Journal of Clinical Immunology

, Volume 9, Issue 3, pp 229–241 | Cite as

In vitro immune responses to hepatitis B surface antigen (pre-S2 and S) following remote infection by hepatitis B virus in humans

  • Thomas R. Cupps
  • Jay H. Hoofnagle
  • Ronald W. Ellis
  • William J. Miller
  • Leonard Seeff
  • Ann Guerrera
  • John L. Gerin
  • Sally A. Haas-Smith
Original Articles


In this report we evaluate the human immune response to hepatitis B surface antigen (HBsAg) following remote infection with hepatitis B virus (HBV). HBsAg-reactive lymphocytes can be readily demonstrated in the peripheral blood of individuals with established immunity following infection with HBV.In vitro stimulation with small doses of plasma-derived HBsAg, yeast-derived HBsAg (S region) or pre-S2 peptide will induce specific IgG to HBsAg (anti-HBs) in the absence of a polyclonal increase in total IgG. The pre-S2 peptide will stimulate, in a T cell-dependent fashion, thein vitro production of anti-HBs with specificity for the S domain. This anti-HBs production is mediated by pre-S2-stimulated soluble T-cell factors. Peripheral blood mononuclear cells from individuals with established immunity proliferate to the yeast-derived HBsAg but not to the plasma-derived HBsAg or pre-S2 peptide. The chronic HBsAg carriers do not produce anti-HBs following stimulation with HBsAg regardless of the source or component of antigen used. Different study protocols failed to demonstrate HBsAg-specific responses in the peripheral blood mononuclear cells of chronic carriers.

