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Comparative study of the invasiveness of Salmonella serotypes Typhimurium, Choleraesuis and Dublin for Caco-2 cells, HEp-2 cells and rabbit ileal epithelia.
J Med Microbiol. 2000 Jun; 49(6):503-511.JM

Abstract

Patterns of invasiveness of Salmonella serotypes Typhimurium, Choleraesuis and Dublin in Caco-2 cells (without centrifugation) were compared with previously published studies of the rabbit ileal invasion assay (RIIA) and (where relevant) a HEp-2 cell invasion assay. Optimal conditions for the use of Caco-2 cell monolayers in bacterial invasion assays were defined. Centrifuge-assisted attachment of bacteria to cells was not used routinely as this increased the invasiveness of known hypo-invasive strains and detachment of Caco-2 cells. Inocula with too high bacterial numbers resulted in rapid acidification of media and detachment of the monolayers. The invasiveness of Typhimurium strains TML, WAKE, WII8, LT7, SL1027 and M206 in Caco-2 cells reflected that seen in the RIIA. The invasiveness of Choleraesuis strain A50 was similar to that in the RIIA except that bacteria grown at 37 degrees C and used without storage at 4 degrees C were slightly more invasive than those grown at 37 degrees C and stored at 4 degrees C before use. Dublin strain 3246 showed no apparent temperature-regulated invasiveness in Caco-2 cells, in contrast to the results observed in the RIIA. Dublin strain 3246 did not cleave tight junctions in the Caco-2 cell monolayer as it did in rabbit ileal epithelia both in vitro and in vivo. Three TnphoA insertion LPS mutants of Typhimurium TML were uniformly hypo-invasive in both Caco-2 cells and the RIIA; in contrast, they were differentially invasive in HEp-2 cells. Three smooth TnphoA insertion mutants of Typhimurium TML (invH, invG and pagC) were hypo-invasive in both the Caco-2 and HEp-2 cell invasion assays but not in the RIIA.

Authors+Show Affiliations

No affiliation info availableMicrobial Molecular Genetics and Cell Biology Group, School of Biological Sciences, University of Birmingham, Edgbaston, Birmingham B15 2TT and *Department of Physiology, Medical School, University of Birmingham, Edgbaston, Birmingham B15 2TT.No affiliation info available

Pub Type(s)

Comparative Study
Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

10847203

Citation

Bolton, Alex J., et al. "Comparative Study of the Invasiveness of Salmonella Serotypes Typhimurium, Choleraesuis and Dublin for Caco-2 Cells, HEp-2 Cells and Rabbit Ileal Epithelia." Journal of Medical Microbiology, vol. 49, no. 6, 2000, pp. 503-511.
Bolton AJ, Osborne MP, Stephen J. Comparative study of the invasiveness of Salmonella serotypes Typhimurium, Choleraesuis and Dublin for Caco-2 cells, HEp-2 cells and rabbit ileal epithelia. J Med Microbiol. 2000;49(6):503-511.
Bolton, A. J., Osborne, M. P., & Stephen, J. (2000). Comparative study of the invasiveness of Salmonella serotypes Typhimurium, Choleraesuis and Dublin for Caco-2 cells, HEp-2 cells and rabbit ileal epithelia. Journal of Medical Microbiology, 49(6), 503-511. https://doi.org/10.1099/0022-1317-49-6-503
Bolton AJ, Osborne MP, Stephen J. Comparative Study of the Invasiveness of Salmonella Serotypes Typhimurium, Choleraesuis and Dublin for Caco-2 Cells, HEp-2 Cells and Rabbit Ileal Epithelia. J Med Microbiol. 2000;49(6):503-511. PubMed PMID: 10847203.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Comparative study of the invasiveness of Salmonella serotypes Typhimurium, Choleraesuis and Dublin for Caco-2 cells, HEp-2 cells and rabbit ileal epithelia. AU - Bolton,Alex J, AU - Osborne,Michael P, AU - Stephen,John, PY - 2000/6/10/pubmed PY - 2000/6/17/medline PY - 2000/6/10/entrez SP - 503 EP - 511 JF - Journal of medical microbiology JO - J Med Microbiol VL - 49 IS - 6 N2 - Patterns of invasiveness of Salmonella serotypes Typhimurium, Choleraesuis and Dublin in Caco-2 cells (without centrifugation) were compared with previously published studies of the rabbit ileal invasion assay (RIIA) and (where relevant) a HEp-2 cell invasion assay. Optimal conditions for the use of Caco-2 cell monolayers in bacterial invasion assays were defined. Centrifuge-assisted attachment of bacteria to cells was not used routinely as this increased the invasiveness of known hypo-invasive strains and detachment of Caco-2 cells. Inocula with too high bacterial numbers resulted in rapid acidification of media and detachment of the monolayers. The invasiveness of Typhimurium strains TML, WAKE, WII8, LT7, SL1027 and M206 in Caco-2 cells reflected that seen in the RIIA. The invasiveness of Choleraesuis strain A50 was similar to that in the RIIA except that bacteria grown at 37 degrees C and used without storage at 4 degrees C were slightly more invasive than those grown at 37 degrees C and stored at 4 degrees C before use. Dublin strain 3246 showed no apparent temperature-regulated invasiveness in Caco-2 cells, in contrast to the results observed in the RIIA. Dublin strain 3246 did not cleave tight junctions in the Caco-2 cell monolayer as it did in rabbit ileal epithelia both in vitro and in vivo. Three TnphoA insertion LPS mutants of Typhimurium TML were uniformly hypo-invasive in both Caco-2 cells and the RIIA; in contrast, they were differentially invasive in HEp-2 cells. Three smooth TnphoA insertion mutants of Typhimurium TML (invH, invG and pagC) were hypo-invasive in both the Caco-2 and HEp-2 cell invasion assays but not in the RIIA. SN - 0022-2615 UR - https://www.unboundmedicine.com/medline/citation/10847203/Comparative_study_of_the_invasiveness_of_Salmonella_serotypes_Typhimurium_Choleraesuis_and_Dublin_for_Caco_2_cells_HEp_2_cells_and_rabbit_ileal_epithelia_ L2 - http://jmm.microbiologyresearch.org/pubmed/content/journal/jmm/10.1099/0022-1317-49-6-503 DB - PRIME DP - Unbound Medicine ER -