E E Hepatitis E SARS 29, Lancet. E A B Enterically-Transmitted Non-A, Hepatitis E. Virus HEV nm. 1.35g/cm s ALT AST HEV HEV

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7850 2004 Hepatitis E Tian-Cheng LI Naokazu TAKEDA Tatsuo MIYAMURA SARS 8 Lancet E E E Hepatitis E VirusHEV E E HEV HEV E 1955 29,000 E E A A B Enterically-Transmitted Non-A, Non-B Hepatitis 1983 Balayan A B A IgG 27 30nm 1.35g/cm 3 176 183s ALT AST A B 1990 Reyes cdna E E 2 208-0011 Department of Virology, II, National Instituite of Infectious Diseases (Gakuen 4-7-1, Musashi-murayama, Tokyo) 6

79 HEV water-borne hepatitis virus HEV HEV HEV HCV HBV E HEV HEV HEV 50nm HEV HEV E 30nm 50nm 1991 Tam HEV HEV 4 Genotype 90 Genotype 1G 1 81 82 Genotype 2G 2 G2 10 10 E US1 US2 1112 78 80 S1 Genotype 3G 3 13 5159 U Genotype 4G 4 14 E G3 G4 1517 HEV 7.2Kb RNA HEV ORF2 7

80 5 cap 3 7,200 HEV 3 ORF1ORF3 ORF2 5 18 5 27 5,000 ORF1 Y X RNA RNA 3 2,000 ORF2 72 kda ORF3 ORF1 ORF2 IgG IgM RNA RT-PCR IgM E 1 20 21 E E HEV HEV RNA HEV E 1920 HEV FRhK42BSA549 HEV HEV E A E HEV 15 50 4 HAV E 0 10 AST ALT HEV HEV 7.5kb RNA ORF1 ORF2 ORF3 MT: Y: YPro: PR: X: X H: RdRp: RNA RNA HEV E AST ALT IgM IgG IgM 3 IgG HEVRNA HEVRNA 8

81 ALT AST HEV RT-PCR ELISA RT-PCR ORF1 N 500 ORF2 500 RNA HEV RNA RNA HEV ELISA 2223 111 23 24nm Virus-like particlevlp 24 VLP ELISA HEV IgM IgG IgG IgM E 900 IgG ELISA A B C 0 70 A C 29 IgG B 2 IgG 0.42/534 30 IgG A B C 1.914.1 3.5 3 9

82 HEV 5.4 25 100nm 23.7nm 1.285g/cm 3 100nm E 10 98 A B C IgG IgM 23 23.5 IgG IgM E HEV A B C E HEV 32 3 9.4% 10 3 30% HEV 56 17 30.4% E RNA ORF1 ORF2 RT- PCR 3 G3 17 G1 G4 3 G1 87 A B C 11 13% E 15 HEV A B C 1 E 40 ALT AST (1,000 IU/L ) G3 G4 G3 G4 HEV 60% 7% 40% 87% 90% HEV 26 HEV 1.9% 27 HEV IgG 10

83 13.3 31.5% 36% 2829 HEV 30 31 HEV E E DNA DNA Gene gun ORF2 HEV HEV 32 ORF2 56kDa HEV G1 G2G3 HEV 33 56kDa VLP BALB/c IgGIgM IgA 34 IgA HEV HEV VLP IgG 35 VLP 1997 Meng S1 13 E US1 US2 HEV 20 96% 44%36%9% 44 94% 29 62%42 67% 12% IgG 3637 HEV 26.7% 8% 38 HEV HEV HEV 100% 39 HEV HEV 50 60 HEV 4041 HEV G3 G4 HEV 11

84 HEV HEV HEV 42 G1 G2 HEV G3 G4 G1 G2 HEV HEV HEV HEV G3 E HEV HEV E E HEV E E HEV Tei S., et al.: Zoonotic transmission of hepatitis E virus from deer to human beings. Lancet. (9381): 371-373, 2003. Takahashi K., et al.: Full-genome nucleotide sequence of a hepatitis E virus strain that may be indigenous to Japan. Virology (1): 9-12, 2001. Purcell R.H.: Hepatitis E virus, In B. N. Fields, D. M. Knipe, P. M. Howley, R. M. Chanock, J. L. Melnick, T. P. Monath, B. Roizman, and S. E. Straus (ed.), Fields virology, 3rd ed. Lippincott-Raven Publishers, Philadelphia. 1996. p. 2831-2843. Jameel S., et al.: Enteric non-a, non-b hepatitis: epidemics, animal transmission, and hepatitis E virus detection by the polymerase chain reaction. J Med Virol, (4): 263-270, 1992. Balayan M.S., et al.: Evidence for a virus in non-a, non-b hepatitis transmitted via the fecal-oral route. Intervirology : 23-31, 1983. Bradley D., et al.: Aetiological Agent of Enterically Transmitted non-a, non-b Hepatitis. J. gen. Virol., : 731-738, 1988. Reyes G.R., et al.: Isolation of a cdna from the virus responsible for enterically transmitted Non-A, Non-B hepatitis. Science (4948), 1335-1339, 1990. Chauhan A., et al.: Hepatitis E virus transmission to a volunteer. Lancet. (8838):149-150, 1993. Tam A.W., et al.: Hepatitis E virus (HEV): molecular cloning and sequencing of the full-length viral genome. Virology (1), 120-131, 1991. 10Maila H.T., Bowyer S.M., Swanepoel R.: Identification of a new strain of hepatitis E virus from an outbreak in Namibia in 1995. J. Gen. Virol : 89-95, 2004. 11Schlauder GG., et al.: The sequence and phylogenetic analysis of a novel hepatitis E virus isolated from a patient with acute hepatitis reported in the United States. J Gen Virol (3): 447-456, 1998. 12Erker J.C., et al.: A hepatitis E virus variant from the United States: molecular characterization and transmission in cynomolgus macaques. J Gen Virol. (3): 681-690,1999. 13Meng X.J., et al.: A novel virus in swine is closely related to the human hepatitis E virus. Proc. Natl. Acad. Sci. USA : 9860-9865, 1997. 14Wang Y., et al.: A divergent genotype of hepatitis E virus in Chinese patients with acute hepatitis. J Gen Virol : 169-177, 1999. 15Mizuo H., et al.: Polyphyletic strains of hepatitis E virus are responsible for sporadic cases of acute hepatitis in Japan. J. Clin. Microbiol (9):3209-3218, 2002. 16Takahashi K., et al.: Genetic heterogeneity of hepatitis E virus recovered from Japanese patients with acute spo- 12

