heparitin-sulfate and Glomerulonephritis--Membranoproliferative

The high IgA (HIGA) strain of ddY mice represents an inbred model of IgA nephropathy that shows mesangioproliferative glomerulonephritis with mesangial IgA deposition. In this study, aggravation of glomerulonephritis in HIGA mice through lipopolysaccharide (LPS)-triggered activation of coagulation was investigated.. Twelve-week-old HIGA and BALB/c mice were intraperitoneally injected with LPS twice at an interval of 3 days, and kidney specimens were collected 7 days after the second LPS injection. In an intervention experiment, the factor Xa inhibitor danaparoid was injected intraperitoneally every day for 7 days after the first LPS injection.. LPS injection induced macrophage infiltration and cellular proliferation in the mesangium together with fibrin deposition and monocyte chemoattractant protein 1 mRNA expression, as well as antigen deposition of tissue factor, factor V, factor X, and protease-activated receptor 2. These phenomena were obvious in HIGA mice when compared to BALB/c mice. Interestingly, toll-like receptor 4 was intensely expressed in HIGA mice before LPS injection and subsequently decreased. Danaparoid treatment significantly ameliorated proteinuria, cellular proliferation, and fibrin deposition.. The present data suggest that tissue factor and factor V induction by LPS may in part accelerate mesangioproliferative glomerulonephritis through activation of factor X and downstream proinflammatory and procoagulant mechanisms.

Topics: Animals; Anticoagulants; Blood Coagulation; Blotting, Western; Chemokine CCL2; Chondroitin Sulfates; Dermatan Sulfate; Factor V; Factor X; Female; Fibrin; Gene Expression; Glomerular Mesangium; Glomerulonephritis, Membranoproliferative; Heparitin Sulfate; Immunoglobulin A; Immunohistochemistry; Injections, Intraperitoneal; Lipopolysaccharides; Mice; Mice, Inbred BALB C; Microscopy, Fluorescence; Proliferating Cell Nuclear Antigen; Receptor, PAR-2; Reverse Transcriptase Polymerase Chain Reaction; Thromboplastin; Toll-Like Receptor 4

Recently we showed that antinuclear autoantibodies complexed to nucleosomes can bind to heparan sulphate (HS) in the glomerular basement membrane (GBM) via the histone part of the nucleosome. Histones have been identified in glomerular deposits in human and murine lupus nephritis. In addition, a decreased HS staining in the GBM was found, most probably due to masking by deposition of antibodies complexed to nucleosomes.. In this study we first investigated whether histones or nucleosomes could be identified in glomerular deposits in human lupus nephritis, and secondly whether the presence of these nuclear components was correlated with absence of HS staining. Kidney biopsies of SLE patients (11 with diffuse proliferative glomerulonephritis (DPGN) and six with membranous glomerulonephritis (MGN)) and non-SLE glomerular diseases were stained for histones. DNA, nucleosomes, IgG and HS.. Using a polyclonal anti-H3 1 21 antiserum, histones were detected in all patients with DPGN and in two of six patients with SLE-MGN (P < 0.01). Using a monoclonal antihistone antibody, histones were stained in three patients with DPGN, but in none of the biopsies with MGN. Using nucleosome specific monoclonal antibodies, nucleosomes were detected in five patients with DPGN, in two patients with MGN, but in none of the biopsies with non-SLE glomerulonephritis. HS staining was nearly absent in DPGN, whereas staining was only moderately reduced in patients with MGN and controls (P = 0.001).. Using polyclonal and monoclonal antihistone antisera, histones were identified in all patients with DPGN and their presence was associated with a decrease of HS staining. Nucleosomes were identified in five of 11 patients with DPGN and in two of six patients with MGN. This is the first demonstration of nucleosomes in glomerular deposits in SLE nephritis.

Topics: Adolescent; Adult; Animals; Antibodies, Monoclonal; Antigen-Antibody Complex; DNA; Female; Fluorescent Antibody Technique, Indirect; Glomerulonephritis, Membranoproliferative; Glomerulonephritis, Membranous; Heparitin Sulfate; Histones; Humans; Immunoglobulin G; Kidney Glomerulus; Lupus Nephritis; Male; Mice; Middle Aged; Nucleosomes

Most glomerular pathologies are associated with alterations of the matrix compartment. Using reagents directed against the alpha 1/alpha 2 and alpha 3 chains of type IV collagen [alpha 1/alpha 2(IV), alpha 3(IV)], laminin, heparan sulphate proteoglycan (HPG), fibronectin, collagen I, and collagen III, we investigated the modifications of the glomerular matrix components in several human glomerular lesions compared with normal kidney. In type I membranous glomerulonephritis (MGN) (nine cases), we did not observe alterations in the matrix component distribution. In MGN types II and III (five cases), the spikes and chainettes were made of the alpha 3(IV) chain, laminin, and HPG, while the alpha 1/alpha 2(IV) chains were localized along the subendothelial side of the glomerular basement membrane (GBM). In focal and segmental glomerulosclerosis (six cases), fibronectin, alpha 1/alpha 2(IV) chains, laminin, and small amounts of interstitial collagens were detected within the collapsed capillary loops; the newly formed matrix material between the podocytes and the GBM contained the alpha 1/alpha 2(IV) chains, laminin, and HPG but not the alpha 3(IV) chain. In crescentic glomerulonephritis (six cases), fibronectin was the most abundant and, in purely cellular crescents, the unique component. A basement membrane-like network containing laminin, HPG, alpha 1/alpha 2(IV) chains, and interstitial collagens developed in a second step between the crescent cells. Interstitial collagens were present in the crescent framework, even when the integrity of Bowman's capsule was preserved.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Adult; Collagen; Extracellular Matrix Proteins; Fibronectins; Glomerulonephritis; Glomerulonephritis, Membranoproliferative; Glomerulonephritis, Membranous; Glomerulosclerosis, Focal Segmental; Heparan Sulfate Proteoglycans; Heparitin Sulfate; Humans; Immunoenzyme Techniques; Kidney Glomerulus; Laminin; Proteoglycans

To clarify whether heparan sulfate proteoglycans (HSPG) on glomerular basement membrane (GBM) are produced by mesangial cell in vivo, the number of anionic sites (AS) on lamina rara externa of GBM and the changes in urinary albumin excretion rate (UAE) were observed on mesangiolysis in rats induced by monoclonal anti-rat Thy1 antibody. Mesangiolysis was remarkable at day 3 after the administration of anti-rat Thy1 antibody, whereas GBM as well as epithelial cell remained well preserved, and detached endothelial cell was observed. The number of AS on GBM was significantly (p < 0.001) decreased in anti-rat Thy1 antibody treated rats (18.6 +/- 0.4/1000nm), when compared with that of normal control rats (22.8 +/- 0.4/1000nm). Furthermore, UAE was markedly (p < 0.05) increased at day 3 (1692.6 +/- 627.5 micrograms/24h) compared with the value before (188.2 +/- 30.6 micrograms/24h) administration of anti-rat Thy1 antibody. From these results, it can be concluded that some part of AS on GBM may be produced by the mesangial cell.

Topics: Animals; Anions; Antilymphocyte Serum; Basement Membrane; Glomerular Mesangium; Glomerulonephritis, Membranoproliferative; Heparan Sulfate Proteoglycans; Heparitin Sulfate; Kidney Glomerulus; Male; Proteoglycans; Rats; Rats, Wistar; T-Lymphocytes

Extracellular matrix expansion is frequently noted in mesangioproliferative renal diseases. This study investigates the role of immunologic factors in glomerular matrix accumulation. The gene expression of type I and IV collagen, laminin and s-laminin was examined in the rat model of mesangial proliferative glomerulonephritis induced with anti-Thy 1.1 antibody. Northern analysis was performed on glomerular RNA isolated one, three and five days after disease induction and at day 3 following prior complement depletion. Tissue was immunostained for the protein products of these genes as well as for heparan sulfate proteoglycan, entactin and PCNA (a marker of cell proliferation) at days 1, 3, 5, 14, 21 and 42. A seven- to ten-fold increase of collagen IV and laminin mRNA as well as de novo expression of collagen I mRNA occurred at days 3 and 5 corresponding to the time of maximal proliferation. S-laminin mRNA levels only increased three-fold. With the exception of s-laminin, mesangial staining for all examined matrix proteins increased to a maximum at day 5 and decreased thereafter. Focal alterations of the glomerular architecture and matrix persisted at day 42. Complement depletion prevented the histological abnormalities as well as the increased expression of matrix proteins at day 3. These findings indicate that immunologic injury in the mesangium may result in overproduction of extracellular matrix components and may ultimately contribute to the development of glomerulosclerosis.

