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Biomedical subjects

J Leceta

Publications and source records attributed to J Leceta.

At least 37 records · Page 2Linked to original sources
PubMed1999

Vasoactive intestinal peptide and pituitary adenylate cyclase-activating polypeptide inhibit endotoxin-induced TNF-alpha production by macrophages: in vitro and in vivo studies.

Vasoactive intestinal peptide (VIP) is a neuropeptide synthesized by immune cells that can modulate several immune aspects, including the function of cells involved in the inflammatory response, such as macrophages and monocytes. The production and release of cytokines by activated phagocytes are important events in the pathogenesis of ischemia-reperfusion injury. There is abundant evidence that the proinflammatory cytokine TNF-alpha is an important mediator of shock and organ failure complicating Gram-negative sepsis. VIP has been shown to attenuate the deleterious consequences of this pathologic phenomenon. In this study we have investigated the effects of VIP and the structurally related neuropeptide pituitary adenylate cyclase-activating polypeptide (PACAP38) on the production of TNF-alpha by endotoxin-activated murine peritoneal macrophages. Both neuropeptides rapidly and specifically inhibit the LPS-stimulated production of TNF-alpha, exerting their action through the binding to VPAC1 receptor and the subsequent activation of the adenylate cyclase system. VIP and PACAP regulate the production of TNF-alpha at a transcriptional level. In vitro results were correlated with an inhibition of both TNF-alpha expression and release in endotoxemic mice in vivo. The immunomodulatory role of VIP in vivo is supported by the up-regulation of VIP release in serum and peritoneal fluid by LPS and proinflammatory cytokines such as TNF-alpha, IL-1beta, and IL-6. These findings support the idea that under toxicity conditions associated with high LPS doses, VIP and PACAP could act as protective mediators that regulate the excessive release of TNF-alpha to reduce inflammation or shock.

Animals↗
PubMed1999

Vasoactive intestinal peptide (VIP) and pituitary adenylate cyclase-activation polypeptide (PACAP) protect mice from lethal endotoxemia through the inhibition of TNF-alpha and IL-6.

The neuropeptides vasoactive intestinal peptide (VIP) and pituitary adenylate cyclase-activating polypeptide (PACAP) down-regulate cytokine production. Because human septic shock involves excessive cytokine production, the effect of VIP/PACAP was investigated in a high endotoxemia murine model. Both peptides protect against endotoxin-induced lethality and prevent septic shock-associated histopathological alterations. VIP/PACAP reduce serum and peritoneal TNF-alpha and IL-6, suggesting that the protective effect is exerted by inhibiting the production of endogenous TNF-alpha/IL-6. Consistent with this mechanism, VIP does not protect against septic shock induced by exogenous TNF-alpha. The immunomodulatory role of VIP in vivo is supported by the appearance of high levels of VIP in serum and peritoneal fluid following LPS administration. Thus, the neuropeptides VIP/PACAP protect from the lethal effect of high endotoxemia, presumably by down-regulating TNF-alpha and IL-6 production, and may offer an alternative in the treatment of human septic shock syndrome.

Animals↗
PubMed1999

Regulation of VIP production and secretion by murine lymphocytes.

Vasoactive intestinal peptide (VIP) is a neuropeptide present in the lymphoid microenvironment with a multiplicity of actions. Two sources for VIP have been described in the immune system, the terminals present in central and peripheral lymphoid organs and the immune cells. Although VIP is synthesized by lymphocytes, there is no evidence demonstrating that VIP is released, and which stimuli are able to induce VIP production and secretion. In this study, we demonstrated for the first time, that agents that mediate important immune functions, such as proliferation and antigenic stimulation (Con A, LPS, and anti-TCR antibody), inflammation (LPS, TNFalpha, IL-6 and IL-1beta) or apoptosis (dexamethasone) induce the production and release of VIP to the lymphoid microenvironment. We conclude that VIP is produced and secreted by lymphocytes and propose that during an immune response, the timely release of VIP within the lymphoid organs and peritoneum should influence the differentiation and/or downregulation of the ongoing response.

Animals↗
PubMed1998

VIP and PACAP enhance IL-6 release and mRNA levels in resting peritoneal macrophages: in vitro and in vivo studies.

Vasoactive intestinal peptide (VIP), a neuropeptide produced by lymphocytes has been previously reported to modulate cytokine expression in T lymphocytes. In this study, we investigated the effects of VIP and of the structurally related neuropeptide pituitary adenylate cyclase-activating polypeptide (PACAP38) on the production of IL-6 in unstimulated murine peritoneal macrophages. Both neuropeptides stimulate rapidly, specifically, and similarly the production of IL-6, exerting their action through two different receptor/signal transduction systems, i.e., primarily through the binding to VIP1/PACAP receptor followed by adenylate cyclase activation, and partially through the activation of protein kinase C following binding to PACAP-R. VIP and PACAP38 regulate the production of IL-6 at a transcriptional level, affecting the de novo synthesis of this cytokine. The stimulatory in vitro effect correlates with the stimulation of IL-6 expression and release in vivo. These studies suggest that VIP/PACAP play a role in immune system homeostasis, participating in the intricate cytokine network and controlling local immune responses. In addition, the understanding of the factors that regulate the expression and release of IL-6 by macrophages is important for the elucidation of the role of IL-6 in health and disease.

Animals↗
PubMed1998

Vasoactive intestinal peptide and pituitary adenylate cyclase-activating polypeptide modulate endotoxin-induced IL-6 production by murine peritoneal macrophages.

Vasoactive intestinal peptide (VIP) is a neuropeptide synthesized by immune cells that can modulate several immune aspects, including the function of cells involved in the inflammatory response, such as macrophages and monocytes. Production and release of cytokines by activated mononuclear phagocytes is an important event in the pathogenesis of ischemia-reperfusion injury. VIP has been shown to attenuate the deleterious consequences of this pathologic phenomenon. We have investigated the effects of VIP and PACAP38 on the production of interleukin-6 (IL-6), a proinflammatory cytokine, by endotoxin-activated murine macrophages. Both neuropeptides exhibit a dual effect on the IL-6 production by peritoneal macrophages. Whereas VIP and PACAP inhibit with similar dose-response curves the release of IL-6 from macrophages stimulated with a LPS dose range from 100 pg/mL to 10 microg/mL, both neuropeptides enhance IL-6 secretion in unstimulated macrophages and in macrophages stimulated with very low LPS concentrations (1-10 pg/mL). The inhibition on LPS-induced IL-6 production is specific, presumably mediated through a subtype of the PACAP-R. VIP and PACAP regulate the production of IL-6 at a transcriptional level. These results were correlated with an inhibition on both IL-6 expression and release in endotoxemic mice in vivo. These findings support the idea that in the absence of stimulation or in the presence of low doses of LPS, VIP and PACAP could play a role in immune system homeostasis. However, under toxicity conditions associated with high LPS doses, VIP and PACAP could act as protective mediators that regulate the excessive release of IL-6 in order to reduce inflammation or shock.

Adenylyl Cyclases↗
PubMed1996

Vasoactive intestinal peptide and pituitary adenylate cyclase-activating polypeptides (PACAP27) and PACAP38) protect CD4+CD8+ thymocytes from glucocorticoid-induced apoptosis.

In the present study, the effects of vasoactive intestinal peptide (VIP) and the pituitary adenylate cyclase-activating polypeptides, PACAP27 and PACAP38, in a concentration range from 10(-13) to 10(-6) mol/L were studied in vitro on the spontaneous and dexamethasone (DEX)-induced apoptosis in rat thymocytes. The results show that VIP and both PACAPs inhibit significantly and in a similar way the DNA fragmentation characteristic of glucocorticoid-induced apoptosis and increase the cell survival of thymocytes, with a maximal effect observed at 10(-8) to 10(-9) mol/L. This study showed the ability of the VIP-receptor (VIP-R) antagonist [N-Ac-Tyr1,D-Phe2]-GRF(1-29) amide to partially reverse the inhibitory effect of VIP and both PACAPs on DEX-induced apoptosis, providing evidence for a specific VIP1-R-mediated response and supporting the involvement of a single receptor for the three neuropeptides. Phenotypic analysis showed that VIP, PACAP27, and PACAP38 protect predominantly CD4+CD8+ thymocytes from glucocorticoid-induced apoptosis. These findings suggest that these neuropeptides could be involved in intrathymic T-cell maturation.

Animals↗
PubMed1996

Characterization of gene expression of VIP and VIP1-receptor in rat peritoneal lymphocytes and macrophages.

