1887

Abstract

A 315 amino acid recombinant segment of the GB virus C (GBV-C) E2 envelope glycoprotein (E2-315) was expressed and secreted from CHO cells. E2-315 was purified by affinity chromatography using a monoclonal antibody directed to a FLAG sequence genetically engineered onto the C terminus of the recombinant protein. The secreted protein had a molecular mass of 48–56 kDa and was shown to be -glycosylated. Amino acid sequencing confirmed the expected N-terminal sequence. Purified E2-315 was used to develop an ELISA for detection of E2 antibodies in human sera. Antibodies to GBV-C E2 appeared to be directed toward conformational epitopes since human sera reactivity was detected in ELISA using native E2-315, but it was extremely weak or non-existent with denatured E2 protein. The use of an ELISA which can detect human GBV-C E2 antibodies will be important in further understanding of the clinical significance and epidemiology of GBV-C.

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1997-08-01
2024-03-28
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References

  1. Alter M. J., Margolis H. S., Krawczynski K., Judson F. N., Mares A., Alexander W. J., Hu P. Y., Miller J. K., Gerber M. A., Sampliner R. E., Meeks E. L., Beach M. J. 1992; The natural history of community acquired hepatitis C in the United States. New Engiand Journai of Medicine 327:1899–1905
    [Google Scholar]
  2. Alter H. J., Nakatsuji Y., Melpolder J., Wages J., Wesley R., Shih J.W.-K., Kim J. P. 1997a; The incidence of transfusion-associated hepatitis G virus infection and its relation to liver disease. New Engiand Journai of Medicine 336:747–754
    [Google Scholar]
  3. Alter M. J., Gallagher M., Morris T. T., Moyer L. A., Meeks E. L., Krawczynski K., Kim J. P., Margolis H. S. 1997b; Acute non-A-E hepatitis in the United States and the role of hepatitis G infection. New Engiand Journai of Medicine 336:741–746
    [Google Scholar]
  4. Becher P., Shannon A. D., Tautz N., Thiel H. -J. 1994; Molecular characterization of border disease virus, a pestivirus from sheep. Viroiogy 198:542–551
    [Google Scholar]
  5. Buti M., Jardi R., Rodriguez-Frias F., Quer J., Esteban R., Guardia J. 1994; Etiology of acute sporadic hepatitis in Spain: the role of hepatitis C and E viruses. Journai of Hepatoiogy 20:589–592
    [Google Scholar]
  6. Chien D. Y., Choo Q. -L., Ralston 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]
  7. Dawson G. J., Chau K. H., Cabal C. M., Yarbough P. O., Reyes G. R., Mushahwar I. K. 1992; Solid-phase enzyme-linked immunosorbent assay for hepatitis E virus IgG and IgM class antibodies utilizing recombinant proteins and synthetic peptides. Journai of Viroiogicai Methods 38:175–186
    [Google Scholar]
  8. Dawson G. J., Mimms L. T., Desai S. 1995; Recent advances in the molecular biology of the hepatitis viruses. Current Hepatoiogy 15:85–144
    [Google Scholar]
  9. Dawson G. J., Schlauder G. G., Pilot-Matias T. J., Thiele D., Leary T. P., Murphy P., Rosenblatt J. E., Simons J. N., Martinson F. E. A., Gutierrez R. A., Lentino J. R., Pachucki C., Muerhoff A. S., Widell A., Tegtmeier G., Desai S., Mushahwar I. K. 1996; Prevalence studies of GB virus-C infection using reverse transcriptase-polymerase chain reaction. Journal of Medicai Viroiogy 50:97–103
    [Google Scholar]
