Human peripheral blood mononuclear cell (PBMC) proliferative responses to live respiratory syncytial (RS) virus, formalin-inactivated RS (Fl-RS) virus, RS virus F (fusion) protein, and RS virus G (attachment) protein were assessed. All donors responded to challenge with whole RS virus antigens and F and G proteins. F protein responses elicited higher levels of response than equivalent concentrations of G protein in nine out of ten adult RS-seropositive donors. Stimulation of PBMC induced low levels of interleukin 2 (IL-2), interferon gamma (IFN-gamma), IL-4, and IL-10 production. Human RS virus-specific T cell lines were generated from peripheral blood cultures following in vitro stimulation with RS virus antigens. All lines generated were shown to be MHC class II restricted. Characterisation of the lines was carried out by determining the levels of IL-2, IFN-gamma, IL-4, and IL-10 in culture supernatants. T cell lines enriched for RS virus-specific cells provided a more sensitive system than PBMC cultures for the detection of cytokines. The pattern of cytokine production varied for the individual lines, and the detection of TH1 and TH2 cytokines was dependent on the nature of the stimulating RS virus antigen. Live RS virus induced a TH1 pattern of cytokines (IL-2 and IFN-gamma), whereas FI-RS virus induced the production of both TH1 and TH2 cytokines. In addition, TH lines specific for individual RS virus proteins produced different cytokine profiles. F protein-specific lines generated TH1-type cytokines (IL-2 and IFN-gamma), whereas G protein-specific lines generated TH2-type cytokines (IL-4 and IL-10).