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A Varas

Publications and source records attributed to A Varas.

36 records · Page 2Linked to original sources

Macrophage-lymphocyte cell clusters in the hypothalamic ventricle of some elasmobranch fish: ultrastructural analysis and possible functional significance.

BACKGROUND: Previous studies have demonstrated the existence of lympho-haemopoietic tissue in the meninges and choroid plexuses of various primitive vertebrates, including the stingray Dasyatis akajei and in early human embryos. In the present study, we extend these results analyzing macrophage-lymphocyte cell clusters found in the floor of the hypothalamic ventricle of several specimens of elasmobranchs. METHODS: After aseptical isolation of the brain from several specimens of smooth dogfish Triakis scyllia, cloudy dogfish Scyliorhinus torazame, gummy shark Mustelus manazo, and stingray Dasyatis akajei their hypothalamic regions were processed routinely by light, scanning, and transmission electron microscopy. RESULTS: The study of serial histological sections demonstrated that the macrophage-lymphocyte cell clusters proceeded from the meningeal lymphohaemopoietic tissue, reaching the ventricular lumen along large blood vessels. In this tissue, macrophages, different sized lymphocytes, lymphoblasts, granulocytes, monocytes, and developing and mature plasma cells were closely packed among a meshwork of fibroblastic reticular cell processes. It never invaded the brain parenchyma. A cell layer of glial elements and a continuous basement membrane interposed between the lymphoid tissue and the neural elements although some macrophages had migrated across the ependymal cell layer. In the ventricular lumen very irregular macrophages with long cell processes and containing abundant engulfed material of unknown origin formed big cell clusters with neighboring lymphocytes, lymphoblasts, and plasma cells, similar to those described during the immune response. Moreover, electron lucent cells which resembled the antigen-presenting cells of higher vertebrates established intimate surface cell contacts with the surrounding lymphocytes. In the third ventricle of several specimens of gummy shark, Mustelus manazo, morphologically similar cell clusters appeared but these were not connected to the meningeal lympho-haemopoietic tissue. No intraventricular cell aggregates were found in the stingray brain. CONCLUSIONS: Although we cannot rule out that these macrophage-lymphocyte cell clusters represent a permanent structure in the elasmobranch brain they rather seem to be only established after specific stimulation for preventing the entrance of noxious, foreign materials into the elasmobranch brain parenchyma.

Animals

Gamma/delta T cells in fetal, neonatal, and adult rat lymphoid organs.

In the present study, we have analyzed the appearance and maturation of gamma/delta T cells, recognized with a new mAb V65, in the central and peripheral lymphoid organs of fetal, neonatal, and adult Wistar rats. Cytofluorometrical analysis demonstrated the first V65+ gamma/delta T cells in the thymus of 16-17-day embryonic rats, although by immunohistology, they were identified only in 19-day rat embryos in both the cortico-medullary border and thymic medulla. Phenotypically, gamma/delta thymocytes from fetal and neonatal thymus expressed CD3, CD2, and CD5, but only 60-80% were CD8+ and approximately 40-50% expressed the alpha chain (p55) of the IL-2R. In the periphery, the immunohistological study identified for the first time gamma/delta T cells in the splenic white pulp and the gut of 21-day fetal rats, where they occurred within the epithelium as well as in the lamina propria. After birth, gamma/delta lymphocytes appeared in the skin, where they were present as dendritic epidermal T cells in increasing numbers during postnatal life. Whereas these gamma/delta T cells formed the predominant T-cell population in the rat skin, gamma/delta T cells in peripheral lymphoid organs, BALT, or the gut only represented a minor T-cell population. These results are discussed in comparison to gamma/delta T cells of other vertebrate species.

Animals

T-cell development in early partially decapitated chicken embryos.

We have evaluated the immunohistological and cytofluorometric changes that occur in the thymus of chicken embryos partially decapitated at 33-38 hr of incubation (DCx embryos) in an attempt to analyze possible neuroendocrinological influences on T-cell differentiation and, indirectly, the ontogeny of the so-called neuroendocrine-immune network. The thymus of DCx embryos shows important variations that profoundly and selectively affect different T-cell subsets, but not the nonlymphoid cell components of thymic stroma. These modifications include the accumulation of cell precursors, mainly DN (CD4-CD8-) cells and immature CD8lowCD4- cells, which expand but do not differentiate, resulting in an extreme decline of both DP (CD4+ CD8+) cells and TcR alpha beta-expressing cells. Accordingly, both subcapsulary and outer cortex increase in size, whereas the deep cortex and principally the thymic medulla almost disappear in DCx embryos. In contrast, other T-cell subsets of DCx embryos, largely CD8highCD4- cells and TcR gamma delta-expressing cells do not undergo significant variations throughout thymic ontogeny.

