Journal
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS
Volume 380, Issue 3, Pages 575-580Publisher
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.bbrc.2009.01.164
Keywords
Antibody microarrays; Micropatterned co-cultures; Mucosal immune system; T-lymphocytes; Epithelial cells; HIV enteropathy
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Funding
- NIH [R21 DE01 8097, R01 Al 43274]
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Gut-associated lymphoid tissue is a major target and reservoir of human immunodeficiency virus (HIV)infected T-cells. Our studies seek to recapitulate, in vitro, interactions between HIV-infected T-lymphocytes and intestinal epithelial cells in order to investigate the mechanisms underlying the disruption of normal epithelial cell and barrier function. Here, we describe a novel approach for creating co-cultures of healthy or HIV-infected T-lymphocytes (Jurkat) and human intestinal epithelial (HT-29) cells where both cell types are positioned on the same surface in a price spatial configuration (micropattern). This co-culture method simplified observation/monitoring of the two cell types and was particularly suited for laser microdissection-based retrieval of the desired cells for downstream gene expressions Studies. DNA microarray analysis of epithelial cells retrieved from co-cultures with HIV-1-infected vs. uninfected Jurkat cells revealed that epithelial cells from HIV-infected co-cultures exhibited gene expression patterns consistent with disruption of epithelial barrier formation. Overall, the micropatterned co-culture system described here is envisioned as a valuable new tool for delineating how HIV and other infections contribute to dysfunction of mucosal epithelium. (C) 2009 Elsevier Inc. All rights reserved.
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