Skip to main content
Journal of Virology logoLink to Journal of Virology
. 1995 Jul;69(7):4407–4412. doi: 10.1128/jvi.69.7.4407-4412.1995

Occurrence of antibodies reactive with more than one variant of the putative envelope glycoprotein (gp70) hypervariable region 1 in viremic hepatitis C virus-infected patients.

E Scarselli 1, A Cerino 1, G Esposito 1, E Silini 1, M U Mondelli 1, C Traboni 1
PMCID: PMC189182  PMID: 7539508

Abstract

The hepatitis C virus (HCV) is a frequent cause of chronic liver disease. A mechanism proposed as being responsible for virus persistence is evasion of the host immune response through a high mutation rate in crucial regions of the viral genome. We have sequenced the hypervariable region 1 (HVR1) of the virus isolated from three serum samples, collected during 18 months of follow-up, from an asymptomatic HCV-infected patient. A synthetic peptide of 27 amino acids, corresponding to the HVR1 sequence found to be predominant in both the second and third samples, was used as the antigen for detection of antibodies by enzyme-linked immunosorbent assay (ELISA). We observed reactivity against this HVR1 sequence in the first serum sample before the appearance of the viral isolate in the bloodstream; the reactivity increased in the second and third samples while the cognate viral sequence became predominant. Moreover, our results show that antibodies from all three samples recognize a region mapping at the carboxyl-terminal part of the HVR1 and are cross-reactive with the HVR1 sequence previously found in the same patient. The presence of anti-HVR1 antibodies was investigated in a further 142 HCV patients: 121 viremic and 21 nonviremic. Two synthetic peptides were used, the first corresponding to the sequence derived from the patient described above and the second one synthesized according to the sequence of the HCV BK strain. A high frequency of positive reactions against both HVR1 variants was detected in the samples from the viremic individuals. Finally, antibodies cross-reactive with both variants were shown to be present by competitive ELISA in 6 of 10 viremic patients. The potential negative implications of this observation for the host are discussed.

Full Text

The Full Text of this article is available as a PDF (226.8 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Alter H. J., Purcell R. H., Shih J. W., Melpolder J. C., Houghton M., Choo Q. L., Kuo G. Detection of antibody to hepatitis C virus in prospectively followed transfusion recipients with acute and chronic non-A, non-B hepatitis. N Engl J Med. 1989 Nov 30;321(22):1494–1500. doi: 10.1056/NEJM198911303212202. [DOI] [PubMed] [Google Scholar]
  2. Chien D. Y., Choo Q. L., Ralston R., Spaete R., Tong M., Houghton M., Kuo G. Persistence of HCV despite antibodies to both putative envelope glycoproteins. Lancet. 1993 Oct 9;342(8876):933–933. doi: 10.1016/0140-6736(93)91983-s. [DOI] [PubMed] [Google Scholar]
  3. Choo Q. L., Kuo G., Ralston R., Weiner A., Chien D., Van Nest G., Han J., Berger K., Thudium K., Kuo C. Vaccination of chimpanzees against infection by the hepatitis C virus. Proc Natl Acad Sci U S A. 1994 Feb 15;91(4):1294–1298. doi: 10.1073/pnas.91.4.1294. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Choo Q. L., Weiner A. J., Overby L. R., Kuo G., Houghton M., Bradley D. W. Hepatitis C virus: the major causative agent of viral non-A, non-B hepatitis. Br Med Bull. 1990 Apr;46(2):423–441. doi: 10.1093/oxfordjournals.bmb.a072408. [DOI] [PubMed] [Google Scholar]
  5. Farci P., Alter H. J., Govindarajan S., Wong D. C., Engle R., Lesniewski R. R., Mushahwar I. K., Desai S. M., Miller R. H., Ogata N. Lack of protective immunity against reinfection with hepatitis C virus. Science. 1992 Oct 2;258(5079):135–140. doi: 10.1126/science.1279801. [DOI] [PubMed] [Google Scholar]
