Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: UMLS:C0010346 (Crohn's disease)
 21,615 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

We have developed an enzymatic technique for isolating human intestinal mucosal lymphoid cells. This method was found to be superior to mechanical methods in regard to cell yield and survival. It is based on treating mucosa with serum-free solutions containing collagenase and deoxyribonuclease, followed by isolating the lymphoid cells through centrifugation steps involving fetal calf serum and ficoll-hypaque. Exposure of peripheral blood lymphocytes to the components of the enzymatic solution did not appreciably alter their uptake of tritiated thymidine in the presence or absence of mitogens. Application of the method to derive lymphoid cells from Crohn's disease, ulcerative colitis, and normal intestinal mucosa has shown that gut mucosal lymphocytes from inflammatory bowel disease (1) exceed the number of those from normal mucosa by a factor of 3 to 5; (2) show different degrees of tritiated thymidine uptake, spontaneously and in response to mitogens, depending upon the time they are harvested during the dissociation process; (3) are better stimulators than responders in the allogeneic mixed lymphocyte reaction; (4) generate suppressor cell activity comparable to that of peripheral blood lymphocytes; (5) cannot, in contrast to peripheral blood lymphocytes, generate antibody-dependent cell mediated cytotoxicity; and (6) produce an average of 5 times more IgM than equal numbers of peripheral blood lymphocytes.
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PMID:Gut mucosal lymphocytes in inflammatory bowel disease: isolation and preliminary functional characterization. 15 97

Many patients with Crohn's disease have suppressor T cells circulating in the peripheral blood that are potent inhibitors of immunoglobulin synthesis in vitro. The purpose of this study was to examine the regulatory effects of T cells isolated from the lamina propria of patients with Crohn's disease or of patients with other diseases. Lamina propria cells were isolated from surgically resected intestine by sequential ethylenediaminetetraacetic acid and collagenase incubations. T cells were purified from lamina propria either by anti-F(ab)2' affinity columns or by a panning technique using monoclonal anti-T cell antibodies. Changes in the pokeweed mitogen-stimulated synthesis of immunoglobulin M, immunoglobulin G, and immunoglobulin A by normal peripheral blood indicator lymphocytes, as measured by sandwich enzyme-linked immunosorbent assay, was used as an index of help and suppression. Helper T-cell activity was sought by coculturing lamina propria T cells with normal peripheral blood cells plus pokeweed mitogen. Helper T-cell activity was found among lamina propria T-cell populations of patients and controls to a roughly comparable extent; help was provided for all isotypes. Suppressor T-cell activity was sought by coculturing lamina propria T-cells with normal B cells plus irradiated normal T cells plus pokeweed mitogen. No significant suppressor T-cell activity was observed in these cocultures, whether the cells were obtained from control intestine, the grossly uninvolved margin of Crohn's disease intestine, or actively inflamed Crohn's disease intestine. We conclude that suppressor T-cell activity of the sort found previously in the peripheral blood of patients with Crohn's disease is not demonstrable in the intestinal lesions of Crohn's disease. Helper T-cell activity is the predominant regulatory activity in both control and inflamed intestinal lamina propria.
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PMID:T cell-B cell regulation in the intestinal lamina propria in Crohn's disease. 286 Nov 39

There are no known intestinal cell phenotypic markers characteristic of the transmural idiopathic inflammatory disease of the small bowel. To address this issue, we developed monoclonal antibodies specific for Crohn's disease tissue by generating a library of hybridomas specific for intestine following murine immunization with intestinal epithelial cells from a patient with Crohn's disease. The epithelial cells were initially harvested from a fresh operative specimen by gentle scraping and digestion with 1% collagenase on ice. Cells were then washed, evaluated for viability, and polytron homogenized. After immunization and subsequent fusion, hybridoma cell lines producing distinct antibody-binding patterns were identified by indirect immunoepifluorescence (IIEF). Wells representing distinct patterns were subcloned and carried to limiting dilution. Of the multiple hybridomas studied, the predominant target antibodies were goblet cell and glycoprotein-like molecules. Patterns of antibody binding identified by IIEF included: brush border, goblet cell, surface glycoprotein, enterocyte membranes, basilar crypt inclusions, circumferential goblet cell, and a heterogeneous goblet cell glycoprotein pattern. Molecular weight determination and cross-reactivity with various human tissues were studied by immunoblotting following sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis. Several hybridomas were specific for the intestine but were not specific for Crohn's disease.
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PMID:Generation of monoclonal antibodies to involved ileum of Crohn's disease: characterization of a panel of antibodies with goblet cell membrane and brush border-specific reactivity. 304 35

