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. 2016 Oct 28;5(12):e1250991. doi: 10.1080/2162402X.2016.1250991

Figure 2.

Figure 2.

Intra-melanoma DP T cells mediate memory B-cell proliferation through CD40L engagement. Intra-melanoma SP CD4+, DP and SP CD8+ CD40L− T-cell lines were co-cultured with CFSE-labeled B cells for 4 d at a 1:1 ratio on anti-CD3-coated microwells, and B-cell proliferation was evaluated by flow cytometry gated on CD19+ cells. (A) Representative CFSE dilution profiles of total (CD19+), naive (CD19+CD27−) and memory (CD19+CD27+) B-cell fractions after co-cultured with CD3-activated SP CD4+, DP and SP CD8+ CD40L− T cells. B cell proliferation was expressed as the percentage of CFSE dilution. Results of total (B), naive or memory (C) B-cell proliferation stimulated with each T-cell subpopulations derived from the three patients M125, M265 and M305 are gathered on a single graph (n = 3 independent experiments with three different B-cell donors). (D) Memory B-cell proliferation stimulated with DP T cells derived from M125, M265 and M305 patients in the presence of either recombinant human CD40 Fcγ1 (black bars), control IgG1 Fc (gray bars) or without treatment (white bars). Results are normalized to the non-treated control condition (n = 3 independent experiments with three different B-cell donors).