In the present report we show the gene expression pattern of VIP and VIP1 receptor in two peritoneal cell populations, macrophages and lymphocytes by reverse transcription (RT) and polymerase chain reaction (PCR). Only in the lymphoid cells we have obtained a specific VIP cDNA product of 458 bp identical in size to the one obtained from cerebral cortex. On the other hand, we have obtained in both peritoneal populations lymphocytes and macrophages, a specific VIP1 receptor cDNA product of 311 bp identical in size to that obtained from lung. These results have been confirmed by Southern blot hybridization. Our findings suggest an autocrine/paracrine action of VIP in peritoneal microenvironment, supporting an immunoregulatory role for this neuropeptide.

Animals↗
PubMed1996

Differential VIP and VIP1 receptor gene expression in rat thymocyte subsets.

Nervous, endocrine, and immune systems share a large number of regulatory molecules including hormones and neuropeptides. Vasoactive intestinal peptide (VIP) plays an important role in a variety of immunological functions. In the present report, we sorted purified thymocytes of the four major thymic subsets defined by CD4 and CD8 phenotypes. We demonstrate by reverse transcription (RT) and polymerase chain reaction (PCR) both VIP and VIP1 receptor gene expression in double positive (CD4+CD8+) and single positive (CD4+CD8-, CD4-CD8+) thymocyte subsets. Double negative thymocytes (CD4-CD8-) lack VIP and VIP1 receptor gene expression.

Animals↗
PubMed1996

Expression of vasoactive intestinal peptide in lymphocytes: a possible endogenous role in the regulation of the immune system.

Experimental evidence is accumulating showing that vasoactive intestinal peptide (VIP) acts as an immunoregulatory peptide. Findings from our laboratory and others indicate that cells of the immune system are able to produce VIP. We have detected immunoreactivity for VIP in lymphocytes by immunohistochemical methods at specific locations of both central and peripheral lymphoid organs. Double immunofluorescence staining and flow cytometry analysis indicate that both T and B lymphocytes contain VIP that has been proved to be mostly VIP 1-28 by high-performance liquid chromatography and radioimmunoassay. VIP has been also demonstrated by 'in situ' hybridization and reverse transcription followed by polymerase chain reaction. We have also detected induction of VIP in splenic lymphocytes after mitogenic stimulation. Lymphocytes should be sensitive to the endogenously produced VIP because we have also detected VIP receptor expression in different populations of lymphocytes. All this evidence indicates that VIP is an endogenous autocrine modulator of immune function.

Animals↗
PubMed1994

Gene expression of VIP receptor in rat lymphocytes.

In the present report we analyze by reverse transcription (RT) and polymerase chain reaction (PCR) the expression of vasoactive intestinal peptide receptors (VIP-R) in rat thymocytes and peripheral T and B lymphocytes. We have obtained a specific VIP-R cDNA product of 311 bp identical in size to that obtained from lung. These results have been confirmed by Southern hybridization analysis. VIP receptor gene expression in central and peripheral lymphoid organs support the notion that VIP is a immunomodulatory peptide.

Adjuvants, Immunologic↗
PubMed1994

Vasoactive intestinal peptide (VIP) mRNA expression in rat T and B lymphocytes.

Different evidence suggests that VIP has immunoregulatory functions and may be secreted by different cells involved in inflammatory and immune responses. In the present study, we demonstrate by reverse transcription (RT) and polymerase chain reaction (PCR) VIP gene expression in rat thymocytes and T and B cells derived from spleen and lymph nodes. We have obtained a specific VIP cDNA product of 458 bp identical in size to that obtained from cerebral cortex. These results have been confirmed by Southern blot analysis. VIP message has also been detected in the T-T hybridoma YH-1633 and in a non-immune cell line, the pheochromocytoma PC 12. VIP gene expression in central and peripheral lymphoid organs suggests that VIP may be a T and B cell-derived cytokine involved in T-cell differentiation and in cell immune responses.

Animals↗
PubMed1994

Lymphoid cell subpopulations containing vasoactive intestinal peptide in the rat.

In the present study we describe the cell types containing immunoreactive vasoactive intestinal peptide (IR-VIP) in rat thymus, spleen, and lymph nodes. Indirect immunofluorescence staining and flow cytometry indicated that all lymphoid organs studied contained VIP-positive cells, with the spleen and lymph nodes having a higher proportion than the thymus. Vasoactive intestinal peptide was found in both lymphocytes and nonlymphoid cells, lymphocytes predominating among VIP-positive cells. Double immunofluorescent staining and flow cytometry showed that all lymphoid subpopulations identified contained variable proportions of VIP-positive lymphocytes. Immunocytochemical staining of cell suspensions for both light and electron microscopy showed the cytoplasmic localization of the IR-VIP. These findings, coupled to our previous results, are consistent with the idea that VIP may have a lymphoid origin and could be active in local immune responses.

