Colon Epithelial Protein Research Articles

Protective effects of glycine against lipopolysaccharide-induced intestinal apoptosis and inflammation.

Intestinal dysfunction is commonly observed in humans and animals. Glycine (Gly) is a functional amino acid with anti-inflammatory and anti-apoptotic properties. The objective of this study was to test the protective effects of Gly against lipopolysaccharide (LPS)-induced intestinal injury. 28 C57BL/6 mice with a body weight (BW) of 18 ± 2g were randomly assigned into four groups: CON (control), GLY (orally administered Gly, 5g/kg BW/day for 6days), LPS (5mg/kg BW on day 7, i. p.), and GLY + LPS (Gly pretreatment and LPS administration). Histological alterations, inflammatory responses, epithelial cell apoptosis, and changes of the intestinal microbiota were analyzed. Results showed that, compared with the CON group, mice in the LPS treatment group showed decreased villus height, increased crypt depth, and decreased ratio of villus height to crypt depth, which were significantly attenuated by Gly. Neither LPS nor Gly treatment altered morphology of the distal colon tissues. LPS increased the apoptosis of jejunum and colon epithelial cells and protein abundance of cleaved caspase3 in the jejunum, which were markedly abrogated by Gly. LPS also elevated the mRNA levels of Toll-like receptor 4 (TLR4), myeloid differentiation factor 88 (MYD88), pro-inflammatory cytokines, and chemokines in the jejunum and colon. These alterations were significantly suppressed by Gly. In addition, Gly supplementation attenuated infiltration of CD4+, CD8+ T-lymphocytes, CD11b+ and F4/80+ macrophages in the colon. Furthermore, Gly increased the relative abundance of Mucispirillum, Lachnospiraceae-NK4A136-group, Anaerotruncus, Faecalibaculum, Ruminococcaceae-UCG-014, and decreased the abundance of Bacteroides at genus level. Supplementation with Gly might be a nutritional strategy to ameliorate LPS-induced intestinal injury in mice.

Predictors of recurrent specialized intestinal metaplasia after complete laser ablation.

The aim of this study was to determine whether specialized intestinal metaplasia recurs after complete laser ablation and to evaluate the persistence of colon epithelial protein in esophageal mucosa after laser ablation as a predictor of recurrence. A total of 31 patients with specialized intestinal metaplasia (Barrett's esophagus) underwent laser photoablation. Investigators without knowledge of treatment status evaluated serial hematoxylin and eosin-stained slides, Alcian blue-stained slides, and immunohistochemistry for the detection of colon epithelial protein (mAb Das-1). Endoscopic ablation of specialized intestinal epithelium was accomplished in 21 patients after 6.5 +/- 1.2 laser sessions. Complications included one perforation, one UGI bleed and one stricture. Of eight post-laser recurrences, seven were successfully re-ablated; one developed adenocarcinoma requiring esophageal resection. Cardia-type mucosa was present by biopsy at the time of complete ablation in all eight recurrent cases despite a normal endoscopic appearance. Colon epithelial protein was detected in all 31 patients before ablation, six of 21 completely ablated patients before they recurred and all eight recurrences. Only two of 15 patients, colon epithelial protein negative at the time of complete ablation, developed recurrent Barrett's esophagus. Thus, cardia-type mucosa and persistent colon epithelial protein staining after complete ablation of specialized intestinal epithelium were predictors of future recurrence (p < 0.001). Specialized intestinal epithelium was ablated by neodymium:yttrium-aluminum-garnet laser but recurred in eight of 21 (38%) of patients. Colon epithelial protein was present in all primary (31 of 31) and all recurrent (eight of eight) Barrett's esophagus. Recurrent specialized intestinal metaplasia may be deep to squamous epithelium. Replacement of specialized intestinal mucosa by cardia-type mucosa and persistence of colonic epithelial protein are predictors of recurrent specialized intestinal mucosa before its endoscopic or histological detection. Laser ablation of Barrett's epithelium is an investigational intervention that should be restricted to research protocols.

