Micropatterned co-cultures of T-lymphocytes and epithelial cells as a model of mucosal immune system.
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.
Stybayeva
Gulnaz
G
Department of Biomedical Engineering, University of California, 451 East Health Sciences Drive, Davis, CA 95616, USA.
Zhu
He
H
Ramanculov
Erlan
E
Dandekar
Satya
S
George
Michael
M
Revzin
Alexander
A
D018920:Coculture Techniques; D004847:Epithelial Cells / Q000276:immunology; D015658:HIV Infections / Q000235:genetics / Q000276:immunology*; D015497:HIV-1; D006801:Humans; D018928:Immunity, Mucosal; D007413:Intestinal Mucosa / Q000276:immunology*; D019169:Jurkat Cells; D008954:Models, Biological; D020411:Oligonucleotide Array Sequence Analysis; D013601:T-Lymphocytes / Q000276:immunology* / Q000821:virology*