Monoclonal Antibodies Against Gingival Components

1988 ◽  
Vol 2 (2) ◽  
pp. 240-244
Author(s):  
K. Uobe ◽  
S. Wada ◽  
M. Wato ◽  
T. Nishikawa ◽  
A. Tanaka ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Epithelial Cells ◽  
Basement Membrane ◽  
Immunoelectron Microscopy ◽  
Squamous Epithelium ◽  
Periodate Oxidation ◽  
Basement Membranes ◽  
Whole Cells ◽  
Stratified Squamous Epithelium ◽  
Tissue Reactions

The aim of this study was to produce and characterize monoclonal antibodies against human gingival epithelial cells and gingival fibroblasts. By using these whole cells as immunogens, we were able to generate a large number of monoclonal antibodies reacting with tissue antigens, in particular antibodies that reacted with desmosomes (MoAbs 7 and 8) and basement membrane (MoAb FB-1) antigens. MoAbs 7 and 8 produced from epithelial cells stained cell membranes of epithelium and desmosomes, respectively, as shown by light and immunoelectron microscopy. The epitopes to which MoAbs 7 and 8 were reactive were stable against various treatments; only periodate oxidation abolished the tissue reactions with MoAb 8. Extraction of gingiva with SDS or NP-40, SDS-PAGE analysis, and Western blotting showed that the MoAb 8 identified an antigen with a molecular weight of 100,000 daltons. MoAb FB-1 produced from fibroblasts immunolabeled the basement membranes. The FB-1 antigen was clearly different from any of the known ubiquitous basement membrane components, such as type IV collagen, laminin, and fibronectin. Examination of the distribution and localization of the antigen showed that it was present in the basement membrane beneath stratified squamous epithelium. FB-1 did not react with any of types I-VI collagens in ELISA, but immunoelectron microscopy showed that the antigen reacting with FB-1 was present in the lamina fibroreticularis of the basement membrane and was comprised of collagen-like fibers. These results suggest the possible existence of a new collagen other than types I-VI in the basement membrane beneath stratified squamous epithelium.

scholarly journals Die Kaliumhydrogenphosphat-induzierte Nephropathie des Hundes. I. Pathogenese der Tubulusatrophiee [Potassium Hydrogen Phosphate Induced Nephropathy in the Dog. I. Pathogenesis of Tubular Atrophy—author's trans.]

1980 ◽  
Vol 17 (6) ◽  
pp. 699-719 ◽  
Author(s):  
P. Schneider ◽  
G. Pappritz ◽  
R. Müller-Peddinghaus ◽  
M. Bauer ◽  
H. Lehmann ◽  
...  
Keyword(s):  
Acid Phosphatase ◽  
Epithelial Cells ◽  
Basement Membrane ◽  
Basement Membranes ◽  
Tubular Epithelial Cells ◽  
Beagle Dogs ◽  
Lysosomal Degradation ◽  
Tubular Basement Membrane ◽  
Tubular Atrophy ◽  
Study Results

A nephropathy with severe tubular atrophy was observed in Beagle dogs after oral administration of K2HPO4 for 14 or 38 weeks. We describe the complete lysosomal degradation of atrophying tubular epithelial cells. During two experiments of 14 and 38 weeks duration, respectively, a total of 15 Beagle dogs received 0.8 g K2HPO4/kg body weight daily with their food. All dogs were examined clinically at regular intervals. Renal biopsies were taken in the fourth week from beagles of the 14-week study. Results were compared with those of control dogs. At the end of the experiments the animals were killed and necropsies done. Different stains and histochemical reactions were applied to paraffin sections of the kidneys. Acid phosphatase and β-glucuronidase were found on cryostat sections. Kidneys fixed by perfusion of five Beagles from the 38-week study and three Beagles of the 14-week study, and from five control dogs, were examined electron microscopically. Ultrahistochemically, acid phosphatase was demonstrated. Clinically, the dogs in both experiments vomited, were cachectic, and had elevated creatinine and blood urea nitrogen. Morphologically, qualitatively identical changes were seen, but the renal damage was most marked at 38 weeks. There were disseminated tubular atrophy (usually of the proximal tubules), focal scar tissue and nephrocalcinosis. The following pathogenesis was established for the lesions of the proximal tubule: Tubular atrophy begins with loss of differentiation of epithelial cells. Enzyme histochemistry, ultrahistochemistry and electron microscopy show an increase in autophagic vacuoles and autophagolysosomes. The lysosomal bodies showing fusion enclose large parts of the cytoplasm as the process continues. Complete lysosomal degradation of epithelial cells and extrusion of large lysosomes into the tubular lumen follow. After complete enzymatic digestion of the intratubular detritus, the residue is empty, convoluted and collapsed tubular basement membrane. Atrophic tubular epithelial cells have many organelle-free zones at their base, which contain fine filamentous material resembling that of the basement membrane. The degradation processes described here may explain why clinically the urinary sediment contains few cylinders and epithelial cells and why proteinuria decreases significantly toward the end of the experiment. So far, it is not clear whether the tubular basement membrane is synthesized by the tubular cells, by fibroblasts or by both cell types. The presence of basement membrane-like material in tubular epithelial cells and in parietal epithelial cells of the glomerulus favors the view that epithelial cells produce the basement membranes and that increased production of basement membrane-like material is a sign of loss of differentiation.

scholarly journals Extracellular matrix of lymphoid tissues in the chick.

1989 ◽  
Vol 37 (5) ◽  
pp. 757-763 ◽  
Author(s):  
A Colombatti ◽  
A Poletti ◽  
A Carbone ◽  
D Volpin ◽  
G M Bressan
Keyword(s):  
Extracellular Matrix ◽  
Epithelial Cells ◽  
Basement Membrane ◽  
Basement Membranes ◽  
Intestinal Lumen ◽  
Lymphoid Tissues ◽  
White Pulp ◽  
Extracellular Matrix Components ◽  
Coarse Fibers ◽  
Distribution Of Components

We describe the immunohistochemical distribution of components of the extracellular matrix of the chick lymphoid system. In the thymus, basement membranes of epithelial cells bordering the lobules were intensely stained by laminin antibodies; fibronectin antibodies labeled the capsule and the septal matrix, and similar reactivity was seen with tropoelastin and gp 115 antibodies. No positivity was detected with any of the antibodies within the cortical parenchymal cells. Laminin was not detected in the medullary parenchyma, whereas fibronectin was present as coarse fibers. Tropoelastin and gp 115 appeared as a finer and more diffuse meshwork. In the bursa, laminin antibodies outlined the epithelial cells separating the cortex from the medulla. Fibronectin, tropoelastin, and gp 115 antibody stained the interfollicular septa and the cortical matrix, although to a different extent. Laminin was also detected in association with the interfollicular epithelium (IFE) basement membrane, whereas no staining was found underneath the follicle-associated epithelium (FAE). FAE cells not only lack a proper basement membrane but are also not separated from medullary lymphocytes by any of the other extracellular matrix components were investigated. Consequently, medullary lymphocytes are not sequestered, and can come easily into contact with antigens present in the intestinal lumen. All four antibodies stained the spleen capsule and spleen blood vessels, tropoelastin and gp 115 antibodies giving the strongest reactivity. A fine trabecular staining pattern was detected with gp 115 antibodies in the white pulp.

Laminin α-chain expression and basement membrane formation by MLE-15 respiratory epithelial cells

2002 ◽  
Vol 282 (5) ◽  
pp. L1004-L1011 ◽  
Author(s):  
Nguyet M. Nguyen ◽  
Yushi Bai ◽  
Katsumi Mochitate ◽  
Robert M. Senior
Keyword(s):  
Epithelial Cells ◽  
Basement Membrane ◽  
Critical Role ◽  
Basement Membranes ◽  
Alveolar Type ◽  
Type Ii Cells ◽  
Type Ii ◽  
Respiratory Epithelial Cells ◽  
Alveolar Type Ii Cells ◽  
Respiratory Epithelial

Basement membranes have a critical role in alveolar structure and function. Alveolar type II cells make basement membrane constituents, including laminin, but relatively little is known about the production of basement membrane proteins by murine alveolar type II cells and a convenient system is not available to study basement membrane production by murine alveolar type II cells. To facilitate study of basement membrane production, with particular focus on laminin chains, we examined transformed murine distal respiratory epithelial cells (MLE-15), which have many structural and biochemical features of alveolar type II cells. We found that MLE-15 cells produce laminin-α5, a trace amount of laminin-α3, laminins-β1 and -γ1, type IV collagen, and perlecan. Transforming growth factor-β1 significantly induces expression of laminin-α1. When grown on a fibroblast-embedded collagen gel, MLE-15 cells assemble a basement membrane-like layer containing laminin-α5. These findings indicate that MLE-15 cells will be useful in modeling basement membrane production and assembly by alveolar type II cells.

scholarly journals Expression of extracellular matrix components is regulated by substratum.

