Lipopolysaccharide activates distinct signaling pathways in intestinal epithelial cell lines expressing toll-like receptors

Elke Cario, Ian M. Rosenberg, Steven L. Brandwein, Paul L. Beck, Hans Christian Reinecker, Daniel K. Podolsky

Research output: Contribution to journalArticle

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Abstract

LPS elicits several immediate proinflammatory responses in peripheral blood leukocytes via a recently described pathway including CD14, Toll-like receptors (TLR), serine-threonine kinases, and NF-κB transcription factor. However, the functional responses of intestinal epithelial cells (IEC) to stimulation with LPS are unknown. Expression of mRNA and protein for CD14 and TLRs were assessed by RT-PCR, immunoblotting, and immunohistochemistry in mouse and human IEC lines. LPS-induced activation of signaling pathways (p42/p44 mitogen-activated protein kinase (MAPK), c-Jun NH2-terminal kinase (JNK), p38, p65, NF-κB) were assessed by immunoblotting and gel shifts. CD14 mRNA and protein expression were not detectable in IEC. However, human TLR2, TLR3, and TLR4 mRNA were present in IEC. TLR4 protein was expressed in all cell lines; however, TLR2 protein was absent in HT29 cells. Immunofluorescent staining of T84 cells demonstrated the cell-surface presence of the TLRs. LPS-stimulation of IEC resulted in activation (> 1.5-fold)of the three members of the MAPK family. In contrast, LPS did not significantly induce activation of JNK and p38 in CMT93 cells, p38 in T84 cells and MAPK and JNK in HT29 cells. Downstream, LPS activated NF-κB in IEC in a time-, dose-, and serum-dependent manner. IEC express TLRs that appear to mediate LPS stimulation of specific intracellular signal transduction pathways in IEC. Thus, IEC may play a frontline role in monitoring lumenal bacteria.

Original languageEnglish (US)
Pages (from-to)966-972
Number of pages7
JournalJournal of Immunology
Volume164
Issue number2
StatePublished - Jan 15 2000

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Toll-Like Receptors
Lipopolysaccharides
Epithelial Cells
Cell Line
JNK Mitogen-Activated Protein Kinases
HT29 Cells
Mitogen-Activated Protein Kinases
Immunoblotting
Messenger RNA
Proteins
Protein-Serine-Threonine Kinases
Mitogen-Activated Protein Kinase 1
Signal Transduction
Leukocytes
Transcription Factors
Gels
Immunohistochemistry
Staining and Labeling
Bacteria
Polymerase Chain Reaction

ASJC Scopus subject areas

  • Immunology

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Lipopolysaccharide activates distinct signaling pathways in intestinal epithelial cell lines expressing toll-like receptors. / Cario, Elke; Rosenberg, Ian M.; Brandwein, Steven L.; Beck, Paul L.; Reinecker, Hans Christian; Podolsky, Daniel K.

In: Journal of Immunology, Vol. 164, No. 2, 15.01.2000, p. 966-972.

Research output: Contribution to journalArticle

Cario, Elke ; Rosenberg, Ian M. ; Brandwein, Steven L. ; Beck, Paul L. ; Reinecker, Hans Christian ; Podolsky, Daniel K. / Lipopolysaccharide activates distinct signaling pathways in intestinal epithelial cell lines expressing toll-like receptors. In: Journal of Immunology. 2000 ; Vol. 164, No. 2. pp. 966-972.
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