No abstract

The vertebrate immune system uses two kinds of antigen-specific receptors, the immunoglobulin molecules of B cells and the antigen receptors of T cells. T-cell receptors are formed by a combination of two different polypeptide chains, alpha and beta (refs 1-3). Three related gene families are expressed in T cells, those encoding the T-cell receptor, alpha and beta, and a third, gamma (refs 4-6), whose function is unknown. Each of these polypeptide chains can be divided into variable (V) and constant (C) regions. The V beta regions are encoded by V beta, diversity (D beta) and joining (J beta) gene segments that rearrange in the differentiating T cell to generate V beta genes. The V gamma regions are encoded by V gamma, J gamma and, possibly, D gamma gene segments. Studies of alpha complementary DNA clones suggest that alpha-polypeptides have V alpha and C alpha regions and are encoded by V alpha and J alpha gene segments and a C alpha gene. Elsewhere in this issue we demonstrate that 18 of 19 J alpha sequences examined are distinct, indicating that the J alpha gene segment repertoire is much larger than those of the immunoglobulin (4-5) or beta (14) gene families. Here we report the germline structures of one V alpha and six J alpha mouse gene segments and demonstrate that the structures of the V alpha and J alpha gene segments and the alpha-recognition sequences for DNA rearrangement are similar to those of their immunoglobulin and beta-chain counterparts. We also show that the J alpha gene-segment organization is strikingly different from that of the other immunoglobulin and rearranging T-cell gene families. Eighteen J alpha gene segments map over 60 kilobases (kb) of DNA 5' to the C alpha gene.

The T-cell specific, rearranging gamma-chain genes bear striking resemblance to T-cell receptor and immunoglobulin genes, but the role of gamma remains unknown. A central problem is to understand the conditions under which gamma RNA is expressed in cells. The transcription of gamma is abundant in T cells of fetal thymi, but is negligible in peripheral T cells of adults, suggesting that gamma is involved in development of the T-cell repertoire. However, gamma RNA was originally cloned from established lines of cytotoxic T cells (CTLs) derived from adult mice and this expression has been ascribed to non-physiological cell growth. Possibly consistent with this, most of the gamma RNA derives from genes rearranged abortively at the V gamma-J gamma junction of immunoglobulin genes, where V is the variable segment and J the joint segment. Here, we report the detailed analysis of gamma transcription in T cells of adult mice, and find that transcription may occur in T cells with a broad range of surface phenotypes; that it is predominantly of a single V gamma-C gamma unit (where C is the constant region); and that in cells freshly explanted from animals it can be of productively rearranged genes.

Northern analysis, hybridization in situ and cDNA sequence analysis have been used to demonstrate that the induction of T cell gamma-gene expression is a general occurrence when primary splenic T cells of adult mice are cultured in short-term mixed lymphocyte reactions (MLR). Splenic T cells from nine strains of mice examined in eleven different MLR all showed significant induction of gamma-RNA, even when the primary T cell response was to only a three amino acid mismatch in a major histocompatibility complex class I antigen. In MLR examined in detail, the expression is highly enriched for in CD3+ "double-negative" T cells (lacking both CD4 and CD8 expression). A cDNA sequence analysis, constituting the first such analysis of any size of gamma-gene transcripts from circulating, peripheral cells of adult mice, revealed transcription to be frequently of productively rearranged genes. These genes display extensive junctional diversity.