  6. Fox G., Parry N. R., Barnett P. V., McGinn B., Rowlands D. J., Brown F. The cell attachment site on foot-and-mouth disease virus includes the amino acid sequence RGD (arginine-glycine-aspartic acid). J Gen Virol. 1989 Mar;70(Pt 3):625–637. doi: 10.1099/0022-1317-70-3-625. [DOI] [PubMed] [Google Scholar]
  7. Garson J. A., Tuke P. W., Makris M., Briggs M., Machin S. J., Preston F. E., Tedder R. S. Demonstration of viraemia patterns in haemophiliacs treated with hepatitis-C-virus-contaminated factor VIII concentrates. Lancet. 1990 Oct 27;336(8722):1022–1025. doi: 10.1016/0140-6736(90)92487-3. [DOI] [PubMed] [Google Scholar]
  8. Gorny M. K., Xu J. Y., Karwowska S., Buchbinder A., Zolla-Pazner S. Repertoire of neutralizing human monoclonal antibodies specific for the V3 domain of HIV-1 gp120. J Immunol. 1993 Jan 15;150(2):635–643. [PubMed] [Google Scholar]
  9. Higashi Y., Kakumu S., Yoshioka K., Wakita T., Mizokami M., Ohba K., Ito Y., Ishikawa T., Takayanagi M., Nagai Y. Dynamics of genome change in the E2/NS1 region of hepatitis C virus in vivo. Virology. 1993 Dec;197(2):659–668. doi: 10.1006/viro.1993.1641. [DOI] [PubMed] [Google Scholar]
  10. Hijikata M., Kato N., Ootsuyama Y., Nakagawa M., Ohkoshi S., Shimotohno K. Hypervariable regions in the putative glycoprotein of hepatitis C virus. Biochem Biophys Res Commun. 1991 Feb 28;175(1):220–228. doi: 10.1016/s0006-291x(05)81223-9. [DOI] [PubMed] [Google Scholar]
  11. Hsu H. H., Donets M., Greenberg H. B., Feinstone S. M. Characterization of hepatitis C virus structural proteins with a recombinant baculovirus expression system. Hepatology. 1993 May;17(5):763–771. [PubMed] [Google Scholar]
  12. Inoue Y., Suzuki R., Matsuura Y., Harada S., Chiba J., Watanabe Y., Saito I., Miyamura T. Expression of the amino-terminal half of the NS1 region of the hepatitis C virus genome and detection of an antibody to the expressed protein in patients with liver diseases. J Gen Virol. 1992 Aug;73(Pt 8):2151–2154. doi: 10.1099/0022-1317-73-8-2151. [DOI] [PubMed] [Google Scholar]
  13. Javaherian K., Langlois A. J., McDanal C., Ross K. L., Eckler L. I., Jellis C. L., Profy A. T., Rusche J. R., Bolognesi D. P., Putney S. D. Principal neutralizing domain of the human immunodeficiency virus type 1 envelope protein. Proc Natl Acad Sci U S A. 1989 Sep;86(17):6768–6772. doi: 10.1073/pnas.86.17.6768. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Kato N., Ootsuyama Y., Ohkoshi S., Nakazawa T., Sekiya H., Hijikata M., Shimotohno K. Characterization of hypervariable regions in the putative envelope protein of hepatitis C virus. Biochem Biophys Res Commun. 1992 Nov 30;189(1):119–127. doi: 10.1016/0006-291x(92)91533-v. [DOI] [PubMed] [Google Scholar]
  15. Kato N., Ootsuyama Y., Tanaka T., Nakagawa M., Nakazawa T., Muraiso K., Ohkoshi S., Hijikata M., Shimotohno K. Marked sequence diversity in the putative envelope proteins of hepatitis C viruses. Virus Res. 1992 Feb;22(2):107–123. doi: 10.1016/0168-1702(92)90038-b. [DOI] [PubMed] [Google Scholar]
  16. Kato N., Sekiya H., Ootsuyama Y., Nakazawa T., Hijikata M., Ohkoshi S., Shimotohno K. Humoral immune response to hypervariable region 1 of the putative envelope glycoprotein (gp70) of hepatitis C virus. J Virol. 1993 Jul;67(7):3923–3930. doi: 10.1128/jvi.67.7.3923-3930.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Kelker H. C., Schlesinger D., Valentine F. T. Immunogenic and antigenic properties of an HIV-1 gp120-derived multiple chain peptide. J Immunol. 1994 Apr 15;152(8):4139–4148. [PubMed] [Google Scholar]
