1887

Abstract

Hepatitis C virus (HCV) is the aetiological agent responsible for most cases of non-A non-B hepatitis. Hepatitis C is a disease of clinical importance because of its high infection rate in blood donors and its persistence as chronic infections which may lead to cirrhosis and hepatocellular carcinoma in the long term. The variability of the HCV genome has posed difficulties in serological detection and vaccine design. The recent advance in phage technology offers a means of cloning human anti-HCV antibodies of a defined specificity that may have potential therapeutic use. We now report the generation of a phage display library using the V genes of a HCV-infected patient and the V genes of two non-immune individuals. From this library we were able to obtain specific IgG single-chain Fvs (scFvs) that recognize viral core and envelope proteins by selection on synthetic peptides derived from the core sequence PKARRPEGRTWAQPG and the envelope E2 sequence RPIDDFDQGWGPITY. The specificity of the scFvs was demonstrated by their specific reactions with homologous peptides in ELISA and the specific blocking of scFv binding by homologous peptides, in a dose-dependent manner, in inhibition ELISA. The binding of the anticore 4c2 to homologous peptide was blocked by HCV-positive human sera in an antibody-concentration-dependent manner, suggesting that the scFv recognizes a similar if not identical epitope to those of one or more of the polyclonal antibodies present in the sera.

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1996-10-01
2021-10-24
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References

  1. Akatsuka T., Donets M., Scaglione L., Ching W. M., Shih J. W. K., Di Bisceglie A. M., Feinstone S. M. 1993; B-cell epitopes on the hepatitis C virus nucleocapsid protein determined by human monospecific antibodies. Hepatology 18:503–510
    [Google Scholar]
  2. Burioni R., Williamson R. A., Sanna P. P., Bloom F. E., Burton D. R. 1994; Recombinant human Fab to glycoprotein D neutralizes infectivity and prevents cell-to-cell transmission of herpes simplex viruses 1 and 2 in vitro . Proceedings of the National Academy of Sciences, USA 91:355–359
    [Google Scholar]
  3. Burton D. R., Pyati J., Koduri R., Sharp S. J., Thornton G. B., Parren P. W. H. I., Sawyer L. S. W., Hendry R. M., Dunlop N., Nara P. L., Lamacchia M., Garratty E., Stiehm E. R., Bryson Y. J., Cao Y., Moore J. P., Ho D. D., Barbas C. F. III 1994; Efficient neutralization of primary isolates of HIV-1 by a recombinant human monoclonal antibody. Science 266:1024–1027
    [Google Scholar]
  4. Chan S.-W., Simmonds P., McOmish F., Yap P.-L., Mitchell R., Dow B., Follett E. 1991; Serological responses to infection with three different types of hepatitis C virus. Lancet 338:1391
    [Google Scholar]
  5. Chien D. Y., Choo Q. L., Raison R., Spaete R., Tong M., Houghton M., Kuo G. 1993; Persistence of HCV despite antibodies to both putative envelope glycoproteins. Lancet 342:933
    [Google Scholar]
  6. Choo Q. L., Kuo G., Weiner A. J., Overby L. R., Bradley D. W., Houghton M. 1989; Isolation of a cDNA clone derived from a blood-borne non-A, non-B viral hepatitis genome. Science 244:359–362
    [Google Scholar]
  7. Choo Q. L., Richman K. H., Han J. H., Berger K., Lee C., Dong C., Gallegos C., Coit D., Medina-Selby A., Barr P. J., Weiner A. J., Bradley D. W., Kuo G., Houghton M. 1991; Genetic organization and diversity of the hepatitis C virus. Proceedings of the National Academy of Sciences, USA 88:2451–2455
    [Google Scholar]
  8. Choo Q.-L., Kuo G., Ralston R., Weiner A., Chien D., Van Nest G., Han J., Berger K., Thudium K., Kuo C., Kansopon J., Mefarland J., Tabrizi A., Ching K., Moss B., Cummins L. B., Houghton M., Muchmore E. 1994; Vaccination of chimpanzees against infection by the hepatitis C virus. Proceedings of the National Academy of Sciences, USA 91:1294–1298
    [Google Scholar]
  9. Cook G. P., Tomlinson I. M. 1995; The human immunoglobulin VH repertoire. Immunology Today 16:237–242
    [Google Scholar]
  10. Crowe J. E. Jr, Murphy B. R., Chanock R. M., Williamson R. A., Barbas C. F. III, Burton D. R. 1994; Recombinant human respiratory syncytial vims (RSV) monoclonal antibody Fab is effective therapeutically when introduced directly into the lungs of RSV-infected mice. Proceedings of the National Academy of Sciences, USA 91:1386–1390
    [Google Scholar]
