Extensive genomic diversity has been observed among hepatitis E virus (HEV) strains. However, the implication of the genetic heterogeneity on HEV antigenic properties is uncertain. In this study, monoclonal antibodies (Mabs) against truncated ORF2-encoded proteins (aa452？617, designated p166 proteins) derived from HEV strains of Burma (genotype 1a, p166Bur), Pakistan (1b, p166Pak) and Morocco (1c, p166Mor) were raised and used for identification of HEV antigenic diversity. Six Mabs reacted to these 3 p166 proteins as well as p166 proteins constructed from strains derived from Mexico (genotype？2), US？(genotype？3) and China (genotype 4), indicating the existence of pan？genotypic epitopes. Two Mabs, 1B5 and 6C7, reacted with p166Bur and p166Mor, but not p166Pak or p166s derived from genotypes 2, 3, and 4, indicating that these 2 Mabs recognized strain-specific HEV epitopes. Both the common and specific epitopes could not be mapped by 23 synthetic peptides spanning the p166Bur sequence, suggesting that they are confirmation？dependent. Comparative sequence analysis showed that p166Bur and p166Mor shared an identical aa sequence along their entire lengths, whereas for p166Pak the aas occupying positions 606 and 614 are different from aas at corresponding positions of p166Bur and p166Mor. Reactivity between 1B5 and p166Bur was abrogated with mutation of p166Bur/A606V, whereas p166Pak acquired the reactivity to 1B5 with mutation of p166Pak/V606A. However, mutations of p166Bur/L614M and P166Pak/M614L did not affect the immunoreactivity. Therefore, the aa occupying position 606 plays a critical role in maintaining the antigenicity of the HEV p166 proteins.
References
[1]
Dalton, HR; Bendall, R; Ijaz, S; Banks, M. Hepatitis E: An emerging infection in developed countries. Lancet Infect. Dis？2008, 8, 698–709, doi:10.1016/S1473-3099(08)70255-X. 18992406
[2]
Kamar, N; Selves, J; Mansuy, JM; Ouezzani, L; Peron, JM; Guitard, J; Cointault, O; Esposito, L; Abravanel, F; Danjoux, M; Durand, D; Vinel, JP; Izopet, J; Rostaing, L. Hepatitis E virus and chronic hepatitis in organ-transplant recipients. N. Eng. J. Med？2008, 358, 811–817, doi:10.1056/NEJMoa0706992.
[3]
Pan, JS; Zhang, K; Zhou, JY; Wu, C; Zhuang, H; Zhou, YH. Application of truncated immunodominant polypeptide from hepatitis E virus (HEV) ORF2 in an assay to exclude nonspecific binding in detecting anti-HEV immunoglobulin M. J. Clin. Microbiol？2010, 48, 779–784, doi:10.1128/JCM.01671-09. 20042615
[4]
Emerson, SU; Anderson, D; Arankalle, A; Meng, XJ; Purdy, M; Schlauder, GG; Tsarev, SA. Hepevirus. In Virus Taxonomy: Eighth Report of the International Committee on Taxonomy of Viruses; Fauquet, CM, Mayo, MA, Maniloff, J, Desselberger, U, Ball, LA, Eds.; Elsevier Academic Press: London, UK, 2004; pp. 851–855.
