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. 2015 Mar 12;5:8925. doi: 10.1038/srep08925

Figure 3. Env-specific CD4+ T cells frequency and cytokine secretion.

Figure 3

Frequencies of cytokine-producing CD4+ T cells (IFN-γ, IL-2, TNF-α) after in vitro stimulation of PBMCs with 15-mer peptides spanning YU2 gp140-F. (A) Gating strategy from one representative animal (K17: in the AbISCO+CpG group) sampled two weeks after the second immunization is shown. Living, single lymphocytes (upper panels), were gated on CD3 and CD4 (middle panels) and then independently for IFN-γ, IL-2 or TNF-α positive populations (lower panels). (B) Kinetics of the frequency of cytokine producing Env-specific CD4+ T cells for each animal two weeks after the first, second and third immunization. Data from six individual animals were plotted after medium subtraction. (C) Total cytokine-producing Env-specific CD4+ T cells were subdivided in seven populations based on the production of one, two or three cytokines (IFN-γ, IL-2, TNF-α). Data from two weeks after the first and the second immunizations (n = 12) after medium subtraction are shown and analyzed with Mann-Whitney test. NS = non-significant.