B型肝炎ワクチン応答とHBV排除の免疫ゲノム解析に基づくキャリア化阻止戦略の構築

2019 Fiscal Year Annual Research Report

• Project/Area Number: 17H04168
• Research Institution: National Center for Global Health and Medicine
• Principal Investigator: 考藤 達哉 国立研究開発法人国立国際医療研究センター, その他部局等, 研究センター長 (80372613)
• Co-Investigator(Kenkyū-buntansha): 西田 奈央 国立研究開発法人国立国際医療研究センター, その他部局等, 上級研究員 (50456109) ; 田中 純子 広島大学, 医系科学研究科(医), 教授 (70155266) ; 小松 陽樹 東邦大学, 医学部, 准教授 (80424711)
• Project Period (FY): 2017-04-01 – 2020-03-31
• Keywords: B型肝炎ワクチン / 母子感染 / 水平感染 / 免疫 / 遺伝要因 / ウイルスゲノム / カンボジア / 乾燥濾紙血法（DBS）

Outline of Annual Research Achievements

HBワクチンによるHBs抗体誘導機序を明らかにするために、B型肝炎患者のPBMCを用いてHBs抗原・HBc抗原特異的B細胞の検出系を確立した。濾胞性ヘルパーT細胞（TFH）-B細胞系列を効果的に活性化する免疫アジュバントの探索を目的として、健康成人・B型慢性肝炎患者のPBMCからTFHを誘導するIn vitro培養系を樹立した。TLRリガンドによる形質細胞様樹状細胞の賦活がTFH誘導に重要であり、アジュバントとして期待できることを明らかにした。
カンボジア4,541人を対象にした乾燥濾紙血法(DBS)による検討ではHBs抗原陽性率は小児0.5%、母親4.7%であった。HBs抗原陽性/HBV DNA陽性の母70名と児12名に関する系統樹解析では、genotype C（80.49％）をカンボジア全体で認め、genotype B（19.51％）はベトナムとの国境近傍に認めた。S遺伝子変異を伴う17名の母親から出生した児のうち、HBワクチン接種既往がある児12名にはS遺伝子変異を認めず、接種既往がない児５名にはS gene 変異を認めた。
カンボジア人HBV感染患者497検体からDBS法で抽出したゲノムDNAを用いて、HLA-DPB1タイピング（PCR-Luminex法）及び疾患関連遺伝子（CXCL13等）SNPsのタイピングを実施した。疾患抵抗性アリルHLA-DPB1*04:01と、疾患感受性アリルHLA-DPB1*13:01を同定した。疾患関連SNPsは有意な差を認めなかった。
HBワクチンによる母子感予防不成功2家族において、臍帯と爪からHBVワクチンエスケープ変異株を検出した。HBVキャリア児に対するペグインターフェロン療法施行中のサイトカイン動態を検討した。著効例は非著効例と比べて血清CXCL9, 10, 11, 13が治療開始24週までに高値を示す傾向があることを見出した。

