Skip to main content
Infection and Immunity logoLink to Infection and Immunity
. 1995 Apr;63(4):1462–1467. doi: 10.1128/iai.63.4.1462-1467.1995

Association with MDCK epithelial cells by Salmonella typhimurium is reduced during utilization of carbohydrates.

D A Schiemann 1
PMCID: PMC173175  PMID: 7890410

Abstract

Association of Salmonella typhimurium with MDCK epithelial cells in monolayers, represented primarily by intracellular bacteria after 30 min of contact, with centrifugation followed by vigorous washing, was measured during aerobic and anaerobic growth of the bacteria in brain heart infusion broth. Cell association was greatest during a short period in the late log phase of growth under aerobic conditions. At this time, the pH of the growth medium was changing from acid to alkaline and glucose (0.2% initially) was exhausted. Addition of excess glucose (0.5%) to brain heart infusion broth, which was not exhausted before the bacteria entered the stationary phase of growth, in which cell association dropped sharply, resulted in repression of cell association by the bacteria. The repressive effect of glucose on cell association could not be reversed by exogenous cyclic AMP in the bacterial growth medium. Under anaerobic conditions, the effect of glucose on cell association by the bacteria was not as great and the glucose was not exhausted before the bacteria entered the stationary phase. When S. typhimurium was grown in a rich but carbohydrate-free medium, cell association by the bacteria increased earlier in the growth cycle under both aerobic and anaerobic conditions. The addition of glucose and certain other utilizable carbohydrates to this medium caused a repression of cell association by S. typhimurium that was greater under aerobic growth conditions. These results show that cell association by S. typhimurium, which is accompanied by rapid internalization (cell invasion), is the same under aerobic and anaerobic conditions if the bacteria are grown to the log phase in a carbohydrate-free medium. This suggests that prior reports of greater cell invasion by S. typhimurium during anaerobic growth may have arisen from the use of media containing carbohydrates which were found to be more repressive during aerobic growth of the bacteria.

Full Text

The Full Text of this article is available as a PDF (245.2 KB).