A sensitive 4 h 51Cr-release cytotoxicity assay has been developed using as targets colonic epithelial cells obtained by Dispase-collagenase digestion of resected mucosa or colonoscopic biopsies. Peripheral blood mononuclear cells (MNC) from most healthy donors showed low, but significant levels of cytotoxicity for normal epithelial cell target cells of 8.7 (4.4) % (mean (SD] and similar levels were found in 14 ulcerative colitis (6.5 (4.4) %) and 16 Crohn's disease (6.2 (5.2) %) patients. Neither drug therapy nor disease activity influenced the results. The sensitivity of colonic epithelial cells isolated from inflamed and histologically normal mucosa to lysis by peripheral blood MNC from a single donor was not affected by the underlying disease. Anti-epithelial cell activity did not correlate with anti-K562 activity and the cytotoxic cell was plastic non-adherent and Leu-11b-. None of 15 MNC populations isolated from mucosa of normal, tumour bearing, or chronically inflamed intestine exhibited significant lysis of colonic epithelial cells despite killing of K562 target cells in 10. Lymphokine activated killer (LAK) cells, generated by interleukin-2 stimulation in vitro of nine intestinal and seven peripheral blood MNC populations, exhibited high levels of lysis of K562 cells but, on every occasion, failed to lyse colonic epithelial cells. These data indicate that spontaneously cytotoxic or LAK cells are unlikely to play a role in the generation of colonic epithelial cell injury by direct cytotoxicity in inflammatory bowel disease.
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PMID:Lysis of colonic epithelial cells by allogeneic mononuclear and lymphokine activated killer cells derived from peripheral blood and intestinal mucosa: evidence against a pathogenic role in inflammatory bowel disease. 326 5

The leakiness of the cell membranes of colonic epithelial cells isolated by the collagenase/Dispase technique from normal or diseased colons was assessed in a 4 h 51Cr release assay. Cells from normal, adenoma bearing or cancer bearing colons showed 51Cr release of 8% or less in almost all of 46 cell populations tested. In contrast, cells from mucosa affected by ulcerative colitis [11.9 (4.3%) n = 23] or Crohn's disease [8.4 (2.7%) n = 18] released significantly more 51Cr than the non-inflamed groups. Values are expressed as mean (SD). Overall, release values were greater in ulcerative colitis than Crohn's disease (p less than 0.01). In Crohn's disease, cells obtained from histologically inflamed mucosa released significantly more 51Cr [9.7 (2.5%) n = 11] than those from non-inflamed mucosa [6.4 (1.5%) n = 7, p less than 0.02] whereas, in ulcerative colitis, abnormal release values were found in 8 of 13 cell populations isolated from mucosa showing no histological evidence of active disease. In five patients with distal ulcerative colitis, cells from mucosa not apparently involved demonstrated normal 51Cr release in four of five studies despite abnormal release from cells from involved mucosa suggesting that a diffuse abnormality of the colonic epithelial cell is not usually present. These data indicate that chronic mucosal inflammation per se is associated with abnormalities of the colonic epithelial cell but that, in ulcerative colitis, the abnormality remains in many patients with quiescent disease. Identification of the local factors responsible for such an abnormality may contribute to an understanding of the pathogenesis of ulcerative colitis.
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PMID:Ulcerative colitis--a disease characterised by the abnormal colonic epithelial cell? 337 20

Lamina proprial lymphocytes (LPL), isolated by an EDTA-collagenase technique from patients with various colonic diseases, were investigated for LMIF release in vitro. On stimulation with the preparation of Kunin antigen, macrophage-depleted LPL from patients with severely or moderately active ulcerative colitis showed LMIF release which was significantly greater than that observed using LPL from patients with mild colitis or from those with other diseases of the large bowel, including Crohn's disease. Results similar to those obtained with LPL were found with the corresponding peripheral blood lymphocytes (PBL) stimulated by the preparation of Kunin antigen. In contrast, nonspecific stimulation in vitro with Concanavalin A showed no differences in LMIF releases by the LPL or PBL in the various disease groups. It is suggested that hypersensitivity to Kunin antigen may have pathogenic significance in ulcerative colitis.
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PMID:Leukocyte migration inhibitory factor (LMIF) release by human colonic lymphocytes. 627 Oct 90

Mononuclear cells were isolated from the mucosa and submucosa of small intestine and colon of 22 subjects with localized, anatomically remote disease and four subjects with Crohn's disease (nine specimens) by sequential treatment with EDTA and collagenase. The effects of isolation techniques on cell yields and viability were examined. Secretion of specific IgA, IgM and IgG antibodies to common faecal Escherichia coli strains by individual mononuclear cells was studied using a haemolytic plaque assay. A majority of specific antibody secreting cells secreted IgA antibody. This response was greatest and most consistent in the distal colon but extended from stomach to rectum. There was no evidence of a primary defect in IgA antibody response in the few subjects with Crohn's disease available for study.
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PMID:Escherichia coli antibody-secreting cells in the human intestine. 680 77

