Figure 2

Induction of CD4⁺CD25⁺Foxp3⁺IFNγ⁺and CD4⁺CD25⁺CD127^-IFNγ⁺PBL and cell proliferation in the presence of PMA/Ionomycin and recombinant proteins against cell surface molecules . (a, b) PBL of 5 healthy control individuals (HC1-HC5) were incubated in medium or stimulated with PMA/Ionomycin for 16 h in the presence of recombinant rCD178, rCD152, rCD279, rCD28, and rCD95 in final concentrations of 0.1 μg/ml and 1 μg/ml. Then, assay was measured using four-color fluorescence flow cytometry. Proportions of CD4⁺CD25⁺Foxp3⁺IFNγ⁺ and CD4⁺CD25⁺CD127^-IFNγ⁺ PBL were calculated as proportion of all cultured CD4⁺ PBL. rCD28 reduced induction of CD4⁺CD25⁺Foxp3⁺IFNγ⁺ PBL (p < 0.05). Higher concentrations of recombinant proteins tend to increase iTreg induction, especially of rCD152 and rCD95 (p < 0.05). In the presence of rCD95, CD4⁺CD25⁺CD127^-IFNγ⁺ PBL increased strongly (with vs without rCD95: p < 0.05). (c) PBL of 5 healthy control individuals (HC1-HC5) were incubated in medium or stimulated with PHA for 3 days in the presence of recombinant rCD178, rCD152, rCD279, rCD28, and rCD95 in final concentrations of 0.1 μg/ml and 1 μg/ml. Then, proportion of CFSE^low PBL was determined using flow cytometry. rCD178 and rCD95 reduced cell proliferation strongly (p < 0.05). Data are given as mean±SD.