Skip to main content
PMC home page
Infection and Immunity logoLink to Infection and Immunity
. 1995 Jun;63(6):2255–2261. doi: 10.1128/iai.63.6.2255-2261.1995

Circumvention of outer surface protein A immunity by host-adapted Borrelia burgdorferi.

S W Barthold 1, E Fikrig 1, L K Bockenstedt 1, D H Persing 1
PMCID: PMC173294  PMID: 7768606

Abstract

Outer surface protein A (OspA), which is abundantly expressed in cultured Borrelia burgdorferi, appears to be down-regulated or masked following low-dose infection, and OspA immunization did not prevent infection, dissemination, or disease development with host-adapted spirochetes. Seroconversion of mice to B. burgdorferi OspA depended on dose and viability of inoculated spirochetes. Mice inoculated with > 10(4) live spirochetes and > 10(7) heat-killed spirochetes seroconverted to OspA, but mice inoculated with fewer spirochetes did not seroconvert to OspA at 2 weeks after inoculation. Growth temperature of spirochetes was not a factor for infectious dose or seroconversion to OspA. Spirochetes grown at 30, 34, or 38 degrees C had the same median infectious dose. Growth temperature did not influence infectious dose when mice were inoculated intraperitoneally or intradermally and did not influence dose-related immunologic recognition of OspA. Mice hyperimmunized with recombinant OspA-glutathione S-transferase (GT) fusion protein or GT (controls) were challenged by syringe inoculation with 10(3) spirochetes or by transplantation of infected skin from syngenic mice infected for 2 or 8 weeks. OspA-GT-immunized mice resisted syringe challenge but developed disseminated infections following transplantation of infected skin. Identical results were obtained in mice passively immunized with hyperimmune serum to OspA-GT or GT and then challenged by syringe or infected skin transplant. The number of spirochetes in infected skin, determined by quantitative PCR directed toward both plasmid and genomic targets, was less than the syringe challenge dose.

Full Text

The Full Text of this article is available as a PDF (454.3 KB).

