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. 1993 Jan;61(1):81–90. doi: 10.1128/iai.61.1.81-90.1993

Role of attached lipid in immunogenicity of Borrelia burgdorferi OspA.

L F Erdile 1, M A Brandt 1, D J Warakomski 1, G J Westrack 1, A Sadziene 1, A G Barbour 1, J P Mays 1
PMCID: PMC302690  PMID: 8418068

Abstract

OspA is a protective antigen of the Lyme disease spirochete Borrelia burgdorferi. Expression of the full-length B. burgdorferi B31 OspA gene in Escherichia coli produces a protein that is processed posttranslationally by signal peptidase II and contains an attached lipid moiety. The recombinant OspA lipoprotein has been purified by detergent extraction and ion-exchange chromatography. Priming and boosting with OspA lipoprotein, either with no adjuvant or adsorbed to alum, elicited a strong, dose-dependent immunoglobulin G response. Serum from vaccinated mice inhibited spirochetal growth in vitro. Mice immunized twice with as little as 0.4 micrograms of OspA lipoprotein were protected against an intradermal challenge with 10(4) infectious spirochetes. The ability of the purified recombinant lipoprotein to induce a strong protective response in the absence of toxic adjuvants makes it an excellent candidate antigen for a human vaccine against Lyme disease. By contrast, no serum immunoglobulin G or growth inhibitory response to OspA nonlipoprotein was seen at any dose. The difference in immunogenicities of the lipoprotein and nonlipoprotein forms of OspA is not due to any difference in the antigenicities of the two proteins. These results suggest that posttranslational lipid attachment is a critical determinant of the immunogenicity of the OspA protein.

