Immune responses to a mouse fibroblast line, transfected with a plasmid that causes expression of the rabies virus glycoprotein under the control of a simian virus 40 early promoter, were studied. Transfected cells were shown to be recognized in vitro by a panel of monoclonal antibodies directed to conformational epitopes of the different antigenic sites of the glycoprotein. They stimulated rabies virus glycoprotein-specific T helper cells in the presence of antigen-presenting cells and were, furthermore, recognized by rabies virus-induced cytolytic T cells. In vivo, immunization of H-2-compatible mice with the transfected cell line led to a rabies virus glycoprotein-specific antibody response, and to protection against a subsequent challenge with live virus. We propose this procedure, i.e. use of cell lines transfected with plasmids expressing a viral protein under a mammalian promoter, as a simple and inexpensive method to screen individual viral proteins for their ability to elicit immune responses, including T helper cells, cytolytic T cells, antibodies, and protection against viral challenge.