heparitin-sulfate and Granuloma

Heparan sulfate (HS) is a glycosaminoglycan that is anchored to the outside of cell membranes. Under ordinary circumstances, it is not present in the interstitium, but under certain circumstances, mainly in the setting of inflammation and tissue repair, HS can be shed from the cell surface into the interstitium in a regulated fashion. Under these circumstances, interstitial HS seems to have an immunomodulatory function because of its binding of many cytokines. However, it is not known which cell types present at an inflammatory site are responsible for this shedding.. We have investigated the presence of interstitial HS by immunohistochemistry in various inflammatory skin diseases characterized by different compositions of the inflammatory infiltrate.. Strong interstitial HS immunoreactivity was present only in diseases with a predominantly histiocytic infiltrate but not in diseases with a predominantly lymphocytic or neutrophilic infiltrate.. This indicates that histiocytes have a direct or indirect role in the HS shedding process. In the well-formed granulomas of sarcoidosis, interstitial HS immunoreactivity was spatially associated with the fibrotic ring at the periphery of the granulomas, but not with the center harboring the histiocytes. This suggests that histiocytes can stimulate fibroblasts to shed HS into the interstitium.

Topics: Cell Communication; Fibroblasts; Granuloma; Heparitin Sulfate; Histiocytes; Humans; Immunohistochemistry; Inflammation; Retrospective Studies; Skin Diseases

Connective tissue cells (myofibroblasts) from liver inflammatory granulomatous reactions to schistosome eggs are able to sustain a long-term proliferation of myeloid cells, both in vivo and in vitro. We have addressed the question of the molecular mechanisms involved in control of this extramedullar stroma-dependent production of inflammatory cells. Heparan sulfate proteoglycans (HSPGs) were purified from granuloma-derived connective tissue cells and bound to plastic or collagen substrate. Their ability to bind recombinant murine granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin-3 (IL-3), to stimulate proliferation of the FDC-P1 myeloid cell lineage, and to modify growth factor activity was monitored. The specificity of this stroma cell-derived glycosaminoglycan interaction with the myeloid growth factors was analyzed by comparing other glycosaminoglycans and sulfated polysaccharides. HSPGs could act as an artificial myelopoietic stroma; they were both required and sufficient for binding and presenting GM-CSF and IL-3 in biologically active form. Moreover, they were able to mediate an increase in the specific growth-promoting activity of GM-CSF and IL-3. This was specific for stroma-derived heparan sulfate and heparin, since heparan sulfate derived from other cells, other glycosaminoglycans and related molecules had no effect. These results indicate that HSPGs can stimulate and control the in situ proliferation of myeloid cells, modifying in both quantitative and qualitative terms the composition of inflammatory cell infiltrates in hepatic granulomas.

Topics: Animals; Cell Division; Granulocyte-Macrophage Colony-Stimulating Factor; Granuloma; Growth Substances; Hematopoiesis; Heparan Sulfate Proteoglycans; Heparitin Sulfate; Interleukin-3; Liver; Mice; Mice, Inbred C3H; Proteoglycans; Schistosomiasis mansoni

An important pathological outcome of schistosomiasis is hepatic fibrosis, with a significant deposit of collagens and proteoglycans. In this study, hepatic and granuloma-associated glycosaminoglycans (GAGs) were analyzed both quantitatively and qualitatively at the acute stage of murine infection with Schistosoma mansoni. The effects of IFN gamma, which has been successfully used for reducing collagen deposition in the liver during schistosomiasis, were also analyzed in granulomas and the surrounding liver parenchyma. Acute schistosomiasis resulted in a 4.4-fold increase in total hepatic GAG content, from which granulomatous GAGs--mainly chondroitin sulfates A/C and B--represented only one sixth of total GAGs amount. Therefore, the increase was found predominantly in the parenchyma. In this compartment, qualitative changes were also induced with a marked increase in the proportion of chondroitin sulfates A/C balanced by a decrease in the proportion of heparan sulfate and dermatan sulfate. IFN gamma reduced parenchymal GAG content by 47%. Qualitatively, the cytokine increased the proportion of heparan sulfate and reduced the quantity of chondroitin sulfates A/C by half in this compartment. By contrast, IFN gamma had neither quantitative nor qualitative effect on fibroinflammatory granulomas. In these structures, the absence of heparan sulfate--which is suspected to mediate IFN gamma activity--might explain these observations.

