Skip to main content
NCBI home page
Infection and Immunity logoLink to Infection and Immunity
. 1994 Jun;62(6):2285–2288. doi: 10.1128/iai.62.6.2285-2288.1994

Toxicity of lipopolysaccharide and of soluble extracts of Salmonella typhimurium in mice immunized with a live attenuated aroA salmonella vaccine.

P Mastroeni 1, B Villarreal-Ramos 1, J A Harrison 1, R Demarco de Hormaeche 1, C E Hormaeche 1
PMCID: PMC186509  PMID: 8188350

Abstract

Mice immunized intravenously 10 days earlier (but not those immunized 2 months earlier) with an attenuated Salmonella typhimurium SL3261 aroA live vaccine and tested for delayed-type hypersensitivity by injection of crude Salmonella extracts in the footpad can die within 24 to 48 h of an unexplained allergic reaction. The lethal reaction could be prevented by prior administration of anti-tumor necrosis factor alpha serum. Injection of lipopolysaccharide (LPS) (either purified phenol-water-extracted [Westphal] LPS or protein-rich trichloracetic acid-extracted [Boivin] LPS) was also lethal for mice immunized 10 days before. An LPS-rich crude Salmonella extract was more toxic than one which contained less LPS, suggesting that LPS may have been involved in the lethal reactions to crude antigens. Mild alkaline hydrolysis removes O-linked acyl groups from lipid A and eliminates many toxic effects of LPS; however, both Boivin LPS and Westphal LPS remained toxic for immunized mice after alkaline hydrolysis. In contrast, alkaline hydrolysis of crude whole Salmonella extracts (which caused marked protein degradation) reduced the lethal toxicity of the extracts, especially for an LPS-rich preparation. Mice immunized orally with the live vaccine did not show hypersensitivity to either LPS or crude extracts. The results suggest that the lethal reaction to crude Salmonella antigens in mice immunized 10 days earlier is complex, that tumor necrosis factor alpha is involved, and that allergic reactions to crude antigens (but not to LPS alone) can be reduced by mild alkaline hydrolysis.

