bromochloroacetic-acid and Glomerulonephritis

To examine the cellular components at different stages of the crescent formation in four most common types of human crescentic glomerulonephritis (CGN), including anti-GBM disease (GBM-CGN), crescentic IgA nephropathy (IgA-CGN), ANCA associated pauci-immune CGN (ANCA-CGN) and crescentic lupus glomerulonephritis (LN-CGN).. Renal biopsy specimens of patients with GBM-CGN (n = 10), IgA-CGN (n = 12), ANCA-CGN (n = 12), and LN-CGN (n = 11) were selected. Immunohistochemistry was adopted to identify the cellular components using different cell markers including cytokeratin (PEC), CD68 (macrophage), nestin (podocyte), podocalyxin (podocyte), CD3 (lymphocyte), CD15 (neutrophil) and PCNA.. There were different subtypes of cell components identified during the formation of a cellular crescent in 4 different types of human CGN. Mainly of PEC 11.4 (0.0, 95.0)%, macrophage 8.0 (0.0, 35.0)% and podocyte 5.5 (0.0, 22.0)% and their constitutive percentages were different among various CGNs (P < 0.01). In all the CGNs studied, there were 50% of cells were negative to all the cell markers adopted for this expeiment. Podocalyxin positive cells 0.5 (0.0, 9.6)% were significantly less than nestin positive cells 5.5 (0.0, 22.0)% in all CGNs. PCNA positive cells were 44.7 (16.7, 83.3)% in the cellular crescent of all CGNs and co-localized with nestin (38/45 cases), CK (42/45 cases) or CD68 (24/45 cases).. PEC, macrophage and podocyte might play important roles in the formation of crescents. The staining disparity of nestin and podocalyxin indicates that podocyte dedifferentiation may occur during the crescent formation. PEC, podocytes and macrophages may participate in the formation of crescent in common CGNs through active cellular proliferation.

Topics: Anti-Glomerular Basement Membrane Disease; Antibodies, Antineutrophil Cytoplasmic; Antigens, CD; Antigens, Differentiation, Myelomonocytic; Cell Proliferation; Epithelial Cells; Glomerulonephritis; Glomerulonephritis, IGA; Humans; Intermediate Filament Proteins; Keratins; Lupus Nephritis; Macrophages; Nerve Tissue Proteins; Nestin; Podocytes; Proliferating Cell Nuclear Antigen; Sialoglycoproteins

Topics: Adult; Biomarkers; Cell Dedifferentiation; Disease Progression; Glomerulonephritis; HIV Seronegativity; Humans; Intellectual Disability; Keratins; Ki-67 Antigen; Kidney Failure, Chronic; Kidney Glomerulus; Macrophages; Male; Podocytes; Renal Dialysis

Tubulo-interstitial fibrosis in dogs may result from primary injury to the interstitium or develop secondary to other renal diseases. As in human renal pathology, tubular epithelial cells (TEC) are believed to actively participate in the mechanisms of renal fibrosis. In this study, we examined the changes in the tubular epithelial component in two specific canine diseases. Immunohistochemistry showed the expression of the epithelial marker cytokeratin, the smooth muscle marker alpha-SMA, the mesenchymal marker vimentin and PCNA in 20 dogs with membranous glomerulonephritis and membrano-proliferative glomerulonephritis. Results showed that the loss of the epithelial marker in TEC was directly correlated to the grade of tubulo-interstitial disease present and independent of the type of glomerulonephritis. Varying degrees of vimentin positivity were detected in tubular epithelium in areas of inflammation, and low numbers of scattered alpha-SMA-positive cells were also observed. Immunohistochemistry showed that epithelial tubular cells lose their cytokeratin staining characteristics and transdifferentiate into cells exhibiting key mesenchymal immunophenotypic feature of vimentin-positive staining in both diseases investigated. The integrity of the tubular basement membrane is likely to be fundamental in maintaining the epithelial phenotype of TEC. Animal models provide opportunities for investigating the pathogenesis of renal fibrosis in humans.

Topics: Actins; Animals; Cell Differentiation; Dogs; Epithelial Cells; Female; Fibrosis; Glomerulonephritis; Keratins; Kidney Tubules; Male; Vimentin

According to recent reports, deformity and size of mesothelial cells reflect mesothelial condition. We applied flow cytometry (FCM) to the analyzation of mesothelial cells in peritoneal dialysis effluent (PDE) and the relationship between the period of peritoneal dialysis (PD) and peritoneal function. Eighteen patients treated for two to 89 months by PD were selected. Their dialysate: plasma creatinine ratio (D/P creatinine)was 0.67 +/- 0.086 (0.53 to 0.87). Overnight PDE was drained and centrifuged. The cell population of peritoneal cells identified by anti-cytokeratin, CD14 and 45 antibodies was studied by FCM. Cytokeratin positive cells were identified as mesothelial cells, distinct from macrophages, granulocytes or lymphocytes. The forward scatter (FSC) of cytokeratin positive cells, fluorescence intensity of cytokeratin and percentage of cytokeratin-positive cells in PDE were 395.6 +/- 55.5 (298.31 to 527.72), 333.9 +/- 272.9 (67.55 to 1,071.95), and 6.75 +/- 6.1% (0.44 to 21.14), respectively. There was a positive correlation between D/P creatinine and FSC, and a negative correlation between D/P creatinine and cytokeratin fluorescence intensity or the percentage of cytokeratin positive cells. However, there was no correlation between the period of PD and FSC, cytokeratin fluorescence intensity or the percentage of cytokeratin-positive cells. It was suggested that the alteration of mesothelial cells is not necessarily influenced by the period of PD, but influences peritoneal function. It was found that the analysis of cell population by FCM reflects the morphological and functional changes in the peritoneum of patients on PD.

Topics: Adult; Aged; Cell Separation; Chronic Disease; Dialysis Solutions; Epithelial Cells; Female; Flow Cytometry; Glomerulonephritis; Humans; Keratins; Male; Middle Aged; Peritoneal Dialysis; Peritoneum

Connective tissue growth factor (CTGF) has recently been recognized as an important profibrotic factor and is up-regulated in various renal diseases with fibrosis. The present study describes the sequential localization of CTGF mRNA and its association with transforming growth factor (TGF)-beta1 in human crescentic glomerulonephritis (CRGN). Furthermore, we examined the phenotype of CTGF-expressing cells using serial section analysis. Kidney biopsy specimens from 18 CRGN patients were examined using in situ hybridization and immunohistochemistry. CTGF mRNA was expressed in the podocytes and parietal epithelial cells (PECs) in unaffected glomeruli. In addition, it was strongly expressed in the cellular and fibrocellular crescents, particularly in pseudotubule structures. Serial sections revealed that the majority of CTGF mRNA-positive cells in the crescents co-expressed the epithelial marker cytokeratin, but not a marker for macrophages. Moreover, TGF-beta1, its receptor TGF-beta receptor-I, and extracellular matrix molecules (collagen type I and fibronectin) were co-localized with CTGF mRNA-positive crescents. Our results suggest that CTGF is involved in extracellular matrix production in PECs and that it is one of the mediators promoting the scarring process in glomerular crescents.

Topics: Activin Receptors, Type I; Adult; Aged; Biopsy; Child, Preschool; Collagen Type I; Connective Tissue Growth Factor; Epithelial Cells; Female; Fibronectins; Gene Expression; Glomerulonephritis; Humans; Immediate-Early Proteins; Immunohistochemistry; In Situ Hybridization; Intercellular Signaling Peptides and Proteins; Keratins; Kidney; Kidney Glomerulus; Male; Middle Aged; Protein Serine-Threonine Kinases; Receptor, Transforming Growth Factor-beta Type I; Receptors, Transforming Growth Factor beta; RNA, Messenger; Transforming Growth Factor beta; Transforming Growth Factor beta1

