Recombinant vaccinia viruses were constructed that expressed the complete env gene of feline immunodeficiency virus with or without the nucleotide sequence encoding the cleavage site between the surface (SU) protein and the transmembrane (TM) protein. The removal of this cleavage site resulted in the expression of a 150K protein that was processed into a 130K protein and was not cleaved into the SU and the TM proteins. Removal of the cleavage site also facilitated incorporation of the SU protein in immune-stimulating complexes (iscoms). Antibody responses to both an SU and a TM peptide representing two immunodominant B cell epitopes were measured. These were higher in cats immunized with iscoms prepared from the cleavage site-deleted envelope protein than in cats immunized with iscoms prepared from the native envelope protein or immunized with the envelope protein and the adjuvant Quil A.