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)

Long-term (greater than 2 years) topical, conjunctival application of fluoresceinyl ovalbumin (FL-OA) induced allergic conjunctivitis-like lesions and hyperplasia of conjunctival-associated lymphoid tissue (CALT) in guinea pigs. Single-cell suspensions of CALT and spleen were prepared by collagenase digestion and cultured with or without FL-OA or lipopolysaccharide; the culture supernatants were assayed for IgG, IgA, IgM, and IgE antibody. Absolute values (ng Ab protein/ml) of anti-FL-OA IgG subclasses (IgG1 and IgG2) were measured using purified preparations of IgG1 and IgG2 anti-FL-OA antibody standards in an enzyme-linked immunosorbent assay. Immunohistochemical studies were performed using frozen sectioned CALT tissues as well as cultured single cells. IgG1, IgG2, IgA, IgM, but not IgE, anti-FL-OA antibodies were detected in the culture supernatants of both CALT and spleen. IgG- and IgA-secreting plasma cells were demonstrated in immunoperoxidase-stained CALT and single-cell cultures. The ratio of IgG1 to IgG2 isotypes produced by CALT in vitro was significantly higher than that produced by spleen and also that found in serum. These findings indicated that a site-specific regulation of antibody isotypes may exist within the hyperplastic CALT induced by the long-term topical exposure to FL-OA.
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PMID:Experimental allergic conjunctivitis: production of different isotypes of antibody by conjunctival-associated lymphoid tissue in culture. 327 15

During routine lower gastrointestinal endoscopy of children for suspected chronic inflammatory bowel disease, it is possible to visualize lymphoid follicles (Peyer's patches) in the last few centimeters of the terminal ileum. Biopsy specimens have been taken from these Peyer's patches and the lymphoid cells have been isolated by collagenase digestion. The mean cell yield of Peyer's patch cells was 1.7 x 10(8) lymphocytes/g tissue. Isolated Peyer's patch cells were 26%-54% CD3+ (pan T cells), 14%-34% CD4+ (helper/inducer T cells), and 9%-17% CD8+ (suppressor/cytotoxic T cells). Twenty six percent to 48% of the Peyer's patch cells were B cells; less than 1% contained cytoplasmic immunoglobulin A. When stimulated in vitro with phytohemagglutinin or pokeweed mitogen, cells from Peyer's patches proliferated. The successful isolation of functional cells from Peyer's patches will now allow studies to be done at the afferent limb of mucosal immunoregulation in humans.
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PMID:Selective biopsy of human Peyer's patches during ileal endoscopy. 350 86

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

Equine immunoglobulin was detected along the glomerular basement membrane of three human homograft recipients who had been treated with equine anti-lymphocyte globulin. Anti-lymphocyte globulins, given these patients, were obtained by immunization of horses with lymphocytes from human spleens and/or lymph nodes and contained glomerular basement membrane-reactive antibodies. Quantitative paired-label isotope experiments (in rats) demonstrated that 30-170 mug/ml of kidney-fixing antibodies were present in these preparations. The anti-lymphocyte globulins formed a line of identity with a sheep anti-human glomerular basement serum when reacted against collagenase-solubilized human glomerular basement membrane in double diffusion in agar. The renal fixation of these antibodies was blocked by absorption with human glomerular basement membrane, but not by buffy-coat leukocytes, indicating that they were directed specifically toward antigens in the basement membrane and were not cross-reacting anti-lymphocyte antibodies. Anti-lymphocyte globulin preparations for human use were studied for glomerular basement membrane-reactive antibodies by a direct immunofluorescent assay in rats. Anti-lymphocyte globulin from 13 of 20 horses, and 7 of 10 serum pools from horses immunized with lymphocytes derived from solid lymphoid organs (spleen, thymus, lymph node, tonsil), contained glomerular basement membrane-reactive antibodies. Sera from 18 horses injected with thoracic duct cells or cultured lymphoblasts had no glomerular basement membrane-reactive antibodies. An equine anti-human thymus serum containing glomerular basement membrane-reactive antibodies, which produced fatal glomerulonephritis in monkeys, was shown to cause both immediate and delayed glomerular injury in monkeys after intravenous injection. The reaction of this antibody with glomerular basement membrane in vivo was associated with little complement deposition in spite of the fact that the antibody could fix complement. This lack of glomerular complement fixation resulted from almost complete in vivo decomplementation of the monkeys receiving this anti-lymphocyte globulin.
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PMID:Glomerular basement membrane--reactive antibody in anti-lymphocyte globulin. 499 86

