Large cortical thymocytes from C57BL/6-Tla(a) mice have been prepared rapidly and in high yield by a combination of centrifugal elutriation and differential binding to peanut agglutinin (PNA)-coated plates. The cells in these lymphoblast-rich fractions were clearly distinct from the majority of thymocytes, with up to 70 percent in the S or G(2) + M phases of the cell cycle and an average rate of [(35)S]methionine incorporation per cell up to 20 times higher than that of the majority population.
The populations of cells resolved in this fractionation were characterized by monitoring their rates of synthesis of specific glycoproteins, thymus- leukemia antigen (TL) and the Lyt-2, Lyt-3 complex (Lyt-2/3), relative to their total protein synthesis. Cells that bound to PNA synthesized high levels of Lyt-2/3, consistent with their identification as cortical thymocytes. Those that failed to bind made little or no Lyt-2/3, as expected for medullary cells, The fraction of dividing lymphoblasts that bound to PNA was enriched in cortical thymocyte precursors, including all the large cells detectably active in synthesizing Lyt-2. It differed sharply from the small cortical cells, however, in the synthesis of TL. Although both populations displayed abundant surface TL, the TL glycoprotein was produced actively in fractions containing dividing cells but made at a drastically reduced rate by the nondividing majority of cortical thymocytes. Thus, TL seems to be made at a narrowly circumscribed stage of early thymocyte development that is correlated with rapid proliferation. In most of the descendants of such blast cells, the TL glycoprotein is presumably retained on the cell surface as long as no substantial membrane turnover takes place. Ongoing TL synthesis may therefore serve as a marker for a unique developmental state which terminates rapidly in normal differentiation but may be extended by agents that give rise to TL(+) thymic lymphomas.