Prevention of OVA-IC–mediated OT-II (CD4) and OT-I (CD8) activation by IVIg. Equal numbers of bone marrow–derived dendritic cells (BMDCs) and OT-II (top panels) or OT-I (bottom panels) cells were incubated for 3 days in the presence of 2.5 μg/mL of OVA-IC prepared as described previously,1 with or without 10 mg/mL of IVIg (Gamunex). The background activation was established in the absence of OVA-IC (left panels). After incubation, the cells were recovered, washed and labeled with either anti–CD4-FITC or anti–CD8-PE and anti–CD69-APC (eBioscience). T-cell activation was determined by measuring the expression of CD69 on CD4- (top panels) or CD8- (bottom panels) gated cells by flow cytometry (Accuri C6 flow cytometer). The data were analyzed using the FCS Express 4.0 software (De Novo) and are representative of 4 independent experiments done with different lots of BMDCs.