Predictors of recurrent specialized intestinal metaplasia after complete laser ablation

Objectives The aim of this study was to determine whether specialized intestinal metaplasia recurs after complete laser ablation and to evaluate the persistence of colon epithelial protein in esophageal mucosa after laser ablation as a predictor of recurrence. Methods A total of 31 patients with specialized intestinal metaplasia (Barrett’s esophagus) underwent laser photoablation. Investigators without knowledge of treatment status evaluated serial hematoxylin and eosin–stained slides, Alcian blue–stained slides, and immunohistochemistry for the detection of colon epithelial protein (mAb Das-1). Results Endoscopic ablation of specialized intestinal epithelium was accomplished in 21 patients after 6.5 ± 1.2 laser sessions. Complications included one perforation, one UGI bleed and one stricture. Of eight post-laser recurrences, seven were successfully re-ablated; one developed adenocarcinoma requiring esophageal resection. Cardia-type mucosa was present by biopsy at the time of complete ablation in all eight recurrent cases despite a normal endoscopic appearance. Colon epithelial protein was detected in all 31 patients before ablation, six of 21 completely ablated patients before they recurred and all eight recurrences. Only two of 15 patients, colon epithelial protein negative at the time of complete ablation, developed recurrent Barrett’s esophagus. Thus, cardia-type mucosa and persistent colon epithelial protein staining after complete ablation of specialized intestinal epithelium were predictors of future recurrence (p < 0.001). Conclusions Specialized intestinal epithelium was ablated by neodymium:yttrium-aluminum-garnet laser but recurred in eight of 21 (38%) of patients. Colon epithelial protein was present in all primary (31 of 31) and all recurrent (eight of eight) Barrett’s esophagus. Recurrent specialized intestinal metaplasia may be deep to squamous epithelium. Replacement of specialized intestinal mucosa by cardia-type mucosa and persistence of colonic epithelial protein are predictors of recurrent specialized intestinal mucosa before its endoscopic or histological detection. Laser ablation of Barrett’s epithelium is an investigational intervention that should be restricted to research protocols.

Externalization of tropomyosin isoform 5 in colon epithelial cells.

Ulcerative colitis (UC) is associated with autoantibody response to a cytoskeletal protein, human tropomyosin (hTM) isoform-5 (hTM5). Because hTM5 is an intracellular protein, it may remain inaccessible to the autoantibodies. Therefore, we have investigated the possibility of externalization of hTM5 in colon epithelial cells. Freshly isolated colonic and small intestinal epithelial cells and LS-180 colon cancer cell line were examined for surface expression of hTM5 by flow cytometric analysis using hTM isoform-specific MoAbs. The extracellular release of hTM5 was determined by Western blot and radioimmunoprecipitation analyses. Physical association of hTM5 with a membrane-associated colon epithelial protein (CEP) was examined by co-immunoprecipitation of hTM5 with anti-CEP MoAb, and CEP with anti-hTM5 MoAb. Cell surface expression of hTM5 was observed in colonic epithelial and LS-180 cells but not in small intestinal epithelial cells. LS-180 cells spontaneously released hTM5 as well as CEP into the culture medium that was significantly stimulated by a calcium ionophore, A23187, but inhibited by phorbol-12-myristate-13-acetate, monensin and methylamine. Co-immunoprecipitation experiments revealed that hTM5 forms a complex with CEP. We conclude that hTM5 is externalized in colon but not in small intestinal epithelial cells. The physical association of hTM5 with CEP suggests a possible chaperone function of CEP in the transport of hTM5, a putative target autoantigen in UC.

Colon epithelial cellular protein induces oral tolerance in the experimental model of colitis by trinitrobenzene sulfonic acid

Rectal administration of trinitrobenzene sulfonic acid (TNBS) produces chronic co-litis in experimental animals. However, the role of epithelial cellular protein(s) in this model is unknown. We examined whether oral tolerance can be induced in this model with colon epithelial cell proteins and whether it is organ specific. Rats were fed five times with extracts of LS-180 human colon cancer cells or HT 1080 human fibroblast cells. Syngeneic normal rat colon or small intestinal extracts were fed to separate groups of rats. After oral feedings, each rat received TNBS by enema. Rats were killed 15 days later, and the following were measured: gross and histologic disease score, weight, thickness, and myeloperoxidase values of colon and serum interferon-γ (IFN-γ) and transforming growth factor-β (TGF-β) levels. Rectal TNBS alone produced severe colitis with a 26% mortality rate. Rats fed LS-180 or rat colon extract before TNBS enema were protected, as evidenced by reductions in mortality rate, disease scores, and myeloperoxidase values. However, rats fed HT 1080 or small intestine extract lacked such protection. To examine the possible mechanism of the oral tolerance, T lymphocytes from mesenteric lymph nodes and spleen of LS-180 extract-fed rats were passively transferred to naive rats, and this was followed by TNBS enema. These rats showed clear protection. Protected animals had low IFN-γ and high TGF-β levels. This study demonstrates that cellular protein(s) from human colon epithelial cells, but not from human fibroblasts, can induce oral tolerance in experimental colitis. This oral tolerance is mediated by primed mesenteric and splenic T lymphocytes. (J Lab Clin Med 2001;138:257-69)

This month in J Lab Clin Med: Issue highlights for October 2001

This month in J Lab Clin Med: Issue highlights for October 2001

A colon epithelial protein(s) induces oral tolerance in a dose dependent manner in the trinitrobenzene sulfonic acid (TNBS) model of colitis in rats

A colon epithelial protein(s) induces oral tolerance in a dose dependent manner in the trinitrobenzene sulfonic acid (TNBS) model of colitis in rats

A colon epithelial protein(s) induces oral tolerance in a dose dependent manner in the trinitrobenzene sulfonic acid (TNBS) model of colitis in rats

A colon epithelial protein(s) induces oral tolerance in a dose dependent manner in the trinitrobenzene sulfonic acid (TNBS) model of colitis in rats

Colon epithelial protein(s)can induce oral tolerance with altered cytokine responses in the 2,4,6-trinitrobenzene sulfonic acid (TNBS) induced model of experimental colitis

Colon epithelial protein(s)can induce oral tolerance with altered cytokine responses in the 2,4,6-trinitrobenzene sulfonic acid (TNBS) induced model of experimental colitis

Co-expression of human tropomyosin isoform 5 and a novel colon epithelial protein (cep) in the colon epithelium and extracolonic organs commonly involved in ulcerative colitis

Co-expression of human tropomyosin isoform 5 and a novel colon epithelial protein (cep) in the colon epithelium and extracolonic organs commonly involved in ulcerative colitis

Ultrastructural localization of a novel colon epithelial protein associated with ulcerative colitis: [formula omitted]∗, N. Gujral∗, K.M. Das∗, P.S. Amenta+. Departments of Pathology+ and Medicine∗, Robert Wood Johnson Medical School, New Brunswick, NJ

Ultrastructural localization of a novel colon epithelial protein associated with ulcerative colitis: [formula omitted]∗, N. Gujral∗, K.M. Das∗, P.S. Amenta+. Departments of Pathology+ and Medicine∗, Robert Wood Johnson Medical School, New Brunswick, NJ

A shared and unique peptide in the human colon, eye, and joint detected by a monoclonal antibody

Background/Aims: Extraintestinal manifestations of inflammatory bowel disease include involvement of the eye and joint. However, the pathogenesis of ocular and joint complications is unknown. The aim of the current study is to examine the presence of cross-reactive peptide in the colon and ocular and joint tissue. Methods: Using a murine monoclonal antibody (7E12H12; immunoglobulin M isotype) developed against a colon epithelial protein, various parts of the eyes (n = 10) and joints (n = 8) were examined. A sensitive immunoperoxidase assay was used. Results: In the ocular and joint tissues, the reactivity of 7E12H12 monoclonal antibody was exclusively present in the nonpigmented epithelial cells of the ciliary processes and chondrocytes, respectively. Other structures of the eyeball, such as the sclera, cornea, iris, and retina, and the synovium of the joints did not react with 7E12H12 monoclonal antibody. Colonic epithelial cells examined in parallel also reacted with the monoclonal antibody. Conclusions: The colon epithelial cells share a unique peptide with the nonpigmented ciliary epithelial cells and chondrocytes. Future studies of the peptide(s) expressing this common epitope shared by the colon and extracolonic organs known to be involved in colitis may provide further understanding of the pathogenesis of extraintestinal complications in patients with ulcerative colitis.

Ulcerative Colitis Serum Recognizes the M&lt;sub&gt;r&lt;/sub&gt; 40K Protein on Colonic Adenocarcinoma Cells for Antibody-Dependent Cellular Cytotoxicity

Antibodies in ulcerative colitis (UC) serum promote antibody-dependent cellular cytotoxicity (ADCC) against colon cancer cells. The target molecules on the cells recognized by the antibody are unknown. This study examines the role of the M(r) 40K colon epithelial protein in this process. Serum from 29 UC patients, but not from 19 patients with Crohn's disease, significantly (p < 0.001) induced more cytotoxicity than did normal serum against DLD-1 colon cancer cells which are known to express the M(r) 40K protein. Peripheral blood lymphocytes but not polymorphonuclear leukocytes showed ADCC. Anti-M(r) 40K monoclonal antibodies significantly (p < 0.001) inhibited the UC-serum-induced ADCC, but not the spontaneous cytotoxicity. These results suggest the involvement of the M(r) 40K protein in ADCC induced by UC serum.