85 radic hepatitis. J Infect Dis. (9): 1342-1345. 2002. 17Okamoto H., et al.: Analysis of the complete genome of indigenous swine hepatitis E virus isolated in Japan. Biochem Biophys Res Commun (5): 929-936, 2001. 18Reyes GR., et al.: Molecular organization and replication of hepatitis E virus (HEV). Arch Virol [Suppl], : 15-25, 1993. 19Berke T., Matson, DO.: Reclassification of the Caliciviridae into distinct genera and exclusion of hepatitis E virus from the family on the basis of comparative phylogenetic analysis. Arch Virol (7): 1421-1436, 2000. 20ICTV approved Virus Orders, Families and Genera. (http://www.ictvdb.iacr.ac.uk/ictv/fr-fst-g.htm) 21Khuroo M.S., et al.: Incidence and severity of viral hepatitis in pregnancy. Am J Med : 252-255, 1981. 22Tsarev S.A., et al.: ELISA for antibody to hepatitis E virus (HEV) based on complete open reading frame 2 protein expressed in insect cells: identification of HEV infection in primates. J. Infect. Dis (2): 369-378, 1993. 23Purdy M.A., et al.: Expression of a hepatitis E virus (HEV)-trpE fusion protein containing epitopes recognized by antibodies in sera from human cases and experimentally infected primates. Microbiol Immunol, (6): 615-621, 1992. 24T-C Li., et al.: Expression and self-assembly of empty virus-like par ticles of hepatitis E virus. J. Virol : 7207-7213, 1997. 25T-C Li., et al.: An empty virus-like particlebased enzymelinked immunosorbent assay for antibodies to hepatitis E virus,.j Med Virol : 327-333, 2000. 26Takahashi M., et al.: Swine hepatitis E virus strains in Japan form four phylogenetic clusters comparable with those of Japanese isolates of human hepatitis E virus. J Gen Virol. (Pt 4): 851-862, 2003. 27Yazaki Y., et al.: Sporadic acute or fulminant hepatitis E in Hokkaido, Japan, may be food-borne, as suggested by the presence of hepatitis E virus in pig liver as food. J Gen Virol (9): 2351-2357, 2003. 28Hirano M., et al.: Evidence for widespread infection of hepatitis E virus among wild rats in Japan.Hepatol Res. (1): 1-5, 2003. 29Hirano M., et al.: Prevalence of antibody against hepatitis E virus in various species of non-human primates: evidence of widespread infection in Japanese monkeys (Macaca fuscata). Jpn J Infect Dis. (1): 8-11, 2003. 30Matsuda H., et al.: Severe hepatitis E virus infection after ingestion of uncooked liver from a wild boar. J Infect Dis. (6): 944, 2003. 31Tamada Y., et al.: Consumption of wild boar linked to cases of hepatitis E. J Hepatol, in Press, 2004. 32Kamili S., et al.: Protective efficacy of hepatitis e virus DNA vaccine administered by gene gun in the cynomolgus macaque model of infection. J Infect Dis. (2): 258-264, 2004. 33Purcell R.H., et al.: Pre-clinical immunogenicity and ef ficacy trial of a recombinant hepatitis E vaccine. Vaccine. (19-20): 2607-2615, 2003. 34T-C Li., et al.: Oral administration of hepatitis E virus-like particles induces a systemic and mucosal immune response in mice. Vaccine : 3476-3484, 2001. 35T-C Li., et al.: Protection of cynomolgus monkeys against HEV infection by oral administration of recombinant hepatitis E virus-like particles. Vaccine. (3-4): 370-377, 2004. 36Meng X.J.: Novel strains of hepatitis E virus identified from humans and other animal species: is hepatitis E a zoonosis?j Hepatol (5): 842-845, 2000. 37Kabrane-Lazizi Y., et al.: Evidence for widespread infection of wild rats with hepatitis E virus in the United States. Am J Trop Med Hyg (2): 331-335, 1999. 38Meng X.J., et al.: Prevalence of antibodies to hepatitis E virus in veterinarians working with swine and in normal blood donors in the United States and other countries.j Clin Microbiol. (1): 117-122, 2002. 39Nishizawa T., et al.: Characterization of Japanese swine and human hepatitis E virus isolates of genotype IV with 99 % identity over the entire genome. J Gen Virol. (5): 1245-1251, 2003. 40Haqshenas G., et al.: Genetic identification and characterization of a novel virus related to human hepatitis E virus from chickens with hepatitis-splenomegaly syndrome in the United States. J Gen Virol. : 2449-2462, 2001. 41Haqshenas G., et al.: The putative capsid protein of the newly identified avian hepatitis E virus shares antigenic epitopes with that of swine and human hepatitis E viruses and chicken big liver and spleen disease virus. J. Gen. Virol : 2201-2209, 2002. 42Meng XJ., et al.: Genetic and experimental evidence for cross-species infection by swine hepatitis E virus. J Virol (12): 9714-9721, 1998. 13