M. Torroba

Time‐course expression of Toll‐like receptors 2 and 4 in inflammatory bowel disease and homeostatic effect of VIP

Toll‐like receptor 2 (TLR2) and ‐4 mediate signals from a great variety of bacterial gut products, giving the host a panel of microbe‐recognizing receptors. Under homeostatic conditions, TLRs act as protective receptors of the intestinal epithelium. When homeostasis is disrupted in diseases such as inflammatory bowel disease, TLR2 and ‐4 are deregulated. Our study demonstrates, by using a trinitrobenzene sulfonic acid‐induced colitis model of Crohn’s disease, the constitutive expression and the up‐regulation of TLR2 and ‐4 at messenger and protein levels in colon extracts, as well as in macrophages, dendritic cells, and lymphocytes from mesenteric lymphoid nodes. Vasoactive intestinal peptide (VIP) treatment induced a decrease of TLR2 and ‐4 expressions approaching ethanol control levels. Our results suggest that VIP modulation of TLR2 and ‐4 could be explained by two possible mechanisms. The first one would be the secondary reduction of TLR2 and ‐4 caused by the VIP‐mediated decrease of inflammatory mediators such as interleukin‐1β and interferon‐ γ, which synergize with bacterial products, contributing to the amplification of TLR presence in the intestine. The other possible mechanism would involve a VIP‐mediated decrease of nuclear factor‐κB, which would cause a direct down‐regulation of TLR expression. In summary, the resultant physiological effect is the decrease of TLR2 and ‐4 expressions to homeostatic levels. Our study describes for the first time the role of a peptide present in the gut microenvironment as an effective modulator of the initial steps of acute inflammation, acting at local and systemic levels and leading to the restoration of the homeostasis lost after an established inflammatory/autoimmune disease.

VIP and Tolerance Induction in Autoimmunity

Abstract:  Vasoactive intestinal peptide (VIP) is a potent anti‐inflammatory agent with immunoregulatory properties, skewing the immune response to a Th2 pattern of cytokine production. Here, we studied the effect of treatment with VIP in the development of diabetes in nonobese diabetic (NOD) mice, an animal model of type 1 diabetes. Mice treated with VIP from 4 weeks of age did not develop diabetes and showed milder insulitis than nontreated mice. The protective mechanism of VIP was associated with a reduction in the circulating levels of Th1 cytokines. In the pancreas of VIP‐treated animals, regulatory T cell markers predominate, as indicated by the upregulation of FoxP3 and transforming growth factor‐β (TGF‐β), and the downregulation of the transcription factor, T‐bet. These findings indicate that VIP restores tolerance to pancreatic islets by promoting the local differentiation and function of regulatory T cells.

Effect of VIP on TLR2 and TLR4 Expression in Lymph Node Immune Cells During TNBS-Induced Colitis

Abstract:  Toll‐like receptors (TLRs) are a family of pattern recognition receptors (PRRs), which recognize numerous molecules collectively named pathogen‐associated molecular patterns, with an essential role in inflammatory conditions and connecting innate and acquired immune responses. Moreover, a new function of TLRs in the intestinal mucosa has been described. Under homeostatic conditions, TLRs act to protect the intestinal epithelium; but when homeostasis is disrupted, TLRs appear deregulated. Disruption of intestinal homeostasis occurs in disorders, such as Crohns disease (CD). Trinitrobenzene sulfonic acid (TNBS)‐induced colitis is a murine model of human CD and vasoactive intestinal polypeptide (VIP) exerts a beneficial effect, by decreasing both inflammatory and autoimmune components of the disease. Recently, we have demonstrated the constitutive expression of TLR2 and TLR4 at mRNA and protein levels in colon extracts and their upregulation in TNBS‐treated mice as well as the effect of VIP treatment, approaching control levels. However, the systemic effect is little known. The present results demonstrate a beneficial role of VIP, restoring homeostatic conditions through the regulation of both lymphoid cell traffic and TLR2/4 expression on macrophages (MØ), dendritic cells (DCs), and CD4 and CD8 T lymphocytes.

Ultrastructural changes in the thymus of the turtle Mauremys caspica in relation to the seasonal cycle

SummaryChanges in the ultrastructure of the thymus of the turtle Mauremys caspica, with special reference to its non-lymphoid components, were studied in relation to the seasonal cycle. The thymic cortex contains framework-forming epithelial-reticular cells and free macrophages, while the medulla includes, in addition, mature and presumptive pro-interdigitating cells. The ultrastructural features of these cells are generally similar to those described for non-lymphoid components of the mammalian thymus. The turtle thymus undergoes cortical involution in spring, with recovery periods in May–June and during autumn. A moderate involution occurs in winter. At the beginning of spring, cortical (but not medullary) epithelial-reticular cells show degenerative changes, probably related to high levels of circulating testosterone. In spring and autumn, mature interdigitating cells are absent, but macrophages, monocytes, and pro-interdigitating cells are found. During May–June, the cortical epithelial-reticular population recovers and macrophages, monocytes, and interdigitating cells are actively phagocytic. In summer, the epithelial-reticular cells in both cortex and medulla display normal ultrastructural features; mature and immature interdigitating cells are absent and some macrophages are detected occasionally. The results suggest that non-lymphoid components of the reptilian thymus can play a role in governing T-lymphocyte differentiation, and that the thymic cortex and medulla exhibit different cycles of seasonal activity.

Changes in the thymus and spleen of the turtle Mauremys caspica after testosterone injection: a morphometric study.

To confirm a possible role of sex hormones in governing the seasonal variations affecting the reptilian lymphoid organs, a morphometric analysis was carried out on the thymus and spleen of turtles, Mauremys caspica, intraperitoneally injected with a single dose of testosterone propionate (TP) at the third week of June when physiological levels of testosterone are low. At 4 and 6 weeks, control turtles show an apparent lymphocyte mobilization both from thymus and spleen with a decrease in the percentage of thymic cortex, numbers of cortical lymphocytes, and mitotic index, but increased numbers of medullary lymphocytes. In the thymic cortex of treated turtles, there is a decrease in the same parameters but they occur in the first 2 weeks, whereas the medullary lymphocytes also undergo reduction at 4 and 6 weeks. In addition, the number of reticuloepithelial cells per area unit decreases at 2 weeks in the thymic cortex of treated turtles. These results are discussed from the point of view of a biphasic effect of testosterone on turtle lymphoid organs: In the first weeks, the changes observed could be attributed to the high levels of testosterone; after 4 and 6 weeks, variations are dependent both on long-term effects of testosterone and on those induced by the high values of corticosterone occurring in summer.

Morphological, histochemical, and ultrastructural characterization of the accessory cells of neuromasts in the salamander Pleurodeles waltlii

SummaryThe ultrastructural and histochemical features of the accessory cells of the neuromast of the salamander P. waltlii have been examined. Three types of accessory cells, supporting, mantle, and basal, were found, but only the first 2 are considered in this article. Supporting cells characterized by a highly dilated endoplasmic reticulum occur among and surrounding sensory cells. Mantle cells, morphologically different from the supporting cells, surround the remainder of the neuromast. Both types of accessory cells exhibit histochemically different secretory materials. Our morphological results suggest that both accessory cells contribute to the formation of cupular material.

Macrophages and epithelial cells of the thymus gland: an ultrastructural study in the natterjack, Bufo calamita

In the present study, the ultrastructure of the stromal components, basically epithelial elements and macrophages, of the thymus of adult natterjacks, Bufo calamita has been analyzed. A network of stellate epithelial-reticular cells joined together by desmosomes, constitutes the main component of the thymic parenchyma in both cortex and medulla. In the medulla pale, electron-lucent epithelial cells, sometimes showing surface interdigitations, are striking elements. Moreover, uni- and multicellular epithelial cysts appear in the thymic medulla as well as granulated cells of possible endocrine significance. Remarkably, isolated or grouped gland cells whose morphology and cytoplasmic content resemble that of the skin glands, were occasionally found. Finally, macrophages, multinucleated giant cells and dendritic-like cells, the latter intimately associated to lymphocytes, occur in the thymus of Bufo calamita. The most remarkable morphologic characteristics of all those non-lymphoid cell types, as well as their possible functional significance are comparatively discussed with available information on the amphibian and higher vertebrate thymic cytoarchitecture.

Fine structure and histochemistry of the ampullary organ of the urodele amphibian Pleurodeles.

A morphological study by light and electron microscopy on the lateral line system of the urodele amphibian Pleurodeles waltii demonstrates the presence of sensory organs other than neuromasts in the head. From their morphology, they have been called ampullary organs. The ampullary organs occur in the bottom of a groove and consist of three different types of cells: sensory, supporting and mantle cells. Histochemical analysis indicates that the last two are secretory cells, probably involved in the production of the material filling the ampulla and the groove.