Panel A shows histograms. The horizontal axis is labeled Cell Trace and the vertical axis is labeled Modal. Panel B shows histograms. The horizontal axis is labeled Cell Trace and the vertical axis is labeled Modal. Panel C shows scatter plots representing the percentages of TCR V delta 2 cells within T cells at day 0 and 3.5 in the presence or absence of HMBPP. The horizontal axis is labeled Days after expansion and the vertical axis is labeled percent of V delta 2 positive within CD3 positive. Panel D shows flow cytometry plots of CD39, ICOS, CD137, and CD69 expression on TCR V delta 2 cells from P1 and a healthy control cultured in the presence or absence of HMBPP. The horizontal axis is labeled Cell Trace and the vertical axis is labeled with the respective markers. Panels E, F, G, and H show line graphs representing the activation of TCR V delta 2 cell lines derived from three healthy controls versus P1 challenged with THP-1 cells exposed to increasing doses of HMBPP. The horizontal axis is labeled HMBPP concentration in nanomolar and the vertical axis is labeled with the respective cytokines interferon gamma, tumor necrosis factor alpha, interleukin 4, and interleukin 13 in nanograms per milliliter.
P1-derived TCR Vδ2 cells expand more efficiently in the presence of HMBPP. (A) Proliferation of PBMC-derived TCR Vδ2 cells after 3.5 days in the presence (red) or absence (black) of a suboptimal dose of HMBPP (0.4 nM). Representative cell trace histograms represent samples from HC105, HC106, HC107, and P1. (B) Proliferation of PBMC-derived TCR Vδ2 cells after 3.5 days in the presence of 1 µg/ml of phytohemagglutinin. Representative cell trace histograms are shown for samples from HC105, HC106, HC107 (black), and P1 (red). (C) Percentages of TCR Vδ2 cells within T cells at days 0 and 3.5 in the presence or absence of HMBPP (0.4 nM). The data correspond to one experiment. (D) Representative flow cytometry plots of CD39, ICOS, CD137, and CD69 expression on TCR Vδ2 cells from P1 and HC105 expanded in the presence (red) or absence (black) of HMBPP (0.4 nM). (E–H) Activation of TCR Vδ2 cell lines derived from HC105, HC106, HC107 (black), and P1 (red) challenged with THP-1 cells exposed to increasing doses of HMBPP. Dose–response curves of (E) IFN-γ, (F) TNF-α, (G) IL-4, and (H) IL-13 release mean ± SD of triplicate independent cultures. Representative plot of one of two experiments.