Selected References

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

  1. Aguero-Rosenfeld M. E., Nowakowski J., McKenna D. F., Carbonaro C. A., Wormser G. P. Serodiagnosis in early Lyme disease. J Clin Microbiol. 1993 Dec;31(12):3090–3095. doi: 10.1128/jcm.31.12.3090-3095.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Appel M. J., Allan S., Jacobson R. H., Lauderdale T. L., Chang Y. F., Shin S. J., Thomford J. W., Todhunter R. J., Summers B. A. Experimental Lyme disease in dogs produces arthritis and persistent infection. J Infect Dis. 1993 Mar;167(3):651–664. doi: 10.1093/infdis/167.3.651. [DOI] [PubMed] [Google Scholar]
  3. Armstrong A. L., Barthold S. W., Persing D. H., Beck D. S. Carditis in Lyme disease susceptible and resistant strains of laboratory mice infected with Borrelia burgdorferi. Am J Trop Med Hyg. 1992 Aug;47(2):249–258. doi: 10.4269/ajtmh.1992.47.249. [DOI] [PubMed] [Google Scholar]
  4. Assous M. V., Postic D., Paul G., Névot P., Baranton G. Western blot analysis of sera from Lyme borreliosis patients according to the genomic species of the Borrelia strains used as antigens. Eur J Clin Microbiol Infect Dis. 1993 Apr;12(4):261–268. doi: 10.1007/BF01967256. [DOI] [PubMed] [Google Scholar]
  5. Barbour A. G. Isolation and cultivation of Lyme disease spirochetes. Yale J Biol Med. 1984 Jul-Aug;57(4):521–525. [PMC free article] [PubMed] [Google Scholar]
  6. 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]
  7. Barthold S. W. Antigenic stability of Borrelia burgdorferi during chronic infections of immunocompetent mice. Infect Immun. 1993 Dec;61(12):4955–4961. doi: 10.1128/iai.61.12.4955-4961.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Barthold S. W., Bockenstedt L. K. Passive immunizing activity of sera from mice infected with Borrelia burgdorferi. Infect Immun. 1993 Nov;61(11):4696–4702. doi: 10.1128/iai.61.11.4696-4702.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Barthold S. W., Sidman C. L., Smith A. L. Lyme borreliosis in genetically resistant and susceptible mice with severe combined immunodeficiency. Am J Trop Med Hyg. 1992 Nov;47(5):605–613. doi: 10.4269/ajtmh.1992.47.605. [DOI] [PubMed] [Google Scholar]
  10. Barthold S. W., de Souza M. S., Janotka J. L., Smith A. L., Persing D. H. Chronic Lyme borreliosis in the laboratory mouse. Am J Pathol. 1993 Sep;143(3):959–971. [PMC free article] [PubMed] [Google Scholar]
  11. Bergström S., Garon C. F., Barbour A. G., MacDougall J. Extrachromosomal elements of spirochetes. Res Microbiol. 1992 Jul-Aug;143(6):623–628. doi: 10.1016/0923-2508(92)90120-d. [DOI] [PubMed] [Google Scholar]
  12. Burkot T. R., Wirtz R. A., Luft B., Piesman J. An OspA antigen-capture enzyme-linked immunosorbent assay for detecting North American isolates of Borrelia burgdorferi in larval and nymphal Ixodes dammini. J Clin Microbiol. 1993 Feb;31(2):272–278. doi: 10.1128/jcm.31.2.272-278.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Champion C. I., Blanco D. R., Skare J. T., Haake D. A., Giladi M., Foley D., Miller J. N., Lovett M. A. A 9.0-kilobase-pair circular plasmid of Borrelia burgdorferi encodes an exported protein: evidence for expression only during infection. Infect Immun. 1994 Jul;62(7):2653–2661. doi: 10.1128/iai.62.7.2653-2661.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. 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]
  15. Dorward D. W., Huguenel E. D., Davis G., Garon C. F. Interactions between extracellular Borrelia burgdorferi proteins and non-Borrelia-directed immunoglobulin M antibodies. Infect Immun. 1992 Mar;60(3):838–844. doi: 10.1128/iai.60.3.838-844.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Dorward D. W., Schwan T. G., Garon C. F. Immune capture and detection of Borrelia burgdorferi antigens in urine, blood, or tissues from infected ticks, mice, dogs, and humans. J Clin Microbiol. 1991 Jun;29(6):1162–1170. doi: 10.1128/jcm.29.6.1162-1170.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Dressler F., Whalen J. A., Reinhardt B. N., Steere A. C. Western blotting in the serodiagnosis of Lyme disease. J Infect Dis. 1993 Feb;167(2):392–400. doi: 10.1093/infdis/167.2.392. [DOI] [PubMed] [Google Scholar]
  18. 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]
