1887

Abstract

Healthy carriers of hepatitis C virus (HCV) infection exhibit a specific antibody response against all HCV antigens, which could play a role in disease control. Generation of panels of human antibodies may permit a thorough characterization of this response and further identify particular antibodies with potential clinical value. To this effect, we have established a human phage-display antibody library from a patient exhibiting a high antibody response against HCV antigens and no clinical symptoms of disease. This library was screened against a recombinant core antigen [amino acids (aa) 1–119] produced in .. Two recombinant Fab-carrying phages (rFabCs) were isolated and characterized. Both rFabC3 and rFabC14 recognize aa 1–48 on core antigen, but rFabC14 is competed out by a synthetic peptide, C (aa 1–20), at much lower concentrations than rFabC3. In order to identify more precisely the recognition sites of these antibodies, we produced soluble forms of the rFabs (sFabs), and used them to pan a random phage-display peptide library. A single peptide sequence, QLITKPL, was identified with sFabC3, while two equally represented sequences, HAFPHLH and SAPSSKN, were isolated using sFabC14. The QLITKPL sequence was partially localized between aa 8 and 14 of core protein, but no clear homology was found for the two sFabC14 peptides. However, we confirmed the specificity of these peptides by competition experiments with sFabC14.

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2000-02-01
2019-12-07
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References

  1. Akatsuka, T., Donets, M., Scaglione, L., Ching, W. M., Shih, J. W., Di, B. & Feinstone, S. M. ( 1993; ). B-cell epitopes on the hepatitis C virus nucleocapsid protein determined by human monospecific antibodies. Hepatology 18, 503-510.[CrossRef]
    [Google Scholar]
  2. Ames, R. S., Tornetta, M. A., Jones, C. S. & Tsui, P. ( 1994; ). Isolation of neutralizing anti-C5A monoclonal antibodies from a filamentous phage monovalent Fab display library. Journal of Immunology 152, 4572-4581.
    [Google Scholar]
  3. Aujame, L., Geofroy, F. & Sodoyer, R. ( 1997; ). High affinity human antibodies by phage display. Human Antibodies 8, 155-168.
    [Google Scholar]
  4. Barany, G. & Merrifield, R. B. ( 1980; ). Peptides: analysis, synthesis, biology. In The Peptides, pp. 1-284. Edited by M. T. Gross. New York:Academic Press.
  5. Barbas, C. F.III, Kang, A. S., Lerner, R. A. & Benkovic, S. J. ( 1991; ). Assembly of combinatorial antibody on phage surfaces: the gene III site. Proceedings of the National Academy of Sciences, USA 88, 7978-7982.[CrossRef]
    [Google Scholar]
  6. Bukh, J., Purcell, R. H. & Miller, R. H. ( 1992; ). Sequence analysis of the 5′ noncoding region of hepatitis C virus. Proceedings of the National Academy of Sciences, USA 89, 4942-4946.[CrossRef]
    [Google Scholar]
  7. Bukh, J., Purcell, R. H. & Miller, R. H. ( 1994; ). Sequence analysis of the core gene of 14 hepatitis C virus genotypes. Proceedings of the National Academy of Sciences, USA 91, 8239-8243.[CrossRef]
    [Google Scholar]
  8. Burton, D. R. & Barbas, C. F. ( 1994; ). Human antibodies from combinatorial libraries. Advances in Immunology 57, 191-280.
    [Google Scholar]
  9. Cerino, A. & Mondelli, M. U. ( 1991; ). Identification of an immunodominant B cell epitope on the HCV non structural region defined by human monoclonal antibodies. Journal of Immunology 147, 2092-2096.
    [Google Scholar]
  10. Cerino, A., Boender, P., La, M. N., Rosa, C., Habets, W. & Mondelli, M. U. ( 1993; ). A human monoclonal antibody specific for the N terminus of the hepatitis C virus nucleocapsid. Journal of Immunology 151, 7005-7015.