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Selected References

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

  1. Aberer E., Brunner C., Suchanek G., Klade H., Barbour A., Stanek G., Lassmann H. Molecular mimicry and Lyme borreliosis: a shared antigenic determinant between Borrelia burgdorferi and human tissue. Ann Neurol. 1989 Dec;26(6):732–737. doi: 10.1002/ana.410260608. [DOI] [PubMed] [Google Scholar]
  2. Barbour A. G. Laboratory aspects of Lyme borreliosis. Clin Microbiol Rev. 1988 Oct;1(4):399–414. doi: 10.1128/cmr.1.4.399. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Barbour A. G., Tessier S. L., Todd W. J. Lyme disease spirochetes and ixodid tick spirochetes share a common surface antigenic determinant defined by a monoclonal antibody. Infect Immun. 1983 Aug;41(2):795–804. doi: 10.1128/iai.41.2.795-804.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. 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]
  5. Bessler W. G., Suhr B., Bühring H. J., Muller C. P., Wiesmüller K. H., Becker G., Jung G. Specific antibodies elicited by antigen covalently linked to a synthetic adjuvant. Immunobiology. 1985 Sep;170(3):239–244. doi: 10.1016/S0171-2985(85)80095-4. [DOI] [PubMed] [Google Scholar]
  6. Biesert L., Scheuer W., Bessler W. G. Interaction of mitogenic bacterial lipoprotein and a synthetic analogue with mouse lymphocytes. Isolation and characterization of binding proteins. Eur J Biochem. 1987 Feb 2;162(3):651–657. doi: 10.1111/j.1432-1033.1987.tb10687.x. [DOI] [PubMed] [Google Scholar]
  7. Brandt M. E., Riley B. S., Radolf J. D., Norgard M. V. Immunogenic integral membrane proteins of Borrelia burgdorferi are lipoproteins. Infect Immun. 1990 Apr;58(4):983–991. doi: 10.1128/iai.58.4.983-991.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Burman N., Bergström S., Restrepo B. I., Barbour A. G. The variable antigens Vmp7 and Vmp21 of the relapsing fever bacterium Borrelia hermsii are structurally analogous to the VSG proteins of the African trypanosome. Mol Microbiol. 1990 Oct;4(10):1715–1726. doi: 10.1111/j.1365-2958.1990.tb00549.x. [DOI] [PubMed] [Google Scholar]
  9. Chamberlain N. R., Brandt M. E., Erwin A. L., Radolf J. D., Norgard M. V. Major integral membrane protein immunogens of Treponema pallidum are proteolipids. Infect Immun. 1989 Sep;57(9):2872–2877. doi: 10.1128/iai.57.9.2872-2877.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Cooke W. D., Dattwyler R. J. Spirochetal arthritis, including Lyme disease. Curr Opin Rheumatol. 1990 Aug;2(4):622–627. doi: 10.1097/00002281-199002040-00012. [DOI] [PubMed] [Google Scholar]
  11. Defoort J. P., Nardelli B., Huang W., Ho D. D., Tam J. P. Macromolecular assemblage in the design of a synthetic AIDS vaccine. Proc Natl Acad Sci U S A. 1992 May 1;89(9):3879–3883. doi: 10.1073/pnas.89.9.3879. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Deres K., Schild H., Wiesmüller K. H., Jung G., Rammensee H. G. In vivo priming of virus-specific cytotoxic T lymphocytes with synthetic lipopeptide vaccine. Nature. 1989 Nov 30;342(6249):561–564. doi: 10.1038/342561a0. [DOI] [PubMed] [Google Scholar]
  13. Duffy J. Lyme disease. Ann Allergy. 1990 Jul;65(1):1–13. [PubMed] [Google Scholar]
  14. Dunn J. J., Lade B. N., Barbour A. G. Outer surface protein A (OspA) from the Lyme disease spirochete, Borrelia burgdorferi: high level expression and purification of a soluble recombinant form of OspA. Protein Expr Purif. 1990 Nov;1(2):159–168. doi: 10.1016/1046-5928(90)90011-m. [DOI] [PubMed] [Google Scholar]
  15. Fikrig E., Barthold S. W., Kantor F. S., Flavell R. A. Long-term protection of mice from Lyme disease by vaccination with OspA. Infect Immun. 1992 Mar;60(3):773–777. doi: 10.1128/iai.60.3.773-777.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Fikrig E., Barthold S. W., Kantor F. S., Flavell R. A. Protection of mice against the Lyme disease agent by immunizing with recombinant OspA. Science. 1990 Oct 26;250(4980):553–556. doi: 10.1126/science.2237407. [DOI] [PubMed] [Google Scholar]
  17. Fikrig E., Telford S. R., 3rd, Barthold S. W., Kantor F. S., Spielman A., Flavell R. A. Elimination of Borrelia burgdorferi from vector ticks feeding on OspA-immunized mice. Proc Natl Acad Sci U S A. 1992 Jun 15;89(12):5418–5421. doi: 10.1073/pnas.89.12.5418. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Holmes D. S., Quigley M. A rapid boiling method for the preparation of bacterial plasmids. Anal Biochem. 1981 Jun;114(1):193–197. doi: 10.1016/0003-2697(81)90473-5. [DOI] [PubMed] [Google Scholar]