Selected References

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

  1. Altmeyer R. M., McNern J. K., Bossio J. C., Rosenshine I., Finlay B. B., Galán J. E. Cloning and molecular characterization of a gene involved in Salmonella adherence and invasion of cultured epithelial cells. Mol Microbiol. 1993 Jan;7(1):89–98. doi: 10.1111/j.1365-2958.1993.tb01100.x. [DOI] [PubMed] [Google Scholar]
  2. Baloda S. B., Faris A., Krovacek K. Cell-surface properties of enterotoxigenic and cytotoxic Salmonella enteritidis and Salmonella typhimurium: studies on hemagglutination, cell-surface hydrophobicity, attachment to human intestinal cells and fibronectin-binding. Microbiol Immunol. 1988;32(5):447–459. doi: 10.1111/j.1348-0421.1988.tb01405.x. [DOI] [PubMed] [Google Scholar]
  3. Behlau I., Miller S. I. A PhoP-repressed gene promotes Salmonella typhimurium invasion of epithelial cells. J Bacteriol. 1993 Jul;175(14):4475–4484. doi: 10.1128/jb.175.14.4475-4484.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Betts J., Finlay B. B. Identification of Salmonella typhimurium invasiveness loci. Can J Microbiol. 1992 Aug;38(8):852–857. doi: 10.1139/m92-138. [DOI] [PubMed] [Google Scholar]
  5. Carroll J. J., Smith N., Babson A. L. A colorimetric serum glucose determination using hexokinase and glucose-6-phosphate dehydrogenase. Biochem Med. 1970 Sep;4(2):171–180. doi: 10.1016/0006-2944(70)90093-1. [DOI] [PubMed] [Google Scholar]
  6. Ernst R. K., Dombroski D. M., Merrick J. M. Anaerobiosis, type 1 fimbriae, and growth phase are factors that affect invasion of HEp-2 cells by Salmonella typhimurium. Infect Immun. 1990 Jun;58(6):2014–2016. doi: 10.1128/iai.58.6.2014-2016.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Finlay B. B., Falkow S. Salmonella interactions with polarized human intestinal Caco-2 epithelial cells. J Infect Dis. 1990 Nov;162(5):1096–1106. doi: 10.1093/infdis/162.5.1096. [DOI] [PubMed] [Google Scholar]
  8. Finlay B. B., Fry J., Rock E. P., Falkow S. Passage of Salmonella through polarized epithelial cells: role of the host and bacterium. J Cell Sci Suppl. 1989;11:99–107. doi: 10.1242/jcs.1989.supplement_11.8. [DOI] [PubMed] [Google Scholar]
  9. Finlay B. B., Gumbiner B., Falkow S. Penetration of Salmonella through a polarized Madin-Darby canine kidney epithelial cell monolayer. J Cell Biol. 1988 Jul;107(1):221–230. doi: 10.1083/jcb.107.1.221. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Finlay B. B., Heffron F., Falkow S. Epithelial cell surfaces induce Salmonella proteins required for bacterial adherence and invasion. Science. 1989 Feb 17;243(4893):940–943. doi: 10.1126/science.2919285. [DOI] [PubMed] [Google Scholar]
  11. Francis C. L., Starnbach M. N., Falkow S. Morphological and cytoskeletal changes in epithelial cells occur immediately upon interaction with Salmonella typhimurium grown under low-oxygen conditions. Mol Microbiol. 1992 Nov;6(21):3077–3087. doi: 10.1111/j.1365-2958.1992.tb01765.x. [DOI] [PubMed] [Google Scholar]
  12. 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]
  13. Galán J. E., Curtiss R., 3rd Distribution of the invA, -B, -C, and -D genes of Salmonella typhimurium among other Salmonella serovars: invA mutants of Salmonella typhi are deficient for entry into mammalian cells. Infect Immun. 1991 Sep;59(9):2901–2908. doi: 10.1128/iai.59.9.2901-2908.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Galán J. E., Curtiss R., 3rd Expression of Salmonella typhimurium genes required for invasion is regulated by changes in DNA supercoiling. Infect Immun. 1990 Jun;58(6):1879–1885. doi: 10.1128/iai.58.6.1879-1885.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Giannella R. A., Washington O., Gemski P., Formal S. B. Invasion of HeLa cells by Salmonella typhimurium: a model for study of invasiveness of Salmonella. J Infect Dis. 1973 Jul;128(1):69–75. doi: 10.1093/infdis/128.1.69. [DOI] [PubMed] [Google Scholar]
  16. Ginocchio C., Pace J., Galán J. E. Identification and molecular characterization of a Salmonella typhimurium gene involved in triggering the internalization of salmonellae into cultured epithelial cells. Proc Natl Acad Sci U S A. 1992 Jul 1;89(13):5976–5980. doi: 10.1073/pnas.89.13.5976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Groisman E. A., Ochman H. Cognate gene clusters govern invasion of host epithelial cells by Salmonella typhimurium and Shigella flexneri. EMBO J. 1993 Oct;12(10):3779–3787. doi: 10.1002/j.1460-2075.1993.tb06056.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Horiuchi S., Inagaki Y., Okamura N., Nakaya R., Yamamoto N. Type 1 pili enhance the invasion of Salmonella braenderup and Salmonella typhimurium to HeLa cells. Microbiol Immunol. 1992;36(6):593–602. doi: 10.1111/j.1348-0421.1992.tb02059.x. [DOI] [PubMed] [Google Scholar]
  19. Isberg R. R., Voorhis D. L., Falkow S. Identification of invasin: a protein that allows enteric bacteria to penetrate cultured mammalian cells. Cell. 1987 Aug 28;50(5):769–778. doi: 10.1016/0092-8674(87)90335-7. [DOI] [PubMed] [Google Scholar]
