Figure 3

Manipulation of MHC class II and LFA-1 expression does not affect DMHT HLA-DR surface expression of (A) K562 and K562/DC-SIGN cells that were transfected with the construct pWT-CIITA compared with (B) Raji and Raji/DC-SIGN cells that were transfected with the construct pDN-CIITA (300–1130). On all histograms, the gray curve represents staining with an isotype control antibody, whereas the filled black curve represents HLA-DR MAb staining. The mean fluorescence intensity is shown in the inset of the histograms. (C) Expression of DC-SIGN, HLA-DR, and LFA-1 in donor cells. Raji/DC-SIGN and K562/DC-SIGN cells that co-express either HLA-DR, LFA-1, or both were singly stained with PE-conjugated isotype control Ab or MAb against DC-SIGN, HLA-DR, or LFA-1. Antibody staining (FL2) is depicted by the histogram plots along the x axis. (D) Transmission of HIV-Luc/ADA by DC-SIGN-expressing donor cells that were manipulated for HLA-DR and LFA-1 expression. The HIV-1 capture and transmission assay was performed as described for Figure 1. Hut/CCR5 cells were used as targets and DC-SIGN-negative parental cells were used as controls. +, positive expression; -, negative. Each data set represents the mean of three separate wells of infected cells. One representative experiment out of three is shown. cps, counts per second.