Species-specificity of glial vimentin as revealed by immunocytochemical studies with the Vim 3B4 and V9 monoclonal antibodies.
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Abstract:
Two monoclonal antibodies directed against vimentin, Vim 3B4 and V9 could distinguish between vimentins originating from certain species, when tested on cell lines (Bohn et al, 1992). Our comparative immunohistochemical studies in the rat and chicken brain with the same antibodies suggest the coexistence of two vimentin forms in the glial cells of both species. One of these forms bearing the epitope present in the respective non-glial cell lines is present in astrocytes and Bergmann glia independently of the ontogenic state of the animal. The other epitope appeared also mutually in both species, albeit its expression was more restricted. These patterns suggest that in these two species, the expression of the different vimentin forms might be differently regulated.Keywords:
Neuroglia
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We describe reversible changes of intermediate filaments of fibroblastic cells associated with changes in the functional state of the cells. The changes are revealed by com- paring the immunofluorescence patterns given by a monoclonal antibody and a polyclonal serum, both recognizing vimentin. The state of the filaments depends on culture density; this effect can- not be attributed to the nutritional state of the cells, their growth rate, or substances released into the medium. It seems to depend mainly on the aggregation of filaments during strong cell move- ments. The possible significance of these findings for the func- tional role of intermediate filaments is discussed.
Polyclonal antibodies
Immunofluorescence
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Two monoclonal antibodies directed against vimentin, Vim 3B4 and V9 could distinguish between vimentins originating from certain species, when tested on cell lines (Bohn et al, 1992). Our comparative immunohistochemical studies in the rat and chicken brain with the same antibodies suggest the coexistence of two vimentin forms in the glial cells of both species. One of these forms bearing the epitope present in the respective non-glial cell lines is present in astrocytes and Bergmann glia independently of the ontogenic state of the animal. The other epitope appeared also mutually in both species, albeit its expression was more restricted. These patterns suggest that in these two species, the expression of the different vimentin forms might be differently regulated.
Neuroglia
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A benign pelvic soft tissue tumor from a 50 year old woman was examined by immunohistochemistry and electron microscopy. The tumor cells had abundant eosinophilic cytoplasm with a hyaline appearance, which was filled with large aggregates of intermediate‐sized filaments (IF). The cells were positively immunostained by antibodies against cytokeratin, vimentin, desmin, glial fibrillary acidic protein, and neurofilament proteins. This case represents an extreme example of the simultaneous expression of IF by neoplastic cells, and exemplifies the limited applicability of immunohistochemical detection of IF antigens for pathological diagnosis of neoplasms. Acta Pathol Jpn 41: 65–72, 1991.
Desmin
Neurofilament
Intermediate Filament Protein
Hyaline
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Fifty formalin-fixed, paraffin-embedded meningiomas were studied with a panel of ten antibodies directed against epithelial antigens, intermediate filaments, and neuroectodermal antigens. Twenty-five (50%) of these were stained with monoclonal antibodies (MoAb) to epithelial membrane antigen (EMA), 12 (24%) with keratin antibodies, 9 (18%) for vimentin, and 4 (8%) for S100 protein. Monoclonal antibodies to Leu-7, desmin, neurofilament 200 kDa, and glial fibrillary acidic protein (GFAP) all failed to stain any of the 50 neoplasms. Overall, 34 (68%) meningiomas stained with one or more antibodies. Sixteen (32%) failed to stain at all. The dual epithelial and mesenchymal nature of meningiomas is supported in this study. Applications to diagnostic surgical pathology and histogenetic implications are discussed.
Desmin
Neurofilament
Stain
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Paratope
Isotype
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Polyclonal antibodies
Epitope mapping
Linear epitope
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To prepare monoclonal antibodies against oh(8)dG and to evaluate the relationship between Hp infection and oxidative DNA damage by detecting oh8dG in gastric mucosa.BALB/C mice were immunized with BSA-oh(8)dG conjugate, monoclonal antibodies were prepared by hybridoma technique, the biological characteristics of antibodies were analysed by competitive ELISA, Western blot and immunohistochemistry.Two strains of hybridoma cell were obtained. ELISA and Western blot indicated that the antibodies were fairly specific for oh(8)dG. In immunohistochemistry,the positive rate of oh(8)dG expression in Hp positive tissues and Hp negative tissues was 55% and 5%, respectively(P<0.01).The prepared antibodies can specially recognize oh(8)dG and immunohistochemistry with the monoclonal antibodies showed Hp infection can increase oh(8)dG level in gastric mucosa.
Conjugate
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Six canine, one feline and one equine granular cell tumours (GCTs) were investigated electron microscopically and immunohistochemically. The tumours were tested for reactivity with monoclonal antibodies against vimentin and desmin and with polyclonal antibodies against cytokeratin, S-100 protein, glial fibrillary acidic protein (GFAP) and neuron specific enolase (NSE). All GCTs were characterized by their PAS positive cytoplasmic granules in light microscopy, which in electron microscopy appeared as lysosome-like granules. In each case two canine GCTs were stained by the antibody against cytokeratin, vimentin and S-100 protein. Cells of the equine GCT showed reactivity with the antiserum against S-100 protein. In the feline GCT no reactivity with any of the antibodies tested was observed. These differences of the immunohistochemical reactions of GCTs suggest a nonuniform histogenesis of GCTs in domestic animals. The reactivity of the tumour cells with the antiserum against NSE is discussed.
Polyclonal antibodies
Enolase
Desmin
Histogenesis
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The expression of some types of cytokeratin and vimentin in the liver was studied by using monoclonal antibodies to these types of intermediary filaments. Necrotic samples from livers without pathological finding were examined. A panel of 5 monoclonal antibodies, prepared in Czechoslovakia, to cytokeratins Nos. 7,8,18, and 19, as well as to vimentin was used. The findings confirmed the characteristic expression of cytokeratins 8 and 18 in hepatocytes and of cytokeratins 7, 8, 18, and 19 in epithelial cells of intrahepatic bile canaliculi, that monoclonal antibodies prepared in Czechoslovakia (LAMO, Brno and VUKEO, Brno) can be successfully used in immunochemical studies of intermediary filaments in the liver.
Bone canaliculus
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