  11. Farci P., Alter H. J., Wong D. C., Miller R. H., Govindarajan S., Engle R., Shapiro M., Purcell R. H. 1994; Prevention of hepatitis C vims infection in chimpanzees after antibody-mediated in vitro neutralization. Proceedings of the National Academy of Sciences, USA 91:7792–7796
    [Google Scholar]
  12. Griffiths A. D., Malmqvist M., Marks J. D., Bye J. M., Embleton M. J., McCafferty J., Baier M., Holliger K. P., Gorick B. D., Hughes-Jones N. C., Hoogenboom H. R., Winter G. 1993; Human anti-self antibodies with high specificity from phage display libraries. EMBO foumal 12:725–734
    [Google Scholar]
  13. Holmes E. C., Simmonds P., Cha T.-A., Chan S.-W., McOmish F., Irvine B., Beall E., Yap P. L., Kolberg J., Urdea M. S. 1994; Derivation of a rational nomenclature for hepatitis C vims by phylogenetic analysis of the NS-5 region. In Viral Hepatitis and Liver Disease pp 57–62 Edited by Nishioka K., Suzuki H., Mishiro S., Oda T. Tokyo: Springer-Verlag;
    [Google Scholar]
  14. Hoogenboom H. R., Winter G. 1992; By-passing immunisation: human antibodies from synthetic repertoires of germline V(H) gene segments rearranged in vitro . foumal of Molecular Biology 227:381–388
    [Google Scholar]
  15. Hoogenboom H. R., Griffiths A. D., Johnson K. S., Chiswell D. J., Hudson P., Winter G. 1991; Multi-subunit proteins on the surface of filamentous phage: methodologies for displaying antibody (Fab) heavy and light chains. Nucleic Acids Research 19:4133–4137
    [Google Scholar]
  16. Inchauspe G., Zebedee S. L., Lee D., Sugitani M., Nasoff M. S., Prince A. M. 1991; Genomic stmcture of the human prototype strain H of hepatitis C virus: comparison with American and Japanese isolates. Proceedings of National Academy of Sciences, USA 88:10292–10296
    [Google Scholar]
  17. Kumar U., Cheng D., Thomas H., Monjardino J. 1992; Cloning and sequencing of the structural region and expression of putative core gene of hepatitis C virus from a British case of chronic sporadic hepatitis. Journal of General Virology 73:1521–1525
    [Google Scholar]
  18. Kuppers R., Fischer U., Rajewsky K., Gause A. 1992; Immunoglobulin heavy and light chain gene sequences of a human CD5 positive immunocytoma and sequences of four novel V(H)III germline genes. Immunology Letters 34:57–62
    [Google Scholar]
  19. Laemmli U. K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685
    [Google Scholar]
  20. Lautner-Rieske A., Huber C., Meindl A., Pargent W., Schable K. F., Thiebe R., Zocher I., Zachau H. G. 1992; The human immunoglobulin К locus. Characterization of the duplicated A regions. European Journal of Immunology 22:1023–1029
    [Google Scholar]
  21. McCafferty J., Griffiths A. D., Winter G., Chisweli D. J. 1990; Phage antibodies: filamentous phage displaying antibody variable domains. Nature 348:552–554
    [Google Scholar]
  22. Marks J. D., Hoogenboom H. R., Bonnert T. P., McCafferty J., Griffiths A. D., Winter G. 1991a; By-passing immunization: human antibodies from V-gene libraries displayed on phage. Journal of Molecular Biology 222:581–597
    [Google Scholar]
  23. Marks J. D., Tristem M., Karpas A., Winter G. 1991b; Oligonucleotide primers for polymerase chain reaction amplification of human immunoglobulin variable genes and design of family-specific oligonucleotide probes. European Journal of Immunology 21:985–991
    [Google Scholar]
  24. Marks J. D., Ouwehand W. H., Bye J. M., Finnern R., Gorick B. D., Voak D., Thorpe S. J., Hughes-Jones N. C., Winter G. 1993; Human antibody fragments specific for human blood group antigens from a phage display library. Bio /Technology 11:1145–1149
    [Google Scholar]
  25. Munro S., Pelham H. R. B. 1986; An Hsp-like protein in the ER:identity with the 78 kD glucose regulated protein and immunoglobulin heavy chain binding protein. Cell 46:291–300
    [Google Scholar]
  26. Okamoto H., Kanai N., Mishiro S. 1992a; Full-length nucleotide sequence of a Japanese hepatitis C virus isolate (HC-J1). with high homology to USA isolates. Nucleic Acids Research 20:6410
    [Google Scholar]
  27. Okamoto H., Kojima M., Okada S., Yoshizawa H., lizuka H., Tanaka T., Muchmore E. E. A. P. D., Ito Y., Mishiro S. 1992b; Genetic drift of hepatitis C virus during an 8.2-year infection in a chimpanzee: variability and stability. Virology 190:894–899
    [Google Scholar]
  28. Okamoto H., Kurai K., Okada S., Yamamoto K., lizuka H., Tanaka T., Fukuda S., Tsuda F., Mishiro S. 1992c; Full-length sequence of a hepatitis C virus genome having poor homology to reported isolates: comparative study of four distinct genotypes. Virology 188:331–341
