Skip to main content
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1982 Dec;79(24):7871–7875. doi: 10.1073/pnas.79.24.7871

Specificity of antibodies elicited by a synthetic peptide having a sequence in common with a fragment of a virus protein, the hepatitis B surface antigen.

A R Neurath, S B Kent, N Strick
PMCID: PMC347451  PMID: 6185953

Abstract

We predicted the localization of a major hepatitis B surface antigen (HBsAg) determinant within residues 135-155 [Neurath, A.R., Strick, N. & Oleszko, W.R. (1981) J. Virol. Methods 3, 115-125]. A peptide corresponding to this sequence (P135-155) was synthesized, linked to macromolecular carriers, and used to immunize rabbits. In accordance with published data on shorter peptides within the same amino acid sequence, antibodies to HBsAg were elicited. A detailed analysis of the immune response to P135-155 revealed the following: A heterogeneous population of IgG and IgM antibodies reacting with P135-155 was detected in the antisera by a variety of radioimmunoassays and enzyme-linked fluorescence immunoassays. Only a subpopulation of these antibodies reacted with HBsAg. The equilibrium constant (K) for the reaction of the antibodies with HBsAg (K = 4 X 10(5) M-1) was approximately two orders of magnitude lower than K for the reaction with P135-155 and was below K for the reaction between HBsAg and antibodies elicited by HBsAg (K greater than 10(7) M-1). Preimmunization with P135-155 did not result in an enhanced response to subsequent immunization with HBsAg. Peptides more accurately mimicking determinants on HBsAg may have to be synthesized for possible application in antiviral prophylaxis.

