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Infection and Immunity, April 2007, p. 1765-1770, Vol. 75, No. 4
0019-9567/07/$08.00+0     doi:10.1128/IAI.01442-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Induction of Monocyte Chemotactic Protein 1 in Colonic Epithelial Cells by Entamoeba histolytica Is Mediated via the Phosphatidylinositol 3-Kinase/p65 Pathway{triangledown}

Srinivas J. Kammanadiminti,{dagger} Indranil Dey, and Kris Chadee*

Department of Microbiology and Infectious Diseases, University of Calgary, 3330 Hospital Drive NW, Calgary T2N 4N1, Alberta, Canada

Received 7 September 2006/ Returned for modification 17 October 2006/ Accepted 21 January 2007

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 µg/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 I{kappa}B kinase activity. Moreover, inhibiting the classical pathway of NF-{kappa}B 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 NF{kappa}B p65 subunit. SAP induced phosphorylation of p65 and enhanced NF-{kappa}B 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.

* Corresponding author. Mailing address: Department of Microbiology and Infectious Diseases, University of Calgary, 3330 Hospital Dr. NW, Calgary T2N 4N1, Alberta, Canada. Phone: (403) 210-3975. Fax: (403) 270-2772. E-mail: kchadee{at}ucalgary.ca.

{triangledown} Published ahead of print on 5 February 2007.

Editor: W. A. Petri, Jr.

{dagger} Present address: Cangene Corporation, 155 Innovation Drive, Winnipeg R3T 5Y3, Manitoba, Canada.

Infection and Immunity, April 2007, p. 1765-1770, Vol. 75, No. 4
0019-9567/07/$08.00+0     doi:10.1128/IAI.01442-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



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