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. 1991 Jun;59(6):1911–1915. doi: 10.1128/iai.59.6.1911-1915.1991

Mapping antibody-binding domains of the major outer surface membrane protein (OspA) of Borrelia burgdorferi.

W H Schubach 1, S Mudri 1, R J Dattwyler 1, B J Luft 1
PMCID: PMC257942  PMID: 2037351

Abstract

The major outer surface membrane protein of Borrelia burgdorferi, OspA, is one of several antigens recognized by sera from some patients in the chronic phase of Lyme borreliosis. We have expressed the OspA open reading frame in Escherichia coli and generated a series of deletion constructs of the gene and expressed them as trpE fusion proteins in E. coli. These constructs were used to identify antibody-binding sites of both rabbit antiserum and mouse monoclonal antibodies (MAbs) directed against OspA. All antibodies tested failed to bind to a fusion protein containing the first 61 amino acids of OspA, suggesting that the amino-terminal domain of OspA is unexposed to the cell surface. The binding site for one MAb, 184.1, was identified in a region centered around amino acid 61, while the binding site for MAb 105.5 was identified in a region centered around amino acids 214 to 217. Sera from two patients which were reactive to OspA identified distinct epitopes that lie between those recognized by our MAbs.

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Selected References

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

  1. Barbour A. G., Burgdorfer W., Grunwaldt E., Steere A. C. Antibodies of patients with Lyme disease to components of the Ixodes dammini spirochete. J Clin Invest. 1983 Aug;72(2):504–515. doi: 10.1172/JCI110998. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Barbour A. G., Hayes S. F., Heiland R. A., Schrumpf M. E., Tessier S. L. A Borrelia-specific monoclonal antibody binds to a flagellar epitope. Infect Immun. 1986 May;52(2):549–554. doi: 10.1128/iai.52.2.549-554.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bergström S., Bundoc V. G., Barbour A. G. Molecular analysis of linear plasmid-encoded major surface proteins, OspA and OspB, of the Lyme disease spirochaete Borrelia burgdorferi. Mol Microbiol. 1989 Apr;3(4):479–486. doi: 10.1111/j.1365-2958.1989.tb00194.x. [DOI] [PubMed] [Google Scholar]
  4. Coleman J. L., Benach J. L. Isolation of antigenic components from the Lyme disease spirochete: their role in early diagnosis. J Infect Dis. 1987 Apr;155(4):756–765. doi: 10.1093/infdis/155.4.756. [DOI] [PubMed] [Google Scholar]
  5. Craft J. E., Fischer D. K., Shimamoto G. T., Steere A. C. Antigens of Borrelia burgdorferi recognized during Lyme disease. Appearance of a new immunoglobulin M response and expansion of the immunoglobulin G response late in the illness. J Clin Invest. 1986 Oct;78(4):934–939. doi: 10.1172/JCI112683. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Craft J. E., Grodzicki R. L., Steere A. C. Antibody response in Lyme disease: evaluation of diagnostic tests. J Infect Dis. 1984 May;149(5):789–795. doi: 10.1093/infdis/149.5.789. [DOI] [PubMed] [Google Scholar]
  7. Greene R. T., Walker R. L., Nicholson W. L., Heidner H. W., Levine J. F., Burgess E. C., Wyand M., Breitschwerdt E. B., Berkhoff H. A. Immunoblot analysis of immunoglobulin G response to the Lyme disease agent (Borrelia burgdorferi) in experimentally and naturally exposed dogs. J Clin Microbiol. 1988 Apr;26(4):648–653. doi: 10.1128/jcm.26.4.648-653.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Grodzicki R. L., Steere A. C. Comparison of immunoblotting and indirect enzyme-linked immunosorbent assay using different antigen preparations for diagnosing early Lyme disease. J Infect Dis. 1988 Apr;157(4):790–797. doi: 10.1093/infdis/157.4.790. [DOI] [PubMed] [Google Scholar]
  9. Jameson B. A., Wolf H. The antigenic index: a novel algorithm for predicting antigenic determinants. Comput Appl Biosci. 1988 Mar;4(1):181–186. doi: 10.1093/bioinformatics/4.1.181. [DOI] [PubMed] [Google Scholar]
  10. Jiang W., Luft B. J., Munoz P., Dattwyler R. J., Gorevic P. D. Cross-antigenicity between the major surface proteins (ospA and ospB) and other proteins of Borrelia burgdorferi. J Immunol. 1990 Jan 1;144(1):284–289. [PubMed] [Google Scholar]
  11. Johnson R. C., Kodner C., Russell M. Passive immunization of hamsters against experimental infection with the Lyme disease spirochete. Infect Immun. 1986 Sep;53(3):713–714. doi: 10.1128/iai.53.3.713-714.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Koerner T. J., Hill J. E., Myers A. M., Tzagoloff A. High-expression vectors with multiple cloning sites for construction of trpE fusion genes: pATH vectors. Methods Enzymol. 1991;194:477–490. doi: 10.1016/0076-6879(91)94036-c. [DOI] [PubMed] [Google Scholar]
  13. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  14. Luft B. J., Dattwyler R. J. Lyme borreliosis. Curr Clin Top Infect Dis. 1989;10:56–81. [PubMed] [Google Scholar]
  15. Luft B. J., Jiang W., Munoz P., Dattwyler R. J., Gorevic P. D. Biochemical and immunological characterization of the surface proteins of Borrelia burgdorferi. Infect Immun. 1989 Nov;57(11):3637–3645. doi: 10.1128/iai.57.11.3637-3645.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Saiki R. K., Gelfand D. H., Stoffel S., Scharf S. J., Higuchi R., Horn G. T., Mullis K. B., Erlich H. A. Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science. 1988 Jan 29;239(4839):487–491. doi: 10.1126/science.2448875. [DOI] [PubMed] [Google Scholar]
  17. Schaible U. E., Kramer M. D., Eichmann K., Modolell M., Museteanu C., Simon M. M. Monoclonal antibodies specific for the outer surface protein A (OspA) of Borrelia burgdorferi prevent Lyme borreliosis in severe combined immunodeficiency (scid) mice. Proc Natl Acad Sci U S A. 1990 May;87(10):3768–3772. doi: 10.1073/pnas.87.10.3768. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Schmitz J. L., Schell R. F., Hejka A. G., England D. M. Passive immunization prevents induction of Lyme arthritis in LSH hamsters. Infect Immun. 1990 Jan;58(1):144–148. doi: 10.1128/iai.58.1.144-148.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Schutzer S. E., Coyle P. K., Belman A. L., Golightly M. G., Drulle J. Sequestration of antibody to Borrelia burgdorferi in immune complexes in seronegative Lyme disease. Lancet. 1990 Feb 10;335(8685):312–315. doi: 10.1016/0140-6736(90)90606-6. [DOI] [PubMed] [Google Scholar]

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