The mucosa associated lymphoid tissues of the intestinal lamina propria or the bronchial mucosa, respectively, represent a separated and well defined immunologic compartment. Due to highly specialized functions, subpopulations of lymphoid cells are distributed unevenly between the compartments and unique regulatory mechanisms developed to sustain integrity of mucosal surfaces. With this study we investigate whether an omentum associated lymphoid tissue exists and whether it partakes in immunologic processes involved in the perpetuative intestinal inflammation in Crohn's disease. Mononuclear cells from surgically resected omentum tissue (10 patients with Crohn's disease, 10 patients with malignomas or inflammatory control diseases (diverticulitis)) were isolated by collagenase digestion and subsequent serial density centrifugation. Phenotypic analysis was carried out by immunofluorescent labeling with a panel of monoclonal antibodies as well as peanut agglutinin. Most interestingly, the percentage of CD4-T(Helper) cells among omentum mononuclear cells was decreased in comparison with peripheral blood mononuclear cells, whereas the percentage of monocytes/macrophages and of natural killer cells appeared to be increased. In comparison with normal peripheral blood a higher percentage of normal omentum mononuclear cells were activated. It thus appears that a defined immunologic compartment exists which is different in its cellular composition from peripheral blood as well as from intestinal mucosa associated lymphoid tissue and which may be called omentum associated lymphoid tissue. In Crohn's disease subpopulations of omentum mononuclear cells did not change in number, however immunologic activation increased further and appears to be highly increased in comparison to both omentum cells from disease specificity controls (diverticulitis) and Crohn's disease peripheral blood cells. We conclude that omentum associated lymphoid tissue may be described as an unique immunologic compartment which may play a role in activating events in chronic intestinal inflammation in Crohn's disease. Further studies will address functional characteristics of the omentum associated lymphoid tissue and will investigate regulatory mechanisms which may contribute to the inflammatory process in Crohn's disease.
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PMID:[Activation of the major omentum-associated lymphoid tissue in Crohn disease]. 770 65

Butyrate promotes epithelial cell healing and improves symptoms when administered rectally in patients with distal ulcerative colitis (UC). It was hypothesized that butyrate may enhance healing in patients with UC by stimulating colonocyte proliferation and/or protein production. Mucosa from the descending colon was obtained from patients with UC (n = 5), Crohn's disease (n = 8), diverticulitis (n = 6), and cancer (normal tissue 10 cm from tumor; n = 10). Epithelial cells were isolated using dispase/collagenase and differential sedimentation and incubated for 4 hr at 37 degrees C with either Na butyrate (10 mM) or NaCl (10 mM). Protein synthesis was assessed by [14C]leucine incorporation and proliferation was determined with [3H]thymidine. Mean viability and purity were >88%. Spontaneous proliferation was significantly increased in UC when compared to diverticulitis and normal controls. Butyrate significantly increased protein synthesis in UC epithelial cells when compared to saline control. The therapeutic effects of butyrate in patients with UC may be due to its use by epithelial cells as a metabolic fuel to increase protein production and promote healing.
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PMID:Butyrate increases colonocyte protein synthesis in ulcerative colitis. 804 Nov 40

Very little is currently known regarding the underlying mechanisms involved in the etiology of intestinal strictures in chronic inflammatory bowel disease. The deposition of extracellular matrix components, especially collagens, is thought to play an important role in the etiology of intestinal strictures. The main goal of this study was therefore to investigate the collagen metabolism in patients with inflammatory bowel disease using the in situ hybridization technique. We determined de novo synthesis of the (pro)-collagen mRNA transcripts alpha 1I, alpha 1III, alpha 1IV, alpha 2V as well as the collagen degrading enzyme mRNA transcripts collagenase type I and IV in Crohn's disease and ulcerative colitis and compared the rate of expression semiquantitatively to healthy controls. We found a significant increase of all (pro)-collagen transcripts tested in Crohn's disease and ulcerative colitis as compared to healthy control tissues, indicating an increased de novo synthesis of all collagens in both inflammatory bowel diseases. However, we observed a significant difference in the expression of the collagenase mRNA transcripts between Crohn's disease and ulcerative colitis. Compared to healthy control subjects we were unable to detect a significant difference in the expression of collagenase type I and IV in Crohn's disease; in contrast, we observed a significant increase in the rate of expression for the collagenases in ulcerative colitis as compared to controls or biopsies from patients with Crohn's disease.(ABSTRACT TRUNCATED AT 250 WORDS)
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PMID:[Indications for different collagen metabolism in Crohn disease and ulcerative colitis]. 849 73


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