Skip to main content
PMC home page
The Journal of Clinical Investigation logoLink to The Journal of Clinical Investigation
. 1985 Oct;76(4):1338–1347. doi: 10.1172/JCI112108

Structural analysis of hepatitis B surface antigen by monoclonal antibodies.

E Ben-Porath, J R Wands, R A Marciniak, M A Wong, L Hornstein, R Ryder, M Canlas, A Lingao, K J Isselbacher
PMCID: PMC424071  PMID: 2414317

Abstract

A method has been developed for the analysis of hepatitis B surface antigen (HBsAg) antigenic structure at the molecular level that creates "fingerprints" or "signatures" of various hepatitis B viral (HBV) strains. This technique employs high affinity IgM and IgG monoclonal antibodies (anti-HBs) directed against distinct and separate determinants on HBsAg. In performing this antigenic structural analysis, separate binding curves for different monoclonal anti-HBs are generated by measuring immunoreactivity in serial dilutions of HBsAg-positive serum by radioimmunoassay. Since the HBsAg concentration in serum is unknown, the binding profiles of groups of samples are aligned by an iterative least-squares procedure to generate the numerical signature characteristic of the viral strain. The numerical signatures are then displayed on a computer-graphic plot. The signature profiles of HBsAg subtypes are a true reflection of their antigenic structure, and in vertical and horizontal transmission studies the molecular characteristics of the viral epitopes are conserved. By signature analysis we found substantial antigenic heterogeneity among the ayw3 strain both in the U.S. and France, as well as in populations of the Far East and Africa. Populations in Ethiopia, Gambia, and the Philippines were infected with two antigenically distinct HBV strains. In some newly identified HBV strains, it was found that epitopes identified by some monoclonal antibodies were absent or substantially reduced, which suggested that a genetic mutation may have occurred. Thus this study suggests that there is far more antigenic heterogeneity in HBV than previously recognized. These variants are antigenically distinct from each other at the epitope level, and were heretofore unrecognized by polyvalent anti-HBsAg antibodies.