Full text

PDF
2285

Selected References

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

  1. Collins F. M., Mackaness G. B. Delayed hypersensitivity and arthus reactivity in relation to host resistance in salmonella-infected mice. J Immunol. 1968 Nov;101(5):830–845. [PubMed] [Google Scholar]
  2. Freudenberg M. A., Galanos C. Tumor necrosis factor alpha mediates lethal activity of killed gram-negative and gram-positive bacteria in D-galactosamine-treated mice. Infect Immun. 1991 Jun;59(6):2110–2115. doi: 10.1128/iai.59.6.2110-2115.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Galdiero F., de L'ero G. C., Benedetto N., Galdiero M., Tufano M. A. Release of cytokines induced by Salmonella typhimurium porins. Infect Immun. 1993 Jan;61(1):155–161. doi: 10.1128/iai.61.1.155-161.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. 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]
  5. Hormaeche C. E., Fahrenkrog M. C., Pettifor R. A., Brock J. Acquired immunity to Salmonella typhimurium and delayed (footpad) hypersensitivity in BALB/c mice. Immunology. 1981 Jul;43(3):547–554. [PMC free article] [PubMed] [Google Scholar]
  6. Hormaeche C. E., Joysey H. S., Desilva L., Izhar M., Stocker B. A. Immunity conferred by Aro- Salmonella live vaccines. Microb Pathog. 1991 Feb;10(2):149–158. doi: 10.1016/0882-4010(91)90075-l. [DOI] [PubMed] [Google Scholar]
  7. Hormaeche C. E. Natural resistance to Salmonella typhimurium in different inbred mouse strains. Immunology. 1979 Jun;37(2):311–318. [PMC free article] [PubMed] [Google Scholar]
  8. KOTELKO K., STAUB A. M., TINELLI R. [Immunochemical study on Salmonella. VIII. Role of O acetyl groupings in the specificity of the O:5 factor]. Ann Inst Pasteur (Paris) 1961 May;100:618–637. [PubMed] [Google Scholar]
  9. Kawakami M., Hara Y., Osawa N. Experimental salmonellosis: hypersensitivity to cell wall lipopolysaccharide and anti-infectious resistance of mice infected with Salmonella. Infect Immun. 1971 Nov;4(5):519–524. doi: 10.1128/iai.4.5.519-524.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Killar L. M., Eisenstein T. K. Delayed-type hypersensitivity and immunity to Salmonella typhimurium. Infect Immun. 1986 May;52(2):504–508. doi: 10.1128/iai.52.2.504-508.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Killar L. M., Eisenstein T. K. Differences in delayed-type hypersensitivity responses in various mouse strains in the C3H lineage infected with Salmonella typhimurium, strain SL3235. J Immunol. 1984 Sep;133(3):1190–1196. [PubMed] [Google Scholar]
  12. Lugtenberg B., Van Alphen L. Molecular architecture and functioning of the outer membrane of Escherichia coli and other gram-negative bacteria. Biochim Biophys Acta. 1983 Mar 21;737(1):51–115. doi: 10.1016/0304-4157(83)90014-x. [DOI] [PubMed] [Google Scholar]
  13. Mastroeni P., Arena A., Costa G. B., Liberto M. C., Bonina L., Hormaeche C. E. Serum TNF alpha in mouse typhoid and enhancement of a Salmonella infection by anti-TNF alpha antibodies. Microb Pathog. 1991 Jul;11(1):33–38. doi: 10.1016/0882-4010(91)90091-n. [DOI] [PubMed] [Google Scholar]
  14. Mastroeni P., Villarreal-Ramos B., Demarco de Hormaeche R., Hormaeche C. E. Delayed (footpad) hypersensitivity and Arthus reactivity using protein-rich antigens and LPS in mice immunized with live attenuated aroA Salmonella vaccines. Microb Pathog. 1993 May;14(5):369–379. doi: 10.1006/mpat.1993.1036. [DOI] [PubMed] [Google Scholar]
  15. Mastroeni P., Villarreal-Ramos B., Hormaeche C. E. Adoptive transfer of immunity to oral challenge with virulent salmonellae in innately susceptible BALB/c mice requires both immune serum and T cells. Infect Immun. 1993 Sep;61(9):3981–3984. doi: 10.1128/iai.61.9.3981-3984.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Mastroeni P., Villarreal-Ramos B., Hormaeche C. E. Role of T cells, TNF alpha and IFN gamma in recall of immunity to oral challenge with virulent salmonellae in mice vaccinated with live attenuated aro- Salmonella vaccines. Microb Pathog. 1992 Dec;13(6):477–491. doi: 10.1016/0882-4010(92)90014-f. [DOI] [PubMed] [Google Scholar]
  17. Matsuura M., Galanos C. Induction of hypersensitivity to endotoxin and tumor necrosis factor by sublethal infection with Salmonella typhimurium. Infect Immun. 1990 Apr;58(4):935–937. doi: 10.1128/iai.58.4.935-937.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. NETER E., WESTPHAL O., LUDERITZ O., GORZYNSKI E. A., EICHENBERGER E. Studies of enterobacterial lipopolysaccharides; effects of heat and chemicals on erythrocyte-modifying, antigenic, toxic and pyrogenic properties. J Immunol. 1956 May;76(5):377–385. [PubMed] [Google Scholar]
  19. Shnyra A., Hultenby K., Lindberg A. A. Role of the physical state of Salmonella lipopolysaccharide in expression of biological and endotoxic properties. Infect Immun. 1993 Dec;61(12):5351–5360. doi: 10.1128/iai.61.12.5351-5360.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Sultzer B. M., Goodman G. W. Endotoxin protein: a B-cell mitogen and polyclonal activator of C3H/HeJ lymphocytes. J Exp Med. 1976 Sep 1;144(3):821–827. doi: 10.1084/jem.144.3.821. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Tacket C. O., Hone D. M., Losonsky G. A., Guers L., Edelman R., Levine M. M. Clinical acceptability and immunogenicity of CVD 908 Salmonella typhi vaccine strain. Vaccine. 1992;10(7):443–446. doi: 10.1016/0264-410x(92)90392-w. [DOI] [PubMed] [Google Scholar]
  22. Villarreal B., Mastroeni P., de Hormaeche R. D., Hormaeche C. E. Proliferative and T-cell specific interleukin (IL-2/IL-4) production responses in spleen cells from mice vaccinated with aroA live attenuated Salmonella vaccines. Microb Pathog. 1992 Oct;13(4):305–315. doi: 10.1016/0882-4010(92)90040-u. [DOI] [PubMed] [Google Scholar]

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

RESOURCES