Skip to main content
Infection and Immunity logoLink to Infection and Immunity
. 1993 May;61(5):1859–1866. doi: 10.1128/iai.61.5.1859-1866.1993

Immune responses to Streptococcus sobrinus surface protein antigen A expressed by recombinant Salmonella typhimurium.

T A Doggett 1, E K Jagusztyn-Krynicka 1, R Curtiss 3rd 1
PMCID: PMC280776  PMID: 8478075

Abstract

In this study, we used a vaccine strain of Salmonella typhimurium to express antigenic determinants of the SpaA antigen of Streptococcus sobrinus, which is involved in the caries-forming process. We cloned either a single repeat (pYA2901) or three tandem repeats (pYA2905) of the 0.48-kb fragment of the spaA gene, which codes for an important component of the SpaA protein, plus a 1.2-kb minor antigenic determinant and measured the resulting immune responses to SpaA in orally immunized BALB/c mice. The single or triple repeat of the spaA gene fragment was inserted into the Asd+ vector pYA292 and was transformed into the S. typhimurium delta cya delta crp vaccine strain chi 4072 containing delta asd in the chromosome. Female BALB/c mice were then orally immunized with two doses of the S. typhimurium containing either of the two SpaA constructs, and the immune responses to the expressed SpaA protein were assessed. Significant serum immunoglobulin G (IgG) anti-SpaA titers were detected in mice immunized with chi 4072(pYA2905) but not chi 4072(pYA2901). Salivary anti-SpaA IgA titers were minimal and were only detected in mice immunized with S. typhimurium expressing the SpaA encoded by pYA2905. Intestinal anti-SpaA IgA titers, however, were detected in both groups of mice, particularly in mice immunized with chi 4072(pYA2905). An oral booster 26 weeks after the initial series of immunizations resulted in increased serum IgG titers in both chi 4072(pYA2901)- and chi 4072(pYA2905)-immunized animals, particularly in the chi 4072(pYA2905)-immunized animals. No anamnestic IgA response was detected in the saliva following the booster immunization.