Full text

PDF
1342

Images in this article

1342Image
on p.1342

Selected References

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

  1. Bar-Shany S., Edwards V. M., Mosley J. W., Bancroft W. H. Subtypes of hepatitis B antigen among Israeli blood donors. Vox Sang. 1973;25(2):105–112. doi: 10.1111/j.1423-0410.1973.tb04818.x. [DOI] [PubMed] [Google Scholar]
  2. Beasley R. P., Hwang L. Y. Postnatal infectivity of hepatitis B surface antigen-carrier mothers. J Infect Dis. 1983 Feb;147(2):185–190. doi: 10.1093/infdis/147.2.185. [DOI] [PubMed] [Google Scholar]
  3. Bernier R. H., Sampliner R., Gerety R., Tabor E., Hamilton F., Nathanson N. Hepatitis B infection in households of chronic carriers of hepatitis B surface antigen: factors associated with prevalence of infection. Am J Epidemiol. 1982 Aug;116(2):199–211. doi: 10.1093/oxfordjournals.aje.a113406. [DOI] [PubMed] [Google Scholar]
  4. Bolton A. E., Hunter W. M. The labelling of proteins to high specific radioactivities by conjugation to a 125I-containing acylating agent. Biochem J. 1973 Jul;133(3):529–539. doi: 10.1042/bj1330529. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bouvier G. L., Williams A. Serotypes of hepatitis B antigen (HBs Ag): the problem of "new" determinants, as exemplified by "t". Am J Med Sci. 1975 Jul-Aug;270(1):165–171. doi: 10.1097/00000441-197507000-00023. [DOI] [PubMed] [Google Scholar]
  6. Couroucé-Pauty A. M., Plançon A., Soulier J. P. Distribution of HBsAg subtypes in the world. Vox Sang. 1983;44(4):197–211. doi: 10.1111/j.1423-0410.1983.tb01885.x. [DOI] [PubMed] [Google Scholar]
  7. Couroucé-Pauty A. M., Soulier J. P. Further data on HBs antigen subtypes - geographical distribution. Vox Sang. 1974;27(6):533–549. doi: 10.1111/j.1423-0410.1974.tb02451.x. [DOI] [PubMed] [Google Scholar]
  8. Couroucé A. M., Lee H., Drouet J., Canavaggio M., Soulier J. P. Monoclonal antibodies to HBsAg: a study of their specificities for eight different HBsAg subtypes. Dev Biol Stand. 1983;54:527–534. [PubMed] [Google Scholar]
  9. Couroucé A. M., Soulier J. P. The a(w) subdeterminants. Bibl Haematol. 1976;42:31–41. doi: 10.1159/000398987. [DOI] [PubMed] [Google Scholar]
  10. Crainic R., Couillin P., Blondel B., Cabau N., Boué A., Horodniceanu F. Natural variation of poliovirus neutralization epitopes. Infect Immun. 1983 Sep;41(3):1217–1225. doi: 10.1128/iai.41.3.1217-1225.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Crainic R., Couillin P., Cabau N., Boue A., Horodniceanu F. Determination of type 1 poliovirus subtype classes with neutralizing monoclonal antibodies. Dev Biol Stand. 1981;50:229–234. [PubMed] [Google Scholar]
  12. Donea-Debroise B., Brocteur J., Andre A., Remacle M. B. The use of radio-immuno-inhibition assay for the study of the y, d and w determinants of hepatitis B surface antigen. Biomedicine. 1979 Nov;30(5):260–264. [PubMed] [Google Scholar]
  13. Fujiyama A., Miyanohara A., Nozaki C., Yoneyama T., Ohtomo N., Matsubara K. Cloning and structural analyses of hepatitis B virus DNAs, subtype adr. Nucleic Acids Res. 1983 Jul 11;11(13):4601–4610. doi: 10.1093/nar/11.13.4601. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Gerhard W., Croce C. M., Lopes D., Koprowski H. Repertoire of antiviral antibodies expressed by somatic cell hybrids. Proc Natl Acad Sci U S A. 1978 Mar;75(3):1510–1514. doi: 10.1073/pnas.75.3.1510. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Humphrey D. D., Kew O. M., Feorino P. M. Monoclonal antibodies of four different specificities for neutralization of type 1 polioviruses. Infect Immun. 1982 May;36(2):841–843. doi: 10.1128/iai.36.2.841-843.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Okada K., Yamada T., Miyakawa Y., Mayumi M. Hepatitis B surface antigen in the serum of infants after deliver from asymptomatic carrier mothers. J Pediatr. 1975 Sep;87(3):360–363. doi: 10.1016/s0022-3476(75)80635-4. [DOI] [PubMed] [Google Scholar]