1990 ◽  
Vol 110 (4) ◽  
pp. 1405-1415 ◽  
Author(s):  
C H Streuli ◽  
M J Bissell
Keyword(s):  
Extracellular Matrix ◽  
Epithelial Cells ◽  
Basement Membrane ◽  
Type I Collagen ◽  
Basement Membranes ◽  
Type I ◽  
Collagen Gels ◽  
Extracellular Matrix Components ◽  
Matrix Components ◽  
Cells Cultured

Reconstituted basement membranes and extracellular matrices have been demonstrated to affect, positively and dramatically, the production of milk proteins in cultured mammary epithelial cells. Here we show that both the expression and the deposition of extracellular matrix components themselves are regulated by substratum. The steady-state levels of the laminin, type IV collagen, and fibronectin mRNAs in mammary epithelial cells cultured on plastic dishes and on type I collagen gels have been examined, as has the ability of these cells to synthesize, secrete, and deposit laminin and other, extracellular matrix proteins. We demonstrate de novo synthesis of a basement membrane by cells cultured on type I collagen gels which have been floated into the medium. Expression of the mRNA and proteins of basement membranes, however, are quite low in these cultures. In contrast, the levels of laminin, type IV collagen, and fibronectin mRNAs are highest in cells cultured on plastic surfaces, where no basement membrane is deposited. It is suggested that the interaction between epithelial cells and both basement membrane and stromally derived matrices exerts a negative influence on the expression of mRNA for extracellular matrix components. In addition, we show that the capacity for lactational differentiation correlates with conditions that favor the deposition of a continuous basement membrane, and argue that the interaction between specialized epithelial cells and stroma enables them to create their own microenvironment for accurate signal transduction and phenotypic function.

scholarly journals Development of mouse mammary gland: identification of stages in differentiation of luminal and myoepithelial cells using monoclonal antibodies and polyvalent antiserum against keratin.

1986 ◽  
Vol 34 (8) ◽  
pp. 1037-1046 ◽  
Author(s):  
A Sonnenberg ◽  
H Daams ◽  
M A Van der Valk ◽  
J Hilkens ◽  
J Hilgers
Keyword(s):  
Monoclonal Antibodies ◽  
Mammary Gland ◽  
Epithelial Cells ◽  
Basement Membrane ◽  
Cell Types ◽  
Mouse Mammary Gland ◽  
Type I ◽  
Basal Cells ◽  
Alveolar Cells ◽  
Luminal Type

The development of the mouse mammary gland was studied immunohistochemically using monoclonal antibodies against cell surface and basement membrane proteins and a polyclonal antibody against keratin. We have identified three basic cell types: basal, myoepithelial, and epithelial cells. The epithelial cells can be subdivided into three immunologically related cell types: luminal type I, luminal type II, and alveolar cells. These five cell types appear at different stages of mammary gland development and have either acquired or lost one of the antibody-defined antigens. The cytoplasmic distribution of several of these antigens varied according to the location of the cells within the mammary gland. Epithelial cells which did not line the lumen expressed antigens throughout the cytoplasm. These antigens were demonstrated on the apical site in situations where the cells lined the lumen. One antigen became increasingly basolateral as the cells became attached to the basement membrane. The basal cells synthesize laminin and deposit it at the cell base. They are present in endbuds and ducts and are probably the stem cells of the mammary gland. Transitional forms have been demonstrated which developmentally link these cells with both myoepithelial and (luminal) epithelial cells.

The Importance of Mesenchymal Factors in the Differentiation of Chick Epidermis

Development ◽  
1961 ◽  
Vol 9 (3) ◽  
pp. 370-384
Author(s):  
C. B. McLoughlin
Keyword(s):  
Epithelial Cells ◽  
Connective Tissue ◽  
Squamous Epithelium ◽  
The Other ◽  
Β Cells ◽  
Preen Gland ◽  
Other Hand ◽  
Stratified Squamous Epithelium