Full text

PDF
7871

Selected References

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

  1. Anilionis A., Wunner W. H., Curtis P. J. Structure of the glycoprotein gene in rabies virus. Nature. 1981 Nov 19;294(5838):275–278. doi: 10.1038/294275a0. [DOI] [PubMed] [Google Scholar]
  2. Arnon R. Chemically defined antiviral vaccines. Annu Rev Microbiol. 1980;34:593–618. doi: 10.1146/annurev.mi.34.100180.003113. [DOI] [PubMed] [Google Scholar]
  3. Arnon R., Sela M. Antibodies to a unique region in lysozyme provoked by a synthetic antigen conjugate. Proc Natl Acad Sci U S A. 1969 Jan;62(1):163–170. doi: 10.1073/pnas.62.1.163. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Charnay P., Mandart E., Hampe A., Fitoussi F., Tiollais P., Galibert F. Localization on the viral genome and nucleotide sequence of the gene coding for the two major polypeptides of the hepatitis B surface antigen (HBs Ag). Nucleic Acids Res. 1979 Sep 25;7(2):335–346. doi: 10.1093/nar/7.2.335. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Czernilofsky A. P., Levinson A. D., Varmus H. E., Bishop J. M., Tischer E., Goodman H. M. Nucleotide sequence of an avian sarcoma virus oncogene (src) and proposed amino acid sequence for gene product. Nature. 1980 Sep 18;287(5779):198–203. doi: 10.1038/287198a0. [DOI] [PubMed] [Google Scholar]
  6. Dreesman G. R., Sanchez Y., Ionescu-Matiu I., Sparrow J. T., Six H. R., Peterson D. L., Hollinger F. B., Melnick J. L. Antibody to hepatitis B surface antigen after a single inoculation of uncoupled synthetic HBsAg peptides. Nature. 1982 Jan 14;295(5845):158–160. doi: 10.1038/295158a0. [DOI] [PubMed] [Google Scholar]
  7. Fiers W., Contreras R., Haegemann G., Rogiers R., Van de Voorde A., Van Heuverswyn H., Van Herreweghe J., Volckaert G., Ysebaert M. Complete nucleotide sequence of SV40 DNA. Nature. 1978 May 11;273(5658):113–120. doi: 10.1038/273113a0. [DOI] [PubMed] [Google Scholar]
  8. Garoff H., Frischauf A. M., Simons K., Lehrach H., Delius H. Nucleotide sequence of cdna coding for Semliki Forest virus membrane glycoproteins. Nature. 1980 Nov 20;288(5788):236–241. doi: 10.1038/288236a0. [DOI] [PubMed] [Google Scholar]
  9. Goldenberg K., Berkowitz J. M., Praissman M. Methods in the quantitation of antigen-antibody reactions measured by radioimmunoassay: application to the gastrin system revealing large and restricted binding energies. J Immunol. 1974 Mar;112(3):1008–1018. [PubMed] [Google Scholar]
  10. Hardy M. M., Moore D. M. Neutralization of foot-and-mouth disease virus. I. Sensitization of the 140S virion by antibody also reactive with the 12S protein subunit. J Gen Virol. 1981 Aug;55(Pt 2):415–427. doi: 10.1099/0022-1317-55-2-415. [DOI] [PubMed] [Google Scholar]
  11. Hopp T. P. A synthetic peptide with hepatitis B surface antigen reactivity. Mol Immunol. 1981 Sep;18(9):869–872. doi: 10.1016/0161-5890(81)90009-2. [DOI] [PubMed] [Google Scholar]
  12. Jou W. M., Verhoeyen M., Devos R., Saman E., Fang R., Huylebroeck D., Fiers W., Threlfall G., Barber C., Carey N. Complete structure of the hemagglutinin gene from the human influenza A/Victoria/3/75 (H3N2) strain as determined from cloned DNA. Cell. 1980 Mar;19(3):683–696. doi: 10.1016/s0092-8674(80)80045-6. [DOI] [PubMed] [Google Scholar]
  13. Kitamura N., Semler B. L., Rothberg P. G., Larsen G. R., Adler C. J., Dorner A. J., Emini E. A., Hanecak R., Lee J. J., van der Werf S. Primary structure, gene organization and polypeptide expression of poliovirus RNA. Nature. 1981 Jun 18;291(5816):547–553. doi: 10.1038/291547a0. [DOI] [PubMed] [Google Scholar]
  14. Küpper H., Keller W., Kurz C., Forss S., Schaller H., Franze R., Strohmaier K., Marquardt O., Zaslavsky V. G., Hofschneider P. H. Cloning of cDNA of major antigen of foot and mouth disease virus and expression in E. coli. Nature. 1981 Feb 12;289(5798):555–559. doi: 10.1038/289555a0. [DOI] [PubMed] [Google Scholar]
  15. Laplanche A., Courouce A. M., Benhamou E., Jungers P., Crosnier J. Responses to hepatitis B vaccine. Lancet. 1982 Jan 23;1(8265):222–222. doi: 10.1016/s0140-6736(82)90784-x. [DOI] [PubMed] [Google Scholar]
  16. Lee C. L., Atassi M. Z. Delineation of the third antigenic site of lysozyme by application of a novel 'surface-simulation' synthetic approach directly linking the conformationally adjacent residues forming the site. Biochem J. 1976 Oct 1;159(1):89–93. doi: 10.1042/bj1590089. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Lerner R. A., Green N., Alexander H., Liu F. T., Sutcliffe J. G., Shinnick T. M. Chemically synthesized peptides predicted from the nucleotide sequence of the hepatitis B virus genome elicit antibodies reactive with the native envelope protein of Dane particles. Proc Natl Acad Sci U S A. 1981 Jun;78(6):3403–3407. doi: 10.1073/pnas.78.6.3403. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Massey R. J., Schochetman G. Topographical analysis of viral epitopes using monoclonal antibodies: mechanism of virus neutralization. Virology. 1981 Nov;115(1):20–32. doi: 10.1016/0042-6822(81)90085-4. [DOI] [PubMed] [Google Scholar]
  19. Massey R. J., Schochetman G. Viral epitopes and monoclonal antibodies: isolation of blocking antibodies that inhibit virus neutralization. Science. 1981 Jul 24;213(4506):447–449. doi: 10.1126/science.6264601. [DOI] [PubMed] [Google Scholar]