  18. Kumar U., Brown J., Monjardino J., Thomas H. C. Sequence variation in the large envelope glycoprotein (E2/NS1) of hepatitis C virus during chronic infection. J Infect Dis. 1993 Mar;167(3):726–730. doi: 10.1093/infdis/167.3.726. [DOI] [PubMed] [Google Scholar]
  19. Kumar U., Monjardino J., Thomas H. C. Hypervariable region of hepatitis C virus envelope glycoprotein (E2/NS1) in an agammaglobulinemic patient. Gastroenterology. 1994 Apr;106(4):1072–1075. doi: 10.1016/0016-5085(94)90770-6. [DOI] [PubMed] [Google Scholar]
  20. Kurosaki M., Enomoto N., Marumo F., Sato C. Rapid sequence variation of the hypervariable region of hepatitis C virus during the course of chronic infection. Hepatology. 1993 Dec;18(6):1293–1299. [PubMed] [Google Scholar]
  21. Lesniewski R. R., Boardway K. M., Casey J. M., Desai S. M., Devare S. G., Leung T. K., Mushahwar I. K. Hypervariable 5'-terminus of hepatitis C virus E2/NS1 encodes antigenically distinct variants. J Med Virol. 1993 Jun;40(2):150–156. doi: 10.1002/jmv.1890400213. [DOI] [PubMed] [Google Scholar]
  22. Miller R. H., Purcell R. H. Hepatitis C virus shares amino acid sequence similarity with pestiviruses and flaviviruses as well as members of two plant virus supergroups. Proc Natl Acad Sci U S A. 1990 Mar;87(6):2057–2061. doi: 10.1073/pnas.87.6.2057. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Mink M. A., Benichou S., Madaule P., Tiollais P., Prince A. M., Inchauspe G. Characterization and mapping of a B-cell immunogenic domain in hepatitis C virus E2 glycoprotein using a yeast peptide library. Virology. 1994 Apr;200(1):246–255. doi: 10.1006/viro.1994.1182. [DOI] [PubMed] [Google Scholar]
  24. Mita E., Hayashi N., Ueda K., Kasahara A., Fusamoto H., Takamizawa A., Matsubara K., Okayama H., Kamada T. Expression of MBP-HCV NS1/E2 fusion protein in E. coli and detection of anti-NS1/E2 antibody in type C chronic liver disease. Biochem Biophys Res Commun. 1992 Mar 31;183(3):925–930. doi: 10.1016/s0006-291x(05)80278-5. [DOI] [PubMed] [Google Scholar]
  25. Müller H. M., Pfaff E., Goeser T., Theilmann L. Genetic variability of German hepatitis C virus isolates. J Med Virol. 1993 Aug;40(4):291–306. doi: 10.1002/jmv.1890400407. [DOI] [PubMed] [Google Scholar]
  26. Nara P. L., Garrity R. R., Goudsmit J. Neutralization of HIV-1: a paradox of humoral proportions. FASEB J. 1991 Jul;5(10):2437–2455. doi: 10.1096/fasebj.5.10.1712328. [DOI] [PubMed] [Google Scholar]
  27. Nishihara T., Nozaki C., Nakatake H., Hoshiko K., Esumi M., Hayashi N., Hino K., Hamada F., Mizuno K., Shikata T. Secretion and purification of hepatitis C virus NS1 glycoprotein produced by recombinant baculovirus-infected insect cells. Gene. 1993 Jul 30;129(2):207–214. doi: 10.1016/0378-1119(93)90270-d. [DOI] [PubMed] [Google Scholar]
  28. Ogata N., Alter H. J., Miller R. H., Purcell R. H. Nucleotide sequence and mutation rate of the H strain of hepatitis C virus. Proc Natl Acad Sci U S A. 1991 Apr 15;88(8):3392–3396. doi: 10.1073/pnas.88.8.3392. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Okamoto H., Sugiyama Y., Okada S., Kurai K., Akahane Y., Sugai Y., Tanaka T., Sato K., Tsuda F., Miyakawa Y. Typing hepatitis C virus by polymerase chain reaction with type-specific primers: application to clinical surveys and tracing infectious sources. J Gen Virol. 1992 Mar;73(Pt 3):673–679. doi: 10.1099/0022-1317-73-3-673. [DOI] [PubMed] [Google Scholar]
  30. Okamoto H., Tokita H., Sakamoto M., Horikita M., Kojima M., Iizuka H., Mishiro S. Characterization of the genomic sequence of type V (or 3a) hepatitis C virus isolates and PCR primers for specific detection. J Gen Virol. 1993 Nov;74(Pt 11):2385–2390. doi: 10.1099/0022-1317-74-11-2385. [DOI] [PubMed] [Google Scholar]