Although there has been extensive research into the mechanisms involved in glomerular crescent formation, it is not yet fully understood how this change may cause renal function impairment. The aim of this study is to identify morphologic changes which may be responsible for this phenomenon. Thirty-eight renal biopsies showing glomerulonephritis with extracapillary proliferation (20 vasculitis-related, 6 idiopathic, 9 due to immune-complex deposition and 3 superimposed on diabetic nephropathy) were considered, and 146 glomeruli in which both crescents and the urinary pole were found at the same time, were studied. The involvement of the urinary pole by cellular crescents was observed in 93.1 and 100% of the glomeruli with segmental or circumferential crescents, respectively. A tridimensional study, for the evaluation of the glomeruli as a whole, was performed on 8 biopsies by means of the step-section technique and disclosed the involvement of the urinary space and a close contact between crescent and tubular cells in all 54 investigated glomeruli. The reported features do not seem to be related to the type of cells which formed the crescent. Indeed, as shown by immunohistochemical study on 10 cases with anti-cytokeratin and anti-CD68 antisera, the crescent localization at the urinary pole had no correlation with the prevalence of epithelial or macrophagic cells. These findings suggest that crescents, due to epithelial proliferation or macrophage clustering, tend to localize at the urinary pole and thus come into close contact with cells of the proximal convoluted tubule: the formation of a sort of plug or a 'glomerular stone' could well explain the block in the urine flow and the consequent impairment of renal function in the acute phase of the disease, even in those cases where crescents are segmental.

Topics: Antibodies, Antineutrophil Cytoplasmic; Antigens, CD; Antigens, Differentiation, Myelomonocytic; Glomerulonephritis; Humans; Immunohistochemistry; Keratins; Kidney Diseases; Kidney Glomerulus; Vasculitis

We report six cases of hyperplastic mesothelial cells located in the sinuses of lymph nodes. All patients but one had a concurrent serosal fluid collection (two pericardial, two pleural, one abdominal) at the time of the lymph node biopsy. All effusions cleared with treatment of the underlying disorder, which included lymphoproliferative processes, congestive heart failure, and inflammatory diseases (Dressler syndrome, vasculitis, and glomerulonephritis). Four cases were associated with vascular prominence of the involved nodal sinuses, a feature that may reflect the cause of the underlying effusion or support the transient persistence of benign mesothelial cells in lymph nodes. Two cases were characterized by distention of the nodal sinuses by sheets of mitotically active mesothelial cells. The differential diagnosis includes metastatic carcinoma, keratin-positive dendritic cells native to lymph nodes, and metastatic malignant mesothelioma. Because the latter shares both clinical and morphological features with cases of benign mesothelial cells in lymph nodes, we believe that this distinction may not always be possible in a given biopsy specimen and therefore that careful clinical follow-up is required in such cases.

Topics: Actins; Adolescent; Adult; Biomarkers, Tumor; Carcinoma; Dendritic Cells; Diagnosis, Differential; Epithelium; Female; Glomerulonephritis; Heart Failure; Humans; Hyperplasia; Immunohistochemistry; Keratins; Lymph Nodes; Lymphoproliferative Disorders; Male; Mesothelioma; Middle Aged; Platelet Endothelial Cell Adhesion Molecule-1; Vasculitis

The expression of cytokeratin and vimentin was studied in the glomerular epithelial cells of canine kidneys with and without glomerular abnormalities. Using ultrastructural, immunogold single and double labelling techniques, cytokeratin and vimentin were found together in the visceral glomerular epithelial cells (vGECs) of abnormal kidneys. In normal kidneys, the vGECs expressed only vimentin, and cytokeratin was found exclusively in parietal glomerular epithelial cells (pGECs). These results confirm previous findings in the same animals, obtained by immunohistological staining techniques.

Topics: Animals; Dog Diseases; Dogs; Epithelium; Female; Glomerulonephritis; Keratins; Kidney Glomerulus; Male; Microscopy, Immunoelectron; Vimentin

Visceral glomerular epithelial cells (vGECs) originate from a mesenchymal blastema and transiently express cytokeratin during embryogenesis. There are no reports of cytokeratin expression in vGECs of mature, normal or damaged, human or other mammalian kidneys in vivo, but in vitro studies have provided evidence of the synthesis of cytokeratin in cultured vGECs. Cytokeratin expression was observed in vGECs in the damaged kidneys of four dogs with spontaneous renal diseases and, by using monoclonal antibodies, type 18 cytokeratin was identified. vGECs are apparently able to (re-) activate in vivo a mechanism for switching on the synthesis of cytokeratin in damaged glomeruli.

Topics: Animals; Dogs; Epithelium; Glomerulonephritis; Keratins; Kidney Failure, Chronic; Kidney Glomerulus; Male

The cellular composition of crescents in diffuse crescentic glomerulonephritis is still controversial. Ten renal biopsies were studied on serial sections by using antikeratin antibodies as specific markers of epithelial cells of Bowman's capsule and both anti-macrophage and anti-lymphocyte antibodies. Semiautomatic morphometry showed that cellular crescents consisted of epithelial cells of Bowman's capsule (24-61%), of macrophages (19-34%) and of unlabelled cells (12-53%). In each biopsy, parietal epithelial cells outnumbered macrophages within crescents.

