ALT-803 stimulates proliferation and activation of human NK cells and T cells in vitro. ALT-803 stimulates proliferation and activation of human NK cells.

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ALT-803 stimulates proliferation and activation of human NK cells and T cells in vitro. ALT-803 stimulates proliferation and activation of human NK cells and T cells in vitro. A, human PBMCs (n = 7 normal donors) were cultured for 7 days in media alone or media containing 0.5 nmol/L ALT-803 or IL15. At the end of the incubation period, changes in absolute cell counts were determined following staining with fluorochrome-conjugated antibodies against CD8a, CD4, CD335, CD19, and CD4/CD25/FOXP3. Bars represent mean ± SE. *, P < 0.05; **, P < 0.01; ***, P < 0.001 comparing ALT-803 vs. media control. B, ALT-803 ranging from 0.07 to 5.5 nmol/L was added to human PBMC cultures (n = 2 donors). After 7 days, changes in CD8+ T-cell counts were assessed. Bars represent mean ± SE. *, P < 0.05; **, P < 0.01 comparing ALT-803 (open bars) vs. media control (closed bar). C, representative normal donor PBMCs were cultured for 5 days in the indicated concentrations of ALT-803. Cells were then harvested, and expression of the cell surface activation markers CD25 and CD69 was determined on CD4+ T cells, CD8+ T cells, and NK cells by flow cytometry. The symbols represent mean fluorescence intensity (MFI) ± SE of triplicates. Similar treatment-related changes were obtained with other human donor PBMCs though control levels of CD25 and CD69 varied between individuals. D, human PBMCs (n = 2 donors) were cultured for 5 days in the indicated concentrations of ALT-803. Following stimulation, cells were harvested and analyzed for intracellular perforin (open squares) and granzyme B (closed triangles) expression in NK cells and CD8+ T cells. The symbols represent the MFI ± SE. *, P < 0.05; **, P < 0.01; ***, P < 0.001 comparing ALT-803 vs. media control. In each case, the results shown are representative of at least two independent experiments. Peter R. Rhode et al. Cancer Immunol Res 2016;4:49-60 ©2016 by American Association for Cancer Research