Skip to main content
Infection and Immunity logoLink to Infection and Immunity
. 1995 Aug;63(8):3213–3217. doi: 10.1128/iai.63.8.3213-3217.1995

Protection of C3H/He mice from experimental Borrelia burgdorferi infection by immunization with a 110-kilodalton fusion protein.

R F Bey 1, M E Larson 1, D E Lowery 1, B W Lee 1, K S Knutson 1, R R Simonson 1, V L King 1
PMCID: PMC173440  PMID: 7622251

Abstract

A 110-kDa Borrelia burgdorferi fusion protein, Escherichia coli expressing the fusion protein, transformed E. coli lacking the fusion protein insert, and lyophilized whole B. burgdorferi bacteria were compared for immunogenicity in C3H/He mice. Immunized mice were challenged with a variety of isolates from the United States or the European isolate P/Gau 3 weeks following the last inoculation. An average of 76.7% of the mice immunized with 25 micrograms of lyophilized whole B. burgdorferi cells were protected from infection, while 60% of the mice immunized with the 110-kDa fusion protein were protected. Whole E. coli bacteria expressing the fusion protein protected 57.7% of immunized mice against experimental challenge. Lower levels of protection occurred in mice challenged with the European isolate than in those challenged with isolates originating from the United States. These results demonstrate the potential of the 110-kDa fusion protein for use as a component of a subunit vaccine for prevention of Lyme borreliosis.

Full Text

The Full Text of this article is available as a PDF (233.3 KB).