Topics: Antibodies, Monoclonal; Epithelium; Glomerulonephritis; Histocytochemistry; Humans; Immunochemistry; Keratins; Kidney; Kidney Glomerulus; Lymphocytes; Macrophages; Parietal Cells, Gastric; Staining and Labeling

A study was made of the cells forming the crescents in human crescentic glomerulonephritis. The investigation was performed using a panel of antibodies with immunoperoxidase techniques in formalin fixed, paraffin embedded renal biopsy material. Some of the cells of glomerular crescents were found to contain cytokeratin intermediate filaments, as did some of the cells of the normal parietal epithelium of Bowman's capsule. Leucocytes were also found in crescents, often in the outer part, and their presence was associated with a mantle of inflammatory cells around the glomerulus. The use of paraffin embedded rather than frozen tissue allowed better histological assessment than has been possible in previous studies. The glomerular crescents appeared to be primarily epithelial in origin, with leucocytes contributing to the overall inflammatory response.

Topics: Antibodies, Monoclonal; Biopsy; Glomerulonephritis; Histocompatibility Antigens; Humans; Keratins; Kidney Glomerulus; Leukocyte Common Antigens; Membrane Proteins; Mucin-1; Muramidase; Nephrectomy

The distribution of intermediate filament proteins (vimentin, desmin, and cytokeratin) was studied by means of immunofluorescence in the normal human and rat glomerulus and in pathologic human glomeruli. Antifibronectin antibodies were used as mesangial markers. In normal human glomeruli, vimentin antibodies stained endothelial cells, podocytes, and mesangial cells; desmin antibodies, surprisingly, stained podocytes. In normal rat glomeruli, the pattern of vimentin staining was the same as in humans, but desmin antibodies stained both mesangial cells and podocytes. In human and rat glomeruli cytokeratin staining was confined to segments of Bowman's capsule. In human pathologic glomeruli, vimentin and desmin antibodies stained the structures that were positive in normal glomeruli, giving a characteristic pattern for each pathologic condition examined. These results are compatible with the mesenchymal origin of podocytes and mesangial cells and suggest that both cells have smooth muscle-like phenotypic features. Mesangial cells may have slightly different differentiation paths in humans and rats, leading to a distinct expression of intermediate filament proteins.

Topics: Animals; Desmin; Female; Fibronectins; Fluorescent Antibody Technique; Glomerulonephritis; Humans; Intermediate Filament Proteins; Keratins; Kidney Glomerulus; Rats; Vimentin

Recent studies have suggested that crescents are primarily of monocytic origin and that epithelial cells are a minor factor in their composition. Frozen sections of renal biopsies from 11 cases of crescentic glomerulonephritis (CGN) and 5 controls (2 acute interstitial nephritis, 1 focal glomerulosclerosis, 1 benign recurrent hematuria, 1 normal kidney) were stained for intracellular cytokeratin (CK) with a mouse monoclonal anti-CK antiserum (PKK1) and nonspecific esterase (NSE) activity. Indirect immunofluorescence with PKK1 antiserum showed that in all biopsies there was positive staining of collecting duct and proximal and distal tubular epithelium but no reactions in blood vessels or interstitium. In control case glomeruli there was no staining of the tuft, including the visceral epithelium. In all cases some parietal epithelium was CK-positive. In 4 CGN biopsies the majority of the crescents showed cytoplasmic staining for CK in more than 50% of the crescent cells. In 2 cases most crescents contained between 10-50% CK-positive cells, whereas in 5 biopsies little or no CK was present in the majority of crescents. In all but one CGN case the majority of crescents contained fewer than 30% NSE-positive cells (monocytes). Electron microscopy demonstrated intermediate filaments in many crescent cells and scattered desmosomes within crescents. The results indicate that epithelial cells, probably of parietal epithelial origin, contribute significantly to crescent formation.

Topics: Antibodies, Monoclonal; Glomerulonephritis; Histocytochemistry; Humans; Keratins; Kidney Glomerulus; Microscopy, Electron