Preparation and characterization of a new monoclonal antibody CAF7 specific for a leukocyte activation antigen

A new murine hybridoma, CAF7, is raised using as an antigen the T-leukemic cell line CEM. It produces a monoclonal antibody (mAb) specific for an activation antigen on human lymphocytes. CAF7 stains monocytes, very weakly resting lymphocytes, and granulocytes, part of the thrombocytes, but not erythrocytes. After activation with phytohemagglutinin (PHA), CAF7 expression on peripheral lymphocytes rises as early as 5 hr post stimulation and 72-hr blasts express it at a considerable level. The cell lines Jurkat, CEM, MOLT4, Raji, Reh, K562, and MONOMAC6 are positive for CAF7. CAF7 immunoprecipitated an antigen, which when subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) under nonreducing conditions migrated as a single band with a molecular weight of 120-140 kD. Under reducing conditions, it appears as two bands at 90-100 and 40 kD. The pattern of expression and the biochemical characteristics of CAF7 do not match any of the clusters defined in the Leukocyte Typing Workshops, but do resemble that of a previously described antigen, 4F2.