  19. Fikrig E., Barthold S. W., Flavell R. A. OspA vaccination of mice with established Borrelia burgdorferi infection alters disease but not infection. Infect Immun. 1993 Jun;61(6):2553–2557. doi: 10.1128/iai.61.6.2553-2557.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. 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]
  21. Fikrig E., Barthold S. W., Marcantonio N., Deponte K., Kantor F. S., Flavell R. A. Roles of OspA, OspB, and flagellin in protective immunity to Lyme borreliosis in laboratory mice. Infect Immun. 1992 Feb;60(2):657–661. doi: 10.1128/iai.60.2.657-661.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. 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]
  23. Garon C. F., Dorward D. W., Corwin M. D. Structural features of Borrelia burgdorferi--the Lyme disease spirochete: silver staining for nucleic acids. Scanning Microsc Suppl. 1989;3:109–115. [PubMed] [Google Scholar]
  24. Gern L., Rais O., Capiau C., Hauser P., Lobet Y., Simoen E., Voet P., Pêtre J. Immunization of mice by recombinant OspA preparations and protection against Borrelia burgdorferi infection induced by Ixodes ricinus tick bites. Immunol Lett. 1994 Mar;39(3):249–258. doi: 10.1016/0165-2478(94)90166-x. [DOI] [PubMed] [Google Scholar]
  25. Gern L., Schaible U. E., Simon M. M. Mode of inoculation of the Lyme disease agent Borrelia burgdorferi influences infection and immune responses in inbred strains of mice. J Infect Dis. 1993 Apr;167(4):971–975. doi: 10.1093/infdis/167.4.971. [DOI] [PubMed] [Google Scholar]
  26. Golde W. T., Burkot T. R., Sviat S., Keen M. G., Mayer L. W., Johnson B. J., Piesman J. The major histocompatibility complex-restricted response of recombinant inbred strains of mice to natural tick transmission of Borrelia burgdorferi. J Exp Med. 1993 Jan 1;177(1):9–17. doi: 10.1084/jem.177.1.9. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Golde W. T., Kappel K. J., Dequesne G., Feron C., Plainchamp D., Capiau C., Lobet Y. Tick transmission of Borrelia burgdorferi to inbred strains of mice induces an antibody response to P39 but not to outer surface protein A. Infect Immun. 1994 Jun;62(6):2625–2627. doi: 10.1128/iai.62.6.2625-2627.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Hinnebusch J., Barbour A. G. Linear- and circular-plasmid copy numbers in Borrelia burgdorferi. J Bacteriol. 1992 Aug;174(16):5251–5257. doi: 10.1128/jb.174.16.5251-5257.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Kalish R. A., Leong J. M., Steere A. C. Association of treatment-resistant chronic Lyme arthritis with HLA-DR4 and antibody reactivity to OspA and OspB of Borrelia burgdorferi. Infect Immun. 1993 Jul;61(7):2774–2779. doi: 10.1128/iai.61.7.2774-2779.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Keller D., Koster F. T., Marks D. H., Hosbach P., Erdile L. F., Mays J. P. Safety and immunogenicity of a recombinant outer surface protein A Lyme vaccine. JAMA. 1994 Jun 8;271(22):1764–1768. [PubMed] [Google Scholar]
  31. Ma B., Christen B., Leung D., Vigo-Pelfrey C. Serodiagnosis of Lyme borreliosis by western immunoblot: reactivity of various significant antibodies against Borrelia burgdorferi. J Clin Microbiol. 1992 Feb;30(2):370–376. doi: 10.1128/jcm.30.2.370-376.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Ma Y., Weis J. J. Borrelia burgdorferi outer surface lipoproteins OspA and OspB possess B-cell mitogenic and cytokine-stimulatory properties. Infect Immun. 1993 Sep;61(9):3843–3853. doi: 10.1128/iai.61.9.3843-3853.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Malawista S. E., Barthold S. W., Persing D. H. Fate of Borrelia burgdorferi DNA in tissues of infected mice after antibiotic treatment. J Infect Dis. 1994 Nov;170(5):1312–1316. doi: 10.1093/infdis/170.5.1312. [DOI] [PubMed] [Google Scholar]
  34. Persing D. H., Rutledge B. J., Rys P. N., Podzorski D. S., Mitchell P. D., Reed K. D., Liu B., Fikrig E., Malawista S. E. Target imbalance: disparity of Borrelia burgdorferi genetic material in synovial fluid from Lyme arthritis patients. J Infect Dis. 1994 Mar;169(3):668–672. doi: 10.1093/infdis/169.3.668. [DOI] [PubMed] [Google Scholar]
