同种T细胞识别机制与潜在应用价值.PPTVIP

Figure 2. The dimer-expanded CD8lowCD28- T cells exhibit suppressive effects on alloreactive T cells. (A) A negative correlation between CD8lowCD28- and IFN-γ-expressing T cells was observed using curve estimation and linear regression analyses in the alloreactive coculture of HLA-2-ve PBLs and T2/Tyr with (dimer treated) and without (non-dimer treated) the Tyr/HLA-A2 dimer. Correlation coefficients and p values are shown for each analysis (n=15). (B) The dimer-expanded CD8lowCD28- T cells showed suppressive effects on alloreactive T cells. The T2/Tyr-primed CD8high and the Tyr/HLA-A2 dimer-expanded CD8lowCD28- T cells were sorted by FACS from non-dimer treated and the dimer treated alloreactive cocultures, respectively. The CD8high T cells (1 × 105 cells/well) were mixed with the CD8lowCD28- T cells at the indicated ratios, and restimulated with the T2/Tyr for 24 h. IFN-γ ELISPOT assays were performed in duplicate and mean values ± SEM from 4 independent experiments are shown (*** for p0.001 and ** for p0.01 by LSD t-test, compared with ELISPOT results of the CD8high). (C) CD8lowCD28- T cells from the non-dimer treated coculture were unable to suppress alloreactive T cells. CD8high and CD8lowCD28- T cells were sorted from non-dimer treated cocultural bulks by the FACS, mixed at ratio of 1:1, then restimulated with T2/Tyr. IFN-γ positive cells were tested for the T-cell alloreactivity, and mean values ± SEM from 4 independent experiments are shown (*** for p0.001 by LSD t-test, compared with ELISPOT results of the CD8high). * Figure 3. The CD8lowCD28- T cells exhibit alloantigen-specific suppression which requires cell-cell contact. Two HLA-A2 restricted peptides, Tyr and HBC, were used to pulse T2 cells and HLA-A2 dimer. Hyperresponsive T2/Tyr- or T2/HBC-primed CD8high T cells were mixed with suppressive Tyr/HLA-A2 dimer- or HBC/HLA-A2 dimer-expanded CD8lowCD28- T cells, and restimulated by T2/Tyr, T2/HBC or E006 (Table Ⅰ). ELISPOT assays for IFN-γ production in