Research Progress Status

令和元年度が最終年度であるため、記入しない。

Strategy for Future Research Activity

令和元年度が最終年度であるため、記入しない。

Research Products

• [Int'l Joint Research] WHO カンボジア/カンボジア王国保健省(カンボジア)
  • Country Name: CAMBODIA
  • Counterpart Institution: WHO カンボジア/カンボジア王国保健省
• [Int'l Joint Research] CDC(米国)
  • Country Name: U.S.A.
  • Counterpart Institution: CDC
• [Int'l Joint Research] ヴィントン医科大学(ベトナム)
  • Country Name: VIET NAM
  • Counterpart Institution: ヴィントン医科大学
• [Journal Article] A prospective trial of vaccine to prevent hepatitis B virus reactivation after hematopoietic stem cell transplantation. (2020)
  • Author(s): Nishikawa K, Kimura K, Kanda Y, Sugiyama M, Kakihana K, Doki N, Ohashi K, Bae SK, Takahashi K, Ishihara Y, Mizuno I, Onishi Y, Onozawa M, Onizuka M, Yamamoto M, Ishikawa T, Inoue K, Kusumoto S, Hashino S, Saito H, Kanto T, Sakamaki H, Mizokami M.
  • Journal Title: Bone Marrow Transplantation Volume: - Pages: -
  • DOI: 10.1038/s41409-020-0833
  • Peer Reviewed
• [Journal Article] Evaluation of the efficiency of dried blood spot-based measurement of hepatitis B and hepatitis C virus seromarkers (2020)
  • Author(s): Yamamoto C, Nagashima S, Isomura M, Ko K, Chuon C, Akita T, Katayama K, Woodring J, Hossain MS, Takahashi K, Tanaka J
  • Journal Title: Scientific Reports Volume: 10 Pages: 3857
  • DOI: 10.1038/s41598-020-60703-1.
  • Peer Reviewed / Open Access
• [Journal Article] Discordance of hepatitis B vaccination policies for healthcare workers between the USA, the UK, and Germany (2020)
  • Author(s): Komatsu Haruki、Klenerman Paul、Thimme Robert
  • Journal Title: Hepatology Research Volume: 50 Pages: 272～282
  • DOI: 10.1111/hepr.13470
  • Peer Reviewed
• [Journal Article] Existence of hepatitis B virus surface protein mutations and other variants: demand for hepatitis B infection control in Cambodia (2020)
  • Author(s): Ko K, Takahashi K, Nagashima S, Yamamoto C, Ork V, Sugiyama A, Akita T, Ohisa M, Chuon C, Shafiqul Hossain M, Mao B, Tanaka J,
  • Journal Title: BMC Infectious Disease Volume: 20 Pages: 305
  • DOI: 10.1186/s12879-020-05025-3.
  • Peer Reviewed / Open Access
• [Journal Article] Immune Determinants in the Acquisition and Maintenance of Anti-HBs in Adults After First-Time Hepatitis B Vaccination. (2019)
  • Author(s): 3.Doi H, Yoshio S, Yoneyama K, Kawai H, Sakamoto Y, Shimagaki T, Aoki Y, Osawa Y, Yoshida H, Kanto T
  • Journal Title: Hepatology Communications Volume: 3 Pages: 812-824
  • DOI: 10.1002/hep4.1357.
  • Peer Reviewed / Open Access
• [Journal Article] Hepatitis B surface antigen seroprevalence among pre- and post-vaccine cohorts in Cambodia, 2017 (2019)
  • Author(s): Ork V, Woodring J, Shafiqul Hossain M, Wasley A, Nagashima S, Yamamoto C, Chuon C, Sugiyama A, Ohisa M, Akita T, Ko K, Mao B, Tanaka J
  • Journal Title: Vaccine Volume: 37 Pages: 5059-5066
  • DOI: 10.1016/j.vaccine.2019.06.073
  • Peer Reviewed
• [Journal Article] Acquisition rate of antibody to hepatitis B surface antigen among medical and dental students in Japan after three-dose hepatitis B vaccination (2019)
  • Author(s): Nagashima S, Yamamoto C, Ko K, Chuon C, Sugiyama A, Ohisa M, Akita T, Katayama K, Yoshihara M, Tanaka J
  • Journal Title: Vaccine Volume: 37 Pages: 145-151
  • DOI: 10.1016/j.vaccine.2018.11.019
  • Peer Reviewed
• [Journal Article] High possibility of hepatocarcinogenesis in HBV genotype C1 infected Cambodians is indicated by 340 HBV C1 full-genomes analysis from GenBank. (2019)
  • Author(s): Chuon C, Takahashi K, Matsuo J, Katayama K, Yamamoto C, Ko K, Hok S, Nagashima S, Akbar SMF, Tanaka J
  • Journal Title: Scientific Reports Volume: 9 Pages: 12186
  • DOI: 10.1038/s41598-019-48304-z
  • Peer Reviewed / Open Access
• [Journal Article] POGLUT1, the Putative Effector Gene Driven by rs2293370 in Primary Biliary Cholangitis Susceptibility Locus Chromosome 3q13.33. (2019)
  • Author(s): Hitomi Y, Ueno K, Kawai Y, Nishida N, et al.
  • Journal Title: Scientific Reports Volume: 9 Pages: 102
  • DOI: 10.1038/s41598-018-36490-1.
  • Peer Reviewed / Open Access
• [Journal Article] NFKB1 and MANBA Confer Disease Susceptibility to Primary Biliary Cholangitis via Independent Putative Primary Functional Variants. (2019)
  • Author(s): Hitomi Y, Nakatani K, Kojima K, Nishida N, Kawai Y, Kawashima M, Aiba Y, Nagasaki M, Nakamura M, Tokunaga K.
  • Journal Title: Cell Mol Gastroenterol Hepatol. Volume: 7 Pages: 515-532
