomolecular Structure and Dynamics

The genome feature of SARS-CoV-2 leads the virus to mutate and creates new variants of concern. Tackling viral mutations is also an important challenge for the development of a new vaccine. Accordingly, in the present study, we undertook to identify B- and T-cell epitopes with immunogenic potential for eliciting responses to SARS-CoV-2, using computational approaches and its tailoring to coronavirus variants. A total of 47 novel epitopes were identified as immunogenic triggering immune responses and no toxic after investigation with in silico tools. Furthermore, we found these peptide vaccine candidates showed a significant binding affinity for MHC I and MHC II alleles in molecular docking investigations. We consider them to be promising targets for developing peptide-based vaccines against SARS-CoV-2. Subsequently, we designed two efficient multi-epitopes vaccines against the SARS-CoV-2, the first one based on potent MHC class I and class II T-cell epitopes of S (FPNITNLCPF–NYNYLYRLFR–MFVFLVLLPLVSSQC), M (MWLSYFIASF–GLMWLSYFIASFRLF), E (LTALRLCAY–LLFLAFVVFLLVTLA), and N (SPRWYFYYL–AQFAPSASAFFGMSR). The second candidate is the result of the tailoring of the first designed vaccine according to three classes of SARS-CoV-2 variants. Molecular docking showed that the protein-protein binding interactions between the vaccines construct and TLR2–TLR4 immune receptors are stable complexes. These findings confirmed that the final multi-epitope vaccine could be easily adapted to new viral variants. Our study offers a shortlist of promising epitopes that can accelerate the development of an effective and safe vaccine against the virus and its adaptation to new variants. Communicated by Ramaswamy H. Sarma.

DOI： 10.1080/07391102.2022.2075468

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Hashem MA, Kayesh MEH, Maetani F, Goto A, Nagata N, Kasori A, Imanishi T, Tsukiyama-Kohara K . Subtype distribution and expression of the koala retrovirus in the Japanese zoo koala population. . Infect Genet Evol. 105297 2022年5月査読国際共著

Ide Y., Kitab B., Ito N., Okamoto R., Tamura Y., Matsui T., Sakoda Y., Tsukiyama-Kohara K. . Characterization of host factors associated with the internal ribosomal entry sites of foot-and-mouth disease and classical swine fever viruses . Scientific Reports12 ( 1 ) 6709 - 6709 2022年4月査読国際誌

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Scientific Reports

Foot-and-mouth disease virus (FMDV) and classical swine fever virus (CSFV) possess positive-sense single-stranded RNA genomes and an internal ribosomal entry site (IRES) element within their 5′-untranslated regions. To investigate the common host factors associated with these IRESs, we established cell lines expressing a bicistronic luciferase reporter plasmid containing an FMDV-IRES or CSFV-IRES element between the Renilla and firefly luciferase genes. First, we treated FMDV-IRES cells with the French maritime pine extract, Pycnogenol (PYC), and examined its suppressive effect on FMDV-IRES activity, as PYC has been reported to have antiviral properties. Next, we performed microarray analysis to identify the host factors that modified their expression upon treatment with PYC, and confirmed their function using specific siRNAs. We found that polycystic kidney disease 1-like 3 (PKD1L3) and ubiquitin-specific peptidase 31 (USP31) were associated with FMDV-IRES activity. Moreover, silencing of these factors significantly suppressed CSFV-IRES activity. Thus, PKD1L3 and USP31 are host factors associated with the functions of FMDV- and CSFV-IRES elements.

DOI： 10.1038/s41598-022-10437-z

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Kayesh M.E.H., Hashem M.A., Kohara M., Tsukiyama-Kohara K. . In vivo Delivery Tools for Clustered Regularly Interspaced Short Palindromic Repeat/Associated Protein 9-Mediated Inhibition of Hepatitis B Virus Infection: An Update . Frontiers in Microbiology13 953218 - 953218 2022年査読国際誌

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Frontiers in Microbiology

Chronic hepatitis B virus (HBV) infection remains a major global health problem despite the availability of an effective prophylactic HBV vaccine. Current antiviral therapies are unable to fully cure chronic hepatitis B (CHB) because of the persistent nature of covalently closed circular DNA (cccDNA), a replicative template for HBV, which necessitates the development of alternative therapeutic approaches. The CRISPR/Cas system, a newly emerging genome editing tool, holds great promise for genome editing and gene therapy. Several in vitro and/or in vivo studies have demonstrated the effectiveness of HBV-specific clustered regularly interspaced short palindromic repeat (CRISPR)/associated protein 9 (CRISPR/Cas9) systems in cleaving HBV DNA and cccDNA. Although recent advances in CRISPR/Cas technology enhance its prospects for clinical application against HBV infection, in vivo delivery of the CRISPR/Cas9 system at targets sites remains a major challenge that needs to be resolved before its clinical application in gene therapy for CHB. In the present review, we discuss CRISPR/Cas9 delivery tools for targeting HBV infection, with a focus on the development of adeno-associated virus vectors and lipid nanoparticle (LNP)-based CRISPR/Cas ribonucleoprotein (RNP) delivery to treat CHB. In addition, we discuss the importance of delivery tools in the enhancement of the antiviral efficacy of CRISPR/Cas9 against HBV infection.