  10. Dille B. J., Surowy T. K., Gutierrez R. A., Coleman P. F., Knigge M. F., Carrick R. J., Aach R. D., Hollinger F. B., Stevens C. E., Barbosa L. H., Nemo G. J., Mosely J. W., Dawson G. J., Mushahwar I. K. 1997; ELISA for the detection of antibodies to the E2 protein of GBV- C. Journai of Infectious Diseases 175:458–461
    [Google Scholar]
  11. Donis R. O., Corapi W., Dubovi E. J. 1988; Neutralizing monoclonal antibodies to bovine diarrhoea virus bind to the 56K to 58K glycoprotein. Journai of Generai Viroiogy 69:77–86
    [Google Scholar]
  12. Dubuisson J., Rice C. M. 1996; Hepatitis C virus glycoprotein folding: disulfide bond formation and association with calnexin. Journai of Viroiogy 70:778–786
    [Google Scholar]
  13. Erker J. C., Simons J. N., Muerhoff A. S., Leary T. L., Chalmers M. L., Desai S. M., Mushahwar I. K. 1996; Molecular cloning and characterization of a GB virus C isolate from a patient with non A-E hepatitis. Journai of Generai Viroiogy 77:2713–2720
    [Google Scholar]
  14. Fagan E. A., David S. E., Tovey G. M., Lloyd G., Smith H. M., Portmann B., Tan K. -C., Zuckerman A. J., Williams R. 1992; Toga virus-like particles in acute liver failure attributed to sporadic non-A, non- B hepatitis and recurrence after liver transplantation. Journai of Medicai Viroiogy 38:71–77
    [Google Scholar]
  15. 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., Purcell R. H. 1992; Lack of protective immunity against reinfection with hepatitis C virus. Science 258:135–140
    [Google Scholar]
  16. Felgner P. L., Gadek T. R., Holm M., Roman R., Chan H. W., Wenz M., Northrop J. P., Ringold G. M., Danielsen M. 1987; Lipofection: a highly efficient, lipid-mediated DNA-transfection procedure. Proceedings of the National Academy of Sciences USA: 847413–7417
    [Google Scholar]
  17. Guirakhoo F., Heinz F. X., Kunz C. 1989; Epitope model of tick- borne encephalitis virus envelope glycoprotein E : analysis of structural properties, role of carbohydrate side chains and conformational changes occurring at acidic pH. Viroiogy 169:90–99
    [Google Scholar]
  18. Hall R. A., Kay B. H., Burgess G. W., Clancy P., Fanning I. D. 1990; Epitope analysis of the envelope and non-structural proteins of Murray Valley encephalitis virus. Journai of Generai Viroiogy 71:2923–2930
    [Google Scholar]
  19. Harada S., Suzuki R., Ando A., Watanabe Y., Yagi S., Miyamura T., Saito I. 1995; Establishment of a cell line constitutively expressing E2 glycoprotein of hepatitis C virus and humoral response of hepatitis C patients to the expressed protein. Journai of Generai Viroiogy 76:1223–1231
    [Google Scholar]
  20. Kuwanda S.K, Hollinger B., Lin H. J., Yarbough P. O., Wiesner R. H., Kaese D., Rakela J. 1994; Non-A, non-B fulminant hepatitis is also non-E and non-C. American Journai of Gastroenteroiogy 89:57–61
    [Google Scholar]
  21. Laemmli U. K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685
    [Google Scholar]
  22. Leary T. P., Muerhoff A. S., Simons J. N., Pilot-Matias T. J., Erker J. C., Chalmers M. L., Schlauder G. G., Dawson G. J., Desai S. M., Mushahwar I. K. 1996; Sequence and genomic organization of GBV- C: a novel member of the Flaviviridae associated with human non A-E hepatitis. Journal of Medicai Viroiogy 48:60–67
    [Google Scholar]