    [Google Scholar]
  11. Chan, S. W., Bye, J. M., Jackson, P. & Allain, J. P. ( 1996; ). Human recombinant antibodies specific for hepatitis C virus core and envelope E2 peptides from an immune phage display library. Journal of General Virology 77, 2531-2539.[CrossRef]
    [Google Scholar]
  12. Chemello, L., Cavalletto, D., Pontisso, P., Bortolotti, F., Donada, C., Donadon, V., Frezza, M., Casarin, P. & Alberti, A. ( 1993; ). Patterns of antibodies to hepatitis C virus in patients with chronic non-A, non-B hepatitis and their relationship to viral replication and liver disease. Hepatology 17, 179-182.[CrossRef]
    [Google Scholar]
  13. Chiba, J., Ohba, H., Matsuura, Y., Watanabe, Y., Katayama, T., Kikuchi, S., Saito, I. & Miyamura, T. ( 1991; ). Serodiagnosis of hepatitis C virus (HCV) infection with an HCV core protein molecularly expressed by a recombinant baculovirus. Proceedings of the National Academy of Sciences, USA 88, 4641-4645.[CrossRef]
    [Google Scholar]
  14. 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.[CrossRef]
    [Google Scholar]
  15. Choo, Q.-L., Weiner, A. J., Overby, L. R., Kuo, G., Houghton, M. & Bradley, D. W. ( 1990; ). Hepatitis C virus: the major causative agent of viral non-A, non-B hepatitis. British Medical Bulletin 46, 423-441.
    [Google Scholar]
  16. Clackson, T., Hoogenboom, H. R., Griffiths, A. D. & Winter, G. ( 1991; ). Making antibody fragments using phage display libraries. Nature 352, 624-628.[CrossRef]
    [Google Scholar]
  17. Engberg, J., Andersen, P. S., Nielsen, L. K., Dziegiel, M., Johansen, L. K. & Albrechtsen, B. ( 1995; ). Phage-display libraries of murine and human antibody Fab fragments. In Antibody Engineering Protocols, pp. 355-376. Edited by S. Paul. Totowa, NJ:Humana Press.
  18. Felici, F., Luzzago, A., Folgori, A. & Cortese, R. ( 1993; ). Mimicking of discontinuous epitopes by phage-displayed peptides. II. Selection of clones recognized by a protective monoclonal antibody against the Bordetella pertussis toxin from phage peptide libraries. Gene 128, 21-27.[CrossRef]
    [Google Scholar]
  19. Ferroni, P., Mascolo, G., Zaninetti, M., Colzani, D., Pregliasco, F., Pirisi, M., Barbone, F. & Gasparini, V. ( 1993; ). Identification of four epitopes in hepatitis C virus core protein. Journal of Clinical Microbiology 31, 1586-1591.
    [Google Scholar]
  20. Folgori, A., Tafi, R., Meola, A., Felici, F., Galfre, G., Cortese, R., Monaci, P. & Nicosia, A. ( 1994; ). A general strategy to identify mimotopes of pathological antigens using only random peptide libraries and human sera. EMBO Journal 13, 2236-2243.
    [Google Scholar]
  21. Goeser, T., Muller, H. M., Ye, J., Pfaff, E. & Theilmann, L. ( 1994; ). Characterization of antigenic determinants in the core antigen of the hepatitis C virus. Virology 205, 462-469.[CrossRef]
    [Google Scholar]
  22. Grakoui, A., McCourt, D. W., Wychowski, C., Feinstone, S. & Rice, C. M. ( 1993a; ). Characterization of the hepatitis C virus-encoded serine protease: determination of proteinase dependent polyprotein cleavage sites. Journal of Virology 67, 2832-2843.
    [Google Scholar]
  23. Grakoui, A., McCourt, D. W., Wychowski, C., Feinstone, S. & Rice, C. M. ( 1993b; ). A second hepatitis C virus encoded proteinase. Proceedings of the National Academy of Sciences, USA 90, 10583-10587.[CrossRef]
    [Google Scholar]
  24. Grakoui, A., Wychowski, C., Lin, C., Feinstone, S. M. & Rice, C. M. ( 1993c; ). Expression and identification of hepatitis C virus polyprotein cleavage products. Journal of Virology 67, 1385-1395.
