Figure 3.

FACS analysis of surface expression of CXCR5 and CD40L by central memory T cells (CD4⁺CD62L⁺CCR7⁺) with negative CD27 expression in day-14 MDLN co-cultures with medium, A375, U87 and MDA-MB-231. MDLN cultures were from a representative patient. Various MDLN co-cultures were stained and analyzed by 11-color FACS. The gating of central memory T cells (CD4⁺CD62L⁺CCR7⁺) was the same as described in Figure 2. The gated central memory T cells were analyzed using FSC vs CD27-AF700 to quantify the CD4⁺CD62L⁺CCR7⁺CD27⁺ and the CD4⁺CD62L⁺CCR7⁺CD27⁻ cell subsets. The gated CD4⁺CD62L⁺CCR7⁺CD27⁻ subsets were analyzed using FSC vs CD40L-PE-Cy5 and FSC vs CXCR5-AF488. Controls were designed in the same way as described in Figure 2.