Figure 4.
Figure 4. Expression of ΔLNGFR does not confer an apparent antiapoptotic effect in LV'VFas+ T cells in vitro. (A) Unmodified and LV'VFas+ T cells are equally susceptible to Fas-mediated apoptosis. T cells were transduced to express the LV'VFas transgene and enriched by immunomagnetic selection for ΔLNGFR expression (▪) or left untransduced (□). Aliquots of T-cell cultures either unmodified or LV'VFas+ were exposed to cyFasL (4-32 ng/mL) or medium alone. After 24 hours, cells were stained with 7-AAD for 15 minutes on ice and examined by flow cytometry. Survival was assessed by both forward/side scatter and staining with 7-AAD. Data are shown for a representative experiment with macaque T cells. (B) Expression of ΔLNGFR by LV'VFas+ T cells does not interfere with the sensitivity to AP1903-induced cell death. Aliquots of unmodified (□) or LV'VFas+ T cells (▪) were exposed to 10 nM AP1903 in the absence or presence of various concentrations of NGF (0-1000 ng/mL). Cells were washed, and the survival of LV'VFas+ T cells as compared to the untreated cells was evaluated 24 hours later by trypan blue exclusion. Data are shown as the mean (± SD) of 3 independent experiments.

Expression of ΔLNGFR does not confer an apparent antiapoptotic effect in LV'VFas+ T cells in vitro. (A) Unmodified and LV'VFas+ T cells are equally susceptible to Fas-mediated apoptosis. T cells were transduced to express the LV'VFas transgene and enriched by immunomagnetic selection for ΔLNGFR expression (▪) or left untransduced (□). Aliquots of T-cell cultures either unmodified or LV'VFas+ were exposed to cyFasL (4-32 ng/mL) or medium alone. After 24 hours, cells were stained with 7-AAD for 15 minutes on ice and examined by flow cytometry. Survival was assessed by both forward/side scatter and staining with 7-AAD. Data are shown for a representative experiment with macaque T cells. (B) Expression of ΔLNGFR by LV'VFas+ T cells does not interfere with the sensitivity to AP1903-induced cell death. Aliquots of unmodified (□) or LV'VFas+ T cells (▪) were exposed to 10 nM AP1903 in the absence or presence of various concentrations of NGF (0-1000 ng/mL). Cells were washed, and the survival of LV'VFas+ T cells as compared to the untreated cells was evaluated 24 hours later by trypan blue exclusion. Data are shown as the mean (± SD) of 3 independent experiments.

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