    [Google Scholar]
  25. Griffiths, A. D., Williams, S. C., Hartley, O., Tomlinson, I. M., Waterhouse, P., Crosby, W. L., Kontermann, R. E., Jones, P. T., Low, N. M., Allison, T. J., Prospero, T. D., Hoogenboom, H. R., Nissim, A., Cox, J. P. L., Harrison, J. L., Zaccolo, M., Gherardi, E. & Winter, G. ( 1994; ). Isolation of high affinity human antibodies directly from large synthetic repertoires. EMBO Journal 13, 3245-3260.
    [Google Scholar]
  26. Harada, S., Watanabe, Y., Takeuchi, K., Suzuki, T., Katayama, T., Takebe, Y., Saito, I. & Miyamura, T. ( 1991; ). Expression of processed core protein of hepatitis C virus in mammalian cells. Journal of Virology 65, 3015-3021.
    [Google Scholar]
  27. Hijikata, M., Mizushima, H., Akagi, T., Mori, S., Kakiuchi, N., Kato, N., Tanaka, T., Kimura, K. & Shimotohno, K. ( 1993; ). Two distinct proteinase activities required for the processing of a putative nonstructural precursor protein of hepatitis C virus. Journal of Virology 67, 4665-4675.
    [Google Scholar]
  28. Hoess, R. H., Mack, A. J., Walton, H. & Reilly, T. M. ( 1994; ). Identification of a structural epitope by using a peptide library displayed on filamentous bacteriophage. Journal of Immunology 153, 724-729.
    [Google Scholar]
  29. Hoogenboom, H. R., Marks, J. D., Griffiths, A. D. & Winter, G. ( 1992; ). Building antibodies from their genes. Immunological Reviews 130, 41-68.[CrossRef]
    [Google Scholar]
  30. Hosein, B., Fang, C. T., Popovsky, M. A., Zhang, M. & Wang, C. Y. ( 1991; ). Improved serodiagnosis of hepatitis C virus infection with synthetic peptide antigen from capsid protein. Proceedings of the National Academy of Sciences, USA 88, 3647-3651.[CrossRef]
    [Google Scholar]
  31. Huse, W. D., Sastry, L., Iverson, S. A., Kang, A. S., Alting-Mees, M., Burton, D. R., Benkovic, S. J. & Lerner, R. A. ( 1989; ). Generation of a large combinatorial library of the immunoglobulin repertoire in phage lambda. Science 246, 1275-1281.[CrossRef]
    [Google Scholar]
  32. Jolivet-Reynaud, C., Dalbon, P., Viola, F., Yvon, S., Paranhos-Baccala, G., Piga, N., Bridon, L., Trabaud, M. A., Battail, N., Sibai, G. & Jolivet, M. ( 1998; ). HCV core immunodominant region analysis using monoclonal antibodies and human sera: characterization of major epitopes useful for antigen detection. Journal of Medical Virology 56, 300-309.[CrossRef]
    [Google Scholar]
  33. Kiyosawa, K. & Furuta, S. ( 1994; ). Hepatitis C virus and hepatocellular carcinoma. Current Studies in Hematology and Blood Transfusion 61, 98-120.
    [Google Scholar]
  34. Kiyosawa, K., Tanaka, E., Sodeyama, T. & Furuta, S. ( 1994; ). Natural history of Hepatitis C. Intervirology 37, 101-107.
    [Google Scholar]
  35. Kohler, G. & Milstein, C. ( 1975; ). Continuous cultures of fused cells secreting antibody of predefined specificity. Nature 256, 495-497.[CrossRef]
    [Google Scholar]
  36. Lane, D. P. & Stephen, C. W. ( 1993; ). Epitope mapping using bacteriophage peptide libraries. Current Opinion in Immunology 5, 268-271.[CrossRef]
    [Google Scholar]
  37. Lerner, R. A., Kang, A. S., Bain, J. D., Burton, D. R. & Barbas, C. F.III ( 1992; ). Antibodies without immunization. Science 258, 1313-1314.[CrossRef]
    [Google Scholar]
  38. McCafferty, J., Griffiths, A. D., Winter, G. & Chiswell, D. J. ( 1990; ). Phage antibodies: filamentous phage displaying antibody variable domains. Nature 348, 552-554.[CrossRef]
    [Google Scholar]
  39. Manabe, S., Fuke, I., Tanishita, O., Kaji, C., Gomi, Y., Yoshida, S., Mori, C., Takamizawa, A., Yosida, I. & Okayama, H. ( 1994; ). Production of nonstructural proteins of hepatitis C virus requires a putative viral protease encoded by NS3. Virology 198, 636-644.[CrossRef]
    [Google Scholar]
  40. Manzini, P., Calvo, P. L., Brunetto, M. R., Baldi, M., Abate, M. L., Oliveri, F., Negro, F., Balzola, F., Saracco, G., Verme, G., Chiba, J., Miyamura, T. & Bonino, F. ( 1993; ). Clinical significance of the antibody to the putative core protein of hepatitis C virus in patients with chronic liver disease. Liver 13, 222-226.