Topics: Animals; Chondroitin Sulfates; Dermatan Sulfate; Extracellular Matrix; Female; Glycosaminoglycans; Granuloma; Heparitin Sulfate; Interferon-gamma; Liver; Liver Diseases, Parasitic; Mice; Schistosomiasis mansoni

1. Connective tissue cells isolated from hepatic granulomas (GR cells), induced in mouse liver tissue by schistosomal infection, are able to sustain myelopoiesis, while other connective tissue cells such as skin fibroblasts (SF) are not. 2. We compared the ability of SF and GR cells to sustain in vitro proliferation of the FDC-P1 myeloid cell line, dependent upon IL-3 or GM-CSF. 3. Only the GR stroma sustained the proliferation of co-cultured FDC-P1 cells. RT-PCR analysis showed that both cell lines expressed the message for GM-CSF, but not for IL-3. We showed that GM-CSF was produced by, and remained bound to the cell layer through heparan sulfate; this growth factor could be released by high-salt treatment in a biologically active form from both cell types. The same activity could be restored to NaCl-treated GR cells, but not to SF, by incubation with recombinant murine GM-CSF. 4. These results indicate that the ability of connective tissue cells to sustain myelopoiesis depends directly upon the capacity of their heparan sulfate-bearing molecules to bind and present the GM-CSF to the target cells in a biologically active form. Alternatively, a yet unidentified set of cell layer-associated molecules may be required for the positive or negative control of the membrane-bound GM-CSF.

Topics: Animals; Bone Marrow Cells; Cell Division; Cell Line; Connective Tissue; Fibroblasts; Granulocyte-Macrophage Colony-Stimulating Factor; Granuloma; Heparan Sulfate Proteoglycans; Heparitin Sulfate; Interleukin-3; Liver Diseases, Parasitic; Mice; Mice, Inbred C3H; Proteoglycans; Schistosomiasis mansoni

1. This paper summarizes our studies on proteoglycans and glycosaminoglycans in the hepatic fibrosis occurring in schistosomiasis. 2. We have compared proteoglycans and glycosaminoglycans isolated from schistosomal fibrotic granulomas with those obtained from the cellular and extracellular compartments of a murine cell line derived from schistosome-induced granulomas, the primary cell line "GR". 3. Our results have shown some biochemical and structural similarities between proteoglycans and glycosaminoglycans extracted from granulomas and those synthesized and secreted by GR cells, suggesting that these cells may be the major cell population involved in synthesis and accumulation of these molecules in the schistosomal periovular granulomas in liver. Furthermore, we have shown that GR cells can function as an extramedullary myelopoietic stroma that mediates a long-term myeloid proliferation through an autocrine mechanism where the interaction between myelopoietic growth factors and cell-surface heparan sulfate proteoglycans was characterized.

Topics: Animals; Cell Line; Connective Tissue; Dermatan Sulfate; Glycosaminoglycans; Granuloma; Heparitin Sulfate; Liver Cirrhosis, Experimental; Mice; Proteoglycans; Schistosomiasis mansoni