    [Google Scholar]
  29. Olee T., Yang P. M., Siminovitch K. A., Olsen N. J., Hillson J., Wu J., Kozin F., Carson D. A., Chen P. P. 1991; Molecular basis of an autoantibody-associated restriction fragment length polymorphism that confers susceptibility to autoimmune diseases. Journal of Clinical Investigation 88:193–203
    [Google Scholar]
  30. Rath S., Stanley C. M., Steward M. W. 1988; An inhibition enzyme immunoassay for estimating relative antibody affinity and affinity heterogeneity. Journal of Immunological Methods 106:245–249
    [Google Scholar]
  31. Reesink H. W., Nydegger U. E., Tegtmeier G. E., Barbara J., Seidl S., Woodfield D. G., Okochi K., Courouce A. M., Van Der Poel C. L. 1992; Blood donor screening or ‘over screening’: how far to go in avoiding transmission of infectious agents?. Vox Sanguinis 63:59–69
    [Google Scholar]
  32. Scarselli E., Cerino A., Esposito G., Silini E., Mondelli M. U., Traboni C. 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. Journal of Virology 69:4407–4412
    [Google Scholar]
  33. Schier R., Bye J., Apell G., McCall A., Adams G. P., Malmqvist M., Weiner L. M., Marks J. D. 1996; Isolation of high-affinity monomeric human anti-c-erfeB-2 single chain Fv using affinity-driven selection. Journal of Molecular Biology 255:28–43
    [Google Scholar]
  34. Seeff L. B. The NHLBI Study Group 1994; Mortality and morbidity of transfusion-associated type C hepatitis: an NHLBI multicenter study (abstract). Hepatology 20:204A
    [Google Scholar]
  35. Siemoneit K., Da Silva Cardoso M., Wolpl A., Koerner K., Kubanek B. 1994; Isolation and epitope characterization of human monoclonal antibodies to hepatitis C virus core antigen. Hyhridoma 13:9–13
    [Google Scholar]
  36. 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. 1993; Classification of hepatitis C virus into six major genotypes and a series of subtypes by phylogenetic analysis of the NS-5 region. Journal of General Virology 74:2391–2399
    [Google Scholar]
  37. Simmonds P., Alberti A., Alter H. J., Bonino F., Bradley D. W., Brechot C., Brouwer J. T., Chan S. W., Chayama K., Chen D. S., Choo Q. L., Colombo M., Cuypers H. T. M., Date T., Dusheiko G. M., Esteban J. I., Fay O., Hadziyannis S. J., Han J., Hatzakis A., Holmes E. C., Hotta H., Houghton M., Irvine B., Kohara M., Kolberg J. A., Kuo G., Lau J. Y. N., Lelie P. N., Maertens G., Mcomish F., Miyamura T., Mizokami M., Nomoto A., Prince A. M., Reesink H. W., Rice C., Roggendorf M., Schalm S. W., Shikata T., Shimotohno K., Stuyver L., Trepo C., Weiner A., Yap P. L., Urdea M. S. 1994; A proposed system for the nomenclature of hepatitis C viral genotypes. Hepatology 19:1321–1324
    [Google Scholar]
  38. Takamizawa A., Mori C., Manabe S., Murakami S., Fujita J., Onishi E., Andoh T., Yoshida I., Okayama H. 1991; The structure and organization of the hepatitis C virus genome isolated from human carriers. Journal of Virology 65:1105–1113
    [Google Scholar]
  39. Towbin H., Staehelin T., Gordon J. 1979; Electrophoretic transfer of proteins from the polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proceedings of the National Academy of Sciences, USA 76:4350–4354
    [Google Scholar]
  40. Trkola A., Pomales A. B., Yuan H., Korber B., Maddon P. J., Allaway G. P., Katinger H., Barbas C. F. III, Burton D. R., Ho D. D., Moore J. P. 1995; Cross-clade neutralization of primary isolates of human immunodeficiency virus type 1 by human monoclonal antibodies and tetrameric CD4-IgG. Journal of Virology 69:6609–6617
    [Google Scholar]
  41. van Regenmortel M. H. V., Brand J. P., Muller S., Plane S. 1993; Synthetic peptides as antigens. In Laboratory Techniques in Biochemistry and Molecular Biology pp 95–215 Edited by Burdon R. H., Knippenberg P. H.
    [Google Scholar]
  42. Ward E. S., Gussow D., Griffiths A. D., Jones P. T., Winter G. 1989; Binding activities of a repertoire of single immunoglobulin variable domains secreted from Escherichia coli . Nature 341:544–546
    [Google Scholar]
  43. Winter G., Milstein C. 1991; Man-made antibodies. Nature 349:293–299
    [Google Scholar]
  44. Zibert A., Schreier E., Roggendorf M. 1995; Antibodies in human sera specific to hypervariable region 1 of hepatitis C virus can block viral attachment. Virology 208:653–661
    [Google Scholar]
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