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)

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

Spontaneous cell-mediated cytotoxicity (SCMC) and the marker of natural killer (NK) cells mediating SCMC of the human large intestine were studied. Lamina proprial lymphoid cells (LPL) were isolated by sequential dithiothreitol-EDTA-collagenase treatment of the gut specimen. SCMC was measured by the chromium release method. Target cells included P4788 in monolayer, a cell line derived from colon cancer, Chang cells in monolayer, and K562 in suspension. Target cells in monolayer including colon cancer cell line were chosen because they were thought to be more appropriate to assess SCMC for lymphoid cells in the solid organ. While lower compared to cytotoxicities (CT) by peripheral blood lymphoid cells (PBL), define CT were observed in LPL against all three targets. NK cells marker was studied both on LPL by an indirect fluorescent antibody method and on the gut tissue by indirect immunoperoxidase staining using anti HNK-1 monoclonal antibody which defines virtually all NK cells. HNK-1 positive (HNK-1 +) cells were identified in both methods. HNK-1 + cells were observed in the epithelium, lamina propria, and lymph follicle with or without germinal centers. These results clearly demonstrated the presence of SCMC and HNK-1 + cells in the human large bowel.
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PMID:Definite spontaneous cell-mediated cytotoxicity and HNK-1 cells in the human large intestine. 355 47

Pinocytosis and prostaglandin E2 production are two major functions of the mononuclear phagocyte system. The goal of this study was to compare the pinocytosis of horseradish peroxidase and the prostaglandin E2 production between the hepatic and peritoneal resident macrophages in the mouse. Hepatic resident macrophages were isolated by collagenase digestion, differential centrifugation and adherence. Peritoneal resident macrophages were isolated by peritoneal cavity washing followed by adherence. Horseradish peroxidase was endocytosed by hepatic macrophages at a significantly higher rate (118 +/- 12 ng/10(6) cells/60 min) than by peritoneal macrophages (21 +/- 4 ng/10(6) cells/60 min). Prostaglandin E2 production was measured in the culture medium of unstimulated and lymphokine-stimulated hepatic and peritoneal resident macrophages. Prostaglandin E2 concentration in the culture medium of unstimulated peritoneal macrophages was 36.6 +/- 26.8 ng/ml after a 24 h incubation. It was increased by 83 p. 100 in presence of a lymphokine-enriched secondary mixed lymphocyte culture supernatant. In contrast, hepatic macrophages did not produce any significant amount of prostaglandin E2, even if they were incubated in presence of lymphokines. This study shows that hepatic resident macrophages have a higher pinocytic capacity than peritoneal resident macrophages, suggesting that the role of the liver in the clearance of gut-derived antigens is not only due to its portal irrigation but also to the presence of macrophages highly differentiated in their endocytotic properties. The lack of prostaglandin E2 production in hepatic macrophages, in basal conditions as well as after lymphokine-stimulation, suggests that these cells play a minor role in the regulation of the immune response.(ABSTRACT TRUNCATED AT 250 WORDS)
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PMID:[Comparative study of pinocytosis and the production of prostaglandin E2 by hepatic and peritoneal resident macrophages in mice]. 386 Apr 55

Methods have been determined for the isolation, purification and subsequent characterization of separate populations of rat intestinal lymphoid cells, namely intraepithelial (IEL), lamina propria (LPL) and Peyer's patch lymphocytes (PPL). Dissociation of the epithelium from the basement membrane with subsequent release of IEL was achieved by citrate buffer incubation followed by vortex agitation. LPL were released from the remaining tissue by scraping, and PPL were similarly obtained. Some preparations of lamina propria were further subjected to collagenase digestion. After filtration and density gradient centrifugation, average yields of 220 x 10(4) IEL, 54 x 10(4) LPL and 220 x 10(4) PPL per gram of gut were obtained. Immunofluorescence characterization demonstrated that cells bearing the MRC OX8 (T-suppressor) marker predominated in IE1 (73%) and were present in lower concentrations in LPL (26%) and PPL (6%). Cells with the W3/25 (T-helper) marker accounted for a small proportion of each of the lymphocyte preparations. IE1 were unusual in containing a population of cells which were negative for the W3/13 marker for T cells, but were MRC OX8 positive. B lymphocytes were present in PPL (55%) and LPL (31%), but were virtually absent in IEL (less than 1%). Few plasma cells were observed. The techniques described will allow functional investigations to be made and lead to a better understanding of mucosal immunity.
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PMID:Intraepithelial, lamina propria and Peyer's patch lymphocytes of the rat small intestine: isolation and characterization in terms of immunoglobulin markers and receptors for monoclonal antibodies. 704 Feb 14