It is well established that in the developing chick the underlying mesenchyme initiates the appearance of specific epidermal derivatives, e.g. feathers (Sengel, 1956), claws (Cairns & Saunders, 1954), and the preen gland (Gomot, 1958). On the other hand, it is not yet known to what extent the epidermis is independent of mesenchymal intervention for its basic differentiation into a stratified, squamous epithelium. Sobel's (1958) work on the 8-day chick pituitary suggests that the differentiation and multiplication of certain epithelial cells cannot proceed in the absence of mesenchymal elements. She found that the isolated epithelial cells of the hypophysial rudiment survived but were unable to differentiate or multiply; when associated with perichondrial fibroblasts, however, they resumed mitosis and produced typical α and β cells. In the first part of the present investigation, experiments were made to see whether the embryonic epidermis, like the hypophysial epithelium, requires the resence of fibroblasts to enable it to grow and differentiate, or whether it can proliferate, acquire its characteristic squamous structure and keratinize, when isolated and cultivated in the absence of connective tissue.

scholarly journals Localization of α-Dystroglycan on the Podocyte: from Top to Toe

2005 ◽  
Vol 53 (11) ◽  
pp. 1345-1353 ◽  
Author(s):  
Nils P.J. Vogtländer ◽  
Henry Dijkman ◽  
Marinka A.H. Bakker ◽  
Kevin P. Campbell ◽  
Johan van der Vlag ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Monoclonal Antibodies ◽  
Confocal Microscopy ◽  
Cell Membrane ◽  
Basement Membrane ◽  
Immunoelectron Microscopy ◽  
Glomerular Basement Membrane ◽  
Human Kidney ◽  
Charged Membrane ◽  
Apical Localization

α-Dystroglycan (DG) is a negatively charged membrane-associated glycoprotein that links the cytoskeleton to the extracellular matrix. Previously, we described that α-DG covers the whole podocyte cell membrane in the rat. However, our finding was challenged by the description of a strictly basolateral localization in human kidney biopsies, using a different antibody against α-DG. Therefore, we studied the exact localization of glomerular α-DG by using these two antibodies in both species. The studies were performed by using monoclonal antibodies (MoAbs) IIH6 and VIA4.1 in immunofluorescence, confocal microscopy, and immunoelectron microscopy on both rat and human kidney sections, as well as on cultured mouse podocytes. The apical localization of α-DG on podocytes was more dominant than the basolateral localization. The basolateral staining with MoAb VIA4.1 was more pronounced than that of MoAb IIH6. With both MoAbs, the staining in rat kidneys was more prominent, in comparison to human kidneys. We conclude that α-DG is expressed at both the basolateral and apical sides of the podocyte. This localization suggests that α-DG plays a dual role in the maintenance of the unique architecture of podocytes by its binding to the glomerular basement membrane, and in the maintenance of the integrity of the filtration slit, respectively.

scholarly journals Expression of Hornerin in Stratified Squamous Epithelium in the Mouse: A Comparative Analysis with Profilaggrin

2003 ◽  
Vol 51 (4) ◽  
pp. 485-492 ◽  
Author(s):  
Teruhiko Makino ◽  
Mikiro Takaishi ◽  
Masahiko Toyoda ◽  
Masaaki Morohashi ◽  
Nam-ho Huh
Keyword(s):  
Comparative Analysis ◽  
Immunoelectron Microscopy ◽  
Squamous Epithelium ◽  
Mouse Skin ◽  
Structural Features ◽  
Newborn Mouse ◽  
Rt Pcr ◽  
Mouse Tissues ◽  
Stratified Squamous Epithelium ◽  
Novel Protein

We have recently identified a novel protein named hornerin, the structural features of which are most similar to those of profilaggrin, an essential protein for keratinization of epidermal tissues. In this study we examined the expression of hornerin compared with that of profilaggrin in various mouse tissues. Hornerin was expressed in the upper epidermis of newborn mouse skin, as was profilaggrin. In addition, both hornerin and profilaggrin were expressed in the tongue, esophagus, and forestomach. In all four tissues, immunostaining for hornerin and profilaggrin showed a granular pattern, and most of the signals for the two proteins were co-localized on keratohyalin granules. This was confirmed by double immunoelectron microscopy. Within keratohyalin granules, hornerin was detected more frequently in the periphery, whereas profilaggrin was equally distributed. A quantitative RT-PCR revealed that both genes were expressed at highest levels in the forestomach and at the next highest levels in skin. Profilaggrin mRNA was most abundant in the forestomach. In skin, the amount of hornerin mRNA was more than fourfold greater than the amount of profilaggrin mRNA. These results form the basis for a better understanding of possible overlapping and/or differential functions of hornerin and profilaggrin.

