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Infection and Immunity, October 1999, p. 5434-5440, Vol. 67, No. 10
0019-9567/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Infection of Endothelial Cells with Trypanosoma cruzi Activates NF-kappa B and Induces Vascular Adhesion Molecule Expression

Huan Huang,1,* Tina M. Calderon,1 Joan W. Berman,1,2 Vicki L. Braunstein,1 Louis M. Weiss,1,3 Murray Wittner,1 and Herbert B. Tanowitz1,3

Departments of Pathology,1 Microbiology and Immunology,2 and Medicine,3 Albert Einstein College of Medicine, Bronx, New York 10461

Received 29 March 1999/Returned for modification 21 May 1999/Accepted 27 July 1999

Transcriptional activation of vascular adhesion molecule expression, a major component of an inflammatory response, is regulated, in part, by the nuclear factor-kappa B/Rel (NF-kappa B) family of transcription factors. We therefore determined whether Trypanosoma cruzi infection of endothelial cells resulted in the activation of NF-kappa B and the induction or increased expression of adhesion molecules. Human umbilical vein endothelial cells (HUVEC) were infected with trypomastigotes of the Tulahuen strain of T. cruzi. Electrophoretic mobility shift assays with an NF-kappa B-specific oligonucleotide and nuclear extracts from T. cruzi-infected HUVEC (6 to 48 h postinfection) detected two major shifted complexes. Pretreatment with 50× cold NF-kappa B consensus sequence abolished both gel-shifted complexes while excess SP-1 consensus sequence had no effect. These data indicate that nuclear extracts from T. cruzi-infected HUVEC specifically bound to the NF-kappa B consensus DNA sequence. Supershift analysis revealed that the gel-shifted complexes were comprised of p65 (RelA) and p50 (NF-kappa B1). Northern blot analyses demonstrated both the induction of vascular cell adhesion molecule 1 and E-selectin and the upregulation of intercellular adhesion molecule 1 mRNA in HUVEC infected with T. cruzi. Immunocytochemical staining confirmed adhesion molecule expression in response to T. cruzi infection. These findings are consistent with the hypothesis that the activation of the NF-kappa B pathway in endothelial cells associated with T. cruzi infection may be an important factor in the inflammatory response and subsequent vascular injury and endothelial dysfunction that lead to chronic cardiomyopathy.

* Corresponding author. Mailing address: Department of Pathology, Albert Einstein College of Medicine, 1300 Morris Park Ave., Bronx, NY 10461. Phone: (718) 430-2143. Fax: (718) 430-8543. E-mail: Huangh{at}aecom.yu.edu.

Infection and Immunity, October 1999, p. 5434-5440, Vol. 67, No. 10
0019-9567/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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