  20. Metz W. D. Solar energy: unsung potential for wind and biomass. Science. 1978 May 12;200(4342):636–636. doi: 10.1126/science.200.4342.636. [DOI] [PubMed] [Google Scholar]
  21. Müller G. M., Shapira M., Arnon R. Anti-influenza response achieved by immunization with a synthetic conjugate. Proc Natl Acad Sci U S A. 1982 Jan;79(2):569–573. doi: 10.1073/pnas.79.2.569. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Neurath A. R., Prince A. M., Giacalone J. Large-scale purification of hepatitis B surface antigen using affinity chromatography. Experientia. 1978 Mar 15;34(3):414–415. doi: 10.1007/BF01923068. [DOI] [PubMed] [Google Scholar]
  23. Neurath A. R., Strick N. Antibodies as immunological probes for studying the denaturation of HBsAg. J Med Virol. 1981;6(4):309–322. doi: 10.1002/jmv.1890060406. [DOI] [PubMed] [Google Scholar]
  24. Neurath A. R., Strick N. Enzyme-linked fluorescence immunoassays using beta-galactosidase and antibodies covalently bound to polystyrene plates. J Virol Methods. 1981 Oct;3(3):155–165. doi: 10.1016/0166-0934(81)90050-1. [DOI] [PubMed] [Google Scholar]
  25. Neurath A. R., Strick N., Goodman S. Automated enzyme-linked fluorescence immunoassays for antiviral antibodies: a test for anti-HBs. J Virol Methods. 1982 May;4(4-5):297–303. doi: 10.1016/0166-0934(82)90076-3. [DOI] [PubMed] [Google Scholar]
  26. Neurath A. R., Strick N., Huang C. Y., Prince A. M. Antigenic sites related to human serum proteins in HBsAg: isolation from normal human serum of a high-molecular-weight glycoprotein reacting with antialbumin. J Med Virol. 1978;2(3):231–245. doi: 10.1002/jmv.1890020307. [DOI] [PubMed] [Google Scholar]
  27. Neurath A. R., Strick N., Oleszko W. R. Localization of a hepatitis B surface antigen determinant deduced from results of chemical modifications. J Virol Methods. 1981 Sep;3(2):115–125. doi: 10.1016/0166-0934(81)90008-2. [DOI] [PubMed] [Google Scholar]
  28. Neurath A. R., Szmuness W., Stevens C. E., Strick N., Harley E. J. Radioimmunoassay and some properties of human antibodies to hepatitis B core antigen. J Gen Virol. 1978 Mar;38(3):549–559. doi: 10.1099/0022-1317-38-3-549. [DOI] [PubMed] [Google Scholar]
  29. Osler A. G. On the precedence of 19S antibodies in the early immune response. Immunochemistry. 1978 Nov;15(10-11):717–720. doi: 10.1016/0161-5890(78)90098-6. [DOI] [PubMed] [Google Scholar]
  30. Owen J. A., Sigal N. H., Klinman N. R. Heterogeneity of the BALB/c IgM anti-phosphorylcholine antibody response. Nature. 1982 Jan 28;295(5847):347–348. doi: 10.1038/295347a0. [DOI] [PubMed] [Google Scholar]
  31. Pasek M., Goto T., Gilbert W., Zink B., Schaller H., MacKay P., Leadbetter G., Murray K. Hepatitis B virus genes and their expression in E. coli. Nature. 1979 Dec 6;282(5739):575–579. doi: 10.1038/282575a0. [DOI] [PubMed] [Google Scholar]
  32. Prince A. M., Ikram H., Hopp T. P. Hepatitis B virus vaccine: identification of HBsAg/a and HBsAg/d but not HBsAg/y subtype antigenic determinants on a synthetic immunogenic peptide. Proc Natl Acad Sci U S A. 1982 Jan;79(2):579–582. doi: 10.1073/pnas.79.2.579. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Racaniello V. R., Baltimore D. Molecular cloning of poliovirus cDNA and determination of the complete nucleotide sequence of the viral genome. Proc Natl Acad Sci U S A. 1981 Aug;78(8):4887–4891. doi: 10.1073/pnas.78.8.4887. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Sutcliffe J. G., Shinnick T. M., Green N., Liu F. T., Niman H. L., Lerner R. A. Chemical synthesis of a polypeptide predicted from nucleotide sequence allows detection of a new retroviral gene product. Nature. 1980 Oct 30;287(5785):801–805. doi: 10.1038/287801a0. [DOI] [PubMed] [Google Scholar]
  35. Sutcliffe J. G., Shinnick T. M., Verma I. M., Lerner R. A. Nucleotide sequence of Moloney leukemia virus: 3' end reveals details of replications, analogy to bacterial transposons, and an unexpected gene. Proc Natl Acad Sci U S A. 1980 Jun;77(6):3302–3306. doi: 10.1073/pnas.77.6.3302. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Valenzuela P., Gray P., Quiroga M., Zaldivar J., Goodman H. M., Rutter W. J. Nucleotide sequence of the gene coding for the major protein of hepatitis B virus surface antigen. Nature. 1979 Aug 30;280(5725):815–819. doi: 10.1038/280815a0. [DOI] [PubMed] [Google Scholar]
  37. Walter G., Scheidtmann K. H., Carbone A., Laudano A. P., Doolittle R. F. Antibodies specific for the carboxy- and amino-terminal regions of simian virus 40 large tumor antigen. Proc Natl Acad Sci U S A. 1980 Sep;77(9):5197–5200. doi: 10.1073/pnas.77.9.5197. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. 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]
  39. 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]
  40. Wong T. W., Goldberg A. R. Synthetic peptide fragment of src gene product inhibits the src protein kinase and crossreacts immunologically with avian onc kinases and cellular phosphoproteins. Proc Natl Acad Sci U S A. 1981 Dec;78(12):7412–7416. doi: 10.1073/pnas.78.12.7412. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES