DELINEATION OF HUMAN THYMOCYTE DIFFERENTIATION PATHWAYS UTILIZING DOUBLE-STAINING TECHNIQUES WITH MONOCLONAL-ANTIBODIES

Abstract

Three new techniques were introduced for the phenotypic analysis of human thymocytes and T cells. Monoclonal [mouse] antibodies of different subclasses (i.e., IgG1 and IgG2) were used in various combinations and labeled with subclass-specific 2nd layers conjugated to different fluorochromes. Tissue sections were studied with combinations of monoclonal and conventional reagents; anti-mouse Ig-coated macrobead columns were used to separate antibody-tagged cells. With the help of these simple techniques, 4 stages of thymocyte differentiation (prothymocyte, cortical, intermediate and medullary cell types) can be described in detail and the microanatomical relation of T cell subsets with other cell types studied.