Colonic mucosal lymphoid cells, selectively enriched for intraepithelial (IEL) or lamina proprial lymphocytes (LPL), were isolated by sequential EDTA-collagenase treatment of resected human colons. Cytotoxic activities of colonic and peripheral blood lymphoid cells (PBL) were tested in three different assays, using chicken erythrocytes (CRBC) and Chang cells as targets. Antibody-dependent cell-mediated cytotoxicity (ADCC) and PHA-induced cytotoxicity (MICC) for both targets were shown by all the isolates of PBL, as was spontaneous cell-mediated cytotoxicity (SCMC) for Chang cells. However, no SCMC or ADCC for Chang cells was found with LPL, and IEL showed minimal or no activity in either assay. PBL, LPL and IEL demonstrated MICC for Chang cells but, contrasting with PBL and LPL, IEL showed no MCC for CRBC. No significant differences were found between the cytotoxic capabilities of colonic lymphoid cells from patients with inflammatory bowel disease and those from patients with other colonic diseases. Importantly, control studies with PBL showed that SCMC for Chang cells and ADCC for CRBC and Chang cells were reduced by collagenase treatment used in the isolation, of LPL. Also, SCMC for Chang cells was reduced by the treatment of PBL with EDTA. In contrast, neither EDTA nor collagenase reduced MICC for CRBC or Chang cells. Both forms of treatment induced variable degrees of cell losses in the PBL. By analogy, it can be implied that the isolation of intestinal mononuclear cells using EDTA and collagenase may influence some of their cytotoxic activities in vitro. This raises an important caveat in the interpretation of such studies.
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PMID:Human colonic intraepithelial and lamina proprial lymphocytes: cytotoxicity in vitro and the potential effects of the isolation method on their functional properties. 626 95

A microcytotoxicity assay for detection of thyroid-specific complement-dependent cytotoxic antibody (cytotoxic antibody), antibody-dependent cell-mediated cytotoxicity (ADCC) and direct lymphocyte cytotoxicity was developed using human, thyroid epithelial cells as targets. Thyroid tissue was obtained from patient with Graves' disease and was treated with collagenase and then trypsin. The red blood cells, interfollicular fibroblasts and infiltrating lymphoid cells in thyroid tissue from patients with Graves' disease could be removed by this procedure. To obtain entirely single cells as target cells, the suspension of dispersed thyroid epithelial cells was allowed to stand for 1 h at 4 degree C in culture medium to allow cell clumps to settle. For assay of cytotoxic antibody, a mixture of the single target cells, patient's serum and human complement was incubated in microwells for 18 h. After removing the detached cells, remaining target cells in the wells were fixed, stained and counted to assess cytotoxicity. For assay of ADCC and of direct lymphocyte cytotoxicity, target thyroid cells were precultured in the microwells for 18 h. Then effector cells with or without patient's serum were added and cultured further for 24-72 h. Cytotoxicity was assessed as described above. Adherence of effector cells to target thyroid cells sometimes disturbed the enumeration of target cells when the effector/target cell ratio was high. With this microassay system 3 different cytotoxic immune reactions against human thyroid cells could be measured quantitatively at the same time on 2-3 ml blood samples.
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PMID:A microcytotoxicity assay for thyroid-specific cytotoxic antibody, antibody-dependent cell-mediated cytotoxicity and direct lymphocyte cytotoxicity using human thyroid cells. 689 61

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

Attempts were made to isolate adherent phagocytic cells (macrophages) from mouse Peyer's patch cell suspensions. Cell suspensions prepared by teasing apart the Peyer's patches contained no adherent phagocytic cells. However, if Peyer's patch fragments were treated with collagenase to disrupt the tissue matrix, cells prepared in this way contained a subpopulation of adherent phagocytic cells. These cells comprised only 0.1-0.2% of the total nucleated cell population of the Peyer's patch. Similar cells could also be isolated from the Peyer's patches of germ-free mice, but as judged by their ability to ingest opsonized erythrocytes, these cells were less activated than cells from the Peyer's patches of normal mice. Adherent cells from the Peyer's patches of normal mice could present antigen (ovalbumin) to T cells, and Peyer's patches cell suspensions containing adherent cells could be stimulated in vitro to produce an anti-sheep red blood cell plaque-forming cell response in the absence of 2-mercaptoethanol. These studies show that although the frequency of phagocytic adherent cells is extremely low in Peyer's patches, these cells have functions consistent with that of adherent cells in other lymphoid tissues.
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PMID:Isolation and functional characteristics of adherent phagocytic cells from mouse Peyer's patches. 706 73

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


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