Article Figures & Data
Figures
- FIG. 1.
Screening of Rv2903c-derived peptides for binding to HLA-B*3501. RMA-S B*3501 cells were pulsed with 50 μM unlabeled competitor peptide and 1 μM FL-labeled reference peptide. Results are expressed as percent inhibition of the maximum signal of the FL-labeled reference peptide. The vertical dotted line identifies peptides that inhibited the maximum signal by >75%. Positive controls were peptides HIV-1SF2 Nef72–80 (aa 72 to 80) (4T6R), FPVTPRVPL, and EBV TEGU (aa 1974 to 1982), HPLTNNLPL. Peptide Flu-A NP (aa 265 to 273), ILRGSVAHK, was used as a negative control.
- FIG. 2.
HLA-B*3501-restricted cytokine production by activated CD8 T cells in response to mycobacterial peptides. Short-term cultures of PBMC stimulated with peptide Rv2903c (aa 201 to 209) were cocultivated for 6 h with RMA-S B*3501 cells pulsed with a control peptide, HIV-1SF2 Nef72–80 (4T6R) (panels a and c), or the specific peptide (panels b and d). Cytokine secretion was blocked by brefeldin A. Surface markers CD8 and CD69 and intracellular IFN-γ (lower panels) and TNF-α (upper panels) were stained following fixation and permeabilization of cells. Dot plots show log fluorescence intensity. CD8+ bright T cells were gated in the fluorescence-3 channel versus side scatter light plots. Quadrant markers for CD69, IFN-γ, and TNF-α were set using the proper isotype control reagents. Frequencies of positive cells are given as a percentage of the gated CD8+ T cells. Abbreviations: PE, phycoerythrin; FITC, fluorescein isothiocyanate.
