Increased numbers of macrophages in noninflamed gastroduodenal mucosa of patients with Crohn's disease

We investigated immunostained macrophages in the noninflamed mucosa of Crohn's disease patients. Biopsied specimens from endoscopically normal gastroduodenal mucosa of Crohn's disease, ulcerative colitis, and healthy control patients were studied. Sections were examined immunohistochemically using a monoclonal antibody specific for tissue macrophages (CD68). Immunostained mucosal macrophages in the second part of the duodenum, duodenal bulb, gastric antrum, and gastric body of the Crohn's disease group were more numerous than in the ulcerative colitis and control groups. The characteristic findings of Crohn's disease were aggregations, focal subepithelial dense accumulations, and infiltration throughout the mucosa of macrophages not accompanied by a lymphoid infiltrate. The number of macrophages in the gastroduodenal mucosa bore no relationship with the duration of symptoms, clinical activity, or affected site in the intestine. This suggests that the increased number of macrophages in noninflamed mucosa is a histological change characteristic for Crohn's disease that indicates a persistent latent abnormality involving the entire gastrointestinal tract.     

Immunohistochemical study of colonic mucosa macrophages in children with Crohn's disease and ulcerative colitis]

In this research the histological characteristics of the macrophages on the colonic mucosa in Crohn's disease and ulcerative colitis were quantified and analysed. Twelve Crohn's disease, 19 ulcerative colitis and 10 specimen of the rectal mucosa, representing the control group according to the followed model, were studied: I period (PI) = pre-treatment, II period (PII) = up two years of evolution and III period (PIII) = more than two years of evolution. The macrophages were identified in a colonic mucosa by the monoclonal CD68 through the immunoperoxidase method. The macrophages quantification was done by chromatic computer images analysis, that express the area (mm2) used by the CD68 positive cells, in percentage. The percentage of the area used by the macrophages was increased in both diseases, in all the studied periods, when compared with the control group, but without statistic significance. The macrophages' distribution inside the control group mucosa was subepithelial, while in the illness group, it reached all the mucosa that was concentrated on the basis of ulcers and all long the fissures. On the Crohn's disease the CD68 positive cells facilited the identification of the microgranulomas, sometimes unnoticed in the hematoxiline-eosine. Although there was no difference between patients and control group in the macrophages area, the difference in the distribution could suggest the macrophages' participation on the injure in both diseases although they do not permit a differential diagnosis because of the variety of the values. The CD68 did not identify the different functional status of the macrophages, but their position in the mucosa suggest that, in terms of fissures and ulcers, their mainly function should be the phagocitosis and in the other cases, they have been the cells that should show the antigens and that recruit the other inflammatory cells.     

Evaluation of gastroduodenal mucosal lesions in patients with Crohn's disease and ulcerative colitis

Background: Patients with Crohn's disease (CD) are reported to suffer from upper gastroduodenal lesions with varying frequency, although concurrent Helicobacter pylori infection is reported to be low. Methods: A prospective study was carried out on patients diagnosed with CD or ulcerative colitis (UC) in order to evaluate the degree of upper gastroduodenal tract involvement and the prevalence of Helicobacter pylori infection. Results: Gastroduodenal lesions were found in 18 (78%) of 23 CD patients, the location being the stomach in 18 (78%), the duodenal bulb in 16 (70%) and the descending duodenum in 16 (70%). Bamboo joint‐like lesions were found in four cases (17%) in gastric body and cardia. In contrast, gastroduodenal lesions were found in 10 (53%) of 19 UC patients, the location being the stomach in nine (47%), the duodenal bulb in six (32%), and the descending duodenum in three (16%). The H. pylori‐positive rate in patients with CD and UC was 0%, and 11%, respectively. Conclusion: Minute upper gastroduodenal lesions are much more common in CD than in UC patients, especially in the descending duodenum. Accordingly, upper gastrointestinal endoscopy would seem to be a useful means with which to obtain a definitive diagnosis in all suspected IBD cases.     

