Skip to main content
. 2014 Jul 23;5:344. doi: 10.3389/fimmu.2014.00344

Figure 7.

Figure 7

Influenza M1 cross-presentation to CD8+ αβT cells by day 14 γδT cells. (A) Day 14 γδT cells from HLA-A2+ healthy individuals or melanoma patients were treated overnight with influenza M1 protein at concentrations as indicated, washed, and mixed in equal numbers with a HLA-A2-restricted, p58–66-specific CD8+ αβT cell line. In control experiments, day 14 γδT cells were directly pulsed with 0.1 μM M1 peptide p58–66, washed, and then used in the APC assay. After 6 h of incubation, intracellular IFNγ production in M1 p58–66-tetramer + responder cells was determined by flow cytometry. Responder cells cultured in the absence of stimulation or in the presence of PMA and ionomycin served as controls for background and maximal IFNγ responses. (B) Compilation of intracellular IFNγ responses obtained with day 14 γδT cells from PBMC of five HLA-A2+ healthy individuals and four HLA-A2+ melanoma patients. (C) Day 14 γδT cells from HLA-A2+ healthy individuals were treated with 1 μM M1 protein (M1) or not (No Ag) as described in (A) and then mixed with donor matched, CFSE-labeled PBMC at a γδT cell to PBMC ratio of 1:5. Proliferation responses in p58–66-tereamer + CD8 + αβT cells were determined essentially as described in Figure 6. One representative of two independent experiments is shown.