Highly purified populations of lymphocytes were obtained from the murine intestinal mucosa using EDTA-collagenase isolation procedures in combination with discontinuous density centrifugation. Intraepithelial lymphocytes (IEL) were separated from lamina propria lymphocytes (LPL) and, within these two populations, fractions enriched or depleted in gut granular lymphocytes (gGL) were obtained. Using these cells in cytotoxic assays, it was shown that both IEL and LPL possess natural killer (NK) activity, and this was associated with gGL. The major effector cells of gut NK activity appeared to be Thy-1.2+, Lyt-1.1-, and Lyt-2.1-. The susceptibility of gut NK cells to anti-Thy-1.2 plus complement (C) was significantly higher than that of splenic NK cells. In contrast, anti-asialo GM1 and anti-NK-1.2 plus C only slightly affected the gut NK activity. Thus, the phenotype of the gut NK cells appears to be different from the splenic one and provides further evidence for NK heterogeneity and establishes the compartmentalization of one NK subpopulation. Beige mice, deficient in splenic NK activity, also had very low gut NK activity. W/Wv mice, which lack mast cell precursors, had normal numbers of gGL and diminished, but still present, gut and splenic NK activity. This deficiency did not segregate with the genes responsible for the basic hemopoietic stem cell defect, and these results argue against a close ontogenetic relationship between IEL, gGL, and intestinal mucosal mast cells. The relevance of these observations to the cell lineage of the effector cell of gut NK activity is discussed.
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PMID:Characteristics of natural killer cells in the murine intestinal epithelium and lamina propria. 707 24

A technique is described, involving sequential treatment of the human colonic mucosa with EDTA in calcium-magnesium-free medium, and with collagenase, to isolate lymphoid cells enriched for intraepithelial (IEL) or lamina proprial lymphocytes (LPL). The IEL and LPL isolates also contained small numbers of eosinophils, mast cells, neutrophils, and macrophages. Plasma cells were present in the LPL but not in the IEL. The IEL isolates contained approximately equal proportions of T, B, and null cells. In contrast, the LPL suspensions contained 52% of T cells, 22% of B cells, and 26% of null cells. The most prevalent membrane immunoglobulin in the two colonic lymphoid cell suspensions was IgA (IEL--53%; LPL--71%). In colonic tissue sections, the percentages of immunoglobulin-containing cells as well as the proportions of cells containing IgA, both in the epithelial layer and the lamina propria, were similar to those found in the suspensions of lymphocytes stained for membrane immunoglobulin. These and other morphologic and characterization data support the contention that the two colonic lymphoid cell populations, obtained by the isolation procedures, were selectively enriched for intraepithelial or lamina proprial lymphocytes, respectively. Thus, this technique provides an important tool for further studies of the functional properties of the gut-associated lymphoid tissues.
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PMID:Isolation and characterization of colonic intraepithelial and lamina proprial lymphocytes. 738 Feb 3

Microvascular endothelial cells play an important part in inflammation as well as in organ specific leucocyte traffic, and may be functionally different from large vessel endothelium in this respect. This study therefore established a method for isolation and longterm culture of human intestinal microvascular endothelial cells (HIMEC). After dissociation by collagenase/dispase/DNase of mucosal and submucosal tissue obtained from normal adult jejunum, cells were plated and cultured to subconfluence in endothelial serum free medium containing 2.5% fetal calf serum, hydrocortisone, and N6, O2-dibutyryladenosine cyclic monophosphate. Primary cultures were trypsinised and endothelial cells were isolated by paramagnetic beads armed with monoclonal antibody to CD31. Optimal growth conditions for HIMEC cultures were established, allowing up to nine passages (three months in vitro). The cells contained Weibel-Palade bodies, expressed von Willebrand factor, CD31, and VE-cadherin; and bound Ulex Europaeus lectin I. A method to establish longterm cell cultures of HIMEC will facilitate further investigation of the function of intestinal endothelial cells and their participation in physiological and pathological events in the gut.
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PMID:Isolation and longterm culture of human intestinal microvascular endothelial cells. 755 73

