Preparation of monoclonal anti-porcine CD3 antibodies and preliminary characterization of porcine T lymphocytes

Abstract

The CD3-T-cell receptor complex is the clonotypic surface structure by which T lymphocytes recognize foreign antigens and are subsequently activated. Because of the low immunogenicity of the CD3 molecules, anti-CD3 monoclonal antibodies (mAb) are difficult to prepare and have not been available in several species. Following isolation of porcine CD3, 14 anti-porcine CD3 mAb were prepared, which define six groups of CD3-epsilon epitopes, coprecipitate two types of TCR and reveal considerable heterogeneity of CD3 expression amongst lymphocyte subpopulations. Thus, both CD3 positive and negative subpopulations of CD2 or CD8 positive cells were found in the blood. The density of CD3 on CD2+ or CD8+ cells was relatively low and heterogeneous, whereas the CD2-, CD8- or MAC320+ T cells expressed CD3 at a higher and more homogeneous level. Finally, in the thymus, staining with anti-CD3 resolved large thymocytes into two subsets: one expressing a high level of CD3 and the other being negative. In contrast, small thymocytes expressed CD3 at a low and more homogeneous level. Immunohistological studies confirmed the presence of clearly detectable CD3 in thymus medulla and the T-cell regions of peripheral lymphoid tissues. Most of the mAb were mitogenic, when presented to peripheral blood mononuclear cells in immobilized form. The anti-CD3 mAb also induced redirected cytotoxicity which was shown to be Fc receptor dependent.