Full text

PDF
1859

Images in this article

Selected References

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

  1. BACON G. A., BURROWS T. W., YATES M. The effects of biochemical mutation on the virulence of Bacterium typhosum; the loss of virulence of certain mutants. Br J Exp Pathol. 1951 Apr;32(2):85–96. [PMC free article] [PubMed] [Google Scholar]
  2. Brown A., Hormaeche C. E., Demarco de Hormaeche R., Winther M., Dougan G., Maskell D. J., Stocker B. A. An attenuated aroA Salmonella typhimurium vaccine elicits humoral and cellular immunity to cloned beta-galactosidase in mice. J Infect Dis. 1987 Jan;155(1):86–92. doi: 10.1093/infdis/155.1.86. [DOI] [PubMed] [Google Scholar]
  3. Challacombe S. J., Lehner T. Serum and salivary antibodies to cariogenic bacteria in man. J Dent Res. 1976 Apr;55(Spec No):C139–C148. doi: 10.1177/002203457605500306011. [DOI] [PubMed] [Google Scholar]
  4. Chatfield S. N., Dorman C. J., Hayward C., Dougan G. Role of ompR-dependent genes in Salmonella typhimurium virulence: mutants deficient in both ompC and ompF are attenuated in vivo. Infect Immun. 1991 Jan;59(1):449–452. doi: 10.1128/iai.59.1.449-452.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Curtiss R., 3rd, Kelly S. M. Salmonella typhimurium deletion mutants lacking adenylate cyclase and cyclic AMP receptor protein are avirulent and immunogenic. Infect Immun. 1987 Dec;55(12):3035–3043. doi: 10.1128/iai.55.12.3035-3043.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Dagert M., Ehrlich S. D. Prolonged incubation in calcium chloride improves the competence of Escherichia coli cells. Gene. 1979 May;6(1):23–28. doi: 10.1016/0378-1119(79)90082-9. [DOI] [PubMed] [Google Scholar]
  7. Dougan G., Hormaeche C. E., Maskell D. J. Live oral Salmonella vaccines: potential use of attenuated strains as carriers of heterologous antigens to the immune system. Parasite Immunol. 1987 Mar;9(2):151–160. doi: 10.1111/j.1365-3024.1987.tb00496.x. [DOI] [PubMed] [Google Scholar]
  8. Dougan G., Sellwood R., Maskell D., Sweeney K., Liew F. Y., Beesley J., Hormaeche C. In vivo properties of a cloned K88 adherence antigen determinant. Infect Immun. 1986 Apr;52(1):344–347. doi: 10.1128/iai.52.1.344-347.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Ebersole J. L., Steffen M. J. Aging effects on secretory IgA immune responses. Immunol Invest. 1989 Jan-May;18(1-4):59–68. doi: 10.3109/08820138909112227. [DOI] [PubMed] [Google Scholar]
  10. Forester H., Hunter N., Knox K. W. Characteristics of a high molecular weight extracellular protein of Streptococcus mutans. J Gen Microbiol. 1983 Sep;129(9):2779–2788. doi: 10.1099/00221287-129-9-2779. [DOI] [PubMed] [Google Scholar]
  11. Galán J. E., Curtiss R., 3rd Cloning and molecular characterization of genes whose products allow Salmonella typhimurium to penetrate tissue culture cells. Proc Natl Acad Sci U S A. 1989 Aug;86(16):6383–6387. doi: 10.1073/pnas.86.16.6383. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Galán J. E., Nakayama K., Curtiss R., 3rd Cloning and characterization of the asd gene of Salmonella typhimurium: use in stable maintenance of recombinant plasmids in Salmonella vaccine strains. Gene. 1990 Sep 28;94(1):29–35. doi: 10.1016/0378-1119(90)90464-3. [DOI] [PubMed] [Google Scholar]
  13. Germanier R., Fürer E. Immunity in experimental salmonellosis. II. Basis for the avirulence and protective capacity of gal E mutants of Salmonella typhimurium. Infect Immun. 1971 Dec;4(6):663–673. doi: 10.1128/iai.4.6.663-673.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Goldschmidt R. M., Thoren-Gordon M., Curtiss R., 3rd Regions of the Streptococcus sobrinus spaA gene encoding major determinants of antigen I. J Bacteriol. 1990 Jul;172(7):3988–4001. doi: 10.1128/jb.172.7.3988-4001.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Hoiseth S. K., Stocker B. A. Aromatic-dependent Salmonella typhimurium are non-virulent and effective as live vaccines. Nature. 1981 May 21;291(5812):238–239. doi: 10.1038/291238a0. [DOI] [PubMed] [Google Scholar]
  16. Johnson K., Charles I., Dougan G., Pickard D., O'Gaora P., Costa G., Ali T., Miller I., Hormaeche C. The role of a stress-response protein in Salmonella typhimurium virulence. Mol Microbiol. 1991 Feb;5(2):401–407. doi: 10.1111/j.1365-2958.1991.tb02122.x. [DOI] [PubMed] [Google Scholar]
  17. Katz J., Michalek S. M., Curtiss R., 3rd, Harmon C., Richardson G., Mestecky J. Novel oral vaccines: the effectiveness of cloned gene products on inducing secretory immune responses. Adv Exp Med Biol. 1987;216B:1741–1747. [PubMed] [Google Scholar]
  18. Kelly C., Evans P., Bergmeier L., Lee S. F., Progulske-Fox A., Harris A. C., Aitken A., Bleiweis A. S., Lehner T. Sequence analysis of the cloned streptococcal cell surface antigen I/II. FEBS Lett. 1989 Nov 20;258(1):127–132. doi: 10.1016/0014-5793(89)81632-1. [DOI] [PubMed] [Google Scholar]
  19. Lehner T., Haron J., Bergmeier L. A., Mehlert A., Beard R., Dodd M., Mielnik B., Moore S. Local oral immunization with synthetic peptides induces a dual mucosal IgG and salivary IgA antibody response and prevents colonization of Streptococcus mutans. Immunology. 1989 Jul;67(3):419–424. [PMC free article] [PubMed] [Google Scholar]
