Figure 4.

Low γδ T-cell CXCR4 expression is not due to serum-free expansion. Peripheral blood mononuclear cell samples from four individual healthy donors (n = 4 biological replicates) were selected for γδ T cells expanded in serum- or serum-free media. The following parameters were determined: (A) number of γδ T cells (live cell counts multiplied by percent γδ T cells derived from flow, CD3⁺ γδTCR⁺), (B) fold expansion (number γδ T cells divided by number on day 0), and (C) the percentage of γδ T cells by flow cytometry gated on γδTCR⁺ CD3⁺ cells. (D) CXCR4 mean fluorescence intensity (MFI) of γδ T cells expanded in serum- or serum-free media from three healthy peripheral blood mononuclear cells (n = 3 biological replicates) calculated in FlowJo software. Statistics analyzed by paired t-test (p > 0.05 = ns). ns, not significant. (E) Histogram of CXCR4⁺ γδ T cells in unstained γδ T cells, Nalm6 cells as a positive control, peripheral blood mononuclear cells, and four different zoledronate-expanded γδ T cells (n = 4 biological replicates).