HSV-2 infection induces membrane phenotypic changes in macaque DCs. Medium-versus HSV-2–treated (5 pfu per cell or 1 pfu per cell [C, lower row]) immature moDCs were sampled after 4 hours (A-B) or ON reculture and stained for ICP8 combined with Abs against different membrane markers (HLA-DR, CD40, CD80, CD83, CD86) before being assayed by flow cytometry. (A) Representative FACS plots of ICP8 (x-axes) versus HLA-DR (top row) or CD86 (bottom row) from one macaque experiment are shown. Numbers in each plot represent the mean fluorescence intensity (MFI) of the HLA-DR or CD86 expressed by the medium-treated versus HSV-2–infected DCs (gated on the total population). All IgG controls exhibited less than 1 log of staining. (B) MFIs of HLA-DR and CD86 are shown from 2 representative experiments (of 4; animal L534, ▪; animal T602, ▴) for which the complete 4 hours versus ON recultures were performed. MFIs from L534 are derived from panel A. (C) Extended phenotypic analysis of ON-recultured macaque cells exposed to 5 pfu per cell (top row) or 1 pfu per cell (bottom row) was performed and the MFIs of the additional markers calculated. Summarized are the MFIs of HLA-DR, CD40, CD80, CD83, and CD86 expressed by medium-treated cells versus the ICP8^– and ICP8⁺ gated subsets within the HSV-2–infected cells (top row: MFI means ± SEM for 3 to 7 animals, except for the CD40 expression by medium-treated cells, which was performed on the cells from 1 donor; bottom row: MFI means ± SEM from 3 animals). The ICP8⁺ and ICP8^– subsets within the HSV-2–infected cells were defined based on the gate set on medium-cultured DCs stained with the anti-ICP8 Ab. (D) Immature moDCs were exposed to live (HSV, macaque and human) or UV-treated (UV, macaque) HSV-2 (5 pfu per cell or the equivalent) or not (Med, macaque and human) and incubated overnight. The mean (± SEM) MFIs of the indicated markers on the total DC populations are shown for 4 macaque (•) and 5 to 6 human (▵) experiments.