  20. Maskell D. J., Sweeney K. J., O'Callaghan D., Hormaeche C. E., Liew F. Y., Dougan G. Salmonella typhimurium aroA mutants as carriers of the Escherichia coli heat-labile enterotoxin B subunit to the murine secretory and systemic immune systems. Microb Pathog. 1987 Mar;2(3):211–221. doi: 10.1016/0882-4010(87)90022-2. [DOI] [PubMed] [Google Scholar]
  21. McBride B. C., Song M., Krasse B., Olsson J. Biochemical and immunological differences between hydrophobic and hydrophilic strains of Streptococcus mutans. Infect Immun. 1984 Apr;44(1):68–75. doi: 10.1128/iai.44.1.68-75.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Mestecky J., McGhee J. R., Arnold R. R., Michalek S. M., Prince S. J., Babb J. L. Selective induction of an immune response in human external secretions by ingestion of bacterial antigen. J Clin Invest. 1978 Mar;61(3):731–737. doi: 10.1172/JCI108986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Michalek S. M., McGhee J. R., Babb J. L. Effective immunity to dental caries: dose-dependent studies of secretory immunity by oral administration of Streptococcus mutans to rats. Infect Immun. 1978 Jan;19(1):217–224. doi: 10.1128/iai.19.1.217-224.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Michalek S. M., McGhee J. R., Mestecky J., Arnold R. R., Bozzo L. Ingestion of Streptococcus mutans induces secretory immunoglobulin A and caries immunity. Science. 1976 Jun 18;192(4245):1238–1240. doi: 10.1126/science.1273589. [DOI] [PubMed] [Google Scholar]
  25. Poirier T. P., Kehoe M. A., Beachey E. H. Protective immunity evoked by oral administration of attenuated aroA Salmonella typhimurium expressing cloned streptococcal M protein. J Exp Med. 1988 Jul 1;168(1):25–32. doi: 10.1084/jem.168.1.25. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Russell R. R. Wall-associated protein antigens of Streptococcus mutans. J Gen Microbiol. 1979 Sep;114(1):109–115. doi: 10.1099/00221287-114-1-109. [DOI] [PubMed] [Google Scholar]
  27. Schödel F., Will H. Construction of a plasmid for expression of foreign epitopes as fusion proteins with subunit B of Escherichia coli heat-labile enterotoxin. Infect Immun. 1989 Apr;57(4):1347–1350. doi: 10.1128/iai.57.4.1347-1350.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Sjöstedt A., Sandström G., Tärnvik A. Humoral and cell-mediated immunity in mice to a 17-kilodalton lipoprotein of Francisella tularensis expressed by Salmonella typhimurium. Infect Immun. 1992 Jul;60(7):2855–2862. doi: 10.1128/iai.60.7.2855-2862.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Smith R., Lehner T. A radioimmunoassay for serum and gingival crevicular fluid antibodies to a purified protein of Streptococcus mutans. Clin Exp Immunol. 1981 Feb;43(2):417–424. [PMC free article] [PubMed] [Google Scholar]
  30. Srisart P., Reynolds B. L., Rowley D. The correlation between serum IgA antibody levels and resistance to infection with Salmonella typhimurium after oral immunization with various salmonellae. Aust J Exp Biol Med Sci. 1985 Apr;63(Pt 2):177–182. doi: 10.1038/icb.1985.19. [DOI] [PubMed] [Google Scholar]
  31. Stabel T. J., Mayfield J. E., Tabatabai L. B., Wannemuehler M. J. Swine immunity to an attenuated Salmonella typhimurium mutant containing a recombinant plasmid which codes for production of a 31-kilodalton protein of Brucella abortus. Infect Immun. 1991 Sep;59(9):2941–2947. doi: 10.1128/iai.59.9.2941-2947.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Takahashi I., Matsushita K., Nisizawa T., Okahashi N., Russell M. W., Suzuki Y., Munekata E., Koga T. Genetic control of immune responses in mice to synthetic peptides of a Streptococcus mutans surface protein antigen. Infect Immun. 1992 Feb;60(2):623–629. doi: 10.1128/iai.60.2.623-629.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Terleckyj B., Willett N. P., Shockman G. D. Growth of several cariogenic strains of oral streptococci in a chemically defined medium. Infect Immun. 1975 Apr;11(4):649–655. doi: 10.1128/iai.11.4.649-655.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Tite J. P., Gao X. M., Hughes-Jenkins C. M., Lipscombe M., O'Callaghan D., Dougan G., Liew F. Y. Anti-viral immunity induced by recombinant nucleoprotein of influenza A virus. III. Delivery of recombinant nucleoprotein to the immune system using attenuated Salmonella typhimurium as a live carrier. Immunology. 1990 Aug;70(4):540–546. [PMC free article] [PubMed] [Google Scholar]
  35. Tsai C. M., Frasch C. E. A sensitive silver stain for detecting lipopolysaccharides in polyacrylamide gels. Anal Biochem. 1982 Jan 1;119(1):115–119. doi: 10.1016/0003-2697(82)90673-x. [DOI] [PubMed] [Google Scholar]
  36. Yang D. M., Fairweather N., Button L. L., McMaster W. R., Kahl L. P., Liew F. Y. Oral Salmonella typhimurium (AroA-) vaccine expressing a major leishmanial surface protein (gp63) preferentially induces T helper 1 cells and protective immunity against leishmaniasis. J Immunol. 1990 Oct 1;145(7):2281–2285. [PubMed] [Google Scholar]

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

RESOURCES