[5]
Okamoto, H. Genetic variability and evolution of hepatitis E virus. Virus Res？2007, 127, 216–228, doi:10.1016/j.virusres.2007.02.002. 17363102
[6]
Lu, L; Li, CH; Hagedorn, CH. Phylogenetic analysis of global hepatitis E virus sequences: Genetic diversity, subtypes and zoonosis. Rev. Med. Virol？2006, 16, 5–36, doi:10.1002/rmv.482. 16175650
[7]
Zhai, L; Dai, X; Meng, J. Hepatitis E virus genotyping based on full-length genome and partial genomic regions. Virus Res？2006, 120, 57–69, doi:10.1016/j.virusres.2006.01.013. 16472882
[8]
Reyes, GR; Purdy, MA; Kim, JP; Luk, KC; Young, LM; Fry, KE; Bradley, DW. Isolation of a cDNA from the virus responsible for enterically transmitted non-A, non-B hepatitis. Science？1990, 247, 1335–1339, doi:10.1126/science.2107574. 2107574
[9]
Huang, CC; Nguyen, D; Fernandez, J; Yun, KY; Fry, KE; Bradley, DW; Tam, AW; Reyes, GR. Molecular-cloning and sequencing of the Mexico isolate of hepetitis-E virus (HEV). Virology？1992, 191, 550–558, doi:10.1016/0042-6822(92)90230-M. 1448913
[10]
Schlauder, GG; Dawson, GJ; Erker, JC; Kwo, PY; Knigge, MF; Smalley, DL; Rosenblatt, JE; Desai, SM; Mushahwar, IK. 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？1998, 79, 447–456. 9519822
[11]
Wang, YC; Zhang, HY; Ling, R; Li, HM; Harrison, TJ. The complete sequence of hepatitis E virus genotype 4 reveals an alternative strategy for translation of open reading frames 2 and 3. J. Gen. Virol？2000, 81, 1675–1686. 10859372
[12]
Engle, RE; Yu, C; Emerson, SU; Meng, XJ; Purcell, RH. Hepatitis E virus (HEV) capsid antigens derived from viruses of human and swine origin are equally efficient for detecting anti- HEV by enzyme immunoassay. J. Clin. Microbiol？2002, 40, 4576–4580, doi:10.1128/JCM.40.12.4576-4580.2002. 12454155
[13]
Zhou, YH; Purcell, RH; Emerson, SU. An ELISA for putative neutralizing antibodies to hepatitis E virus detects antibodies to genotypes 1, 2, 3, and 4. Vaccine？2004, 22, 2578–2585, doi:10.1016/j.vaccine.2003.12.017. 15193383
[14]
Arankalle, VA; Lole, KS; Deshmukh, TM; Chobe, LP; Gandhe, SS. Evaluation of human (genotype 1) and swine (genotype 4)-ORF2-based ELISAs for anti-HEV IgM and IgG detection in an endemic country and search for type 4 human HEV infections. J. Viral. Hepat？2007, 14, 435–445, doi:10.1111/j.1365-2893.2006.00801.x. 17501765
[15]
Peralta, B; Casas, M; de Deus, N; Martin, M; Ortuno, A; Perez-Martin, E; Pina, S; Mateu, E. Anti-HEV antibodies in domestic animal species and rodents from Spain using a genotype 3-based ELISA. Vet. Microbiol？2009, 137, 66–73, doi:10.1016/j.vetmic.2009.01.006. 19200664
[16]
Wang, YC; Zhang, HY; Li, Z; Gu, WJ; Lan, HY; Hao, W; Ling, R; Li, HM; Harrison, TJ. Detection of sporadic cases of hepatitis e virus (HEV) infection in china using immunoassays based on recombinant open reading frame 2 and 3 polypeptides from HEV genotype 4. J. Clin. Microbiol？2001, 39, 4370–4379, doi:10.1128/JCM.39.12.4370-4379.2001. 11724847
[17]
Herremans, M; Duizer, E; Jusic, E; Koopmans, MPG. Detection of hepatitis E virus-specific immunoglobulin A in patients infected with hepatitis E virus genotype 1 or 3. Clin. Vaccine Immunol？2007, 14, 276–280, doi:10.1128/CVI.00312-06. 17267585
[18]
Bendall, R; Ellis, V; Ijaz, S; Thurairajah, P; Dalton, HR. Serological response to hepatitis E virus genotype 3 infection: IgG quantitation, avidity, and IgM response. J. Med. Virol？2008, 80, 95–101, doi:10.1002/jmv.21033. 18041018
[19]
Jiménez de Oya, N; Galindo, I; Gironés, O; Duizer, E; Escribano, J; Saiz, J. Serological immunoassay for detection of hepatitis E virus on the basis of genotype 3 open reading frame 2 recombinant proteins produced in Trichoplusia ni larvae. J. Clin. Microbiol？2009, 47, 3276–3282, doi:10.1128/JCM.00750-09. 19656986
[20]
Meng, JH; Dai, X; Chang, JC; Lopareva, E; Pillot, J; Fields, HA; Khudyakov, YE. Identification and characterization of the neutralization epitope(s) of the hepatitis E virus. Virology？2001, 288, 203–211, doi:10.1006/viro.2001.1093. 11601892
[21]
Deng, L; Meng, J-h; Zhao, Y; Zhang, H-m; Dai, X. Hepatitis E virus of different genotypes contains multiple-type antigenic epitopes. Acta Microbiol. Sinica？2006, 46, 120–126.