    [Google Scholar]
  41. Marks, J. D., Griffiths, A. D., Malmqvist, M., Clackson, T. P., Bye, J. M. & Winter, G. ( 1992; ). By-passing immunization: building high affinity human antibodies by chain shuffling. Bio/Technology 10, 779-783.[CrossRef]
    [Google Scholar]
  42. Moradpour, D., Wakita, T., Tokushige, K., Carlson, R. I., Krawczynski, K. & Wands, J. R. ( 1996; ). Characterization of three novel monoclonal antibodies against hepatitis C virus core protein. Journal of Medical Virology 48, 234-241.[CrossRef]
    [Google Scholar]
  43. Nasoff, M. S., Zebedee, S. L., Inchauspe, G. & Prince, A. M. ( 1991; ). Identification of an immunodominant epitope within the capsid protein of hepatitis C virus. Proceedings of the National Academy of Sciences, USA 88, 5462-5466.[CrossRef]
    [Google Scholar]
  44. Okamoto, H., Kanai, N. & Mishiro, S. ( 1992; ). Full-length nucleotide sequence of a Japanese hepatitis C virus isolate (HC-J1) with high homology to USA isolates. Nucleic Acids Research 20, 1042-1048.
    [Google Scholar]
  45. Parmley, S. F. & Smith, G. P. ( 1988; ). Antibody-selectable filamentous fd phage vectors: affinity purification of target genes. Gene 73, 305-318.[CrossRef]
    [Google Scholar]
  46. Penin, F., Ladaviere, L., Dalbon, P., Lacoux, X., Deleage, G. & Jolivet, M. (1997). Three-dimensional structural analysis of the immuno-dominant antigenic domain of the HCV nucleocapsid by 1H-NMR. In 4th International Meeting on Hepatitis C Virus and Related Viruses, HCV′97, Kyoto, Japan, p. 10.
  47. Plaisant, P., Burioni, R., Manzin, A., Solforosi, L., Candela, M., Gabrielli, A., Fadda, G. & Clementi, M. ( 1997; ). Human monoclonal recombinant Fabs specific for HCV antigens obtained by repertoire cloning in phage display combinatorial vectors. Research in Virology 148, 165-169.[CrossRef]
    [Google Scholar]
  48. Pujol, F. H., Khudyakov, Y. E., Devesa, M., Leon, G., Blitz, D. L., Monsalve, F., Lambert, S. B., Kalinina, T. Y., Liprandi, F. & Fields, H. A. ( 1996; ). Characterization of the antibody reactivity to synthetic peptides from different parts of the hepatitis C virus genome. Viral Immunology 9, 89-96.[CrossRef]
    [Google Scholar]
  49. Romeo, J. M., Ulrich, P. P., Busch, M. P. & Vyas, G. N. ( 1993; ). Analysis of hepatitis C virus RNA prevalence and surrogate markers of infection among seropositive voluntary blood donors. Hepatology 17, 188-195.[CrossRef]
    [Google Scholar]
  50. Sallberg, M., Ruden, U., Wahren, B. & Magnius, L. O. ( 1992; ). Immunodominant regions within the hepatitis C virus core and putative matrix proteins. Journal of Clinical Microbiology 30, 1989-1994.