Topics: Animals; Antigens, Surface; Antilymphocyte Serum; Complement System Proteins; Extracellular Matrix Proteins; Gene Expression Regulation; Glomerulonephritis, Membranoproliferative; Heparan Sulfate Proteoglycans; Heparitin Sulfate; Immunization, Passive; Kidney Glomerulus; Male; Proteoglycans; Rats; Rats, Inbred Strains; RNA, Messenger; Thy-1 Antigens

Antibodies to glomerular basement membrane, heparan sulfate-proteoglycans are nephrotoxic but possess a weak nephritogenic potential. In order to enhance the nephritogenic potential, the antibodies were intravenously administered into rats presensitized with heterologous rabbit IgG. This resulted in the integration of heterologous and autologous phases, the two phases characteristic of the traditional model of nephrotoxic serum nephritis. The presensitization caused a dramatic shift in the binding characteristics of the heterologous antibodies between the kidney and lymphoid tissues. A proliferative form of immune complex glomerulonephritis associated with a remarkable proteinuric response was observed. In addition, a moderate degree of hematuria was noted as well. The proteinuria was largely complement-dependent and may possibly be cell-mediated as well. The proteinuria became severe with increasing production of host IgG antibodies and with their subsequent sequestration in the glomeruli. The predominant glomerular lesions were in the form of epimembranous/subepithelial immune deposits, which became more frequent with timely increasing titer of host autologous IgG antibodies. These findings indicate that antibodies to heparan sulfate-proteoglycan, an authentic component of the basement membrane, are capable of mediating a glomerular injury with acquisition of nephritogenic potential in an appropriate environment of the host. At present, it seems that this is the sole constituent of the basement membrane whose antibodies are capable of inducing an immune complex nephritis.

Topics: Animals; Antigen-Antibody Complex; Basement Membrane; Binding Sites, Antibody; Chondroitin Sulfate Proteoglycans; Disease Models, Animal; Female; Glomerulonephritis, Membranoproliferative; Glycosaminoglycans; Heparan Sulfate Proteoglycans; Heparitin Sulfate; Immune Complex Diseases; Immunization; Immunoglobulin G; Kidney Glomerulus; Proteoglycans; Rats; Rats, Inbred F344

Administration of antibody, directed against glomerular basement membrane (GBM) heparan sulfate-proteoglycan, into a presensitized rat results in the induction of membranous nephropathy with subepithelial immune-complex deposits. In this investigation, we examined the mechanisms responsible for the formation of subepithelial immune-complex deposits in the anti-HS-PG model. In initial experiments, the intravenously administered radioiodinated antibody was seen exclusively localized in the regions of the glomerular capillary wall where the subepithelial deposits were observed. To determine their exclusive localization in the subepithelial space, kinetics of movement of the intravenously administered antibody was investigated. The antibody localized in the inner layers of the GBM within a few minutes after its administration. It equilibrated in the inner and outer layers of the GBM in a matter of a few hours. Then, after 24 hours, it gradually disappeared from the inner layers of the GBM and persisted in the outer layers only. The ready clearance of the antibody from the inner layers may be related to the differential in the kinetics of lateral intrinsic plasma fluid currents within the GBM. The persistence of heterologous antibody exclusively in the outer layers and the availability of host autologous antibodies probably resulted in the development of immune complex deposits in the subepithelial space. The glomeruli devoid of plasma water currents showed no change in the concentration of the antibody in the inner and outer layers of the GBM or mesangial matrix. Also, no antibody binding was observed with the plasmalemma of either the foot processes or visceral epithelia. The data suggest that the biochemical-biophysical properties of the glomerular capillary wall, in concert with its intraglomerular hemodynamics, most likely played a significant role in the development of subepithelial immune-complex deposits in this model.

Topics: Animals; Antigen-Antibody Complex; Basement Membrane; Binding Sites, Antibody; Chondroitin Sulfate Proteoglycans; Disease Models, Animal; Female; Glomerulonephritis, Membranoproliferative; Glycosaminoglycans; Heparan Sulfate Proteoglycans; Heparitin Sulfate; Immune Complex Diseases; Immunization; Immunoglobulin G; Kidney Glomerulus; Proteoglycans; Rats; Rats, Inbred F344