title: Induction of monocyte chemotactic protein 1 in colonic epithelial cells by Entamoeba histolytica is mediated via the phosphatidylinositol 3-kinase/p65 pathway.
abstract: The role intestinal epithelial cells play in the pathogenesis of amebic colitis is poorly understood. Herein, we demonstrate that secreted and soluble ameba (Entamoeba histolytica) proteins (SAP) induce expression of the chemoattractant monocyte chemotactic protein (MCP) in the colonic epithelial cell lines Caco-2, T84, and LS174T. MCP-1 mRNA induction was both dose and time dependent, with peak induction occurring at 8 h and with 100 mug/ml of SAP. Significant increase in MCP-1 protein expression was observed after 12 h. SAP failed to activate any of the mitogen-activated protein kinase pathways or IkappaB kinase activity. Moreover, inhibiting the classical pathway of NF-kappaB activation did not affect SAP-induced MCP-1 expression. Instead, we find that SAP-induced MCP-1 expression is dependent on posttranslational modification of the NFkappaB p65 subunit. SAP induced phosphorylation of p65 and enhanced NF-kappaB transcriptional activity, which are phosphatidylinositol 3-kinase (PI3 kinase) dependent. Treatment with PI3 kinase inhibitor LY290004 significantly abrogated the activation of Akt, p65, and MCP-1 mRNA induction. We conclude that colonic epithelial cells play a role in the initiation of inflammation by secreting chemokines in response to soluble ameba components.
authors.affiliation: Department of Microbiology and Infectious Diseases, University of Calgary, 3330 Hospital Drive NW, Calgary T2N 4N1, Alberta, Canada.
authors.forename: Srinivas J
authors.initials: SJ
authors.lastname: Kammanadiminti
authors.forename: Indranil
authors.initials: I
authors.lastname: Dey
authors.forename: Kris
authors.initials: K
authors.lastname: Chadee
mesh_terms: D000818:Animals; D015153:Blotting, Western; D002460:Cell Line; D018932:Chemokine CCL2 / Q000096:biosynthesis*; D003106:Colon / Q000276:immunology* / Q000378:metabolism; D004306:Dose-Response Relationship, Immunologic; D004748:Entamoeba histolytica / Q000276:immunology*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004847:Epithelial Cells / Q000276:immunology / Q000378:metabolism / Q000469:parasitology*; D015870:Gene Expression; D006801:Humans; D019869:Phosphatidylinositol 3-Kinases / Q000502:physiology*; D010716:Phosphatidylinositols; D000081082:Phosphoinositide-3 Kinase Inhibitors; D010766:Phosphorylation; D015800:Protozoan Proteins / Q000276:immunology / Q000378:metabolism; D012333:RNA, Messenger / Q000096:biosynthesis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015398:Signal Transduction; D013997:Time Factors; D051996:Transcription Factor RelA / Q000378:metabolism*
chemical_list: D018932:Chemokine CCL2; D004791:Enzyme Inhibitors; D019869:Phosphatidylinositol 3-Kinases; D010716:Phosphatidylinositols; D000081082:Phosphoinositide-3 Kinase Inhibitors; D015800:Protozoan Proteins; D012333:RNA, Messenger; D051996:Transcription Factor RelA