Mucosal Immunology

FIGURE 3

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High frequencies of polyfunctional HIV-specific T cells are associated with preservation of mucosal CD4 T cells in bronchoalveolar lavage

J M Brenchley, K S Knox, A I Asher, D A Price, L M Kohli, E Gostick, B J Hill, C A Hage, Z Brahmi, A Khoruts, H L Twigg III, T W Schacker and D C Douek

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Figure 3.

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Flow cytometric analysis of cytokine production after antigenic stimulation. GI tract lymphocytes from subject 1464 were stimulated with SEB in the presence of brefeldin A for 18 h and then stained for surface and intracellular proteins as described in Methods. Lymphocytes are gated based on characteristic light-scatter properties, single lymphocytes are gated based on forward scatter height vs. forward scatter area, and live T cells are gated based on expression of CD3 without staining for the dead cell dye vivid. CD4 or CD8 T cells are then gated and memory CD4 or CD8 T cells are gated based on the characteristic expression patterns of CD27 and CD45RO. The percentage of memory CD4 and CD8 T cells that produce IFN-gamma, TNF, and IL-2 can then be determined. GI, gastrointestinal.

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