  20. Jones B. D., Lee C. A., Falkow S. Invasion by Salmonella typhimurium is affected by the direction of flagellar rotation. Infect Immun. 1992 Jun;60(6):2475–2480. doi: 10.1128/iai.60.6.2475-2480.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Jones G. W., Rabert D. K., Svinarich D. M., Whitfield H. J. Association of adhesive, invasive, and virulent phenotypes of Salmonella typhimurium with autonomous 60-megadalton plasmids. Infect Immun. 1982 Nov;38(2):476–486. doi: 10.1128/iai.38.2.476-486.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Jones G. W., Richardson L. A. The attachment to, and invasion of HeLa cells by Salmonella typhimurium: the contribution of mannose-sensitive and mannose-resistant haemagglutinating activities. J Gen Microbiol. 1981 Dec;127(2):361–370. doi: 10.1099/00221287-127-2-361. [DOI] [PubMed] [Google Scholar]
  23. Jones G. W., Richardson L. A., Uhlman D. The invasion of HeLa cells by Salmonella typhimurium: reversible and irreversible bacterial attachment and the role of bacterial motility. J Gen Microbiol. 1981 Dec;127(2):351–360. doi: 10.1099/00221287-127-2-351. [DOI] [PubMed] [Google Scholar]
  24. Kihlström E., Edebo L. Association of viable and inactivated Salmonella typhimurium 395 MS and MR 10 with HeLa cells. Infect Immun. 1976 Oct;14(4):851–857. doi: 10.1128/iai.14.4.851-857.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Kihlström E. Infection of HeLa cells with Salmonella typhimurium 395 MS and MR10 bacteria. Infect Immun. 1977 Aug;17(2):290–295. doi: 10.1128/iai.17.2.290-295.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Kihlström E., Nilsson L. Endocytosis of Salmonella typhimurium 395 MS and MR10 by HeLa cells. Acta Pathol Microbiol Scand B. 1977 Oct;85B(5):322–328. doi: 10.1111/j.1699-0463.1977.tb01982.x. [DOI] [PubMed] [Google Scholar]
  27. Kihlström E. The effects of lipopolysaccharides on the association of Salmonella typhimurium with HeLa cells. Scand J Infect Dis Suppl. 1980;Suppl 24:141–143. [PubMed] [Google Scholar]
  28. Kolb A., Busby S., Buc H., Garges S., Adhya S. Transcriptional regulation by cAMP and its receptor protein. Annu Rev Biochem. 1993;62:749–795. doi: 10.1146/annurev.bi.62.070193.003533. [DOI] [PubMed] [Google Scholar]
  29. Kusters J. G., Mulders-Kremers G. A., van Doornik C. E., van der Zeijst B. A. Effects of multiplicity of infection, bacterial protein synthesis, and growth phase on adhesion to and invasion of human cell lines by Salmonella typhimurium. Infect Immun. 1993 Dec;61(12):5013–5020. doi: 10.1128/iai.61.12.5013-5020.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Lee C. A., Falkow S. The ability of Salmonella to enter mammalian cells is affected by bacterial growth state. Proc Natl Acad Sci U S A. 1990 Jun;87(11):4304–4308. doi: 10.1073/pnas.87.11.4304. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Lee C. A., Jones B. D., Falkow S. Identification of a Salmonella typhimurium invasion locus by selection for hyperinvasive mutants. Proc Natl Acad Sci U S A. 1992 Mar 1;89(5):1847–1851. doi: 10.1073/pnas.89.5.1847. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. MacBeth K. J., Lee C. A. Prolonged inhibition of bacterial protein synthesis abolishes Salmonella invasion. Infect Immun. 1993 Apr;61(4):1544–1546. doi: 10.1128/iai.61.4.1544-1546.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Magnusson K. E., Davies J., Grundström T., Kihlström E., Normark S. Surface charge and hydrophobicity of Salmonella, E. coli, Gonococci in relation to their tendency to associate with animal cells. Scand J Infect Dis Suppl. 1980;Suppl 24:135–140. [PubMed] [Google Scholar]
  34. Mintz C. S., Cliver D. O., Deibel R. H. Attachment of Salmonella to mammalian cells in vitro. Can J Microbiol. 1983 Dec;29(12):1731–1735. doi: 10.1139/m83-263. [DOI] [PubMed] [Google Scholar]
  35. Pepe J. C., Miller V. L. The Yersinia enterocolitica inv gene product is an outer membrane protein that shares epitopes with Yersinia pseudotuberculosis invasin. J Bacteriol. 1990 Jul;172(7):3780–3789. doi: 10.1128/jb.172.7.3780-3789.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Schiemann D. A., Shope S. R. Anaerobic growth of Salmonella typhimurium results in increased uptake by Henle 407 epithelial and mouse peritoneal cells in vitro and repression of a major outer membrane protein. Infect Immun. 1991 Jan;59(1):437–440. doi: 10.1128/iai.59.1.437-440.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Tartera C., Metcalf E. S. Osmolarity and growth phase overlap in regulation of Salmonella typhi adherence to and invasion of human intestinal cells. Infect Immun. 1993 Jul;61(7):3084–3089. doi: 10.1128/iai.61.7.3084-3089.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Tavendale A., Jardine C. K., Old D. C., Duguid J. P. Haemagglutinins and adhesion of Salmonella typhimurium to HEp2 and HeLa cells. J Med Microbiol. 1983 Aug;16(3):371–380. doi: 10.1099/00222615-16-3-371. [DOI] [PubMed] [Google Scholar]
  39. Young V. B., Miller V. L., Falkow S., Schoolnik G. K. Sequence, localization and function of the invasin protein of Yersinia enterocolitica. Mol Microbiol. 1990 Jul;4(7):1119–1128. doi: 10.1111/j.1365-2958.1990.tb00686.x. [DOI] [PubMed] [Google Scholar]

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

RESOURCES