HCV and HIV infections impair dendritic cell function. We evaluated the impact of HCV, HIV, and HCV-HIV infection on MDC-NK interactions by analyzing CD3 depleted PBMC for NK cell IFN-gamma in response to IL-12 or poly (I:C). Purified MDC and NK cells were analyzed for TLR ligand-dependent, MDC-dependent NK activity. In HIV infection, IFN-gamma production by CD3 depleted PBMC was reduced in response to poly (I:C), while response to IL-12 was intact in HCV and HIV infections. Poly (I:C) induced activity was dependent on MDC and partially dependent on IL-12, consistent with accessory cell help. In purified MDC-NK co-cultures, MDC dependent NK IFN-gamma and Granzyme B was intact in both HCV and HIV infections, while MDC numerical defects were observed in HIV infection. These data indicate that during viral infection with HIV, accessory cell dependent NK function in the periphery is impaired. This impairment may be related to the identified MDC numerical defect.