  23. Lesniewski R., Okasinski G., Carrick R., VanSant C., Desai S., Johnson R., Scheffel J., Moore B., Mushahwar I. 1995; Antibody to hepatitis C virus second envelope (HCV-E2) glycoprotein: a new marker of HCV infection closely associated with viremia. Journal of Medical Virology 45:415–422
    [Google Scholar]
  24. Lewis J. A., Chang G. -J., Lanciotti R. S., Kinney R. M., Mayer L. W., Trent D. W. 1993; Phylogenetic relationships of dengue-2 viruses. Virology 197:216–224
    [Google Scholar]
  25. Linnen J., Wages J., Zhang-Keck Z. -Y., Fry K. E., Krawczynski K. Z., Alter H., Koonin E., Gallagher M., Alter M., Hadziyannis S., Karayiannis P., Fung K., Nakatsuji Y., Shih J.W.-K., Young L., Piatak M., Hoover C., Fernandez J., Chen S., Zou J. -C., Morris T., Hyams K. C., Ismay S., Lifson J. D., Hess G., Foung S. K. H., Thomas H., Bradley D., Margolis H., Kim J. P. 1996; Molecular cloning and disease association of hepatitis G virus: a transfusion- transmissible agent. Science 271:505–508
    [Google Scholar]
  26. Masuko K., Mitsui T., Iwano K., Yamazaki C., Okuda K., Meguro T., Murayama N., Inoue T., Tsuda F., Okamoto H., Miyakawa Y., Mayumi M. 1996; Infection with hepatitis GB virus C in patients on maintenance hemodialysis. New England Journal of Medicine 334:1485–1490
    [Google Scholar]
  27. Monath T. P., Heinz F. X. 1996; Flaviviruses. In Fields Virology, 3rd edn. pp 961–1034 Fields B. N., Knipe D. M., Howley P. M. Edited by Philadelphia: Lippincott-Raven;
    [Google Scholar]
  28. Muerhoff A. S., Leary T. P., Simons J. N., Pilot-Matias T. J., Dawson G. J., Erker J. C., Chalmers M. L., Schlauder G. G., Desai S. M., Mushahwar I. K. 1995; Genomic organization of GB viruses A and B: two new members of the Flaviviridae associated with GB agent hepatitis. Journal of Virology 69:5621–5630
    [Google Scholar]
  29. Nowak T., Wengler G. 1987; Analysis of disulfides present in the membrane proteins of the West Nile flavivirus. Virology 156:127–137
    [Google Scholar]
  30. Paroli M., Accapezzato D., Barnaba V. 1995; Virus escape from immune control : mechanisms of persistence within the host. Research in Virology 146:267–271
    [Google Scholar]
  31. Peters T., Schlauer H. J., Preisler S., Kopp B., Berthold H., Gerok W., Rasenack J. 1993; Frequency of hepatitis C in acute posttransfusion hepatitis after open-heart surgery: a prospective study in 1,476 patients. Journal of Medical Virology 39:139–145
    [Google Scholar]
  32. Pilot-Matias T. J., Muerhoff A. S., Simons J. N., Leary T. P., Buijk S. L., Chalmers M. L., Erker J. C., Dawson G. J., Desai S. M., Mushahwar I. K. 1996a; Identification of antigenic regions in the GB hepatitis viruses GBV-A, GBV-B and GBV-C. Journal of Medical Virology 48:329–338
    [Google Scholar]
  33. Pilot-Matias T. J., Carrick R. J., Coleman P. F., Leary T. P., Surowy T. K., Simons J. N., Muerhoff A. S., Buijk S. L., Chalmers M. L., Dawson G. J., Desai S. M., Mushahwar I. K. 1996b; Expression of the GB virus C E2 glycoprotein using the Semliki Forest virus vector system and its utility as a serologic marker. Virology 225:282–292
    [Google Scholar]
  34. Rumenapf T., Stark R., Meyers G., Thiel H. -J. 1991; Structural proteins of hog cholera virus expressed by vaccinia virus : characterization and induction of protective immunity. Journal of Virology 65:589–597
    [Google Scholar]
  35. Sallie R., Silva A. E., Purdy M., Smith H., McCaustland K., Tibbs C., Portmann B., Eddleston A., Bradley D., Williams R. 1994; Hepatitis C and E in non-A, non-B fulminant hepatic failure : a polymerase chain reaction and serological study. Journal of Hepatology 20:580–588