Key words

Hepatitis B virus anti-HBs hepatitis B surface antigen (HBsAg) chronic carrier pre-S2 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Hoofnagle JH: Type B hepatitis. Serology, virology and clinical course. Semin Liver Dis 1:6–13, 1981Google Scholar
  2. 2.
    Levo Y, Gorevic PD, Kassab HJ, Zucker-Franklin D, Franklin EC: Association between hepatitis B virus and essential mixed cryoglobulinemia. N Engl J Med 296:1501–1504, 1977Google Scholar
  3. 3.
    Brzosko WJ, Krawcynski K, Nazarewicz T, Morzycka M, Nowoslawski A: Glomerulonephritis associated with hepatitis B surface antigen immune complexes in children. Lancet 2:477–482, 1974Google Scholar
  4. 4.
    Kohler PF, Cronin RE, Hammond WS, Olin D, Carr RI: Chronic membranous glomerulonephritis caused from hepatitis B antigen-antibody immune complexes. Ann Intern Med 81:448–451, 1974Google Scholar
  5. 5.
    Sergent JS, Lockshin MD, Christian CL, Gocke DJ: Vasculitis with hepatitis B antigenemia. Long-term observations in nine patients. Medicine (Baltimore) 55:1–18, 1976Google Scholar
  6. 6.
    Duffy J, Lidsky MD, Sharp JT, Davis JS, Person DA, Hollinger FB, Min KW: Polyarthritis, polyarteritis and hepatitis B. Medicine (Baltimore) 55:19–37, 1976Google Scholar
  7. 7.
    Sherlock S, Fox RA, Niazi SP, Scheuer PJ: Chronic liver disease and primary liver-cell cancer with hepatitis-associated (Australia) antigen in serum. Lancet 1:1243–1247, 1970Google Scholar
  8. 8.
    Bolin TD, Davis AE, Liddelow AG: Liver disease and cell-mediated immunity in hepatitis-associated antigen (HAA) carriers. Gut 14:365–368, 1973Google Scholar
  9. 9.
    Hanson RG, Cooksley WGE, Jones EA, Hoofnagle JH: Immunoregulation of synthesis of antibody to hepatitis B surface antigen in chronic type B heptitis. Hepatology 2:727–733, 1982Google Scholar
  10. 10.
    Dusheiko GM, Hoofnagle JH, Cooksley WG, James SP, Jones EA: Synthesis of antibodies to hepatitis virus by cultured lymphocytes from chronic hepatitis B surface antigen carriers. J Clin Invest 71:1104–1113, 1983Google Scholar
  11. 11.
    Barnaba V, Valesini G, Levrero M, Clarazaccari M, Van Dyke A, Falco M, Musca A, Balsano F: Immunoregulation of the in vitro anti-HBs antibody synthesis in chronic HBsAg carriers and in recently boosted anti-hepatitis vaccine recipients. Clin Exp Immunol 60:259–266, 1985Google Scholar
  12. 12.
    Nowicki M, Tong M, Bohman R: Alterations in the immune response of nonresponders to the hepatitis B vaccine. J Infect Dis 152:1245–1248, 1985Google Scholar
  13. 13.
    Nowicki M, Tong M, Nair P, Stevenson D: Impaired antibody synthesis in patients with chronic hepatitis B infection. Hepatology 6:180–185, 1986Google Scholar
  14. 14.
    Celis E, Kung P, Chang T: Hepatitis B virus-reactive human T lymphocyte clones: antigen specificity and helper function for antibody synthesis. J Immunol 132:1511–1516, 1984Google Scholar
  15. 15.
    Cupps T, Gerin J, Purcell R, Goldsmith P, Fauci A: In vitro antigen-induced antibody responses to hepatitis B surface antigen in man: Kinetic and cellular requirements. J Clin Invest 74:1204–1213, 1984Google Scholar
  16. 16.
    Hellstrom U, Sylvan S, Lundberg P: Regulatory functions of T- and accessory-cells for hepatitis B surface antigen induced specific antibody production and proliferation of human peripheral blood lymphocytes. J Cllin Lab Immunol 16:173–181, 1985Google Scholar
  17. 17.
    Gerin JL, Faust RM, Holland PV: Biophysical characterization of the adr subtype of hepatitis B antigen and preparation of anti-r sera in rabbits. J Immunol 115:100–105, 1975Google Scholar
  18. 18.
    Lowry OH, Rosebrough NJ, Farr AL, Randall RJ: Protein measurement with folin phenol reagent. J Biol Chem 193:265–275, 1951Google Scholar
  19. 19.
    Wampler DE, Lehman ED, Boyer J, McAleer WJ, Scolnick EM: Multiple chemical forms of hepatitis B surface antigen produced in yeast. Proc Natl Acad Sci USA 82:6830–6834, 1985Google Scholar
  20. 20.
    Neurath AR, Kent SBH, Strick N: Location and chemical synthesis of a pre-S gene coded immunodominant epitope of hepatitis B virus. Science 224:392–395, 1984Google Scholar
  21. 21.
    Boyum A: Isolation of mononuclear cells and granulocytes from human blood. Scand J Clin Invest 21 (Suppl):77–89, 1968Google Scholar
  22. 22.
    Falkoff RJM, Zhu LP, Fauci AS: Separate signals for human B cell proliferation and differentiation in response to Staphy-lococcus aureus. Evidence for a two signal model of B cell activation. J Immunol 129:97–102, 1982Google Scholar
  23. 23.
    Werner CH, Klouda PT, Correa MC, Vassali P, Jeannet M: Isolation of B and T lymphocytes by nylon fiber columns. Tissue Antigens 9:227–229, 1977Google Scholar
  24. 24.
    Lane HC, Depper JM, Greene WC, Whalen G, Waldmann TA, Fauci AS: Qualitative analysis of immune function in patients with the acquired immunodeficiency syndrome. N Engl J Med 313:79–84, 1985Google Scholar
