Skip to main content
PMC home page
Infection and Immunity logoLink to Infection and Immunity
. 1993 Feb;61(2):610–618. doi: 10.1128/iai.61.2.610-618.1993

Oral immunization of mice and swine with an attenuated Salmonella choleraesuis [delta cya-12 delta(crp-cdt)19] mutant containing a recombinant plasmid.

T J Stabel 1, J E Mayfield 1, D C Morfitt 1, M J Wannemuehler 1
PMCID: PMC302771  PMID: 8423090

Abstract

Salmonella choleraesuis chi 3781, an attenuated [delta cya-12 delta(crp-cdt)19] mutant, was electroporated with the plasmid pBA31-R7, which codes for the expression of a 31-kDa protein from Brucella abortus (BCSP31). Recombinant S. choleraesuis chi 3781 stably maintained the pBA31-R7 plasmid and continued to express the cloned protein following recovery of the organism from orally inoculated animals. Unlike previous studies using S. typhimurium chi 4064(pBA31-R7), S. choleraesuis chi 3781(pBA31-R7) was able to colonize both the gut mucosa and deep tissues of both BALB/cByJ mice and crossbred swine. Orally inoculated mice developed serum antibodies to both the cloned 31-kDa protein (rBCSP31) and to S. choleraesuis chi 3781 endotoxin. These mice also developed a local intestinal antibody response to Salmonella endotoxin but not to rBCSP31. Similarly, mice inoculated with recombinant S. choleraesuis chi 3781 did not develop a delayed-type hypersensitivity (DTH) footpad response following injection with rBCSP31; however, these mice did respond to S. choleraesuis chi 3781 soluble antigen. Conversely, orally inoculated swine did not develop significant serum or intestinal antibody responses to cloned protein or Salmonella endotoxin, but DTH responses to both cloned protein and S. choleraesuis chi 3781 soluble antigen were strongly positive. The cell-mediated nature of these DTH responses was confirmed by histological examination. Results suggest that S. choleraesuis chi 3781 may be a suitable choice for further studies of vaccine efficacy in swine, especially for diseases which require cell-mediated immunity for resolution.