  35. Philipp M. T., Aydintug M. K., Bohm R. P., Jr, Cogswell F. B., Dennis V. A., Lanners H. N., Lowrie R. C., Jr, Roberts E. D., Conway M. D., Karaçorlu M. Early and early disseminated phases of Lyme disease in the rhesus monkey: a model for infection in humans. Infect Immun. 1993 Jul;61(7):3047–3059. doi: 10.1128/iai.61.7.3047-3059.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Roehrig J. T., Piesman J., Hunt A. R., Keen M. G., Happ C. M., Johnson B. J. The hamster immune response to tick-transmitted Borrelia burgdorferi differs from the response to needle-inoculated, cultured organisms. J Immunol. 1992 Dec 1;149(11):3648–3653. [PubMed] [Google Scholar]
  37. Schaible U. E., Gern L., Wallich R., Kramer M. D., Prester M., Simon M. M. Distinct patterns of protective antibodies are generated against Borrelia burgdorferi in mice experimentally inoculated with high and low doses of antigen. Immunol Lett. 1993 May;36(2):219–226. doi: 10.1016/0165-2478(93)90056-8. [DOI] [PubMed] [Google Scholar]
  38. 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]
  39. Schaible U. E., Wallich R., Kramer M. D., Gern L., Anderson J. F., Museteanu C., Simon M. M. Immune sera to individual Borrelia burgdorferi isolates or recombinant OspA thereof protect SCID mice against infection with homologous strains but only partially or not at all against those of different OspA/OspB genotype. Vaccine. 1993;11(10):1049–1054. doi: 10.1016/0264-410x(93)90132-h. [DOI] [PubMed] [Google Scholar]
  40. Schoenfeld R., Araneo B., Ma Y., Yang L. M., Weis J. J. Demonstration of a B-lymphocyte mitogen produced by the Lyme disease pathogen, Borrelia burgdorferi. Infect Immun. 1992 Feb;60(2):455–464. doi: 10.1128/iai.60.2.455-464.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Schutzer S. E., Coyle P. K., Dunn J. J., Luft B. J., Brunner M. Early and specific antibody response to OspA in Lyme Disease. J Clin Invest. 1994 Jul;94(1):454–457. doi: 10.1172/JCI117346. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Schwan T. G., Piesman J., Golde W. T., Dolan M. C., Rosa P. A. Induction of an outer surface protein on Borrelia burgdorferi during tick feeding. Proc Natl Acad Sci U S A. 1995 Mar 28;92(7):2909–2913. doi: 10.1073/pnas.92.7.2909. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Schwan T. G., Simpson W. J. Factors influencing the antigenic reactivity of Borrelia burgdorferi, the Lyme disease spirochete. Scand J Infect Dis Suppl. 1991;77:94–101. [PubMed] [Google Scholar]
  44. 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]
  45. Simpson W. J., Cieplak W., Schrumpf M. E., Barbour A. G., Schwan T. G. Nucleotide sequence and analysis of the gene in Borrelia burgdorferi encoding the immunogenic P39 antigen. FEMS Microbiol Lett. 1994 Jun 15;119(3):381–387. doi: 10.1111/j.1574-6968.1994.tb06917.x. [DOI] [PubMed] [Google Scholar]
  46. Stevenson B., Bockenstedt L. K., Barthold S. W. Expression and gene sequence of outer surface protein C of Borrelia burgdorferi reisolated from chronically infected mice. Infect Immun. 1994 Aug;62(8):3568–3571. doi: 10.1128/iai.62.8.3568-3571.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Telford S. R., 3rd, Fikrig E., Barthold S. W., Brunet L. R., Spielman A., Flavell R. A. Protection against antigenically variable Borrelia burgdorferi conferred by recombinant vaccines. J Exp Med. 1993 Aug 1;178(2):755–758. doi: 10.1084/jem.178.2.755. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Whitmire W. M., Garon C. F. Specific and nonspecific responses of murine B cells to membrane blebs of Borrelia burgdorferi. Infect Immun. 1993 Apr;61(4):1460–1467. doi: 10.1128/iai.61.4.1460-1467.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Zöller L., Burkard S., Schäfer H. Validity of western immunoblot band patterns in the serodiagnosis of Lyme borreliosis. J Clin Microbiol. 1991 Jan;29(1):174–182. doi: 10.1128/jcm.29.1.174-182.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  50. de Souza M. S., Fikrig E., Smith A. L., Flavell R. A., Barthold S. W. Nonspecific proliferative responses of murine lymphocytes to Borrelia burgdorferi antigens. J Infect Dis. 1992 Mar;165(3):471–478. doi: 10.1093/infdis/165.3.471. [DOI] [PubMed] [Google Scholar]

Articles from Infection and Immunity are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES