We report the isolation of bcl-x, a bcl-2-related gene that can function as a bcl-2-independent regulator of programmed cell death (apoptosis). Alternative splicing results in two distinct bcl-x mRNAs. The protein product of the larger mRNA, bcl-xL, is similar in size and predicted structure to Bcl-2. When stably transfected into an IL-3-dependent cell line, bcl-xL inhibits cell death upon growth factor withdrawal at least as well as bcl-2. Surprisingly, the second mRNA species, bcl-xS, encodes a protein that inhibits the ability of bcl-2 to enhance the survival of growth factor-deprived cells. In vivo, bcl-xS mRNA is expressed at high levels in cells that undergo a high rate of turnover, such as developing lymphocytes. In contrast, bcl-xL is found in tissues containing long-lived postmitotic cells, such as adult brain. Together these data suggest that bcl-x plays an important role in both positive and negative regulation of programmed cell death.

CD2 is a 50-kDa transmembrane glycoprotein that plays an important role in T and natural killer (NT) lymphocyte functions. CD2 serves as both an adhesion molecule and as a costimulatory molecule through interactions with its ligand, CD58, on antigen presenting or target cells. Consistent with earlier studies using a rat anti-CD2 mAb, we have shown that treatment of alloantigen stimulated T lymphocytes with a humanized mAb, MEDI-507 (IgG1, kappa), induced hyporesponsiveness to subsequent stimulation with alloantigen but not to mitogen (phytohemagglutinin). Fluorescence-activated cell sorting analysis of cells from mixed lymphocyte reaction (MLR) treated with MEDI-507 revealed pronounced deletion of T and NK cells, consistent with lack of proliferation in the MLR. MEDI-507 F(ab')2 fragments did not have inhibitory activity or induce deletion of lymphocytes in the MLR. Removal of the NK cell subset by magnetic bead depletion using anti-CD16 and anti-CD56 mAbs eliminated both the T cell deletion and the inhibitory effect. Reconstitution of NK depleted responder populations using autologous NK cells restored the MEDI-507-mediated deletion activity to levels measured in the original MLR. Formaldehyde-fixed NK cells failed to mediate the MEDI-507-induced deletion effect. Altogether, our studies indicate that activated T cells with MEDI-507 bound to CD2 are preferential targets for autologous NK cells through a nonapoptotic cytotoxic mechanism.

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(6,7). After rearrangement, the VH and VL segments undergo sequence diversification by intrachromosomal gene conversion using families of V pseudogene segments as sequence donors (6-10). Given these differences between mammalian and avian Ig gene diversification, it was of interest to determine the molecular mechanisms for the diversification of chicken We have examined the molecular mechanisms for the generation of diversity in the chicken TCRE locus by identifying the germ-line chicken Do gene segment and by analyzing rearranged chicken TCRJ3 genes isolated from the developing thymus and mature spleen.t We observe that chicken TCR,8 diversity is generated by junctional variability, rather than by gene conversion, as in chicken Ig genes. Further, the junctional diversity increases during development due to increased random base-pair (N-nucleotide) addition. Within the junctional variability, however, most rearranged chicken TCR,8 genes have at least one glycine residue, which is encoded in the germ-line by an evolutionarily conserved Do segment. The implications of a conserved glycine for TCR structure and antigen recognition are discussed.

The DNA sequence motif ATTTGCAT (octamer) or its inverse complement has been identified as an evolutionarily conserved element in the promoter region of immunoglobulin genes. Two major DNA-binding proteins that bind in a sequence-specific manner to the octamer DNA sequence have been identified in mammalian species-a ubiq-

Cortical thymocytes that express a TCR on their surface continue to express mRNA for the recombination activating genes RAG-1 and RAG-2. As the expression of these genes appears to be necessary and sufficient for V(D)J recombination to occur, this finding suggests that productive TCR gene rearrangement may not by itself terminate TCR recombination. To further study this question, we have utilized a polymerase chain reaction-based strategy to isolate and sequence excision products of secondary TCR alpha gene rearrangement events. Our data shows that TCR alpha excision products from TCR+ thymocytes contain in-frame V-J joints and thus support the concept that V(D)J recombinatorial activity may continue even in thymocytes which have undergone productive TCR gene rearrangement.