Expression levels of heat shock proteins in enterocyte-like Caco-2 cells after exposure to Salmonella enteritidis.
The enterocytes of the small intestine are occasionally exposed to pathogenic bacteria, such as Salmonella enteritidis 857, an etiologic agent of intestinal infections in humans. The expression of the heat shock response by enterocytes may be part of a protective mechanism developed against pathogenic bacteria in the intestinal lumen. We aimed at investigating whether S. enteritidis 857 is able to induce a heat shock response in crypt- and villus-like Caco-2 cells and at establishing the extent of the induction. To establish whether S. enteritidis 857 interfered with the integrity of the cell monolayer, the transepithelial electrical resistance (TEER) of filter-grown, differentiated (villus-like) Caco-2 cells was measured. We clearly observed damage to the integrity of the cell monolayer by measuring the TEER. The stress response was screened in both crypt- and villus-like Caco-2 cells exposed to heat (40-43 degrees C) or to graded numbers (10(1)-10(8)) of bacteria and in villus-like cells exposed to S. enteritidis 857 endotoxin. Expression of the heat shock proteins Hsp70 and Hsp90 was analyzed by polyacrylamide gel electrophoresis and immunoblotting with monoclonal antibodies. Exposure to heat or Salmonella resulted in increased levels of Hsp70 and Hsp90 in a temperature-effect or Salmonella-dose relationship, respectively. Incubation of Caco-2 cells with S. enteritidis 857 endotoxin did not induce heat shock gene expression. We conclude that S. enteritidis 857 significantly increases the levels of stress proteins in enterocyte-like Caco-2 cells. However, our data on TEER clearly indicate that this increase is insufficient to protect the cells.
Malago
Joshua J
JJ
Department of Veterinary Pathology, Faculty of Veterinary Medicine, Utrecht University, Yalelaan 1, PO Box 80.158, 3508 TD Utrecht, The Netherlands. j.malago@vet.uu.nl
Koninkx
Jos F J G
JF
Ovelgönne
Hans H
HH
van Asten
Fons J A M
FJ
Swennenhuis
Joost F
JF
van Dijk
Jaap E
JE
D018938:Caco-2 Cells; D017097:Electric Impedance; D018840:HSP70 Heat-Shock Proteins / Q000096:biosynthesis* / Q000235:genetics; D018841:HSP90 Heat-Shock Proteins / Q000096:biosynthesis* / Q000235:genetics; D006358:Hot Temperature; D006801:Humans; D007413:Intestinal Mucosa / Q000378:metabolism; D012480:Salmonella Infections / Q000378:metabolism*; D012477:Salmonella enteritidis
D018840:HSP70 Heat-Shock Proteins; D018841:HSP90 Heat-Shock Proteins