Micropatterned co-cultures of T-lymphocytes and epithelial cells as a model of mucosal immune system |
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Authors: | Gulnaz Stybayeva He Zhu Satya Dandekar Alexander Revzin |
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Institution: | a Department of Biomedical Engineering, University of California, 451 East Health Sciences Drive, Davis, CA 95616, USA b Department of Medical Microbiology and Immunology, School of Medicine, University of California, 451 East Health Sciences Drive, Davis 95616, CA, USA c National Center for Biotechnology of the Republic of Kazakhstan, Astana 010000, Kazakhstan |
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Abstract: | 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. |
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Keywords: | Antibody microarrays Micropatterned co-cultures Mucosal immune system T-lymphocytes Epithelial cells HIV enteropathy |
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