    [Google Scholar]
  36. Schimke R. T. 1988; Gene amplification in cultured cells. Journal of Biological Chemistry 263:5989–5991
    [Google Scholar]
  37. Simons J. N., Pilot-Matias T. J., Leary T. P., Dawson G. J., Desai S. M., Schlauder G. G., Muerhoff A. S., Erker J. C., Buijk S. L., Chalmers M. L., VanSant C. L., Mushahwar I. K. 1995a; Identification of two new flavi-like genomes in the GB hepatitis agent. Proceedings of the National Academy of Sciences USA: 923401–3405
    [Google Scholar]
  38. Simons J. N., Leary T. P., Dawson G. J., Pilot-Matias T. J., Muerhoff A. S., Schlauder G. G., Desai S. M., Mushahwar I. K. 1995b; Isolation of novel virus-like sequences associated with human hepatitis. Nature Medicine 1:564–569
    [Google Scholar]
  39. Spaete R. R., Alexander D., Rugroden M. E., Choo Q. -L., Berger K., Crawford K., Kuo C., Leng S., Lee C., Ralston R., Thudium K., Tung J. W., Kuo G., Houghton M. 1992; Characterization of the hepatitis C virus E2/NS1 gene product expressed in mammalian cells. Virology 188:819–830
    [Google Scholar]
  40. Tassopoulos N. C., Hatzakis A., Delladetsima I., Koutelou M. G., Todoulos A., Miriagou V. 1992; Role of hepatitis C virus in acute non-A, non-B hepatitis in Greece : a five-year prospective study. Gastroenterology 102:969–972
    [Google Scholar]
  41. Thiers V., Lunel F., Valla D., Azar N., Fretz C., Frangeul L., Huraux J. -M., Oplon P., Brechot C. 1993; Posttransfusional anti-HCV- negative non-A, non-B hepatitis (II) : serological and polymerase chain reaction analysis for hepatitis C and hepatitis B viruses. Journal of Hepatology 18:34–39
    [Google Scholar]
  42. Towbin H., Staehelin T., Gordon J. 1979; Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets : procedure and some applications. Proceedings of the National Academy of Sciences USA: 764350–4354
    [Google Scholar]
  43. Weiland E., Stark R., Haas B., Rumenapf T., Meyers G., Thiel H. -J. 1990; Pestivirus glycoprotein which induces neutralizing antibodies forms part of a disulfide-linked heterodimer. Journal of Virology 64:3563–3569
    [Google Scholar]
  44. Weiland E., Ahl R., Stark R., Weiland F., Thiel H. -J. 1992; A second envelope glycoprotein mediates neutralization of a pestivirus, hog cholera virus. Journal of Virology 66:3677–3682
    [Google Scholar]
  45. 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., Brunetto M., Barr P. J., Miyamura T., McHutchinson J., Houghton M. 1992; Evidence for immune selection of hepatitis C virus (HCV) putative envelope glycoprotein variants: potential role in chronic HCV infections. Proceedings of the National Academy of Sciences USA: 893468–3472
    [Google Scholar]
  46. Yoshiba M., Okamoto H., Mishiro S. 1995; Detection ofthe GBV- C hepatitis virus genome in serum from patients with fulminant hepatitis of unknown aetiology. Lancet 346:1131–1132
    [Google Scholar]
  47. Zaaijer H. L., Vallari D. S., Cunningham M., Lesniewski R., Reesnick H. W., VanDerPoel C. L., Lelie P. N. 1994; E2 and NS5: new antigens for detection of hepatitis C virus antibodies. Journal of Medical Virology 44:395–397
    [Google Scholar]
  48. Zuckerman A. J. 1996; Alphabet of hepatitis viruses. Lancet 347:558–559
    [Google Scholar]
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