    [Google Scholar]
  51. Sallberg, M., Pumpen, P., Zhang, Z. X., Lundholm, P., Gusars, I., Ruden, U., Wahren, B. & Magnius, L. O. ( 1994; ). Locations of antibody binding sites within conserved regions of the hepatitis C virus core protein. Journal of Medical Virology 43, 62-68.[CrossRef]
    [Google Scholar]
  52. Siemoneit, K., da Silva Cardoso, M., Wölpl, A., Epple, S. & Wintersinger, H. ( 1994a; ). Isotype-specific immune response to a single hepatitis C virus core. Annals of Hematology 69, 129-133.[CrossRef]
    [Google Scholar]
  53. Siemoneit, K., da Silva Cardoso, M., Wölpl, A., Koerner, K. & Kubanek, B. ( 1994b; ). Isolation and epitope characterization of human monoclonal antibodies to hepatitis C virus core antigen. Hybridoma 13, 9-13.[CrossRef]
    [Google Scholar]
  54. 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.[CrossRef]
    [Google Scholar]
  55. Smith, G. P. & Scott, J. K. ( 1993; ). Libraries of peptides and proteins displayed on filamentous phage. Methods in Enzymology 217, 228-257.
    [Google Scholar]
  56. Sodoyer, R., Peubez, I., Pion, C., Dubayle, J., Jacquemot, P., Geoffroy, F., Even, J. & Aujame, L. ( 1997; ). Full-scale ‘naı̈ve’ human antibody repertoires assembled from VH and VL variable regions. Human Antibodies 8, 37-42.
    [Google Scholar]
  57. Tanji, Y., Hijikata, M., Hirowatari, Y. & Shimotohno, K. ( 1994; ). Hepatitis C virus polyprotein processing: kinetics and mutagenic analysis of serine proteinase-dependent cleavage. Journal of Virology 68, 8418-8422.
    [Google Scholar]
  58. van der Poel, C. L. ( 1994; ). Hepatitis C virus infection from blood and blood products. FEMS Microbiology Reviews 14, 241-246.[CrossRef]
    [Google Scholar]
  59. Waterhouse, P., Griffiths, A. D., Johnson, K. S. & Winter, G. ( 1993; ). Combinatorial infection and in vivo recombination: a strategy for making large phage antibody repertoires. Nucleic Acids Research 21, 2265-2266.[CrossRef]
    [Google Scholar]
  60. Williamson, R. A., Burioni, R., Sanna, P. P., Partridge, L. J., Barbas, C. F. III & Burton, D. R. ( 1993; ). Human monoclonal antibodies against a plethora of viral pathogens from single combinatorial libraries. Proceedings of the National Academy of Sciences, USA 90, 4141-4145.[CrossRef]
    [Google Scholar]
  61. Winter, G., Griffiths, A. D., Hawkins, R. E. & Hoogenboom, H. R. ( 1994; ). Making antibodies by phage display technology. Annual Review of Immunology 12, 433-455.[CrossRef]
    [Google Scholar]
  62. Yoshikawa, A., Takahashi, K., Kishimoto, S., Tsuda, F., Akahane, Y., Naito, S., Tanaka, T., Yoshizawa, H., Yamasaki, M., Okamoto, H., Miyakawa, Y. & Mayumi, M. ( 1992; ). Serodiagnosis of hepatitis C virus infection by ELISA antibodies against the putative core protein (p20c) expressed in Escherichia coli. Journal of Immunological Methods 148, 143-150.[CrossRef]
    [Google Scholar]
  63. Yuki, N., Hayashi, N., Hagiwara, H., Naito, M., Ohkawa, K., Kasahara, A., Fusamoto, H., Ohtani, S., Okubo, Y. & Kamada, T. ( 1994; ). Hepatitis C virus antibodies and virus replication in asymptomatic blood donors. Vox Sanguinis 67, 280-285.[CrossRef]
    [Google Scholar]
  64. Zhai, W., Davies, J., Shang, D. Z., Chan, S. W. & Allain, J. P. ( 1999; ). Human recombinant single-chain antibody fragments, specific for the hypervariable region 1 of hepatitis C virus, from immune phage-display libraries. Journal of Viral Hepatitis 6, 115-124.[CrossRef]
    [Google Scholar]
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