The thymus develops from a paired epithelial anlage in the neck. This review considers how ectoderm (vesicula cervicalis) and endoderm (third pharyngeal pouch) contribute to the epithelial stroma of the thymus. Stromal elements of mesodermal origin are capillaries, septae and perivascular spaces and single invading cells. These elements separate the thymus into pseudolobuli. The thymus epithelial space and the perivascular spaces are always separated from each other by a closed, flat epithelial cell layer, with a basal lamina which contributes to the blood-thymus barrier. From the 9th gestational week, prethymic precursor cells from hemopoietic centers, begin to invade the thymus anlage. There they finally mature to committed post-thymic T cells. The thymus microenvironment of postnatal thymus is composed of six different types of epithelial cells and several stromal cells of mesodermal origin. The location of these diverse stationary cells is described, and their functional significance is discussed. Obviously these stromal cell types have a special function in providing the proper environment for T-cell maturation. The function of the thymus includes the maturation and/or selection of antigen specific T-cells. The main issue of intra-thymic T-cell differentiation is the development and expression of T-cell-antigen receptors. The great diversity of these receptors is generated by a rearrangement of the T-cell-receptor-genes in order to furnish the host with a mature T-cell repertoire that is capable of recognizing the world of extrinsic antigens. In a synopsis the manyfold interrelationships between the thymus microenvironment and the developing thymocytes are summarised.

Sympathetic nerves were visualized in sections from rat thymus by immunostaining of tyrosine hydroxylase, the rate‐limiting enzyme of catecholamine biosynthesis, and by glyoxylic acid‐induced fluorescence of catecholamines. Catecholaminergic nerve fibres were detected in close connection to thymic epithelial cells which therefore might be preferred target cells. To evaluate this, rat immunocytochemically defined, cultured thymic epithelial cells were investigated for adrenoceptors and adrenergic effects. In rat cultured thymic epithelial cells mRNA for β1‐ and β2‐adrenoceptors was detected by reverse transcription‐polymerase chain reaction by use of sequence‐specific primers. Specific, saturable binding to the cultivated cells was observed with the β‐adrenoceptor agonist CGP 12177. Adrenaline, noradrenaline or the β‐adrenoceptor agonist, isoprenaline, increased intracellular adenosine 3′: 5′‐cyclic monophosphate (cyclic AMP) levels in cultivated thymic epithelial cells dose‐dependently about 25 fold. The pharmacological properties revealed that this response was mediated by receptors of the β1‐ and the β2‐subtypes. The selective β3‐adrenoceptor agonist BRL 37344 had no effect on cyclic AMP levels. The increase in cyclic AMP was downregulated by preincubation with glucocorticoids like dexamethasone or cortisol which also changed the relative importance of β1‐/β2‐adrenoceptors to the response. Incubation with isoprenaline or the adenylate cyclase activator forskolin decreased basal and serum‐stimulated proliferation of thymic epithelial cells. However, adrenergic stimulation of thymic epithelial cells did not induce interleukin 1 production. Since thymic epithelial cells create a microenvironment which influences the maturation and differentiation of thymocytes to T‐lymphocytes, their observed capacity to respond to catecholamines provides novel evidence for the suggestion that adrenergic stimulation may interfere with the regulation of immune functions. British Journal of Pharmacology (1997) 120, 1401–1408; doi:

A new method for the long-term culture of pure rat thymic epithelial cells was established. The cultures were characterized by immunocytochemistry, electron microscopy and proliferation assays. Non-epithelial thymic cells were eliminated with a reliable and reproducible pre-plating method, by differential trypsin treatment of the cultures and by addition of horse serum to the culture medium instead of fetal calf serum. The final cultures contained more than 95% pure epithelial cells as evidenced by immunostaining for cytokeratin. Ultrastructural studies indicated that these cells are physiologically active epithelial cells with tonofilaments, desmosomes and filopods. The subsets of the thymic epithelial cells in vitro were investigated by comparing their staining pattern with that obtained in situ using several subtype-selective antibodies. Thymic epithelial cells in vitro showed a preferential expression of subcapsular/perivascular and medullary markers. Only few cultivated cells were of cortical origin. In the first to the fourth subcultures, some cells were immunopositive for the thymus hormone/factor thymulin. The proliferation of thymic epithelial cells was stimulated by horse serum and to a lesser extend by fetal calf serum. The adenylate cyclase activators isoproterenol and forskolin, and the glucocorticoid cortisol inhibited the proliferation.

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Normal T cell development depends upon interactions between progenitor cells and the thymic microenvironment. Monoclonal antibodies (Mabs) have been used to define subtypes of thymic epithelium by light microscopy (clusters of thymic epithelial staining [CTES]). We have now used a range of these Mabs together with gold‐coupled reagents in immuno‐electron microscopy to study the fine cellular distribution of the molecules to which the antibodies bind. Anti‐cytokeratin antibodies were used to identify all thymic epithelial cells, while the distribution of MHC class II molecules was revealed with Mabs to shared nonpolymorphic determinants. MR6, a CTES III Mab, shows strong surface labelling of cortical epithelial cells and thymic nurse cells and very weak surface staining of thymocytes, medullary macrophages, and interdigitating cells. Mab 8.18 (CTES V) also labels a cell surface molecule; this is present on Hassall's corpuscles and associated medullary epithelial cells. The molecules detected by Mabs MR6 and 8.18 are therefore located in a position where they are available to interact with external cellular and soluble signals within the thymus. In contrast, Mabs MR10 and MR19 (CTES II) recognise intracellular molecules within subcapsular, perivascular, and medullary epithelium. A similar distribution was seen with Mab 4β, directed against the thymic hormone thymulin, although, in addition to the expected intracellular epithelial staining, large lymphoblasts in the subcapsular zone showed surface positivity, indicating the presence of thymulin bound to surface receptors on these early lymphoid cells. As expected, MHC class II molecules were expressed on some medullary and essentially all cortical epithelial cells. However, although most subcapsular epithelium was class II–negative, some cells did express these MHC molecules on their apical surface and on the surface of their cytoplasmic extensions into the cortex. Interestingly, some cortical epithelial cells surrounding capillaries were positive for both MR6 (CTES III) and for MR10, MR19, and 4β (CTES II). Double‐labelling experiments, using MR6 and MR19 simultaneously, revealed a double‐positive perivascular epithelial cell population in the thymic cortex. The possibility that these cells represent a thymic epithelial progenitor population is discussed. Microsc. Res. Tech. 38:237–249, 1997. © 1997 Wiley‐Liss, Inc.