Human T lymphocyte surface antigens: partial purification and characterization utilizing a high-titer heteroantiserum

A high-titer human T lymphocyte-specific antiserum (A99) has been prepared by immunization of rabbits with a preparation partially purified from membranes of the T cell line, CEM. The specificities recognized by this antiserum included those recognized by TH1 and HTL antisera. Several experiments indicated that these antisera recognized several different proteins. A partial purification of material recognized by A99 serum was obtained by 1) isolation of plasma membranes, 2) detergent solubilization and chromatography on DEAE-cellulose, and 3) affinity chromatography on a wheat germ lectin-Sepharose 4B column. These three steps yielded material about 100-fold purified, and an estimate was made in several ways that on overall purification of 3500-fold would be required to achieve homogeneity. Immunoprecipitation of 125I-labeled 100-fold purified material showed that the major bands recognized by A99 serum had m.w. of 52,000, 96,000 (seen only after reduction), 120,000, 152,000, and 195,000. These data are compared to other published reports of human T lymphocyte-specific proteins.