The jejunum and ileum of 5 day old and adult normal pigs and of 45 day old germ free pigs were used to study the lymphocyte pools in the epithelium and lamina propria by sequential treatments with EDTA, four hours, and 12 hours of collagenase treatment. In adult animals the incubation of the jejunal wall with EDTA resulted in mean (SD) 26.8 (10.9) x 10(6) intraepithelial lymphocytes per g of tissue. The ileal wall gave lower cell yields. After complete digestion of the lamina propria by collagenase a further yield of 35.2 (10.2) x 10(6)/g lymphocytes was achieved. The separation of the gut wall from 5 day old pigs resulted in a 10-fold lower total lymphocyte yield, and the tissue was totally digested after four hours of collagenase treatment. Many eosinophils and mast cells were found in the suspensions from adult animal tissues after the collagenase treatment; 4.7 x 10(6)/g and 4.8 x 10(6)/g, respectively. The suspensions after 12 hour collagenase incubation contained up to 30% plasma cells. Almost all cells isolated by EDTA incubation were CD8+ T cells. After collagenase incubation CD4+ and CD8+ T lymphocytes were found in all animal groups, and in adult animals up to 20% surface Ig+ cells were harvested. When the incorporation of the thymidine analogue bromodesoxyuridine was used to study the lymphocyte production in vivo 3 to 7% lymphocytes in the epithelium were labelled 24 hours later (lamina propria T lymphocytes about 1%). In this study lymphoid as well as non-lymphoid cells have been analysed in mucosal cell suspensions. The absolute cell yield per gram of mucosal tissue is a basis to estimate the pool sizes of intraepithelial and lamina propria lymphocytes.
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PMID:Lymphoid and non-lymphoid cells in the epithelium and lamina propria of intestinal mucosa of pigs. 782 77

Labial salivary glands are found in the majority of insects. They are relatively large, extend back into the thorax, and in Rhodnius, they are cherry red in color due to a pigment derived from traces of hemoglobin absorbed form the gut. In most insects they are acinous shaped, with long excretion channels that present differentiated regions which from salivary reservoirs. The glands may be relatively simple or complexly branched and convoluted. In Rhodnius they are described as being unilobed with no traces of division. The main duct leaves the gland at its anterior extremity. The acini have different kinds of cells but all of them are seen as sources of secretion. Our material has a different shape due to the fact that the animals spent 20 days under starvation conditions. New data are also obtained through treatment with collagenase and HCl. The importance of the study of these glands lies in the fact that it will further understanding of the transmission of Chagas' disease.
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PMID:Scanning electron microscopy of the Rhodnius neglectus (Hemiptera) labial salivary glands after starvation. 825 Feb 70

Monoclonal antibodies (mAbs) were raised by injection of a homogenate of cultured growth cartilage (GC) cells from young rabbit ribs. These mAbs were examined by immunohistochemical staining for their reactivity to paraffin sections of rabbit tissues. The results showed that an mAb reacted preferentially with late hypertrophic and calcified costal GC zones. The mAb also reacted with hypertrophic GC adjacent to bone that existed in sternum and femur, but not to other cartilages, including resting cartilage, articular cartilage, auricular cartilage, nasal cartilage, tracheal cartilage and meniscus cartilage, or with other tissues, including tendon, skin, muscles, lung, liver, heart, thymus, spleen, eye and gut. It reacted with a wider area of the GC zone when the sections were decalcified, although its reactivity with the extended area was much less intensive than that with late hypertrophic and calcified GC zones. On treatment of the sections with bacterial collagenase, neither the reactive area nor its intensity were changed, while when treated with trypsin the reactivity was lost. These results suggest the existence of a certain molecule which distinguishes GC (osteogenic cartilage) from other (non-osteogenic) cartilage. This mAb is a useful probe for distinguishing osteogenic cartilage from non-osteogenic cartilage, and for studying differentiation steps of cartilage cells in endochondral bone formation. The mAb can also be used as a probe for clinical and stored specimens because it reacts with decalcified and paraffin-embedded human specimens.
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PMID:A monoclonal antibody distinguishes growth cartilage from other types of cartilage: a new probe for osteogenic cartilage. 846 88


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