Full text

PDF
610

Images in this article

613Image
on p.613
616Image
on p.616

Selected References

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

  1. Baba T. Cell-mediated immune protection in chickens against Pasteurella multocida. Res Vet Sci. 1984 Mar;36(2):225–230. [PubMed] [Google Scholar]
  2. Berche P., Gaillard J. L., Sansonetti P. J. Intracellular growth of Listeria monocytogenes as a prerequisite for in vivo induction of T cell-mediated immunity. J Immunol. 1987 Apr 1;138(7):2266–2271. [PubMed] [Google Scholar]
  3. Binotto J., MacLachlan P. R., Sanderson K. E. Electrotransformation in Salmonella typhimurium LT2. Can J Microbiol. 1991 Jun;37(6):474–477. doi: 10.1139/m91-078. [DOI] [PubMed] [Google Scholar]
  4. Birnboim H. C., Doly J. A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res. 1979 Nov 24;7(6):1513–1523. doi: 10.1093/nar/7.6.1513. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Breiman R. F., Horwitz M. A. Guinea pigs sublethally infected with aerosolized Legionella pneumophila develop humoral and cell-mediated immune responses and are protected against lethal aerosol challenge. A model for studying host defense against lung infections caused by intracellular pathogens. J Exp Med. 1987 Mar 1;165(3):799–811. doi: 10.1084/jem.165.3.799. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Bretscher P. A., Havele C. Cyclosporin A can switch the immune response induced by antigen from a humoral to a cell-mediated mode. Eur J Immunol. 1992 Feb;22(2):349–355. doi: 10.1002/eji.1830220210. [DOI] [PubMed] [Google Scholar]
  7. Bretscher P. A. Significance and mechanisms of cellular regulation of the immune response. Fed Proc. 1981 Apr;40(5):1473–1478. [PubMed] [Google Scholar]
  8. 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]
  9. Cheers C., Pagram F. Macrophage activation during experimental murine brucellosis: a basis for chronic infection. Infect Immun. 1979 Feb;23(2):197–205. doi: 10.1128/iai.23.2.197-205.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Cheers C. Pathogenesis and cellular immunity in experimental murine brucellosis. Dev Biol Stand. 1984;56:237–246. [PubMed] [Google Scholar]
  11. Cooper G. L., Nicholas R. A., Cullen G. A., Hormaeche C. E. Vaccination of chickens with a Salmonella enteritidis aroA live oral Salmonella vaccine. Microb Pathog. 1990 Oct;9(4):255–265. doi: 10.1016/0882-4010(90)90014-h. [DOI] [PubMed] [Google Scholar]
  12. Curtiss R., 3rd, Goldschmidt R. M., Fletchall N. B., Kelly S. M. Avirulent Salmonella typhimurium delta cya delta crp oral vaccine strains expressing a streptococcal colonization and virulence antigen. Vaccine. 1988 Apr;6(2):155–160. doi: 10.1016/s0264-410x(88)80020-3. [DOI] [PubMed] [Google Scholar]
  13. Dower W. J., Miller J. F., Ragsdale C. W. High efficiency transformation of E. coli by high voltage electroporation. Nucleic Acids Res. 1988 Jul 11;16(13):6127–6145. doi: 10.1093/nar/16.13.6127. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Hanna J., McCracken R., O'Brien J. J. Evaluation of a live Salmonella choleraesuis vaccine by intranasal challenge. Res Vet Sci. 1979 Mar;26(2):216–219. [PubMed] [Google Scholar]
  15. Hassan J. O., Curtiss R., 3rd Control of colonization by virulent Salmonella typhimurium by oral immunization of chickens with avirulent delta cya delta crp S. typhimurium. Res Microbiol. 1990 Sep-Oct;141(7-8):839–850. doi: 10.1016/0923-2508(90)90119-b. [DOI] [PubMed] [Google Scholar]
  16. Jones P. W., Dougan G., Hayward C., Mackensie N., Collins P., Chatfield S. N. Oral vaccination of calves against experimental salmonellosis using a double aro mutant of Salmonella typhimurium. Vaccine. 1991 Jan;9(1):29–34. doi: 10.1016/0264-410x(91)90313-u. [DOI] [PubMed] [Google Scholar]
  17. Kaufmann S. H. Role of T-cell subsets in bacterial infections. Curr Opin Immunol. 1991 Aug;3(4):465–470. doi: 10.1016/0952-7915(91)90004-k. [DOI] [PubMed] [Google Scholar]
  18. Kawahara K., Haraguchi Y., Tsuchimoto M., Terakado N., Danbara H. Evidence of correlation between 50-kilobase plasmid of Salmonella choleraesuis and its virulence. Microb Pathog. 1988 Feb;4(2):155–163. doi: 10.1016/0882-4010(88)90057-5. [DOI] [PubMed] [Google Scholar]
  19. Kelly S. M., Bosecker B. A., Curtiss R., 3rd Characterization and protective properties of attenuated mutants of Salmonella choleraesuis. Infect Immun. 1992 Nov;60(11):4881–4890. doi: 10.1128/iai.60.11.4881-4890.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Koster F. T., McGregor D. D., Mackaness G. B. The mediator of cellular immunity. II. Migration of immunologically committed lymphocytes into inflammatory exudates. J Exp Med. 1971 Feb 1;133(2):400–409. doi: 10.1084/jem.133.2.400. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Kramer T. T., Pardon P., Marly J., Bernard S. Conjunctival and intramuscular vaccination of pigs with a live avirulent strain of Salmonella cholerae-suis. Am J Vet Res. 1987 Jul;48(7):1072–1076. [PubMed] [Google Scholar]