  25. 25.
    Lane HC, Volkman DJ, Fauci AS: In vitro antigen-induced, antigen specific antibody production in man. Specific and polyclonal components, kinetics, and cellular requirements. J Exp Med 154:1043–1057, 1981Google Scholar
  26. 26.
    Volkman DJ, Buescher ES, Gallin JI, Fauci AS: B cell lines as models for inherited phagocytic diseases: Abnormal superoxide generation in chronic granulomatous disease and giant granules in Chediak-Higashi syndrome. J Immunol 133:3006–3009, 1984Google Scholar
  27. 27.
    Volkman DJ, Allyn SP, Fauci AS: Antigen-inducedin vitro antibody production in humans: tetanus toxoid-specified antibody synthesis. J Immunol 129:107–112, 1982Google Scholar
  28. 28.
    Neurath AR, Kent SBH, Strick N, Stark D, Sproul P: Genetic restriction of immune responsiveness to synthetic peptides corresponding to the sequence in the pre-S region of hepatitis B virus (HBV) envelope gene. J Immunol 17:119–125, 1985Google Scholar
  29. 29.
    Milich D, Thornton G, Neurath A, Kent S, Michael M, Tiollais P, Chisari F: Enhanced immunogenicity of the Pre-S region hepatitis B surface antigen. Science 228:1195–1198, 1985Google Scholar
  30. 30.
    Milich D, Peterson D, Leroux-Roels G, Lerner R, Chisari F: Genetic regulation of the immune response to hepatitis B surface antigen (HBsAg). VI. T cell fine specificity. J Immunol 134:4203–4211, 1985Google Scholar
  31. 31.
    Milich M, McLachlan A, Chisari F, Kent S, Thornton G: Immune response to the Pre-S(1)-specific T cell response can bypass nonresponsiveness to the Pre-S(2) and S regions of HBsAg. J Immunol 137:315–322, 1986Google Scholar
  32. 32.
    Milich D, McLachlan A, Chisari F, Thornton G: Nonover-lapping T and B cell determinants on an hepatitis B surface antigen Pre-S(2) region synthetic peptide. J Exp Med 164:532–547, 1986Google Scholar
  33. 33.
    Bianchi L, Gudat F: Immunopathology of hepatitis B.In Progress in Liver Diseases, H Popper, F Schaffner (eds). New York, Grune and Stratton, 1979, Vol 6, pp 371–392Google Scholar
  34. 34.
    Meyer Zum Buschenfelde KH, Hutteroth TH, Arnold W, Hopf V: Immunologic liver injury: The role of hepatitis B viral antigens and liver membrane antigens as targets.In Progress in Liver Diseases, H Popper, F Schaffner, (eds). New York, Grune and Stratton, 1979, Vol 6, pp 407–424Google Scholar
  35. 35.
    Laiwah ACCY, Chaundhuri AKR, Anderson JR: Lymphocyte transformation and leukocyte migration inhibition by Australia antigen. Clin Exp Immunol 15:27–34, 1973Google Scholar
  36. 36.
    DeGast GL, Houwen B, Niewig HO: Specific lymphocyte stimulation by purified, heat-inactivated hepatitis B antigen. Br Med J 4:707–709, 1973Google Scholar
  37. 37.
    Tong MJ, Wallace AM, Peters RL. Reynolds TB: Lymphocyte stimulation in hepatitis B infection. N Engl J Med 293:318–322, 1975Google Scholar
  38. 38.
    Wands JR, Perrotto JL, Alperti E, Isselbacher KJ: Cell-mediated immunity in acute and chronic hepatitis. J Clin Invest 55:921–929, 1975Google Scholar
  39. 39.
    Celis E, Chang T: Antibodies to hepatitis B surface antigen potentiate the response of human T lymphocyte clones to the same antigen. Science 224:297–299, 1984Google Scholar
  40. 40.
    Dudley F, Giustino V, Sherlock S: Cell-mediated immunity in patients positive for hepatitis-associated antigen. Br Med J 4:754–756, 1972Google Scholar
  41. 41.
    Tiku M, Beutner K, Tiku K, Orga P: Cell-mediated immune response to liver tissue antigen and hepatitis B surface antigen after infection with hepatitis B virus in humans. J Infect Dis 138:587–596, 1978Google Scholar
  42. 42.
    Barnaba B, Levero M, Franco A, Zaccari C, Musca A, Balsano F: Antigen specific suppressor T cells from chronic hepatitis B virus (HBV) carriers inhibit the responsiveness to HBsAg of allogenic high-responder lymphocytes. J Clin Lab Immunol 16:137–142, 1985Google Scholar
  43. 43.
    Ellner JJ: Suppressor adherent cells in human tuberculosis. J Immunol 121:2573–2579, 1978Google Scholar
  44. 44.
    Ottesen EA: Modulation of host response in human schistosomiasis. I. Adherent suppressor cells that inhibit lymphocyte proliferative responses to parasite antigens. J Immunol 123:1639–1644, 1979Google Scholar
  45. 45.
    Piessens WF, Ratiwayanto S, Tuti S, Palmieri JH, Piessens PW, Koiman I, Dennis DT: Antigen-specific suppressor cells and suppressor factors in human filariasis withBrugia malayi. N Engl J Med 302:833–837, 1980Google Scholar
  46. 46.
    Chisari FV, Edgington TS: Lymphocyte E rosette inhibitory factor: A regulatory serum lipoprotein. J Exp Med 142:1092–1107, 1975Google Scholar
  47. 47.
    Chisari FV, Routenberg JA, Edgington TS: Mechanisms responsible for defective human T-lymphocyte sheep erythrocyte rosette function associated with hepatitis B virus infection. J Clin Invest 57:1227–1238, 1976Google Scholar
  48. 48.