Isolation of peptides useful for differential diagnosis of Crohn's disease and ulcerative colitis

BACKGROUND: Phage displayed random peptide technology has been utilised to identify binding epitopes of antibodies or receptor ligands. Aim: To isolates peptides from a phage library which are specifically recognised by antibodies in serum from patients with Crohn's disease (CD). METHODS: A phage displayed random peptide library composed of nine amino acids was established and sequentially screened using serum immunogloblin G obtained from CD patients. RESULTS: Five different CD specific peptides were isolated from the phage library. No homology in amino acid sequences was observed among four (CDP-1, -3 to -5) of the five peptides exhibiting different binding characteristics with each CD patient's serum. In contrast, two peptides (CDP-1 and -2) had similar amino acid sequences and similar binding characteristics. Four multiple antigenic peptides (MAP, CDP-1, -3 to -5) were synthesised, and an enzyme linked immunosorbent assay (ELISA) using the four peptides was developed to detect serum antibodies against them. Fifty two of 92 CD patients (56.5%) were detected by ELISA, none of 20 ulcerative colitis (UC) patients, only one of 25 duodenal ulcer patients, and only three of 48 healthy subjects. CONCLUSIONS: ELISA using the four peptides isolated in this study may be useful for the differential diagnosis of CD and UC.     

Ectopic colonic mucosa in ulcerative colitis and in Crohn's disease of the colon

Colectomy specimens from 62 patients (22 with ulcerative colitis, 20 with Crohn's disease of the colon, and 20 with invasive adenocarcinoma [without inflammatory bowel disease]) were reviewed for the presence of ectopic colonic mucosa. One or more foci of ectopic colonic mucosa were found in 16 of the 22 specimens (72 per cent) with ulcerative colitis and in 11 of the 20 specimens (55 per cent) with Crohn's disease of the colon. None of the 20 specimens having adenocarcinoma (without chronic inflammatory bowel disease) had ectopic colonic epithelium. The presence of ectopic colonic mucosa was found to be dependent on the age of the patients (more frequent among younger patients) and on the number of sections per specimen. One adenocarcinoma in a case of long-standing ulcerative colitis had apparently originated in ectopic colonic mucosa. This study was supported by grants from the Karolinska Institute.     

Calprotectin is a stronger predictive marker of relapse in ulcerative colitis than in Crohn's disease

BACKGROUND AND AIMS: The clinical course of inflammatory bowel disease is characterised by a succession of relapses and remissions. The aim of our study was to assess whether the predictive value of faecal calprotectin-a non-invasive marker of intestinal inflammation-for clinical relapse is different in ulcerative colitis (UC) and Crohn's disease (CD). METHODS: Seventy nine consecutive patients with a diagnosis of clinically quiescent inflammatory bowel disease (38 CD and 41 UC) were followed for 12 months, undergoing regular clinical evaluations and blood tests. A single stool sample was collected at the beginning of the study from each patient and the calprotectin concentration was assessed by a commercially available enzyme linked immunoassay. RESULTS: In CD, median calprotectin values were 220.1 mug/g (95% confidence interval (CI) 21.7-418.5) in those patients who relapsed during follow up, and 220.5 mug/g (95% CI 53-388) in non-relapsing patients (p=0.395). In UC, median calprotectin values were 220.6 mug/g (95% CI 86-355.2) and 67 microg/g (95% CI 15-119) in relapsing and non-relapsing patients, respectively (p<0.0001). The multivariate Cox (proportional hazard) regression model, after adjustment for possible confounding variables, showed a twofold and 14-fold increase in the relapse risk, respectively, in those patients with CD and UC in clinical remission who had a faecal calprotectin concentration higher than 150 microg/g. CONCLUSIONS: Faecal calprotectin proved to be an even stronger predictor of clinical relapse in UC than in CD, which makes the test a promising non-invasive tool for monitoring and optimising therapy.     

Soluble ST2: a new and promising activity marker in ulcerative colitis

AIM:To correlate circulating soluble ST2(sST2) levels with the severity of ulcerative colitis(UC) and serum levels of pro-inflammatory cytokines,and to demonstrate the predictive power of sST2 levels for differentiation between active and inactive UC.METHODS:We recruited 153 patients:82 with UC,26 with Crohn's disease(CD) and 43 disease controls [non-inflammatory bowel disease(IBD)].Subjects were excluded if they had diagnosis of asthma,autoimmune diseases or hypertension.The serum levels of sST2 and pro-in...     

Glucosamine synthetase activity of the colonic mucosa in ulcerative colitis and Crohn's disease.

Glucosamine synthetase is the first enzyme in glycoprotein biosynthesis, catalysing the formation of glucosamine-6-phosphate, from which N-acetylglucosamine is formed. The levels of this enzyme in normal human colonic mucosa (in colectomy specimens and rectal biopsies) were found to be 13-8 +/- 4-0 micron mol glucosamine synthesised/h/g wet wt. In the colonic mucosa in ulcerative colitis and Crohn's colitis the enzyme level was diminished when there was loss of epithelial cells in the mucosa, although not when there was just loss of goblet cells. In patients recovering from an acute attack of ulcerative colitis, the enzyme levels rose to a peak above the normal range, an effect which did not occur in patients who did not recover promptly. This recovery peak may be related to the synthesis of gastrointestinal mucus, or immunoglobulin, or the secretory component of IgA, all of which contain large amounts of N-acetylglucosamine.     

