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. 2017 Mar 10;12(3):e0173454. doi: 10.1371/journal.pone.0173454

Fig 6. Recombinant gp41 expressed on murine tumour cells inhibits the effector functions of antigen-primed CD8+ T cells.

Fig 6

a) Analysis of gp41 expression and secretion in stably selected cTRAMP cells. Lysates of empty vector (cTRAMP CL) or gp41 plasmid transfected cTRAMP cells (cTRAMP-gp41 CL) or supernatant of cTRAMP-gp41 cells incubated with NiNTA beads were separated by SDS-PAGE and immunoblotted with the gp41-specific monoclonal antibody 2F5. Similar to transfected 293 cells, gp41 was detected in cell lysates and secreted into the supernatant. b) FACS analysis of pIRESneo vector transfected cTRAMP cells (top) and pIRESneo-gp41 vector transfected cTRAMP-gp41 cells (bottom) using the gp41-specific antibody 5F3 confirms substantial levels of surface expression. c, d) Inhibition of IFNγ and CD25 expression by CD8+ T cells from OT-1 mice co-cultured with pIRESneo vector transfected or gp41 expressing cTRAMP cells pulsed with SIINFEKL peptide. The gating strategy to retrieve viable CD8+ T cells in the mixture is shown in c). Results are representative data from two independent experiments with duplicate measurements.