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TNF- Sensitizes HT-29 Colonic Epithelial Cells to Intestinal Lactobacilli¹

Vance J. McCracken^*,2, Taehoon Chun^*,3, Manuel E. Baldeón^,4, Siv Ahrné, Göran Molin, Roderick I. Mackie^* and H. Rex Gaskins^*,,,5

^* Departments of Animal Sciences and
Veterinary Pathobiology and
Division of Nutritional Sciences, University of Illinois, Urbana, Illinois 61801; and
Laboratory of Food Hygiene, Department of Food Technology, Lund University, Lund, Sweden

The ability of the proinflammatory cytokine tumor necrosis factor- (TNF-) to influence epithelial interleukin (IL)-8 responses to the intestinal bacterium Lactobacillus plantarum 299v was analyzed in the human HT-29 colonic epithelial cell line. In the absence of TNF-, IL-8 mRNA expression was not detectable by Northern blot analysis in HT-29 cells alone or in HT-29 cells co-cultured with L. plantarum 299v. However, TNF- induced IL-8 mRNA expression, and co-culture of TNF--treated HT-29 cells with L. plantarum 299v significantly increased IL-8 mRNA expression above levels induced by TNF- alone in an adhesion-dependent manner. The increase in IL-8 mRNA expression was not observed in TNF--treated HT-29/L. plantarum 299v co-cultures using heat-killed lactobacilli or when L. plantarum adhesion was prevented using mannoside or a trans-well membrane. Paradoxically, IL-8 secretion was decreased in TNF--treated HT-29 cells with L. plantarum 299v relative to cells treated with TNF- alone. TNF--mediated responsiveness to L. plantarum 299v was further investigated by analyzing expression of a coreceptor for bacterial cell wall products CD14. HT-29 cells expressed CD14 mRNA and cell-surface CD14; however, TNF- did not alter CD14 mRNA or cell-surface expression, and blockade of CD14 with monoclonal antibody MY4 did not alter the IL-8 response to L. plantarum 299v in TNF--treated HT-29 cells. These results indicate that although TNF- sensitizes HT-29 epithelial cells to intestinal lactobacilli, the bacteria exert a protective effect by downregulating IL-8 secretion.

Key Words: Lactobacillus • intestinal epithelial cell • interleukin-8 • CD14

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