  19. Howe T. R., LaQuier F. W., Barbour A. G. Organization of genes encoding two outer membrane proteins of the Lyme disease agent Borrelia burgdorferi within a single transcriptional unit. Infect Immun. 1986 Oct;54(1):207–212. doi: 10.1128/iai.54.1.207-212.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Hussain M., Ichihara S., Mizushima S. Accumulation of glyceride-containing precursor of the outer membrane lipoprotein in the cytoplasmic membrane of Escherichia coli treated with globomycin. J Biol Chem. 1980 Apr 25;255(8):3707–3712. [PubMed] [Google Scholar]
  21. Inukai M., Takeuchi M., Shimizu K., Arai M. Mechanism of action of globomycin. J Antibiot (Tokyo) 1978 Nov;31(11):1203–1205. doi: 10.7164/antibiotics.31.1203. [DOI] [PubMed] [Google Scholar]
  22. Johnson R. C., Kodner C., Russell M. Active immunization of hamsters against experimental infection with Borrelia burgdorferi. Infect Immun. 1986 Dec;54(3):897–898. doi: 10.1128/iai.54.3.897-898.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. 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]
  24. Magnarelli L. A., Anderson J. F., Barbour A. G. Enzyme-linked immunosorbent assays for Lyme disease: reactivity of subunits of Borrelia burgdorferi. J Infect Dis. 1989 Jan;159(1):43–49. doi: 10.1093/infdis/159.1.43. [DOI] [PubMed] [Google Scholar]
  25. Metzger J., Jung G., Bessler W. G., Hoffmann P., Strecker M., Lieberknecht A., Schmidt U. Lipopeptides containing 2-(palmitoylamino)-6,7-bis(palmitoyloxy) heptanoic acid: synthesis, stereospecific stimulation of B-lymphocytes and macrophages, and adjuvanticity in vivo and in vitro. J Med Chem. 1991 Jul;34(7):1969–1974. doi: 10.1021/jm00111a008. [DOI] [PubMed] [Google Scholar]
  26. Piesman J., Oliver J. R., Sinsky R. J. Growth kinetics of the Lyme disease spirochete (Borrelia burgdorferi) in vector ticks (Ixodes dammini). Am J Trop Med Hyg. 1990 Apr;42(4):352–357. doi: 10.4269/ajtmh.1990.42.352. [DOI] [PubMed] [Google Scholar]
  27. 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]
  28. Schild H., Deres K., Wiesmüller K. H., Jung G., Rammensee H. G. Efficiency of peptides and lipopeptides for in vivo priming of virus-specific cytotoxic T cells. Eur J Immunol. 1991 Nov;21(11):2649–2654. doi: 10.1002/eji.1830211102. [DOI] [PubMed] [Google Scholar]
  29. Schlesinger M. J. Heat shock proteins. J Biol Chem. 1990 Jul 25;265(21):12111–12114. [PubMed] [Google Scholar]
  30. Schouls L. M., van der Heide H. G., van Embden J. D. Characterization of the 35-kilodalton Treponema pallidum subsp. pallidum recombinant lipoprotein TmpC and antibody response to lipidated and nonlipidated T. pallidum antigens. Infect Immun. 1991 Oct;59(10):3536–3546. doi: 10.1128/iai.59.10.3536-3546.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Simon M. M., Schaible U. E., Kramer M. D., Eckerskorn C., Museteanu C., Müller-Hermelink H. K., Wallich R. Recombinant outer surface protein a from Borrelia burgdorferi induces antibodies protective against spirochetal infection in mice. J Infect Dis. 1991 Jul;164(1):123–132. doi: 10.1093/infdis/164.1.123. [DOI] [PubMed] [Google Scholar]
  32. Singer A., Hodes R. J. Mechanisms of T cell-B cell interaction. Annu Rev Immunol. 1983;1:211–241. doi: 10.1146/annurev.iy.01.040183.001235. [DOI] [PubMed] [Google Scholar]
  33. Stark J. M., Locke J., Heatley R. V. Immunogenicity of lipid-conjugated antigens. I. The influence of chain length and degree of conjugation on induction of antibody in mice. Immunology. 1980 Mar;39(3):345–352. [PMC free article] [PubMed] [Google Scholar]
  34. Stark J. M., Matthews N., Locke J. Immunogenicity of lipid-conjugated antigens. II. Anti-complementary activity and antigen trapping in the spleen. Immunology. 1980 Mar;39(3):353–360. [PMC free article] [PubMed] [Google Scholar]
  35. Studier F. W., Rosenberg A. H., Dunn J. J., Dubendorff J. W. Use of T7 RNA polymerase to direct expression of cloned genes. Methods Enzymol. 1990;185:60–89. doi: 10.1016/0076-6879(90)85008-c. [DOI] [PubMed] [Google Scholar]
  36. Szczepanski A., Benach J. L. Lyme borreliosis: host responses to Borrelia burgdorferi. Microbiol Rev. 1991 Mar;55(1):21–34. doi: 10.1128/mr.55.1.21-34.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Weigel L. M., Brandt M. E., Norgard M. V. Analysis of the N-terminal region of the 47-kilodalton integral membrane lipoprotein of Treponema pallidum. Infect Immun. 1992 Apr;60(4):1568–1576. doi: 10.1128/iai.60.4.1568-1576.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Wilske B., Preac-Mursic V., Schierz G., Kühbeck R., Barbour A. G., Kramer M. Antigenic variability of Borrelia burgdorferi. Ann N Y Acad Sci. 1988;539:126–143. doi: 10.1111/j.1749-6632.1988.tb31846.x. [DOI] [PubMed] [Google Scholar]
  39. Wu H. C., Tokunaga M. Biogenesis of lipoproteins in bacteria. Curr Top Microbiol Immunol. 1986;125:127–157. doi: 10.1007/978-3-642-71251-7_9. [DOI] [PubMed] [Google Scholar]

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