scholarly journals PERIODATE-LYSINE-PARAFORMALDEHYDE FIXATIVE A NEW FIXATIVE FOR IMMUNOELECTRON MICROSCOPY

1974 ◽  
Vol 22 (12) ◽  
pp. 1077-1083 ◽  
Author(s):  
IAN W. MCLEAN ◽  
PAUL K. NAKANE
Keyword(s):  
Endoplasmic Reticulum ◽  
Basement Membrane ◽  
Immunoelectron Microscopy ◽  
Yolk Sac ◽  
Basement Membranes ◽  
Tissue Preservation ◽  
Antibody Technique ◽  
Adequate Tissue ◽  
Basement Membrane Antigen ◽  
Yolk Sac Cells

A new fixative which primarily stabilizes carbohydrate moieties was developed for immunoelectron microscopy. It contains periodate, lysine and paraformaldehyde. Theoretically, the carbohydrates are oxidized by periodate and cross-linked by lysine. The fixative can preserve antigenicity as well as paraformaldehyde and ultrastructure as well as glutaraldehyde. Using this fixative and the peroxidase-labeled antibody technique, basement membrane antigen was localized within the cisternae of endoplasmic reticulum of parietal yolk sac cells and in extracellular basement membranes with adequate tissue preservation, a task which has not been successfully accomplished by conventional fixatives.

scholarly journals Comparison of the ability of basement membranes produced by corneal endothelial and mouse-derived Endodermal PF-HR-9 cells to support the proliferation and differentiation of bovine kidney tubule epithelial cells in vitro.

1984 ◽  
Vol 99 (3) ◽  
pp. 947-961 ◽  
Author(s):  
D Gospodarowicz ◽  
J Lepine ◽  
S Massoglia ◽  
I Wood
Keyword(s):  
Epithelial Cells ◽  
Basement Membrane ◽  
Cell Monolayer ◽  
Basement Membranes ◽  
Low Density ◽  
Kidney Tubule ◽  
Confluent Monolayer ◽  
Bovine Kidney ◽  
Fibroblastic Cells

The proliferation and morphological differentiation of bovine kidney collecting-tubule epithelial cells has been examined as a function of substrata and plasma factors. Collecting kidney tubule explant maintained in vitro gave rise to two distinct cell populations; one was composed mostly of fibroblastic cells whereas the other was epithelioid (EP cells). The proliferation of fibroblastic cells when exposed to serum-supplemented medium was best expressed when cells were maintained on a basement membrane produced by bovine corneal endothelial cells. This basement membrane has a composition, which in previous studies has been shown to favor the proliferation of mesenchymal cells. In contrast, the proliferation of EP cells was best expressed when cells were maintained on a basement membrane produced by the mouse-derived endodermal cell line PF-HR-9 (HR-9-BM). This basement membrane has a biochemical composition very similar to the basement membrane underlying the kidney tubules. Although the fibroblast confluent monolayer maintained on bovine corneal endothelial cell extracellular matrix did not undergo morphogenesis, the confluent monolayer of EP cells maintained on HR-9-BM shows hemicyst formation, suggesting that they were capable of vectorial fluid transport. They also built a complex three-dimensional kidney tubulelike network. Some tubules became grossly visible and floated into the tissue culture medium, remaining tethered to the cell monolayer at either end of the tubule. On an ultrastructural level, the tubules consisted of cells held together with junctional complexes arranged so as to form a lumen. The smallest lumen were bordered by 2-3 cells, and the largest ones by 8-15 cells. The lumens of the larger tubules did contain granular fibrillar and amorphous debris. Low-density EP cell cultures maintained on HR-9-BM could be induced to proliferate at a rate approaching that of cultures exposed to serum when they were exposed to medium supplemented with high-density lipoprotein (HDL, 750 micrograms protein/ml) and transferrin (50 micrograms/ml). When exposed to HDL concentrations equal or lower than 250 micrograms protein/ml, low-density cultures proliferated at a slow rate and readily formed tubulelike structures. This observation indicates that EP cells do not need to reach confluence to undergo morphogenesis, and that HDL, which in the presence of transferrin supports the cell proliferation, can favor their differentiation into tubulelike structures once its concentration becomes limiting for mitogenesis.

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