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T lymphocytes are produced in the thymus from precursors originating in the haemopoietic tissues. On entering the thymus, they undergo a programme of proliferation, T-cell receptor (TCR) gene rearrangement, differentiation and repertoire selection. Although the thymus provides a unique environment for these events, the role of the thymic stroma in regulating specific developmental stages is not well understood. We therefore devised an in vitro system to study the role of individual thymic stromal components in T-cell development. We report here that the development of TCR-CD4-CD8-T-cell precursors into TCR+ cells expressing CD4 and/or CD8 requires the presence of both major histocompatibility complex class II+ epithelial cells and fetal mesenchyme. The requirement for mesenchymal support can be mapped to the initial stages of intrathymic development because the later stages of maturation, from double-positive CD4+CD8+ thymocytes into single-positive CD4+ or CD8+ cells, can be supported by epithelial cells alone. We also show that the requirement for mesenchymal cells can be met by cells of the fibroblast line 3T3 (but not by supernatants from these cells). To our knowledge, these findings provide the first direct evidence that mesenchymal as well as epithelial cells are involved in T-cell development, and suggest that their involvement is stage-specific and likely to be dependent on short-range or contact-mediated interactions.

MHC class II-positive epithelium and mesenchyme cells are both required for T-cell development in the thymus