Animals↗
PubMed1994

Vasoactive intestinal peptide modulation of adherence and mobility in rat peritoneal lymphocytes and macrophages.

In this work, the effects of vasoactive intestinal peptide (VIP) in a concentration range from 10(-13) to 10(-7) M were studied in vitro on two common activities of peritoneal rat lymphocytes and macrophages: adherence and mobility (spontaneous and chemotaxis). The results show that VIP stimulated the adherence of the two cells studied, and increased the macrophage mobility but decreased this activity in lymphocytes. Moreover, a specific protein kinase C (PKC) activator such as phorbol myristate acetate (PMA, 50 ng/ml) also stimulated significantly the adherence and chemotaxis of both macrophages and lymphocytes. By contrast, a PKC inhibitor, retinal (2 x 10(-5) M), decreased significantly these capacities. Macrophages incubated with both VIP and PMA in relation to those incubated with VIP or PMA showed an increase in adherence and chemotaxis, whereas in lymphocytes adherence was also increased but chemotaxis decreased. The incubation with forskolin (10(-5) M), an enhancer of intracellular cAMP levels, produced an inhibitory effect of the chemotaxis activity in both types of cells. VIP prevented this inhibitory effect of forskolin in macrophages but not in lymphocytes. In addition, VIP was chemoattractant for macrophages but not for lymphocytes. The present study proves that VIP proves that VIP has a coronary effect on the two principal and representative types of immune cells in the rat peritoneum: lymphocytes and macrophages, stimulating macrophage chemotaxis through PKC activation and inhibiting lymphocyte chemotaxis through adenylate cyclase activation.

Animals↗
PubMed1993

VIP gene expression in rat thymus and spleen.

Vasoactive intestinal peptide (VIP) is a neuropeptide with immunomodulatory properties. In the present study, we demonstrate VIP gene expression in cells of both thymus and spleen in the rat by in situ hybridization. In thymus sections, hybridization signal for VIP mRNA was found in cells in corticomedullary and medulla regions. In the spleen, cells were labeled at the outer area on the periarteriolar lymphoid sheath of the white pulp. Hybridization signal appeared to be in lymphoid cells. These findings suggest that lymphoid cells might produce VIP, which, if released, could exert a paracrine action on central and peripheral lymphoid organs. We suggest that VIP participates in the bidirectional communication between the nervous and the immune systems.

Animals↗
PubMed1989

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

Changes 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.

Animals↗
PubMed1989

Different sensitivity to the dexamethasone treatment of the lymphoid organs of Rana perezi in two different seasons.

Adult female frogs (Rana perezi) have been used to analyze structural and morphometrical changes which affect lymphoid organs after treatment with a single dose of dexamethasone (DX). Special attention was given to the distinct sensitivity observed during two different periods of the year. Morphological differences were found between adult control frogs sacrificed in November and February. In February, frogs showed degenerated thymic reticular epithelial cells but lesser numbers of cortical pycnotic and mitotic thymocytes than in November. Splenic lymphoid tissue was clearly less well developed in February. No significant variations were found in the morphometrical parameters of jugular bodies whereas the number of lymphocytes of both peripheral blood and bone marrow was significantly higher in February than in November. In contrast, frogs sacrificed in February exhibited less drastic variations as well as a more delayed response to DX-treatment than those of November. The reported differential changes could be attributed to possible distinct levels of circulating sexual and nonsexual steroids during both seasonal periods.

Animals↗
PubMed1988

Effects of neonatal treatment with estrogens on the development of the thymus in rats.

In order to study the effects of sex hormones administered during the neonatal period on the thymic development we have injected 5 days old female rats with a single dose (0.1mg) of estradiol benzoate. The evolution of the thymus gland after treatment was morphometrically analyzed. The thymus of the estrogen-injected animals diminished in size between the 7th and the 15th day, increasing afterwards. The cortex was the most sensitive compartment in the thymus to estrogenic treatment. Both thymic involution and enlargement were associated to changes in the frequency of large lymphoid cells in the subcapsular region and variations in the thymic cortex weights. The observed effects are tentatively attributed to alterations in the thymocyte differentiation probably due to modifications in the secretion of thymic hormones.

Animals↗