[22]
Zhao, Y; Liang, JH; Meng, JH; Zhang, LF; Zhang, HM; Ding, F; Ji, XH. Comparison of B ell antigenic epitopes between hepatitis E virus genotype I, II and genotype III, IV by using monoclonal antibodies. Chin. J. Immunol？2004, 20, 591–595.
[23]
Schofield, D; Purcell, R; Nguyen, H; Emerson, S. Monoclonal antibodies that neutralize HEV recognize an antigenic site at the carboxyterminus of an ORF2 protein vaccine. Vaccine？2003, 22, 257–267, doi:10.1016/j.vaccine.2003.07.008. 14615154
[24]
Meng, JH; Cong, ME; Dai, X; Pillot, J; Purdy, MA; Fields, HA; Khudyakov, YE. Primary structure of open reading frame 2 and 3 of the hepatitis E virus isolated from Morocco. J. Med. Virol？1999, 57, 126–133, doi:10.1002/(SICI)1096-9071(199902)57:2<126::AID-JMV7>3.0.CO;2-O. 9892396
[25]
Meng, XJ; Purcell, RH; Halbur, PG; Lehman, JR; Webb, DM; Tsareva, TS; Haynes, JS; Thacker, BJ; Emerson, SU. A novel virus in swine is closely related to the human hepatitis E virus. Proc. Natl. Acad. Sci. USA？1997, 94, 9860–9865, doi:10.1073/pnas.94.18.9860. 9275216
[26]
Wang, YC; Zhang, HY; Xia, NS; Peng, G; Lan, HY; Zhuang, H; Zhu, YH; Li, SW; Tian, KG; Gu, WJ; Lin, JX; Wu, X; Li, HM; Harrison, TJ. Prevalence, isolation, and partial sequence analysis of hepatitis E virus from domestic animals in China. J. Med. Virol？2002, 67, 516–521, doi:10.1002/jmv.10131. 12115997
[27]
Ji, YL; Zhu, YH; Liang, JR; Wei, XF; Yang, X; Wang, L; Li, LJ; Chang, YB; Tang, RL; Zhuang, H. Swine hepatitis E virus in rural southern China: Genetic characterization and experimental infection in rhesus monkeys (Macaca mulatta). J. Gastroenterol？2008, 43, 565–570, doi:10.1007/s00535-008-2196-3. 18648744
[28]
Kase, JA; Correa, MT; Luna, C; Sobsey, MD. Isolation, detection and characterization of swine hepatitis E virus from herds in Costa Rica. Int. J. Environ. Health Res？2008, 18, 165–176, doi:10.1080/09603120701498311. 18569145
[29]
Xia, HY; Liu, LH; Linde, AM; Belak, S; Norder, H; Widen, F. Molecular characterization and phylogenetic analysis of the complete genome of a hepatitis E virus from European swine. Virus Gene？2008, 37, 39–48, doi:10.1007/s11262-008-0246-9.
[30]
Li, TC; Chijiwa, K; Sera, N; Ishibashi, T; Etoh, Y; Shinohara, Y; Kurata, Y; Ishida, M; Sakamoto, S; Takeda, N; Miyamura, T. Hepatitis E virus transmission from wild boar meat. Emerg. Infect. Dis？2005, 11, 1958–1960, doi:10.3201/eid1112.051041. 16485490
[31]
Nishizawa, T; Takahashi, M; Endo, K; Fujiwara, S; Sakuma, N; Kawazuma, F; Sakamoto, H; Sato, Y; Bando, M; Okamoto, H. Analysis of the full-length genome of hepatitis E virus isolates obtained from wild boars in Japan. J. Gen. Virol？2005, 86, 3321–3326, doi:10.1099/vir.0.81394-0. 16298977
[32]
Martelli, F; Caprioli, A; Zengarini, M; Marata, A; Fiegna, C; Di Bartolo, I; Ruggeri, FM; Delogu, M; Ostanello, F. Detection of Hepatitis E virus (HEV) in a demographic managed wild boar (Sus scrofa scrofa) population in Italy. Vet. Microbiol？2008, 126, 74–81, doi:10.1016/j.vetmic.2007.07.004. 17706898
[33]
Kaci, S; Nockler, K; Johne, R. Detection of hepatitis E virus in archived German wild boar serum samples. Vet. Microbiol？2008, 128, 380–385, doi:10.1016/j.vetmic.2007.10.030. 18068914
[34]
Takahashi, K; Kitajima, N; Abe, N; Mishiro, S. Complete or near-complete nucleotide sequences of hepatitis E virus genome recovered from a wild boar, a deer, and four patients who ate the deer. Virology？2004, 330, 501–505, doi:10.1016/j.virol.2004.10.006. 15567444