Ornithine decarboxylase in colonic mucosa from patients with moderate or severe Crohn's disease and ulcerative colitis.

Polyamines, as well as ornithine decarboxylase (ODC), the enzyme involved in their synthesis, were reported to be closely related to cell proliferation. In Crohn's disease and ulcerative colitis, cell destruction and proliferation increase in the active stage. The aim of the present study was to determine the ODC in both involved and uninvolved areas of the colonic mucosa of active Crohn's disease and ulcerative colitis patients. The patients were divided in two groups, owing to the different level of activity (severe or moderate), by means of clinical endoscopy, laboratory, and histology evaluations. Subjects with suspected disease, but endoscopically unconfirmed, were used as controls. In all ulcerative colitis and Crohn's disease patients the ODC values both in involved and uninvolved mucosa were significantly lower than in controls. In severe Crohn's disease ODC was significantly reduced versus moderate Crohn's disease only in affected tissues. In all ulcerative colitis patients (moderate or severe) the ODC was significantly decreased in involved mucosa compared with uninvolved mucosa. Severe ulcerative colitis showed the significantly lowest ODC. We suggest that the significant decrease of ODC in the bowel mucosa is closely related to the severity of the disease. The highest decrease of ODC in ulcerative colitis patients would be due both to the enhanced cell destruction, and to the feed-back from exogenous increased polyamine production (bowel bacteria, cell desquamation). Therefore ODC would be considered a sensitive index of the inflammatory derangement of the mucosa, especially in acute ulcerative colitis. We conclude that this behaviour may result in an enhanced risk of neoplasia.     

Na,K-ATPase activity in rectal mucosa of children with ulcerative colitis and Crohn's disease

Mucosal Na,K-ATPase activity was studied in rectal biopsy specimens from 19 children with ulcerative colitis (UC) (mean age, 13 years) and 4 children with Crohn's colitis (mean age, 14 years) and compared with biopsy specimens from 12 control children (mean age, 12 years). The Na,K-ATPase activity was significantly decreased in UC with severe rectal inflammation compared with UC in remission or with children with unspecific symptoms and normal mucosa (p less than 0.001, respectively). A higher enzyme activity was shown with age in the group with normal rectal mucosa and no evidence of inflammatory bowel disease (n = 17). The decreased Na,K-ATPase in UC with severe rectal inflammation might contribute to the diarrhoea by impairment of sodium transport.     

Search for a specific marker of mucosal dysplasia in chronic ulcerative colitis

We have tried to identify a histochemical or immunohistochemical marker that would reliably detect mucosal dysplasia in chronic ulcerative colitis. Colonic biopsy specimens were taken from patients with long-standing ulcerative colitis undergoing surveillance colonoscopy. Control tissue was obtained from the margins of colonic resections performed for neoplastic or nonneoplastic diseases. Sections of the specimens were examined by periodic acid-Schiff staining, high iron diamine Alcian blue staining for sialomucins and sulfomucins, and binding of the lectins peanut agglutinin, Ulex europeus agglutinin I, and Ricinis communis I agglutinin. Sections were reacted also with anticarcinoembryonic antigen antibodies to determine whether a nonpolar surface distribution of the antigen was a feature of dysplasia. We found that changes in colonic mucin, characterized by periodic acid-Schiff-positive mucin outside of goblet cells, an increase in binding of peanut agglutinin, and an increase in the relative amount of sialylated mucin, were associated with morphologic dysplasia. However, identical alterations were observed, albeit less commonly, in colitis without dysplasia, particularly in the presence of active inflammation. No abnormality in the surface distribution of carcinoembryonic antigen or Ricinis communis I agglutinin was observed. Thus, we did not identify a reliable corollary test to the histologic diagnosis of mucosal dysplasia in ulcerative colitis. With regard to these histochemical markers, the colonic mucosa appears to respond similarly to inflammation and neoplasia.     

