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. 2010 Feb 17;5(2):e9254. doi: 10.1371/journal.pone.0009254

Figure 5. Impaired IL-17 production by IL-2 expanded effector cells from HIV+ subjects.

Figure 5

IL-17A, IFN-γ, IL-4 and TNFα production was measured from expanded CD4+CD25− effector cell lines. Cells were expanded over six days with irradiated APC, IL-2 and purified anti-CD3 Ab and then re-stimulated for 16 hours with PMA/ION in the presence of 5µg/ml brefeldin. (A) Representative FACS plot showing gating strategy in PBMC of ex vivo IL-17 and IFN-γ production after 4 hours of PMA/ION stimulation from a HIV+ patient and healthy control in the presence of brefeldin A. (B) Representative FACS plots show IL-17 and IFN-γ+ CD4+CD25− cell frequency from expanded cells after 16 hr stimulation with PMA/Ionomycin in the presence of 5µg/ml brefeldin A. (C) Frequency of IL-17 and IFN-γ production from ex vivo CD4+CD45RO+ gated cells within PBMC (D). Frequency of total IL-17 positive cells in expanded CD4+CD45RO+CD25− effector cells from controls (N = 16) and HIV+ (N = 20) subjects. Group differences were determined by Fischer's Exact test and 2-sided p-value reported. (E) Frequency of IFN-γ, TNFα and IL-4 positive cells in expanded cell lines from 4 control (black bars) versus 6 HIV+ (white bar) subjects. (F) Differences in these individual cytokines between control v HIV+ samples was assessed by paired t-test and were non significant.