  31. Rümenapf T., Stark R., Meyers G., Thiel H. J. Structural proteins of hog cholera virus expressed by vaccinia virus: further characterization and induction of protective immunity. J Virol. 1991 Feb;65(2):589–597. doi: 10.1128/jvi.65.2.589-597.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Sakamoto N., Enomoto N., Kurosaki M., Marumo F., Sato C. Sequential change of the hypervariable region of the hepatitis C virus genome in acute infection. J Med Virol. 1994 Jan;42(1):103–108. doi: 10.1002/jmv.1890420119. [DOI] [PubMed] [Google Scholar]
  33. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Silini E., Bono F., Cerino A., Piazza V., Solcia E., Mondelli M. U. Virological features of hepatitis C virus infection in hemodialysis patients. J Clin Microbiol. 1993 Nov;31(11):2913–2917. doi: 10.1128/jcm.31.11.2913-2917.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Simmonds P., Holmes E. C., Cha T. A., Chan S. W., McOmish F., Irvine B., Beall E., Yap P. L., Kolberg J., Urdea M. S. Classification of hepatitis C virus into six major genotypes and a series of subtypes by phylogenetic analysis of the NS-5 region. J Gen Virol. 1993 Nov;74(Pt 11):2391–2399. doi: 10.1099/0022-1317-74-11-2391. [DOI] [PubMed] [Google Scholar]
  36. Spaete R. R., Alexander D., Rugroden M. E., Choo Q. L., Berger K., Crawford K., Kuo C., Leng S., Lee C., Ralston R. Characterization of the hepatitis C virus E2/NS1 gene product expressed in mammalian cells. Virology. 1992 Jun;188(2):819–830. doi: 10.1016/0042-6822(92)90537-y. [DOI] [PubMed] [Google Scholar]
  37. Tam J. P. Synthetic peptide vaccine design: synthesis and properties of a high-density multiple antigenic peptide system. Proc Natl Acad Sci U S A. 1988 Aug;85(15):5409–5413. doi: 10.1073/pnas.85.15.5409. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Taniguchi S., Okamoto H., Sakamoto M., Kojima M., Tsuda F., Tanaka T., Munekata E., Muchmore E. E., Peterson D. A., Mishiro S. A structurally flexible and antigenically variable N-terminal domain of the hepatitis C virus E2/NS1 protein: implication for an escape from antibody. Virology. 1993 Jul;195(1):297–301. doi: 10.1006/viro.1993.1378. [DOI] [PubMed] [Google Scholar]
  39. Weiner A. J., Brauer M. J., Rosenblatt J., Richman K. H., Tung J., Crawford K., Bonino F., Saracco G., Choo Q. L., Houghton M. Variable and hypervariable domains are found in the regions of HCV corresponding to the flavivirus envelope and NS1 proteins and the pestivirus envelope glycoproteins. Virology. 1991 Feb;180(2):842–848. doi: 10.1016/0042-6822(91)90104-j. [DOI] [PubMed] [Google Scholar]
  40. Weiner A. J., Geysen H. M., Christopherson C., Hall J. E., Mason T. J., Saracco G., Bonino F., Crawford K., Marion C. D., Crawford K. A. Evidence for immune selection of hepatitis C virus (HCV) putative envelope glycoprotein variants: potential role in chronic HCV infections. Proc Natl Acad Sci U S A. 1992 Apr 15;89(8):3468–3472. doi: 10.1073/pnas.89.8.3468. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Zhang Y. M., Hayes E. P., McCarty T. C., Dubois D. R., Summers P. L., Eckels K. H., Chanock R. M., Lai C. J. Immunization of mice with dengue structural proteins and nonstructural protein NS1 expressed by baculovirus recombinant induces resistance to dengue virus encephalitis. J Virol. 1988 Aug;62(8):3027–3031. doi: 10.1128/jvi.62.8.3027-3031.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. van Doorn L. J., Capriles I., Maertens G., DeLeys R., Murray K., Kos T., Schellekens H., Quint W. Sequence evolution of the hypervariable region in the putative envelope region E2/NS1 of hepatitis C virus is correlated with specific humoral immune responses. J Virol. 1995 Feb;69(2):773–778. doi: 10.1128/jvi.69.2.773-778.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Virology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES