Abstract
Mucosal tissues of the gastrointestinal and genitourinary tracts serve as major portals of HIV-1 transmission, and recent literature has highlighted the important role of these tissues in pathogenesis. However, our understanding of human mucosal T-cell responses remains limited. We have previously reported methods for isolating, culturing and analyzing mucosal T-lymphocytes obtained from gastrointestinal biopsy tissue. This method of acquiring tissue is minimally invasive and well accepted by patients, and allows sampling of sites that would not otherwise be accessible without surgical intervention. This chapter summarizes the approach currently in use in our laboratory to isolate and study \(\mathrm{CD4}+\) and \(\mathrm{CD8}+\) T-cells from rectal biopsies obtained through flexible sigmoidoscopy. These methods are also applicable, with minor modifications, to small tissue samples obtained from other lymphoid tissues.
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Acknowledgements
The authors thank Drs. Beth Jamieson, Otto Yang, Peter Anton and colleagues at UCLA, and Delandy Young and Timothy Hayes at UC Davis for assistance with protocol development. We also thank Dr. Johnson T. Wong, Harvard University for providing bispecific monoclonal antibodies and Dr. Ron Veazey, Tulane University, for sharing protocols and advice. This work was supported by NIH-NIAID R01 AI-057020 and grant CH05-D-606 from the California Universitywide AIDS Research Program (UARP).
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Shacklett, B.L., Critchfield, J.W., Lemongello, D. (2009). Isolating Mucosal Lymphocytes from Biopsy Tissue for Cellular Immunology Assays. In: Prasad, V.R., Kalpana, G.V. (eds) HIV Protocols. Methods In Molecular Biology™, vol 485. Humana Press. https://doi.org/10.1007/978-1-59745-170-3_23
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DOI: https://doi.org/10.1007/978-1-59745-170-3_23
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