  17. Pereira L., Klassen T., Baringer J. R. Type-common and type-specific monoclonal antibody to herpes simplex virus type 1. Infect Immun. 1980 Aug;29(2):724–732. doi: 10.1128/iai.29.2.724-732.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Sampliner R. E., Loevinger B. L., Tabor E., Gerety R. J. Intrafamilial cluster of hepatitis B virus infection: study of a large family in the United States. Am J Epidemiol. 1981 Jan;113(1):50–54. doi: 10.1093/oxfordjournals.aje.a113065. [DOI] [PubMed] [Google Scholar]
  19. Sandler S. G., Couroucé A. M., Soulier J. P. wVariants of the hepatitis B surface antigen in Israeli populations. Vox Sang. 1978;34(3):149–151. doi: 10.1111/j.1423-0410.1978.tb02457.x. [DOI] [PubMed] [Google Scholar]
  20. Shorey J. A new hepatitis B virus surface antigen. J Infect Dis. 1976 Jan;133(1):1–6. doi: 10.1093/infdis/133.1.1. [DOI] [PubMed] [Google Scholar]
  21. Shorey J., Cheatum D. E., Combes B., Ziff M. Hepatitis B antigen and antibody in patients with systemic lupus erythematosus. Arthritis Rheum. 1974 Sep-Oct;17(5):583–589. doi: 10.1002/art.1780170512. [DOI] [PubMed] [Google Scholar]
  22. Shouval D., Shafritz D. A., Zurawski V. R., Jr, Isselbacher K. J., Wands J. R. Immunotherapy in nude mice of human hepatoma using monoclonal antibodies against hepatitis B virus. Nature. 1982 Aug 5;298(5874):567–569. doi: 10.1038/298567a0. [DOI] [PubMed] [Google Scholar]
  23. Soulier J. P., Couroucé-Pauty A. M. New determinants of hepatitis B antigen (Au or HB antigen). Vox Sang. 1973 Sep;25(3):212–234. doi: 10.1111/j.1423-0410.1973.tb04366.x. [DOI] [PubMed] [Google Scholar]
  24. Stevens C. E., Beasley R. P., Tsui J., Lee W. C. Vertical transmission of hepatitis B antigen in Taiwan. N Engl J Med. 1975 Apr 10;292(15):771–774. doi: 10.1056/NEJM197504102921503. [DOI] [PubMed] [Google Scholar]
  25. Szmuness W., Prince A. M., Hirsch R. L., Brotman B. Familial clustering of hepatitis B infection. N Engl J Med. 1973 Nov 29;289(22):1162–1166. doi: 10.1056/NEJM197311292892203. [DOI] [PubMed] [Google Scholar]
  26. Tong M. J., Weiner J. M., Ashcavai M. W., Redeker A. G., Comparini S., Vyas G. N. A comparative study of hepatitis B viral markers in the family members of Asian and non-Asian patients with hepatitis B surface antigen-positive hepatocellular carcinoma and with chronic hepatitis B infection. J Infect Dis. 1979 Oct;140(4):506–512. doi: 10.1093/infdis/140.4.506. [DOI] [PubMed] [Google Scholar]
  27. Wands J. R., Bruns R. R., Carlson R. I., Ware A., Menitove J. E., Isselbacher K. J. Monoclonal IgM radioimmunoassay for hepatitis B surface antigen: high binding activity in serum that is unreactive with conventional antibodies. Proc Natl Acad Sci U S A. 1982 Feb;79(4):1277–1281. doi: 10.1073/pnas.79.4.1277. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Wands J. R., Carlson R. I., Schoemaker H., Isselbacher K. J., Zurawski V. R., Jr Immunodiagnosis of hepatitis B with high-affinity IgM monoclonal antibodies. Proc Natl Acad Sci U S A. 1981 Feb;78(2):1214–1218. doi: 10.1073/pnas.78.2.1214. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Wands J. R., Marciniak R. A., Isselbacher K. J., Varghese M., Don G., Halliday J. W., Powell L. W. Demonstration of previously undetected hepatitis B viral determinants in an Australian Aboriginal population by monoclonal anti-hbs antibody radioimmunoassays. Lancet. 1982 May 1;1(8279):977–980. doi: 10.1016/s0140-6736(82)91988-2. [DOI] [PubMed] [Google Scholar]
  30. Wands J. R., Wong M. A., Shorey J., Brown R. D., Marciniak R. A., Isselbacher K. J. Hepatitis B viral antigenic structure: signature analysis by monoclonal radioimmunoassays. Proc Natl Acad Sci U S A. 1984 Apr;81(7):2237–2241. doi: 10.1073/pnas.81.7.2237. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Wands J. R., Zurawski V. R., Jr High affinity monoclonal antibodies to hepatitis B surface antigen (HBsAg) produced by somatic cell hybrids. Gastroenterology. 1981 Feb;80(2):225–232. [PubMed] [Google Scholar]
  32. Wiktor T. J., Koprowski H. Monoclonal antibodies against rabies virus produced by somatic cell hybridization: detection of antigenic variants. Proc Natl Acad Sci U S A. 1978 Aug;75(8):3938–3942. doi: 10.1073/pnas.75.8.3938. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Clinical Investigation are provided here courtesy of American Society for Clinical Investigation

RESOURCES