Molecular and antigenic heterogeneity of the rat leukocyte‐common antigen from thymocytes and T and B lymphocytes

Abstract

The molecular forms and antigenic heterogeneity of the leukocyte‐common antigen (L‐CA) of rat lymphocytes have been analyzed. Thymocytes show one main band at 180 kDa, T cells four bands at 180,190, 200 and 220 kDa and B cells one broadband at about 240 kDa. T helper and T cytotoxic cell subsets show the same four bands with some differences in the proportion of each. Four mouse monoclonal antibodies (MRC OX‐1, 28, 29 and 30) reacted with all molecular forms of L‐CA and fell into two sets that were noncompetitive in binding to L‐CA (MRC OX‐1, 28, 29 vs. OX‐30). The antigenic determinants seen by all these antibodies were lost when L‐CA was reduced and alkylated. Three antibodies (MRC OX‐22, 31 and 32) reacted selectively with B cells, T cytotoxic cells and about 2/3 of T helper cells. OX‐22 and OX‐31 competed for binding but were noncompetitive with OX‐32. All these antibodies bound to a subfraction of the 190, 200 and 220‐kDa forms of T cell L‐CA but not at all to the 180‐kDa form of T cells or thymocytes. One antibody bound to B cells only (MRC OX‐33) and precipitated a subfraction of B cell L‐CA. With all the antibodies that did not label thymocytes the antigenic determinants survived reduction and alkylation. Subsequent proteolysis with trypsin then destroyed all determinants except the one reacting with MRC OX‐22 antibody, In this case tryptic peptides retained full antigenic activity which was, however, destroyed by further proteolysis with pronase.