A monoclonal antibody, TcR2, has been shown to recognize an avian homologue of the mammalian alpha/beta T cell receptor (TcR). The TcR2-reactive molecule was found to be a T3-associated heterodimer with relative molecular mass of 90-kDa consisting of disulfide-linked 50-kDa and 40-kDa polypeptides. The sizes of the deglycosylated TcR2 polypeptides differed from those of TcR1, an avian homologue of the mammalian gamma/delta T cell receptor. Immunofluorescence analysis revealed that TcR1 and TcR2 are expressed on separate populations of T cells during their development first in the thymus and then in the periphery. Ontogenetic studies revealed that the TcR1+ thymocytes are generated first and the generation of TcR2+ cells begins approximately 3 days later. While most TcR2+ cells in the thymus expressed both CT4 and CT8, TcR2+ cells in blood and the spleen were either CT4+ or CT8+. The TcR1+ cells in blood and thymus were CT4-CT8-, but the majority of TcR1+ cells in the spleen surprisingly expressed the CT8 marker. The data suggest that TcR1 and TcR2 cells are generated in the thymus as separate T cell sublineages.