Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound   Target Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound

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Query: EC:3.4.24.3 (collagenase)
18,340 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Neutrophil proteases are believed to play a role in the pathogenesis of a variety of human diseases. While many studies of proteases in models of disease have focused on elastase, neutrophils contain several proteases some of which share a high degree of homology. This report describes the production and characterization of an IgG1 murine monoclonal antibody (AHN-10) that reacts with human neutrophil elastase but not with the other major neutrophil neutral proteases: cathepsin G, proteinase 3, collagenase, or the newly purified neutral protease, esterase N. AHN-10 inhibited the elastinolytic activity of purified human neutrophil elastase and could detect elastase in alcohol-fixed cytospin preparations. The epitope recognized by AHN-10 was resistant to treatment with NaIO4, suggesting that the epitope is not a carbohydrate. AHN-10 should be useful for the immunolocalization of neutrophil elastase in tissue specimens and as a stable source of characterized antibody for quantitative identification of neutrophil elastase.
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PMID:Preparation and characterization of a monoclonal antibody that inhibits human neutrophil elastase activity. 341 Dec 32

A transplantable macrophage-like cell line has been obtained and established in W rats. This cell line is in its 85th passage and is perhaps the only established macrophage-like cell line that grows rapidly intraperitoneally in rats. The cells possess some of the typical characteristics of macrophages, such as adherence to glass, phagocytosis, presence of Fc receptors and C3d receptors, la determinants, leukocyte common antigen, lysozyme, non-specific esterase, and glycogen. The tumor also grows as solid when injected sc, intradermally, or intramuscularly. The cells have collagenase, tyrosine-specific protein kinase, and several other hydrolases. Histopathologic and ultrastructural observations suggest it to be a histiocytic tumor. The availability of a macrophage cell line of rat origin provides a useful experimental model to study different macrophage functions at the cellular and molecular level.
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PMID:Development and characterization of a rat histiocyte-macrophage tumor line. 345 75

Primary cultures of adherent rheumatoid synovial cells contain variable proportions of fibroblasts, macrophages, and dendritic cells, as judged by morphological appearance. Comparative studies using various enzymic and histochemical staining procedures showed the dendritic cells to lack many of the characteristic features of macrophages, e.g. the failure to express HLA-DR (Ia) antigen. The dendritic cells and fibroblasts had several similarities, but differed to some extent in their nonspecific esterase activity, phagocytic and proliferative potential. As the proportions of dendritic cells and fibroblasts varied in relation to specific culture conditions, we examined the possibility that these morphologies might represent different functional states rather than distinct cellular origins. Using subcultured synovial fibroblasts with a uniform bipolar appearance, we have shown that exposure to interleukin-1 or mast cell products resulted in a transformation to dendritic morphology. This change in cell shape was prevented by the presence of indomethacin, but was subsequently achieved by the addition of exogenous PGE2. Thus it appears that the latter is the factor that modulates the morphological change of fibroblastic to dendritic cells. This study has also demonstrated the complete and reversible interchange of fibroblast/dendritic morphology, thereby confirming that these different shapes are manifest by the same cell. The changes in phenotypic expression associated with the dendritic appearance include increased production of collagenase, prostaglandin E, and nonspecific esterase, as well as an apparent inability to exhibit phagocytosis and to proliferate in culture. We conclude from our in vitro studies that the phenotypic behaviour of the synovial fibroblast (or synoviocyte) is very variable and dependent to a large extent upon local stimuli, but the identity and hierarchy of such stimulating and suppressive factors in relation to cellular interactions requires further study.
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PMID:Comparative studies of adherent rheumatoid synovial cells in primary culture: characterisation of the dendritic (stellate) cell. 349 52

Dome and dome epithelial cells were selectively dissociated from gut-associated lymphoid tissues of rabbits. Sequential tissue washes in dithiothreitol, EDTA, and collagenase removed the dome epithelium, without disrupting the follicles or villi, and provided a cell suspension containing 74 +/- 6% lymphocytes, 9 +/- 4% columnar epithelial cells, 10 +/- 7% tangible-body macrophages, and 4 +/- 2% M cells (follicle-associated epithelial cells). The last mentioned cells were characterized by transmission electron microscopy as large (20 to 55 microns diameter) cuboidal, round, or oval cells with eccentric nuclei and thin membranous processes surrounding empty vacuoles. The M cells were occasionally joined together by tight junctions. Histochemical and immunocytochemical analyses of M cells with the light microscope showed that they were devoid of immunoglobulins and negative for T-cell antigen and secretory component and had no detectable alkaline phosphatase or endogenous peroxidase activity. The M cells had few vacuoles with faint acid phosphatase activity; nonspecific neutral esterase was abundant. Possible uses for dome and dome epithelial cells are discussed.
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PMID:Dome epithelial M cells dissociated from rabbit gut-associated lymphoid tissues. 354 8

Atherosclerotic aortic intimas of cholesterol-fed rabbits were enzymatically dispersed into single cells by collagenase and elastase. And monocyte-macrophages (M phi) were separated from smooth muscle cells (SMC), using the ability of M phi to adhere to a plastic dish firmly even in the enzyme solutions. Round or oval, heavily lipid-laden cells, so-called foam cells (FC), belonged to the M phi fraction. M phi-FC showed very strong activity for non-specific esterase using alpha-naphthyl butyrate, while SMC showed little or no activity. Some of the FC were large and multinucleated (multinucleated giant foam cells). They also showed positive non-specific esterase staining and are thought to be derived from M phi. M phi-FC synthesized various proliferate in the medium and the number decreased gradually within several days. Some SMC were heavily lipid-laden; however, they retained their original spindle shape. SMC lost lipid droplets gradually as they proliferated to confluence. SMC from atherosclerotic lesions showed higher proliferative activity than those from normal-appearing medias of atherosclerotic aortas or control aortas. Almost no M phi-FC were obtained from the intima-medias of grossly normal portions of atherosclerotic aortas and control aortas. The present method will be useful for studying the role of these cells in the pathogenesis of atherosclerosis.
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PMID:Separation and characterization of macrophages and smooth muscle cells in rabbit atherosclerotic lesions. 366 43

Although some data suggest that macrophages in the reticuloendothelial system (RES) are important sources of thromboxane A2 (TxA2) and prostacyclin (PGI2) during endotoxic shock, we are unaware of data documenting the ability of hepatic macrophages (Kupffer cells) to release either TxA2 or PGI2 when exposed to lipopolysaccharide (endotoxin, LPS). In this study, Kupffer cells were examined for their ability to release prostaglandin E2 (PGE2), TxA2, and PGI2 following stimulation with 0, 1.0, 50.0, and 100.0 micrograms/ml of Escherichia coli LPS. Kupffer cells were obtained from rat livers by enzymatic digestion with 0.05% collagenase followed by enrichment of the macrophage population on the basis of differences in density and adherence among the various cell populations isolated. Based on several criteria (phagocytosis of opsonized sheep erythrocytes, positive staining for esterase and peroxidase, failure to replicate), 95% of adherent cells were Kupffer cells. After 4 days of incubation, cells were stimulated with various doses of LPS for 4 and 8 hr. Prostanoid concentrations in culture supernatants were determined by radioimmunoassay. Increasing doses of LPS significantly (P less than 0.001) increased the concentration of immunoreactive PGE2 (iPGE2) and iTxB2 (the stable metabolite of TxA2). The concentration of i6-keto-PFG1 alpha (stable metabolite of PGI2) increased following stimulation with 1.0 microgram/ml of LPS, but declined as the dose of LPS was increased. The results provide evidence that endotoxin-activated Kupffer cells, like other macrophage populations, release several metabolites of arachidonic acid. Kupffer cell-derived prostanoids, particularly TxA2, may be important mediators of some of the pathophysiologic manifestations of acute endotoxemia.
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PMID:Prostanoid production by lipopolysaccharide-stimulated Kupffer cells. 388 37