[35]
Zhao, CY; Ma, ZR; Harrison, TJ; Feng, RF; Zhang, CT; Qiao, ZL; Fan, JP; Ma, HX; Li, MS; Song, AJ; Wang, YC. A novel genotype of hepatitis E virus prevalent among farmed rabbits in China. J. Med. Virol？2009, 81, 1371–1379, doi:10.1002/jmv.21536. 19551838
[36]
Tei, S; Kitajima, N; Takahashi, K; Mishiro, S. Zoonotic transmission of hepatitis E virus from deer to human beings. Lancet？2003, 362, 371–373, doi:10.1016/S0140-6736(03)14025-1. 12907011
[37]
Meng, JH; Pillot, J; Dai, X; Fields, HA; Khudyakov, YE. Neutralization of different geographic strains of the hepatitis E virus with anti-hepatitis E virus-positive serum samples obtained from different sources. Virology？1998, 249, 316–324, doi:10.1006/viro.1998.9346. 9791023
[38]
Zhou, YH; Chen, ZC; Purcell, RH; Emerson, SU. Positive reactions on Western blots do not necessarily indicate the epitopes on antigens are continuous. Immunol. Cell Biol？2007, 85, 73–78, doi:10.1038/sj.icb.7100004. 17130902
[39]
Yamashita, T; Mori, Y; Miyazaki, N; Cheng, RH; Yoshimura, M; Unno, H; Shima, R; Moriishi, K; Tsukihara, T; Li, TC; Takeda, N; Miyamura, T; Matsuura, Y. Biological and immunological characteristics of hepatitis E virus-like particles based on the crystal structure. Proc. Natl. Acad. Sci. USA？2009, 106, 12986–12991, doi:10.1073/pnas.0903699106. 19620712
[40]
He, J. Molecular detection and sequence analysis of a new hepatitis E virus isolate from Pakistan. J. Viral Hepat？2006, 13, 840–844, doi:10.1111/j.1365-2893.2006.00774.x. 17109684
[41]
Shen, Q; Zhang, W; Cao, X; Mou, J; Cui, L; Hua, X. Cloning of full genome sequence of hepatitis E virus of Shanghai swine isolate using RACE method. Virology？2007, 4, 98, doi:10.1186/1743-422X-4-98.
[42]
Glaser, L; Stevens, J; Zamarin, D; Wilson, IA; Garcia-Sastre, A; Tumpey, TM; Basler, CF; Taubenberger, JK; Palese, P. A single amino acid substitution in 1918 influenza virus hemagglutinin changes receptor binding specificity. Virology？2005, 79, 11533–11536, doi:10.1128/JVI.79.17.11533-11536.2005.
[43]
Shiomi, H; Urasawa, T; Urasawa, S; Kobayashi, N; Abe, S; Taniguchi, K. Isolation and characterisation of poliovirus mutants resistant to heating at 50 degrees C for 30 min. J. Med. Virol？2004, 74, 484–491, doi:10.1002/jmv.20202. 15368512
[44]
Zhang, H; Dai, X; Shan, X; Meng, J. Characterization of antigenic epitopes of the ORF2 protein from hepatitis E virus genotype 4. Virus Res？2009, 142, 140–143, doi:10.1016/j.virusres.2009.02.002. 19428747
[45]
Khudyakov, YE; Lopareva, EN; Jue, DL; Crews, TK; Thyagarajan, SP; Fields, HA. Antigenic domains of the open reading frame 2-encoded protein of hepatitis E virus. J. Clin. Microbiol？1999, 37, 2863–2871. 10449466
[46]
Adler, B; Faine, S. A pomona serogroup-specific, agglutinating antigen in leptospira, identified by monoclonal-antibodies. Pathology？1983, 15, 247–250, doi:10.3109/00313028309083501. 6417619
[47]
Obriadina, A; Meng, JH; Lopareva, EN; Spelbring, J; Burkov, A; Krawczynski, K; Khudyakov, YE; Fields, HA. Antigenic properties of recombinant proteins of hepatitis E virus. In Proceedings of 10th International Symposium on Viral Hepatitis and Liver Disease; International Medical Press: London, UK, 2002; pp. 119–123.