Proteoglycans synthesized in vitro by periovular granulomas isolated from livers of schistosome-infected mice were compared with those produced by granuloma-derived cell lines: the primary cell line GR and the permanent cell line GRX. Proteoglycans were metabolically labelled with 35S-sulfate and extracted with 4 M guanidine-HCl containing 2.0% Triton X-100, in the presence of proteinase inhibitors. The radiolabelled proteoglycans were purified and characterized by anion-exchange, gel-filtration and affinity-column chromatography. Heparan sulfate proteoglycans (HS-PGs) and chondroitin sulfate/dermatan sulfate-containing proteoglycans (CS/DS-PGs) were detected in both the culture medium and the cell-associated fractions obtained from GR cells. More than 90% of the cell-associated HS-PG from these cells contained a hydrophobic portion, as evidenced by their ability to bind to octyl-Sepharose. In contrast, among the secreted proteoglycans, it was the CS/DS-PG and not the HS-PG that bound to this resin. The major fractions of cell-associated and secreted proteoglycans from GRX cells were HS-PGs. Similar to HS-PGs from GR cells, 50% of the cell-associated HS-PG bound to octyl-Sepharose, while only 20% of secreted proteoglycans (HS-PGs) bound to this resin. The proteoglycans purified from the whole granuloma were composed mainly of DS-PG, of a size and hydrophobicity similar to the CS/DS-PG from GR cells. Possible correlations among the structure, secretion, distribution and function of proteoglycans in granulomatous reactions are discussed.

Topics: Animals; Cell Line; Chondroitin Sulfates; Chromatography, Affinity; Chromatography, Gel; Chromatography, Ion Exchange; Connective Tissue; Dermatan Sulfate; Female; Granuloma; Heparitin Sulfate; Liver Diseases, Parasitic; Male; Mice; Mice, Inbred C3H; Proteoglycans; Schistosomiasis mansoni

Sulphated glycosaminoglycans were isolated from schistosome-induced hepatic granuloma and from the pericellular, intracellular and extracellular compartments of two murine cell lines derived from granulomas: the primary cell line GR, and the permanent cell line GRX, established spontaneously from GR. The glycosaminoglycans composition in the whole granuloma was similar to that observed in the intracellular and extracellular compartments of GR cells. This result suggests that GR cells may be the major cell population involved in the synthesis and accumulation of glycosaminoglycans in the granulomas, and play an important role in the process of hepatic fibrosis. The conversion of the primary cell line GR into the established GRX cells did not modify the ratios that prevail among different glycosaminoglycans of the cell surface. However, it decreased the synthesis and secretion of glycosaminoglycans, reduced the proportion of iduronic acid units in the chondroitin sulphate, and increased the proportion of heparan sulphate in intracellular and extracellular pools. These characteristics of the GRX cells are similar to those observed in long-term cultures of smooth-muscle cells. In agreement with the general phenomenon of progressive de-differentiation during in-vitro culture of primary cell lines, these data indicate that the connective tissue cells of liver may belong to the myofibroblastic cell lineage.

Topics: Animals; Cell Line; Chondroitin Sulfates; Female; Glycosaminoglycans; Granuloma; Heparitin Sulfate; Liver Diseases, Parasitic; Male; Mice; Mice, Inbred C3H; Schistosomiasis mansoni

We have characterized sulfated glycosaminoglycans of periovular granulomas induced in mouse liver by experimental infection with Schistosoma mansoni and determined parameters of their synthesis and accumulation by metabolic incorporation of 35S. The major component of glycosaminoglycans isolated from granulomas was dermatan sulfate and the minor component was heparan sulfate. A similar proportion was observed among newly synthesized 35S-labeled glycosaminoglycans, with a slight increase in the relative amount of heparan sulfate. Neither qualitative nor quantitative differences were observed between glycosaminoglycans isolated from granulomas of the acute and the chronic phase of the disease. In contrast, collagen content of granulomas increased eightfold during evolution of the disease from the acute to the chronic phase. It may be concluded that different mechanisms control glycosaminoglycan and collagen synthesis in schistosomal granulomas, as well as the ratio between these components in the extracellular matrix. This is consistent with the loose organization of the extracellular matrix in acute inflammatory reactions and its dense organization in the chronic reactions.

Topics: Animals; Collagen; Dermatan Sulfate; DNA; Electrophoresis, Agar Gel; Female; Glycosaminoglycans; Granuloma; Heparitin Sulfate; Liver; Liver Diseases, Parasitic; Male; Mice; Mice, Inbred C3H; Schistosomiasis mansoni