  • DOI: 10.1016/j.jcmgh.2018.11.006.
  • Peer Reviewed
• [Journal Article] Integrated GWAS and mRNA Microarray Analysis Identified IFNG and CD40L as the Central Upstream Regulators in Primary Biliary Cholangitis. (2019)
  • Author(s): Ueno K, Aiba Y, Hitomi Y, Shimoda S,et al.
  • Journal Title: Hepatology Communication Volume: 4 Pages: 724-738
  • DOI: 10.1002/hep4.1497
  • Peer Reviewed / Open Access
• [Journal Article] Liver disease secondary to congenital heart disease in children. (2019)
  • Author(s): Komatsu H, Inui A, Kishiki K, Kawai H, Yoshio S, Osawa Y, Kanto T, Fujisawa T.
  • Journal Title: Expert Rev Gastroenterol Hepatol. Volume: 49 Pages: 651-666
  • DOI: 10.1080/17474124.2019.
  • Peer Reviewed
• [Journal Article] Loss of fibrocystin promotes interleukin-8-dependent proliferation and CTGF production of biliary epithelium. (2019)
  • Author(s): Tsunoda T, Kakinuma S, Miyoshi M, Kamiya A, Kaneko S, Sato A, Tsuchiya J, Nitta S, Kawai-Kitahata F, Murakawa M, Itsui Y, Nakagawa M, Azuma S, Sogo T, Komatsu H, Mukouchi R, Inui A, Fujisawa T, Nakauchi H, Asahina Y, Watanabe M.
  • Journal Title: Journal of Hepatology Volume: 71 Pages: 143-152
  • DOI: 10.1016/j.jhep.2019.02.024.
  • Peer Reviewed
• [Presentation] Increased frequency of PD-1+CD57+Siglec-7-dysfunctional NK cells in patients with non-alcoholic fatty liver disease. (2019)
  • Author(s): Sakamoto Y, Yoshio S, Kanto T.
  • Organizer: The 10th Asia-Pacific Primary Liver Cancer Expert Meeting,
  • Int'l Joint Research
• [Presentation] Sphingosine-1-phosphate as a mediator of peripheral-dominant liver fibrosis in mice model of congestive hepatopathy. (2019)
  • Author(s): Kawai H, Osawa Y, Sakamoto Y, Tsutsui Y, Yamazoe T, Mori T, Yoshikawa S, Yoshio S, Kanto T.
  • Organizer: The 1st JSH International Liver Conference
  • Int'l Joint Research
• [Presentation] Macrophages and NK cells as a source of biomarkers in patients with NAFLD. (2019)
  • Author(s): Kanto T
  • Organizer: The 1st JSH International Liver Conference
  • Int'l Joint Research / Invited
• [Presentation] Siglec-7 as an indicator of dysregulated NK cells and macrophage inflammation in patients with non-alcoholic fatty liver disease. (2019)
  • Author(s): Sakamoto Y, Yoshio S, Kawai H, Mori T, Aoki Y, Yoshida Y, Arai T, Itokawa N, Osawa Y, Taketomi A, Kanto T.
  • Organizer: AASLD 2019
  • Int'l Joint Research
• [Presentation] Impact of metabolic stress on the pathogenesis of autoimmune liver disease in subjects bearing missense variant of negative immune regulator Lnk/Sh2b3. (2019)
  • Author(s): Taizo Mori, Sachiyo Yoshio, Michitaka Matsuda, Yuzuru Sakamoto, Tomonari Shimagaki, Hironari Kawai, Yosuke Osawa, Tatsuya Kanto.
  • Organizer: International Liver Conference (ILC) 2019
  • Int'l Joint Research
• [Presentation] Epidemiological Assessment of the interventions for elimination of mother-to-child transmission of hepatitis B virus in Japan, (2019)
  • Author(s): Tanaka J, Sugiyama A, Ko K, Yamamoto C
  • Organizer: AASLD The Liver Meeting 2019
  • Int'l Joint Research
• [Presentation] Development of a dried blood spot-based host genome analysis method for hepatitis B-related genes and its clinical application in Cambodia. (2019)
  • Author(s): Setoyama H, Nishida N, Tanaka J, Mizokami M, Sasaki Y, Kanto T.
  • Organizer: AASLD The Liver Meeting 2019
  • Int'l Joint Research
• [Presentation] 18 years follow-up large cohort study on epidemiology of hepatitis C among hemodialysis patients, their long-term prognosis and related risk factors (2019)
  • Author(s): Ko K, Nagashima S, Yamamoto C, Akita T, Ohisa M, Sugiyama A, Katayama K, Takahashi K, Tanaka J,
  • Organizer: 25th International Symposium on Hepatitis C virus and Related Viruses(HCV2019)
  • Int'l Joint Research
• [Presentation] Host factors associated with HBV related HCC in Asian populations (2019)
  • Author(s): Nao Nishida, Masaya Sugiyama, Hiromi Sawai, Yong Poovorawan, Katsushi Tokunaga, and Masashi Mizokami
  • Organizer: AASLD Meeting 2019
  • Int'l Joint Research
• [Remarks] 肝炎・免疫研究センター 肝疾患研究部
  • URL: http://www.ncgmkohnodai.go.jp/kanen_meneki/index.html
• [Patent(Industrial Property Rights)] B型肝炎ワクチンに対する免疫応答性に関わる遺伝子 (2019)
  • Inventor(s): 西田奈央、溝上雅史、徳永勝士
  • Industrial Property Rights Holder: 西田奈央、溝上雅史、徳永勝士
  • Industrial Property Rights Type: 特許
  • Industrial Property Number: 特願2019-200888