Mice infected with an aerosol of influenza virus or immunized with purified UVinactivated whole virus or viral subunits developed delayed-type hypersensitivity (DTH) to influenza virus. The level of DTH was greatly enhanced when mice were injected intraperitoneally 2 days prior to immunization with 200 mg/kg of cyclophosphamide. The reaction peaked at 24 h after elicitation, had the classical DTH histology, and was transferable by immune cells but not by immune serum. DTH induced in cyclophosphamide-pretreated mice persisted for at least 40 days after sensitization. DTH induced by deoxycholate-treated subviral particles (DC particles) or the matrix protein of the influenza virus was type-specific, i.e. DTH induced by D C particles or matrix protein of type A virus cross-reacted only with type A virus and not with type B virus. DTH induced by purified hemagglutinin (HA) subunits, on the other hand, showed subtype specificty. Thus, DTH induced by H A of X31 virus (H3) did not cross-react with PR8 virus (HO). However, DTH induced by HA of one subtype cross-reacted significantly with the variants of the same subtype. Thus, it appears that the effector T cells of DTH do not discriminate between the antigenic variants of influenza virus that are distinguishable by serology. DTH induced by purified, UVinactivated whole virus showed extensive cross-reactivity. This nonspecific reaction was shown to be due to host-derived material, the lipid bilayer. -The theoretical and practical implications of these findings are discussed.