Mouse malignant T-lymphoma CS-21 cells undergo apoptotic cell death in vitro in the absence of lymph node stromal cells but escape apoptosis and proliferate when they are attached to CA-12 stromal cells. A monoclonal antibody raised against CS-21 cell surface molecules (MCS-5) recognized a M(r) 168,000 protein, inhibited binding of CS-21 cells to CA-12 stromal cells, and suppressed apoptosis in CS-21 cells. To identify the M(r) 168,000 protein, we purified it with MCS-5 affinity chromatography and ion exchange chromatography. Partial amino acid sequences of the purified M(r) 168,000 protein were identical to those of CD45, a transmembrane tyrosine phosphatase. The purified protein possessed tyrosine phosphatase activity and was recognized by an anti-CD45 monoclonal antibody. The M(r) 168,000 protein was identified as CD45. To determine the CD45 isoform, we cloned the CD45 gene from the cDNA library of CS-21. Sixteen of 18 clones encoded CD45RO (CD45 lacking exons 4, 5, and 6), and the remainder lacked exons 4, 5, 6, and 7. Like MCS-5, an anti-CD45 monoclonal antibody, also inhibited binding of CS-21 cells to CA-12 cells and suppressed apoptosis in CS-21 cells. Our present results indicate that CD45RO expressed on CS-21 cells mediates adhesion to CA-12 cells and suppression of apoptosis.