Suicide selection of murine T helper clones specific for variable regions of the influenza hemagglutinin molecule

Abstract

A negative selection procedure has been developed to obtain murine T helper clones specific for variable regions of the influenza A hemagglutinin. T cell lines, established from mice primed by intranasal infection with X31 (H3N2) virus, were cross-stimulated with natural variant viruses of known primary sequence (either A/TEXAS/1/77 or A/ENG878/69) and proliferating cells eliminated by treatment with the cell cycle-specific drug 5-bromodeoxyuridine. After two suicide cycles, T cell lines were subtype specific and failed to recognize the natural variants. Clones were established by limiting dilution and their specificity was determined against a panel of viruses. Extensive diversity was evident in the reactivity of clones from individual donors, and two major T cell recognition sites were defined in the globular head region of the hemagglutinin molecule.