T cell early precursors belong to the CD3-CD4-CD8- triple negative (TN) thymocyte population that can be subdivided on the basis of CD44, CD25, and heat-stable Ag (HSA) expression. The kinetics and precursor product relationships of these subsets, as well as of the CD4/8low intermediates, were studied by using pulse labeling with bromodeoxyuridine (BrdUrd). The highest frequencies of DNA-synthesizing cells were found in CD44+CD25+ and CD44-CD25low or CD25- subsets. The major TN cell type (CD44-CD25high), as well as CD44+ CD25-HSAlow early precursors, contained a majority of resting cells. RAG-2-/- mice contained less cells in DNA synthesis than normal mice, and CD44-CD25-/low cells were absent. In female mice transgenic for the anti-HYTCR, CD44-CD25high cells were almost all cycling, but a high percentage of resting cells was found in CD44-CD25- cells. In days following the BrdUrd pulse, there was a reduction in the number of BrdUrd+ cells in most subsets, with the exception of the labeled CD44-CD25high cells that showed a bell-shaped curve. The kinetics and cell size evolution suggest that the majority of these cells do not give rise to CD4+CD8+ cells. In RAG-2-/- cells, the block at the CD44-CD25high stage involved all cells. In TCR transgenic (Tg) mice, no block was seen at the CD44-CD25high stage, suggesting that early expression of a complete TCR receptor precludes the normal selection step. However, another block in the differentiation process was observed at the CD44-CD25- step in TCR Tg mice, suggesting an additional selection point.

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