Intracellular IFN-γ expression in natural killer cells precedes lung CD8+ T cell recruitment during respiratory syncytial virus infection

Natural killer (NK) cells are recruited locally during the initial phases of virus infection and produce cytokines which may affect the subsequent emergence of specific T cells. In this study, cellular responses to primary respiratory syncytial virus (RSV) infection and after vaccination with individual viral proteins were investigated in BALB/c mice using the new NK cell antibody, DX5, Purified DX5(+) cells caused lysis of YAC-1 cell targets. DX5(+) cells did not express CD8, CD45R or MHC class II antigens, A small proportion of DX5(+) cells coexpressed CD4 (10.3%) and CD3 (10.6%). Of the DX5(+)/CD4(+) cells, the majority expressed the alpha/beta T cell receptor and less than 1 % expressed the gamma/delta T cell receptor. During infection with RSV, lung DX5(+)/CD3(-) Nh cells peaked on day 4 of primary infection and were the most numerous subset producing IFN-gamma, as determined by intracellular staining, at this time-point. Less than 1% of the DX5(+) cells secreting IFN-gamma were CD4(+). In the lungs of mice vaccinated with recombinant vaccinia virus expressing individual RSV proteins, increased NK cell cytotoxicity and IFN-gamma production correlated with increased numbers of CD8(+) T cells. Mice with few NK cells subsequently had low CD8(+) T cells and developed lung eosinophilia, IFN-gamma-producing Nh cells therefore form a substantial component of the early cellular response to virus infection with important potential influences on the subsequent development of specific immunity.