  22. 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]
  23. Lagrange P. H., Mackaness G. B., Miller T. E. Influence of dose and route of antigen injection on the immunological induction of T cells. J Exp Med. 1974 Mar 1;139(3):528–542. doi: 10.1084/jem.139.3.528. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Mayfield J. E., Bricker B. J., Godfrey H., Crosby R. M., Knight D. J., Halling S. M., Balinsky D., Tabatabai L. B. The cloning, expression, and nucleotide sequence of a gene coding for an immunogenic Brucella abortus protein. Gene. 1988;63(1):1–9. doi: 10.1016/0378-1119(88)90540-9. [DOI] [PubMed] [Google Scholar]
  25. Morrison D. C., Leive L. Fractions of lipopolysaccharide from Escherichia coli O111:B4 prepared by two extraction procedures. J Biol Chem. 1975 Apr 25;250(8):2911–2919. [PubMed] [Google Scholar]
  26. Nnalue N. A., Stocker B. A. Some galE mutants of Salmonella choleraesuis retain virulence. Infect Immun. 1986 Dec;54(3):635–640. doi: 10.1128/iai.54.3.635-640.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Nnalue N. A., Stocker B. A. Test of the virulence and live-vaccine efficacy of auxotrophic and galE derivatives of Salmonella choleraesuis. Infect Immun. 1987 Apr;55(4):955–962. doi: 10.1128/iai.55.4.955-962.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Nnalue N. A., Stocker B. A. The effects of O-antigen character and enterobacterial common antigen content on the in vivo persistence of aromatic-dependent Salmonella sp. live-vaccine strains. Microb Pathog. 1987 Jul;3(1):31–44. doi: 10.1016/0882-4010(87)90035-0. [DOI] [PubMed] [Google Scholar]
  29. O'Callaghan D., Charbit A. High efficiency transformation of Salmonella typhimurium and Salmonella typhi by electroporation. Mol Gen Genet. 1990 Aug;223(1):156–158. doi: 10.1007/BF00315809. [DOI] [PubMed] [Google Scholar]
  30. Ramshaw I. A., Bretscher P. A., Parish C. R. Regulation of the immune response. II. Repressor T cells in cyclophosphamide-induced tolerant mice. Eur J Immunol. 1977 Mar;7(3):180–185. doi: 10.1002/eji.1830070313. [DOI] [PubMed] [Google Scholar]
  31. Sadoff J. C., Ballou W. R., Baron L. S., Majarian W. R., Brey R. N., Hockmeyer W. T., Young J. F., Cryz S. J., Ou J., Lowell G. H. Oral Salmonella typhimurium vaccine expressing circumsporozoite protein protects against malaria. Science. 1988 Apr 15;240(4850):336–338. doi: 10.1126/science.3281260. [DOI] [PubMed] [Google Scholar]
  32. Schödel F., Enders G., Jung M. C., Will H. Recognition of a hepatitis B virus nucleocapsid T-cell epitope expressed as a fusion protein with the subunit B of Escherichia coli heat labile enterotoxin in attenuated salmonellae. Vaccine. 1990 Dec;8(6):569–572. doi: 10.1016/0264-410x(90)90010-j. [DOI] [PubMed] [Google Scholar]
  33. Sidberry H., Kaufman B., Wright D. C., Sadoff J. Immunoenzymatic analysis by monoclonal antibodies of bacterial lipopolysaccharides after transfer to nitrocellulose. J Immunol Methods. 1985 Feb 11;76(2):299–305. doi: 10.1016/0022-1759(85)90307-2. [DOI] [PubMed] [Google Scholar]
  34. Sixou S., Eynard N., Escoubas J. M., Werner E., Teissié J. Optimized conditions for electrotransformation of bacteria are related to the extent of electropermeabilization. Biochim Biophys Acta. 1991 Jan 17;1088(1):135–138. doi: 10.1016/0167-4781(91)90163-g. [DOI] [PubMed] [Google Scholar]
  35. Stabel T. J., Mayfield J. E., Tabatabai L. B., Wannemuehler M. J. Oral immunization of mice with attenuated Salmonella typhimurium containing a recombinant plasmid which codes for production of a 31-kilodalton protein of Brucella abortus. Infect Immun. 1990 Jul;58(7):2048–2055. doi: 10.1128/iai.58.7.2048-2055.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. 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]
  37. Takashima T., Collins F. M. T-cell-mediated immunity in persistent Mycobacterium intracellulare infections in mice. Infect Immun. 1988 Nov;56(11):2782–2787. doi: 10.1128/iai.56.11.2782-2787.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. 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]
  39. 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]
  40. Weissbluth M., Shulman S. T., Holson B., Lerner C. Salmonella cholera-suis: a distinctive bacterial pathogen. J Pediatr. 1981 Mar;98(3):423–426. doi: 10.1016/s0022-3476(81)80710-x. [DOI] [PubMed] [Google Scholar]

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

RESOURCES