    Neurath AR, Strick N: Radioimmunoassay for albuminbinding sites associated with HBsAg: Correlation of results with the presence of e-antigen in serum. Intervirology 11:128–132, 1979Google Scholar
  49. 49.
    Imai M, Yansae Y, Nojiri T, Migakawa Y, Mayumi M: A receptor for polymerized human and chimpanzee albumins in hepatitis B virus particles co-occurring with HBsAg. Gastroenterology 76:242–247, 1979Google Scholar
  50. 50.
    Machida A, Kishimoto S, Ohnuma H, Miyamoto H, Baba K, Oda K, Nakamura T, Miyakawa Y, Mayumi M: A hepatitis B surface antigen polypeptide (P31) with the receptor for polymerized human as well as chimpanzee albumins. Gastroenterology 85:268–274, 1983Google Scholar
  51. 51.
    Pontisso P, Falcieri E, Schiavon E, Alberti A, Realdi F: Polyalbumin receptors on hepatitis B virus and on 22 nm hepatitis B surface antigen (HBsAg)2 particles. J Med Virol 13:355–360, 1984Google Scholar
  52. 52.
    Yu M, Finlayson J, Shih J: Interactions between various polymerized human albumins and hepatitis B surface antigen. J Virol 55:736–743, 1985Google Scholar
  53. 53.
    Milich D, Papas E, Bhatnager P, Vyas G: Interactions between polymerized human albumin, hepatitis B surface antigen, and complement. I. Binding of polyalbumin to Clq. J Med Virol 7:181–192, 1981Google Scholar
  54. 54.
    Milich D, Papas E, Vyas G: Interactions between polymerized human albumin, hepatitis B surface antigen, and complement. II. involvement of Clq in or near the hepatitis B surface antigen receptor for polyalbumin. J Med Virol 7:193–204, 1981Google Scholar
  55. 55.
    Careoda F, deFranhis R, Monforte A, Vecchi M, Rossi E, Primignani M, Palla M, Dioguardi N: Persistence of circulating HBsAg/IgM complexes in acute viral hepatitis, type B: an early marker of chronic evolution. Lancet 2:358–360, 1982Google Scholar
  56. 56.
    Coursaget P, Courouce A, Barres J, Bouchardeau F, Petat E, Chiron J: Complexes between HBsAg and IgM in serum of patients with viral hepatitis B. Ann Inst Pasteur/Virol 136:141–149, 1985Google Scholar
  57. 57.
    Craxi A, Magrin S, Greco J, Vinci M, Tine F, Raimondo G, Longo G, Pagliaro L: Polyalbumin receptors, hepatitis B surface antigen (HBsAg), and HBsAg/IgM complexes in HBsAg positive patients with and without delta superinfection. J Med Virol 15:383–388, 1985Google Scholar
  58. 58.
    Celis E, Zurawski VR Jr, Chang TW: Regulation of T-cell function by antibodies: Enhancement of the response of human T-cell clones to hepatitis B surface antigen by antigen-specific monoclonal antibodies. Proc Natl Acad Sci USA 81:6846–6850, 1984Google Scholar
  59. 59.
    Celis E, Chang T: Role for monoclonal hepatitis B antibody in hepatitis B immunization programmes. Lancet 1:1219, 1985Google Scholar
  60. 60.
    Peterson LB, Wilner GD, Thomas DW: Proliferating and helper T lymphocytes display distinct specificities in response to human fibronectin B. J Immunol 130:2542–2545, 1983Google Scholar
  61. 61.
    Neurath AR, Kent SBH, Strick N, Taylor P, Stevens CE: Hepatitis B virus contains pre-S gene-encoded domains. Nature 315:154–156, 1985Google Scholar
  62. 62.
    Budkowska A, Dubreuil P, Capel F, Pillot J: Hepatitis B virus pre-S gene-encoded antigenic specificity and anti-pre-S antibody: Relationship between anti-pre-S response and recovery. Hepatology 6:360–368, 1986Google Scholar
  63. 63.
    Hellstrom U, Sylvan S, Kuhns M, Sarin V: Absence of pre-S2 Antibodies in natural hepatitis B virus infection. Lancet 2:889–893, 1986Google Scholar
  64. 64.
    Peters M, Fauci A: Selective activation of antigen-specific human B cells in recently immunized individuals by nonspecific factors in the absence of antigen. J Immunol 130:678–680, 1983Google Scholar
  65. 65.
    Vento S, Rondanelli EG, Ranieri S, O'Brien CJ, Williams R, Eddleston ALWF: Prospective study of cellular immunity to hepatitis B-virus antigens from the early incubation phase of acute hepatitis B. Lancet 2:119–122, 1987Google Scholar

Copyright information

© Plenum Publishing Corporation 1989

Authors and Affiliations

  • Thomas R. Cupps
    • 1
  • Jay H. Hoofnagle
    • 2
  • Ronald W. Ellis
    • 3
  • William J. Miller
    • 3
  • Leonard Seeff
    • 4
  • Ann Guerrera
    • 1
  • John L. Gerin
    • 5
  • Sally A. Haas-Smith
    • 1
  1. 1.Division of Rheumatology, Immunology, and Allergy, Department of MedicineGeorgetown University Medical CenterWashington, DC
  2. 2.Liver Disease Section, Digestive Diseases BranchNational Institute of Diabetes, Digestive and Kidney Diseases, National Institutes of HealthBethesda
  3. 3.Department of Virus and Cell BiologyMerck Sharp & Dohme Research LaboratoriesWest Point
  4. 4.Division of Gastroenterology/Hepatology, Department of MedicineVeterans Administration Medical CenterWashington, DC
  5. 5.Division of Molecular Virology & ImmunologyGeorgetown University Medical CenterRockville

Personalised recommendations