Figure 4

MPLA-tDCs display reduced capability to stimulate allogeneic CD4+ T cell proliferation and IFNγ production. MPLA-tDCs and tDCs show decreased allostimulatory capacity compared to MPLA-matured DCs and similar to iDCs. (A) Alloproliferative response induced by MPLA-tDCs and tDCs was assessed by co-culturing with allogeneic CD4+ T cells stained with CFSE at a 1:2 DC/T cell ratio for 6 days, and proliferation was determined by CFSE dilution through flow cytometry. CD4+ T cells incubated alone or treated with anti-CD3 (OKT3 clone) were used as controls. Representative flow cytometry plots (left) and graphic analysis of DC allostimulatory capacity for all conditions expressed as percentage of CD4+ CFSElow T cells from 7 independent experiments (right) are shown. (B) Production of IFNγ by allogeneic CD4+ T cells was measured on day 6 of culture through intracellular staining after stimulation of cells with PMA/ionomycin for 5 hours and analyzed by flow cytometry. Representative flow cytometry plots (left) and graphic analysis of percentage of IFNγ producing CD4+ CFSElow T cells from 4 independent experiments (right) are shown. Horizontal lines represent median values (*** P < 0.001).