DOI： 10.3389/fmicb.2022.953218

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Kayesh M.E.H. . Toll-like receptor response to hepatitis Bb virus infection and potential of TLR agonists as immunomodulators for treating chronic hepatitis b: An overview . International Journal of Molecular Sciences22 ( 19 ) 2021年9月査読国際誌

Abbadi I. . Non-primate hepacivirus transmission and prevalence: Novel findings of virus circulation in horses and dogs in Morocco . Infection, Genetics and Evolution93 104975 - 104975 2021年9月査読国際誌

ケアシュ・モハメド, 尼子豊, ハシェム・モハメド, 山本直樹, 杉山真也, 溝上雅史, 小原道法, 小原恭子 . B型肝炎ウイルスcccDNAを標的としたCRISPR/Cas9システムの生体内送達法の開発 . 日本癌学会総会記事80回 [J3 - 5] 2021年9月査読国際誌

Honda T, Gomi S, Yamane D, Yasui F, Yamamoto T, Munakata T, Itoh Y, Ogasawara K, Sanada T, Yamaji K, Yasutomi Y, Tsukiyama-Kohara K, Kohara M. . Development and Characterization of a Highly Sensitive NanoLuciferase-Based Immunoprecipitation System for the Detection of Anti-Influenza Virus HA . mSphere6 ( 3 ) e1342-20 2021年5月査読

Honda T. . Development and Characterization of a Highly Sensitive NanoLuciferase-Based Immunoprecipitation System for the Detection of Anti-Influenza Virus HA Antibodies . mSphere6 ( 3 ) 1 - 14 2021年5月査読国際誌

Saito M. . Macrocyclic peptides exhibit antiviral effects against influenza virus HA and prevent pneumonia in animal models . Nature Communications12 ( 1 ) 2654 - 2654 2021年5月査読国際誌

Toll-Like Receptor and Cytokine Responses to Infection with Endogenous and Exogenous Koala Retrovirus, and Vaccination as a Control Strategy . Curr Issues Mol Biol. 43 ( 1 ) 52 - 64 2021年4月査読国際共著

Md Abul Hashem1, 2, 3, Mohammad Enamul Hoque Kayesh1, 2, 4, Fumie Maetani5, Taiki Eiei5, Kyoya Mochizuki5, Shinsaku Ochiai5, Ayaka Ito5, Nanao Ito5, Hiroko Sakurai5, Takayuki Asai5 and Kyoko Tsukiyama-Kohara . Koala retrovirus (KoRV) subtypes and their impact on captive koala (Phascolarctos cinereus) health . Archives of Virologyaccepted 2021年4月査読

Mohammad Enamul Hoque Kayesh 1,2 , Md Abul Hashem 1,3,4 and Kyoko Tsukiyama-Kohara . Toll-Like Receptor Expression Profiles in Koala (Phascolarctos cinereus) Peripheral Blood Mononuclear Cells Infected with Multiple KoRV Subtypes . 11 983 2021年4月査読

Kayesh M.E.H., Hashem M.A., Tsukiyama-Kohara K. . Toll-like receptor expression profiles in koala (Phascolarctos cinereus) peripheral blood mononuclear cells infected with multiple korv subtypes . Animals11 ( 4 ) 2021年4月査読国際誌

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Animals

Toll-like receptors (TLRs), evolutionarily conserved pattern recognition receptors, play an important role in innate immunity by recognizing microbial pathogen-associated molecular patterns. Koala retrovirus (KoRV), a major koala pathogen, exists in both endogenous (KoRV-A) and exogenous forms (KoRV-B to J). However, the expression profile of TLRs in koalas infected with KoRV-A and other subtypes is yet to characterize. Here, we investigated TLR expression profiles in koalas with a range of subtype infection profiles (KoRV-A only vs. KoRV-A with KoRV-B and/or-C). To this end, we cloned partial sequences for TLRs (TLR2–10 and TLR13), developed real-time PCR assays, and determined TLRs mRNA expression patterns in koala PBMCs and/or tissues. All the reported TLRs for koala were expressed in PBMCs, and variations in TLR expression were observed in koalas infected with exogenous subtypes (KoRV-B and KoRV-C) compared to the endogenous subtype (KoRV-A) only, which indicates the implications of TLRs in KoRV infection. TLRs were also found to be differentially expressed in koala tissues. This is the first report of TLR expression profiles in koala, which provides insights into koala’s immune response to KoRV infection that could be utilized for the future exploitation of TLR modulators in the maintenance of koala health.

DOI： 10.3390/ani11040983

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