Phenotypic analysis and maturation potential of mRNA-electroporated DCs.

(A) Immature Mo-DCs (iMo-DCs) were transfected by electroporation with mRNA-encoding EGFP and stained with PE-labeled antibodies specific for CD1a, HLA-DR, and CD86 one day after electroporation (bottom). Untransfected iMo-DCs (top) served as controls and isotype-matched antibodies were used to set quadrants. Results are representative of 3 experiments. (B) iMo-DCs were transfected by electroporation with mRNA-encoding Melan-A and directly stained with a PE-labeled CD80 antibody (bottom) or indirectly stained with a CD83 antibody (top). A representative overlay histogram is shown in which the dashed line represents the control nonelectroporated iMo-DCs, the thin line the electroporated iMo-DCs, and the bold line the electroporated iMo-DCs that were allowed to mature for an additional 24 hours following mRNA electroporation in the presence of TNF-α and LPS. (C) Twelve-day-cultured 34-DCs were transfected by electroporation with mRNA-encoding EGFP and stained with PE-labeled antibodies specific for CD1a, HLA-DR, CD86, and CD80 one day after electroporation (bottom). Untransfected 34-DCs (top) served as controls and isotype-matched antibodies were used to set quadrants. Results are representative of 3 experiments.