Bcl-2 proto-oncogene is expressed in normal lymphoid tissues and its role has been suggested to involve the selection of B and T cells. Here we report three-color flow cytometric studies on the expression of bcl-2 protein in the thymus and periphery. Bcl-2 was expressed in the majority of T and B cells and down-regulated only in the CD1(+), CD3(-to low), CD4(+)8(+) thymocytes, and the B cells that are in cell cycle in the tonsil and lymph node. In contrast, the CD2(+), CD1(-)3(-)4(-)8(-)19(-), early T cells, and mature single positive cells in the thymus, as well as the tonsilar and lymph node T cells, irrespective of their cell cycle status, expressed bcl-2. These results are consistent with the notion that bcl-2 is down-regulated in the T and B cells that are in the process of generating new antigen receptors and subject to subsequent positive selection. (C) 1994 Academic Press, Inc.