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. 2016 Jan 4;5(6):e1123368. doi: 10.1080/2162402X.2015.1123368

Figure 4.

Figure 4.

IMP-3-specific Th cells promote the expansion of A2-IMP-3-SP-specific CTLs. (A) HLA-A2-restricted and IMP-3515–523-SP-specific CTLs (A2-IMP-3-SP-specific CTLs) were generated from a HLA-A2+ HD (HD4), and they were cultured for 7 d with A2-IMP-3-SP (SP) in the presence or absence of IMP-3-LP3 (LP) and LP3-specific Th clone (Th clone). (B) At the end of culture, the frequencies of A2-IMP-3-SP-specific CTLs were measured by HLA-A2/IMP-3515–523 tetramer staining. Representative plots of CD8+ T cells from three independent experiments with similar results are shown. The numbers inside the plots indicate the percentage of CD8+ HLA-A2/ IMP-3515–523 tetramer+T cells. (C) Mean fluorescence intensity of HLA-A2/IMP-3515–523 tetramer-reactive CD8+ T cells. (D) A2-IMP-3-SP-specific CTLs were stimulated with SP in the presence or absence of LP, Th clone, control monoclonal Ab (mAb), and anti-PD-1 mAb. On day 7, CD8+ T cells were analyzed by staining the HLA-A2/IMP-3515–523 tetramer. Representative data are shown from four independent experiments.*p <0.05, **p <0.01. N.S., not significant. (E) Immature DCs were cultured in the presence or absence of autologous IMP-3-LP3-specific Th clones and the cognate peptide. After 48 h of co-culture, the expression of CD40 and CD86 on gated DCs was analyzed. The gray-filled histograms show isotype-matched control staining.