A monoclonal antibody (mAb) generated against sheep T-cell blasts, called I/35 A, blocks sheep autologous E rosetting and competes with purified T11 target structure (TS), the sheep form of LFA3, for binding sites on the sheep T-cell surface. Immunoprecipitation from lysates of surface iodinated sheep T cells identifies the cell surface molecule recognized by mAb I/35 A as a single chain polypeptide migrating as a diffuse band of MW 55,000. From its binding properties and the biochemical nature of the target antigen, we conclude that mAb I/35 A is directed at sheep CD2. This finding makes sheep the first animal model in which the CD2-LFA3 (T11TS) system is defined by mAbs to both receptor and ligand. When analysed by two-colour flow cytometry and by immunohistochemistry, the cellular expression of CD2 in sheep differs significantly to that reported in humans. In peripheral blood, CD2 is found exclusively on CD4+8- and CD4-8+ T cells, while the third, CD4-8- (predominantly SBU-T19+) subset of sheep T cells (around 20% in peripheral blood) is CD2-. In thymus, only low to moderate levels of CD2 expression occurs on 80% of cells. Among these, medullary 'single positive' thymocytes express the highest level of CD2, whereas the CD4-8- 'double negative' population (which in contrast to peripheral CD4-8- T cells contains only very few SBU-T19+ cells) consists of CD2- and weakly positive cells. In peripheral lymphoid organs, CD2+ lymphocytes occur in the T-cell regions of spleen, lymph nodes and jejunal Peyer's patches (JPP). Tissue macrophages found in B-cell follicles of lymph nodes and JPP are also CD2+. The implications of these findings are discussed in terms of the role CD2 plays in the proliferation of immature thymocytes and of the possible importance of CD2/LFA3 interactions in lymphocyte recirculation.