With an increasing interest in the role of the monocyte-macrophage in the pathogenesis of atherosclerosis and as a progenitor of plaque intimal foam cells, a model for the study of foam-cell differentiation in an extravascular environment has been developed. Granulomas were induced in 25 normocholesterolemic (NC) and 28 hypercholesterolemic (HC) rabbits by the subcutaneous injection of 15 ml of 1% carrageenan. Granuloma tissue was harvested at 4, 7, 14, and 28 days and studied by light and transmission electron microscopy. Macrophages and foam cells were isolated by enzymic dispersion with collagenase and cultured for further characterization by scanning electron microscopy, nonspecific esterase (NSE), and oil red O (ORO) staining. Granuloma macrophages from NC rabbits were consistently ORO-negative, contrasting with those from HC rabbits which were strongly ORO-positive, even at 4 and 7 days. With an increasing duration of exposure to hypercholesterolemia, macrophages accumulated increasing amounts of stainable lipid, and in the 28-day HC granulomas, large foam cells distended by lipid inclusions accounted for 70% of the cells present. This model has established that NSE-positive macrophages in HC granulomas accumulate lipid and assume the morphologic characteristics of atheromatous intimal foam cells.
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PMID:Evolution of foam cells in subcutaneous rabbit carrageenan granulomas: I. Light-microscopic and ultrastructural study. 396 33

Antigen-retaining follicular dendritic cells (FDC) have been identified and studied in sections of lymph nodes and spleen, but studies of these cells in culture have been extremely limited. The purpose of this study was to establish techniques to release these fragile cells from mouse lymph nodes in a viable state and to identify these cells routinely in lymph node cell suspensions. FDC were obtained from passively or actively immunized popliteal lymph nodes of mice injected in the footpads with 125I-labeled human serum albumin (HSA) or horseradish peroxidase (HRP). Lymph nodes were removed 1 hr after the footpads had been injected with collagenase. After another hour of incubation in vitro with collagenase, protease, and deoxyribonuclease, FDC were released by gentle teasing and enriched by centrifugation on a low density bovine serum albumin (BSA) or Percoll gradient. Most FDC with the associated radiolabel floated at densities greater than 1.06 g/ml on BSA or Percoll gradients. Slides of the FDC-enriched fraction were prepared, using a cytobucket which allowed the cells to be affixed to glass slides by centrifugation in a less disruptive manner than by cytocentrifugation. FDC that were air-dried and fixed with 3% glutaraldehyde had a characteristic pink acidophilic cytoplasm after Wright's staining, and had a faintly basophilic euchromatic nucleus frequently with peripherally-clumped chromatin. In addition, these cells were large and irregularly shaped (up to 60 micron long). Fixation of FDC with 0.6% paraformaldehyde/ 0.9% glutaraldehyde on poly-L-lysine-coated slides resulted in a preservation of FDC which made possible visualization of long dendritic processes by Nomarski optics. Antigen presence on the cell surface was confirmed by autoradiography and, in the case of HRP, was also visualized enzymatically using diaminobenzidine. In contrast to resident peritoneal macrophages or some contaminating lymph node macrophages present on the same slides, FDC did not phagocytize opsonized sheep red blood cells (SRBC) or adhere to plastic surfaces although they did form rosettes with opsonized SRBC. Cell marker studies indicated FDC have a distinctive phenotype. They were positive for Ia, Fc receptor, and leukocyte common antigen, but negative for Thy-1, Ly-1, Ly-2, endogenous Ig, Mac-1, Mac-2, Mac-3, and F4/80, and negative to weakly positive for nonspecific esterase. Cultured FDC remained viable and retained radiolabeled antigen-antibody complexes on their surfaces and were significantly enriched for FDC.(ABSTRACT TRUNCATED AT 400 WORDS)
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PMID:Follicular dendritic cells in suspension: identification, enrichment, and initial characterization indicating immune complex trapping and lack of adherence and phagocytic activity. 396 23

In situ studies with the mouse macrophage (M phi)-specific antibody, F4/80, have shown that resident M phi in femoral bone marrow (RBMM) form hematopoietic islands with immature myelomonocytic and erythroid cells (Hume, D. A., et al. 1983. J. Exp. Med. 158: 1522). We have isolated these islands (clusters) by collagenase digestion, purified them from single cells by velocity sedimentation, and analyzed their cellular content. The clusters, ranging from 5- to 100 cells, constituted approximately 7% of the total nucleated cells, and greater than 70% contained at least one strongly staining, F4/80+ central M phi. In comparison, less than 26% showed reactivity for alkaline phosphatase, a marker of fibroblastoid reticulum cells. Compared with the nonclustering population, clusters were enriched with RBMM, fibroblastoid cells, and immature hematopoietic cells, but depleted of mature granulocytes and erythrocytes. The RBMM population was purified from other cells in clusters by selective adherence to glass and was compared with resident peritoneal M phi (RPM) for morphology and the presence of antigens, receptors, and enzymes. RBMM spread more extensively than RPM and frequently extended delicate plasma membrane processes. These and subsequent differences were not attributable to the collagenase treatment. Both M phi populations stained positively with antibodies F4/80 and 2.4G2 (Fc receptor IgG1/2b), bore mannosyl/fucosyl receptors, and showed reactivity for acid phosphatase and nonspecific esterase I. In contrast to RPM, RBMM had no detectable Mac-1 antigen (CR3) or complement receptors, but bore higher levels of Fc receptors (IgG2a and IgG2b) and Ia antigens. In addition, RBMM possessed a novel hemagglutinin activity for unopsonized sheep erythrocytes, which was not present on RPM. RBMM showed no respiratory burst activity in response to zymosan particles, but ingested them avidly. The growth properties of clustering and nonclustered populations were compared by measurement of [3H]thymidine incorporation and progenitor assays. Cells in clusters incorporated three- to fourfold more thymidine than nonclustered cells even in the absence of exogenous growth factors, and autoradiography demonstrated that RBMM made contact with proliferating cells. In contrast, the clusters contained over threefold fewer granulocyte/M phi progenitors compared with nonclustering cells. When clusters were cultivated for up to 3 d, there was rapid outgrowth of monocytes and fibroblastoid cells. These studies demonstrate that RBMM bear a distinct morphology and phenotype.(ABSTRACT TRUNCATED AT 400 WORDS)
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PMID:Isolation and characterization of resident stromal macrophages and hematopoietic cell clusters from mouse bone marrow. 403 89

Mononuclear cells were isolated from human peripheral blood by Ficoll-Hypaque centrifugation, and the cells adherent to plastic substrata were cultured in serum-free media supplemented with lactalbumin hydrolysate. These cell cultures, which consisted predominantly of monocyte-macrophages as judged by nonspecific esterase staining, accumulated collagenase in the medium. This collagenase resembled other vertebrate collagenases in that it cleaved native triple-helical type I collagen at a locus 3/4-length away from the amino-terminal end of the molecule. The collagenase activity was inhibited by Na2EDTA, dithiothreitol, and fetal calf serum, while the addition of Ca++ or N-ethylmaleimide enhanced the enzyme activity. The accumulation of collagenase in the culture media was markedly enhanced by the incubation of cells with concanavalin A or phorbol myristic acetate. In the presence of cycloheximide, the levels of collagenase activity were markedly reduced, suggesting that active protein synthesis was required to express the enzyme activity. In additional experiments, monocytes were further purified by counterflow centrifugation-elutriation. The collagenase production was markedly increased in cultures enriched in monocyte-macrophages and devoid of polymorphonuclear leukocytes. The accumulation of collagenase in monocyte cultures incubated for 48 hours in the presence of concanavalin A or phorbol myristic acetate was of the same order of magnitude as in parallel cultures containing the same number of polymorphonuclear leukocytes purified by Ficoll-Hypaque centrifugation and Plasmagel sedimentation.(ABSTRACT TRUNCATED AT 250 WORDS)
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PMID:The production of collagenase by adherent mononuclear cells cultured from human peripheral blood. 609 71


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