Animals

The relevance of cell microenvironments for the appearance of lympho-haemopoietic tissues in primitive vertebrates.

In higher vertebrates, mainly in mammals, a role for the non-lymphoid components of lymphoid organs in governing the maturation and functioning of immune system has been largely demonstrated. In contrast, such a role in the evolution of the vertebrate immune system has only been evidenced indirectly. In the present review we summarize histophysiological results which emphasize the relevance of lympho-haemopoietic stromal elements in the emergence and evolution of vertebrate lymphoid organs. The most primitive vertebrates, the Agnatha, have no true lymphoid organs and, accordingly, their immune responses seem more related to the non-anticipatory defence mechanisms of invertebrates than to the immune responses of vertebrates. So, the appearance and evolution of vertebrate lymphoid organs seems closely related with the emergence of immune capacities. Thymus, spleen and gut-associated lymphoid organs appear early in phylogeny whereas lymph nodes and bone marrow are late phylogenetical adquisitions. However, bone marrowless vertebrates contain numerous organs (i.e., gonads, kidney, brain, etc...), the cell microenvironments of which support lympho-haemopoiesis mimicking the condition of higher vertebrate bone marrow. On the other hand, the lack of germinal centres, another feature of the lymphoid organs of ectothermic vertebrates which impedes the selection of B cells raised after somatic hypermutation, presumably reflects the absence of some of the elements necessary for this organization.

Animals

Ontogeny of rat thymic macrophages. Phenotypic characterization and possible relationships between different cell subsets.

In the present study we combined electron microscopy, immunohistology and primary stromal cell cultures to analyse the ontogeny of rat thymic macrophages (M phi) in an attempt to clarify the relationships between the different macrophage cell subsets described in adult rat thymus. Although phagocytic cells were observed in 15-day-old fetal thymus, monoclonal antibodies (mAb) which recognize different adult macrophage types were unable to identify positive cells until the end of embryonic life. However, our in vitro results from primary thymic stromal cell cultures of 16-day-old fetal rats, and the phenotyping of enriched thymic CD2- cell suspensions, demonstrated that monocyte-like cells which strongly expressed major histocompatibility complex (MHC) class II molecules colonized the embryonic thymus early, giving rise later to distinct macrophage subsets. During the process of maturation, macrophage precursors gradually lost their MHC class II expression, acquired other surface markers (CD45, Thy-1, CD25, CD4, etc.) and increased the acid phosphatase activity. In this respect, ED1+ macrophages, which appeared for the first time in the last stages of embryonic life, consisted of a MHC class II molecule-expressing phagocytic cell population, presumably involved in the elimination of non-selected cortical thymocytes, and of non-phagocytic cells which, in the thymic cortex, might differentiate to ED2+ macrophages throughout ED1+ED2lo/med and ED1+ ED2high intermediate cell stages, observed in vitro in 16-day-old fetal thymic stromal cell cultures. At the end of embryonic life and during the postnatal period the numbers of thymic macrophages increased, particularly in the medulla and corticomedullary border (CMZ), and more slowly in the thymic cortex. This increase was presumably due to the arrival, through perivascular spaces, of new macrophage progenitors, rather than in situ proliferation of pre-existent mature macrophages. The possible function of different thymic macrophage subsets, as well as the relationships between themselves and with their presumptive monocyte-like precursors, are discussed.

Animals

Ontogeny of rat thymic dendritic cells.

In the present study we have combined various in vivo and in vitro approaches to analyse the appearance and development throughout ontogeny and postnatal life of the dendritic cell (DC) populations of rat thymus. The in situ ultrastructural study demonstrated immature interdigitating cells (IDC)/DC in the thymus of 17-day-old embryonic rats, but thymic stromal cell cultures from 16-day-old fetal rats seemed to contain DC precursors which, after several days in culture, produced strongly class II major histocompatibility complex (MHC)-positive, mature DC. According to morphology and class II MHC expression we also defined three different DC populations in the late embryonic rat thymus; two of them, which remained in the adult rat thymus, could represent distinct developmental stages within the IDC/DC lineage. The third cell subset might be involved in a massive process of negative selection, presumably occurring at the end of fetal life in the rat thymus. In supporting the existence of thymic DC subpopulations, we also demonstrated a differential expression of various cell markers, including CD4, CD8, CD25, adhesion molecules and the antigen recognized by OX44 monoclonal antibody (mAb), on thymic DC during both embryonic and adult life. Their possible significance for the attributed functions to thymic DC are discussed extensively.

Animals

Activity of stem cell factor and IL-7 in combination on normal bone marrow B lineage cells.

The production of B cells is regulated by soluble and cell contact signals presumably provided by bone marrow stromal cells. Among these is IL-7, a well characterized proliferative stimulus for a subset of pre-B cells. Stem cell factor (SCF), a stromal cell-derived cytokine with broad hemopoietic effects, has been reported to synergize with IL-7 to drive the proliferation and differentiation of B220- bone marrow cells into B220+ B cell precursors in long term culture. A subsequent report has cast doubt on this result by showing that SCF and IL-7 were incapable of producing mu+ pre-B cells after short term culture. Here, using the cell sorter to assure discrete separation of B220+ and B220- cells followed by soft agar culture to prevent interaction with accessory cells, we demonstrate that the combination of SCF and IL-7 does not stimulate the expansion or differentiation of B220- lymphoid precursors but can act synergistically in the clonal proliferation of B220+ cells.

Animals

[Hepatitis C virus antibodies in chronic non-alcoholic liver disease].

The presence of hepatitis C virus antibodies was studied in 64 patients with non alcoholic liver disease and found in 11 (17%). The greater frequency of positive antibodies was found among patients with cryptogenetic liver disease, specially those without serum auto-antibodies (32%). The antibody was unusually found (0 to 11%) in non alcoholic liver diseases of other etiologies. It is concluded that hepatitis C virus chronic infections may be the etiology of an important number of non alcoholic chronic liver diseases.

Adult

In vitro antigen trapping by gill cells of the rainbow trout: an immunohistochemical study.

An in vitro assay was used to study the involvement of gill cells in the trapping and processing of particulate antigens. Gills were routinely processed for light microscopy after being placed in medium containing either Yersinia ruckeri O-antigen-labelled fluorescent beads, unlabelled fluorescent beads, Y, ruckeri O-antigen or formalin-killed Y. ruckeri, for 0, 30 s, 1, 5 and 30 min. Y. ruckeri formalin-killed cells, Y. ruckeri O-antigen and fluorescent beads labelled with Y. ruckeri O-antigen were taken in by gill epithelial cells as soon as 30 s after administration. In contrast, unlabelled fluorescent beads adhered to the epithelial cell membranes, but did not occur inside the gill cells. These results are discussed principally in relationship with the specificity of antigen trapping.

Animals

Seasonal variations in the immune system of lower vertebrates.

Seasonal variation, affecting the structure and function of the ectotherm immune system, is an excellent 'natural' model of the influence of neuroendocrine rhythms on immunity. In this review, Agustín Zapata, Alberto Varas and Marta Torroba examine the correlations between seasonal changes and circulating steroid levels, and investigate the possibility of a neuroendocrine-immune network in lower vertebrates.

Adrenal Cortex Hormones

Testosterone induces lymphopenia in turtles.

Owing to the possible role of sex steroids in the immune-neuroendocrine interactions found in lower vertebrates, we attempted to delineate the effect of testosterone propionate on peripheral blood (PB) and the lymphoid organs of the turtle Mauremys caspica. A single intraperitoneal injection of 200 micrograms/g body weight produced thymic involution and intense lymphopenia in the spleen and, less severely, in the PB compartment. It is suggested that lymphocyte redistribution may occur among the various compartments of the body as the main effect of hormone-induced lymphocyte redistribution, although the mechanism in reptiles and mammals is not yet understood.

Animals

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.

Animals

Accelerated maturation of the thymic stroma in the progeny of adrenalectomized pregnant rats.

The possible role played by glucocorticoids (GCs) in the development of thymic stromal cell components has been studied in the progeny of adrenalectomized pregnant rats (FAdx), an experimental model which ensures the absence of GCs until the establishment of the fetal hypothalamus-pituitary gland-adrenal gland axis. As previously demonstrated for thymocytes, the lack of GCs early in ontogeny results in an accelerated maturation of the thymic stromal elements. Early expression of specific cell markers for thymic epithelial cell subsets and appearance of a well-established cytokeratin-positive epithelial cytoreticulum confirmed the ultrastructural evidence of a faster maturation of the thymic epithelium in FAdx than in FSham. A similar faster and stronger pattern of both class I and class II molecule expression on the epithelial cells occurred in the former fetuses than in control ones. Changes in the pattern of expression of laminin, but not that of fibronectin, throughout thymic maturation also reflected accelerated maturation. Immunohistochemically identified thymic macrophages appeared late in both FSham and FAdx but in higher numbers in these latter indirectly demonstrating their faster development. Finally, the maturation and turnover of thymic dendritic cells showed a remarkable acceleration in the FAdx. In 15- to 16-day-old FAdx thymuses there was a high number of dendritic cells which sharply decreased in the following days suggesting a massive migration to the periphery and/or in situ cell death. In parallel a new wave of dendritic cell progenitors began to differentiate in the FAdx thymuses but not in the FSham ones. The results are discussed from the view of close relationships known to occur between thymocytes and the stromal components, although a direct effect of GCs cannot be discarded.

Adrenalectomy