Selected References

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

  1. Adam T., Gassmann G. S., Rasiah C., Göbel U. B. Phenotypic and genotypic analysis of Borrelia burgdorferi isolates from various sources. Infect Immun. 1991 Aug;59(8):2579–2585. doi: 10.1128/iai.59.8.2579-2585.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Anderson J. F., Magnarelli L. A., Burgdorfer W., Barbour A. G. Spirochetes in Ixodes dammini and mammals from Connecticut. Am J Trop Med Hyg. 1983 Jul;32(4):818–824. doi: 10.4269/ajtmh.1983.32.818. [DOI] [PubMed] [Google Scholar]
  3. Anzola J., Luft B. J., Gorgone G., Dattwyler R. J., Soderberg C., Lahesmaa R., Peltz G. Borrelia burgdorferi HSP70 homolog: characterization of an immunoreactive stress protein. Infect Immun. 1992 Sep;60(9):3704–3713. doi: 10.1128/iai.60.9.3704-3713.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. 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]
  5. 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]
  6. Burgdorfer W., Barbour A. G., Hayes S. F., Benach J. L., Grunwaldt E., Davis J. P. Lyme disease-a tick-borne spirochetosis? Science. 1982 Jun 18;216(4552):1317–1319. doi: 10.1126/science.7043737. [DOI] [PubMed] [Google Scholar]
  7. Caputa A. C., Murtaugh M. P., Bey R. F., Loken K. I. 110-kilodalton recombinant protein which is immunoreactive with sera from humans, dogs, and horses with Lyme borreliosis. J Clin Microbiol. 1991 Nov;29(11):2418–2423. doi: 10.1128/jcm.29.11.2418-2423.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Ditton H. J., Neuss M., Zöller L. Evidence that Borrelia burgdorferi immunodominant proteins p100, p94 and p83 are identical. FEMS Microbiol Lett. 1992 Jul 15;73(3):217–220. doi: 10.1016/0378-1097(92)90633-y. [DOI] [PubMed] [Google Scholar]
  9. 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]
  10. 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]
  11. 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]
  12. Fuchs R., Jauris S., Lottspeich F., Preac-Mursic V., Wilske B., Soutschek E. Molecular analysis and expression of a Borrelia burgdorferi gene encoding a 22 kDa protein (pC) in Escherichia coli. Mol Microbiol. 1992 Feb;6(4):503–509. doi: 10.1111/j.1365-2958.1992.tb01495.x. [DOI] [PubMed] [Google Scholar]
  13. Grizzle J. E., Starmer C. F., Koch G. G. Analysis of categorical data by linear models. Biometrics. 1969 Sep;25(3):489–504. [PubMed] [Google Scholar]
  14. Hedstrom R., Culpepper J., Harrison R. A., Agabian N., Newport G. A major immunogen in Schistosoma mansoni infections is homologous to the heat-shock protein Hsp70. J Exp Med. 1987 May 1;165(5):1430–1435. doi: 10.1084/jem.165.5.1430. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Howe T. R., Mayer L. W., Barbour A. G. A single recombinant plasmid expressing two major outer surface proteins of the Lyme disease spirochete. Science. 1985 Feb 8;227(4687):645–646. doi: 10.1126/science.3969554. [DOI] [PubMed] [Google Scholar]
  16. 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]
  17. 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]
  18. Kochi S. K., Johnson R. C. Role of immunoglobulin G in killing of Borrelia burgdorferi by the classical complement pathway. Infect Immun. 1988 Feb;56(2):314–321. doi: 10.1128/iai.56.2.314-321.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. 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]
  20. Lam T. T., Nguyen T. P., Montgomery R. R., Kantor F. S., Fikrig E., Flavell R. A. Outer surface proteins E and F of Borrelia burgdorferi, the agent of Lyme disease. Infect Immun. 1994 Jan;62(1):290–298. doi: 10.1128/iai.62.1.290-298.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Lovrich S. D., Callister S. M., Lim L. C., Schell R. F. Seroprotective groups among isolates of Borrelia burgdorferi. Infect Immun. 1993 Oct;61(10):4367–4374. doi: 10.1128/iai.61.10.4367-4374.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Luft B. J., Gorevic P. D., Jiang W., Munoz P., Dattwyler R. J. Immunologic and structural characterization of the dominant 66- to 73-kDa antigens of Borrelia burgdorferi. J Immunol. 1991 Apr 15;146(8):2776–2782. [PubMed] [Google Scholar]
  23. Masuzawa T., Okada Y., Beppu Y., Oku T., Kawamori F., Yanagihara Y. Immunological properties of Borrelia burgdorferi isolated from the Ixodes ovatus in Shizuoka, Japan. Microbiol Immunol. 1991;35(10):913–919. doi: 10.1111/j.1348-0421.1991.tb02030.x. [DOI] [PubMed] [Google Scholar]
  24. Masuzawa T., Okada Y., Yanagihara Y., Sato N. Antigenic properties of Borrelia burgdorferi isolated from Ixodes ovatus and Ixodes persulcatus in Hokkaido, Japan. J Clin Microbiol. 1991 Aug;29(8):1568–1573. doi: 10.1128/jcm.29.8.1568-1573.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Norris S. J., Carter C. J., Howell J. K., Barbour A. G. Low-passage-associated proteins of Borrelia burgdorferi B31: characterization and molecular cloning of OspD, a surface-exposed, plasmid-encoded lipoprotein. Infect Immun. 1992 Nov;60(11):4662–4672. doi: 10.1128/iai.60.11.4662-4672.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Perng G. C., LeFebvre R. B., Johnson R. C. Further characterization of a potent immunogen and the chromosomal gene encoding it in the Lyme disease agent, Borrelia burgdorferi. Infect Immun. 1991 Jun;59(6):2070–2074. doi: 10.1128/iai.59.6.2070-2074.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Preac-Mursic V., Wilske B., Patsouris E., Jauris S., Will G., Soutschek E., Rainhardt S., Lehnert G., Klockmann U., Mehraein P. Active immunization with pC protein of Borrelia burgdorferi protects gerbils against B. burgdorferi infection. Infection. 1992 Nov-Dec;20(6):342–349. doi: 10.1007/BF01710681. [DOI] [PubMed] [Google Scholar]
  28. Probert W. S., LeFebvre R. B. Protection of C3H/HeN mice from challenge with Borrelia burgdorferi through active immunization with OspA, OspB, or OspC, but not with OspD or the 83-kilodalton antigen. Infect Immun. 1994 May;62(5):1920–1926. doi: 10.1128/iai.62.5.1920-1926.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. 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]
  30. 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]
  31. Shinnick T. M. Heat shock proteins as antigens of bacterial and parasitic pathogens. Curr Top Microbiol Immunol. 1991;167:145–160. doi: 10.1007/978-3-642-75875-1_9. [DOI] [PubMed] [Google Scholar]
  32. 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]
  33. Simpson W. J., Schrumpf M. E., Schwan T. G. Reactivity of human Lyme borreliosis sera with a 39-kilodalton antigen specific to Borrelia burgdorferi. J Clin Microbiol. 1990 Jun;28(6):1329–1337. doi: 10.1128/jcm.28.6.1329-1337.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Steere A. C. Lyme disease. N Engl J Med. 1989 Aug 31;321(9):586–596. doi: 10.1056/NEJM198908313210906. [DOI] [PubMed] [Google Scholar]
  35. 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]
  36. Tilly K., Hauser R., Campbell J., Ostheimer G. J. Isolation of dnaJ, dnaK, and grpE homologues from Borrelia burgdorferi and complementation of Escherichia coli mutants. Mol Microbiol. 1993 Feb;7(3):359–369. doi: 10.1111/j.1365-2958.1993.tb01128.x. [DOI] [PubMed] [Google Scholar]
  37. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Wallich R., Helmes C., Schaible U. E., Lobet Y., Moter S. E., Kramer M. D., Simon M. M. Evaluation of genetic divergence among Borrelia burgdorferi isolates by use of OspA, fla, HSP60, and HSP70 gene probes. Infect Immun. 1992 Nov;60(11):4856–4866. doi: 10.1128/iai.60.11.4856-4866.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Wallich R., Moter S. E., Simon M. M., Ebnet K., Heiberger A., Kramer M. D. The Borrelia burgdorferi flagellum-associated 41-kilodalton antigen (flagellin): molecular cloning, expression, and amplification of the gene. Infect Immun. 1990 Jun;58(6):1711–1719. doi: 10.1128/iai.58.6.1711-1719.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Wells S. J., Trent A. M., Robinson R. A., Knutson K. S., Bey R. F. Association between clinical lameness and Borrelia burgdorferi antibody in dairy cows. Am J Vet Res. 1993 Mar;54(3):398–405. [PubMed] [Google Scholar]
  41. Wilske B., Preac-Mursic V., Jauris S., Hofmann A., Pradel I., Soutschek E., Schwab E., Will G., Wanner G. Immunological and molecular polymorphisms of OspC, an immunodominant major outer surface protein of Borrelia burgdorferi. Infect Immun. 1993 May;61(5):2182–2191. doi: 10.1128/iai.61.5.2182-2191.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Young R. A. Stress proteins and immunology. Annu Rev Immunol. 1990;8:401–420. doi: 10.1146/annurev.iy.08.040190.002153. [DOI] [PubMed] [Google Scholar]

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

RESOURCES