Biased JH usage in plasma cell immunoglobulin gene sequences from colonic mucosa in ulcerative colitis but not in Crohn's disease

BACKGROUND: Ulcerative colitis is an inflammatory disease of the colonic and rectal mucosa. Autoantibodies have been observed in ulcerative colitis which may have a role in the pathogenesis of the disease. Evidence also suggests that there is an hereditary predisposition towards the disease, although no individual genes have been identified. AIMS: This is a pilot study of immunoglobulin heavy chain genes (IgH) in ulcerative colitis to determine whether they have any particular genetic characteristics which may lead to a better understanding of the disease aetiology. SUBJECTS: Colonic or rectal tissue was obtained from five children with ulcerative colitis. Tissue was also obtained from five children with Crohn's disease and five children who did not have inflammatory bowel disease as controls. METHODS: B cells and IgD+ B cells were identified by immunohistochemistry on frozen sections. Areas of lamina propria containing plasma cells, and areas of IgD+ B cells were microdissected. The immunoglobulin genes were PCR amplified, cloned, and sequenced. Sequences were analysed for content of somatic mutations and composition of heavy chain. RESULTS: An increase in the use of JH6 and DXP'1, and a decrease in the use of JH4, gene segments in immunoglobulin genes from lamina propria plasma cells, and from virgin IgD+ B cells, was found in patients with ulcerative colitis. These biases were not present in the control groups. CONCLUSIONS: There is a fundamental difference in the immunoglobulin genes from patients with ulcerative colitis. Whether this is caused by a difference in content of immunoglobulin gene segments in the germline or a difference in the recombination mechanism is not known.     

Magnifying chromoscopy, a novel and useful technique for colonoscopy in ulcerative colitis

Ulcerative colitis (UC) is a chronic inflammatory bowel disorder characterized by exacerbations and remissions. The degree of inflammation as assessed by conventional colonoscopy is a reliable parameter of disease activity. However, even when conventional colonoscopy suggests remission and normal mucosal findings, microscopic abnormalities may persist, and relapse may occur later. Patients with long-standing, extensive ulcerative colitis have an increased risk of developing colorectal cancer. Ulcerative colitis-associated colorectal cancer is characterized by an early age at onset, poorly differentiated tumor cells, mucinous carcinoma, and multiple lesions. Early detection of dysplasia and colitic cancer is thus a prerequisite for survival. A relatively new method, magnifying chromoscopy, is thought to be useful for the early detection and diagnosis of dysplasia and colitic cancer, as well as the prediction of relapse.     

Evidence for the isolation of a new virus from ulcerative colitis patients

To assess the possible role of viral infection in the development of ulcerative colitis (UC) and of Crohn's disease (CD), electron-microscopic, physical, and chemical studies were performed on viral agents cultivated in tissue culture following inoculation with surgical specimens obtained from patients with CD, UC, and from control patients with other gastrointestinal diseases. Viral isolates obtained from CD patients were compared with those from UC patients. Each of the agents produced cytopathic change (CPE) in the tissue culture systems used, rabbit ileum (RI), Peking duck, and Riff free chick embryo, and had similar physical and chemical properties including ether resistance, temperature stability, and lack of inhibition by methotrexate. The three tissue culture systems supported viral growth and produced virus-like particles from CD filtrates with similar morphologic appearance, including a central core and an outer coat. Electron-microscopic studies revealed that the particle from CD has a diameter of 60 nm (range 42–71 nm). The agent isolated from UC patients also has a mean particle diameter of 60 nm (range 45–75 nm). The CD and UC agents produced different CPE in RI tissue culture. Agents different from CD-and UC-derived viruses were isolated from one patient with necrotizing enterocolitis and from three patients with carcinoma of the colon. Ileal homogenates of other control patients failed to reveal evidence of viral contaminants and evaluation of the tissue culture systems revealed no adventitious agents. Virus was found in diseased as well as in adjacent histologically normal CD tissue. Guinea pig antibody prepared against CD-derived agents inhibited the growth of each of the CD agents but not of the agents from UC, necrotizing enterocolitis, or colon carcinoma. Antibody directed against the agents from UC failed to inhibit growth of the agents from CD, necrotizing enterocolitis, or colon carcinoma. The current studies therefore suggest that a virus has now been cultivated from UC patients which produced CPE different from the viruses previously cultivated from CD patients and which lacks cross-reactivity with the CD derived virus. Since each filtrate produced viruses in the three different tissue culture systems, these viruses are not tissue culture contaminants but are derived from the human filtrates. These studies also suggest an association between viruses and inflammatory bowel disease but do not establish an etiologic relationship.Supported by a grant from the National Foundation for Ileitis and Colitis.     

Longterm appraisal of the histological appearances of the ileal reservoir mucosa after restorative proctocolectomy for ulcerative colitis.

Between November 1976 and December 1985, 110 patients had restorative proctocolectomy for ulcerative colitis. The histological appearances in the reservoir mucosa were followed up in 60 of 109 survivors over 19-173 months (median 97). The median number of biopsy specimens taken per patient was six with a range of 3-13. These were examined by one pathologist (ICT) unaware of the clinical details using a scoring system previously described to assess the degree of chronic and acute inflammation. There was a significant correlation between the degree of severity of chronic and acute changes (r = 0.6192, p < 0.000001). There was no correlation between the severity of inflammation and the following variables: preoperative duration of disease, presence of cancer or dysplasia in the original operative specimen, extra-alimentary manifestations or the type of reservoir. A significant correlation between severe inflammation and male sex was found (p < 0.035). The 60 patients could be divided into three groups based on the severity and fluctuation of histological inflammation. In group A (n = 27, 45%) chronic changes were minor and acute inflammation was never seen. In group B (n = 25, 42%) chronic changes were more severe and there were transient episodes of acute inflammation. In group C (n = 8, 13%) severe chronic and severe acute inflammation were constantly present. Differentiation of the three groups had clearly occurred within six months from closure of the ileostomy. Patients in group C could be identified on histological criteria within weeks of closure of the ileostomy and were those exclusively at risk of developing chronic pouchitis. Chronic pouchitis never occurred in patients of groups A and B. No case of dysplasia was seen. Histological assessment of the reservoir mucosa with in a few months after closure of the ileostomy seems to define patients who will and who will not subsequently develop pouchitis.     

Enhanced production of interleukin 1-beta by mononuclear cells isolated from mucosa with active ulcerative colitis of Crohn's disease.

IL1-beta production by mononuclear cells isolated from normal and active inflammatory bowel disease mucosa was studied. Significantly more IL1-beta was produced spontaneously by mononuclear cells from the inflamed mucosa compared with those from normal colonic mucosa (median 190 pg/ml (range 45-700) v 20 pg/ml (0-165)). Stimulation with lipopolysaccharide enhanced IL1-beta production by mononuclear cells from active inflammatory bowel disease mucosa but not those from normal mucosa. Depleting the mononuclear cells of macrophages, by panning with monoclonal antibody 3C10, reduced the amount of IL1-beta produced. Enhanced IL1-beta production from the inflamed mucosa may play an important role in the mediation of many inflammatory responses. The enhanced production appears to be the result of a recruited population of cells.     

Identification of a prodromal period in Crohn's disease but not ulcerative colitis

OBJECTIVES: Irritable bowel syndrome, a common gastrointestinal diagnosis, has not been clearly studied in inflammatory bowel disease. Some of the residual symptoms in subjects treated with Crohn's disease and ulcerative colitis are thought to be related to irritable bowel syndrome. The aims of this study were 1) to describe the duration and nature of complaints before the diagnosis of Crohn's disease and ulcerative colitis (prodromal period), and 2) to determine the role of IBS in this prodromal period. METHODS: A total of 66 patients with confirmed inflammatory bowel disease were enrolled in the study. The subjects received a questionnaire to ascertain the nature and duration of symptoms preceding the diagnosis of Crohn's disease or ulcerative colitis, including features described under the Rome criteria for irritable bowel syndrome. RESULTS: Of the 66 subjects analyzed, 45 had Crohn's disease and 21 had ulcerative colitis. The prodromal period was 7.7 +/- 10.7 yr for Crohn's disease and 1.2 +/- 1.8 yr for ulcerative colitis (p < 0.05). Once patients meeting the Rome criteria for irritable bowel syndrome during the prodrome were excluded, the duration of the prodromal period (non-IBS) for ulcerative colitis dropped to 0.8 +/- 1.3 yr compared to 6.9 +/- 9.8 yr in the Crohn's disease group (p < 0.05). The symptoms of the non-IBS prodrome in subjects with Crohn's disease were bloating, diarrhea, stomach pain, heartburn, fever, weight loss, and fatigue. Further analysis demonstrated that subjects whose Crohn's disease initially began as colonic disease had a longer prodrome than with small bowel. In the non-IBS Crohn's group, there was also a correlation between the age at the time of diagnosis and the duration of prodrome (r = 0.67, p < 0.0001). CONCLUSIONS: There is a significant prodromal period before the time of diagnosis of Crohn's disease that is not found in ulcerative colitis even after exclusion of subjects with IBS.