Distribution of lymphocyte subpopulations and plasma cells in the colonic mucosa of children with ulcerative colitis

The distribution of lymphocyte subpopulations and plasma cells of the colonic and rectal mucosae were studied in eight children with ulcerative colitis and 12 healthy controls. In four patients the examinations were also carried out 3 months after the beginning of treatment. No difference in the number of intraepithelial lymphocytes was found between the patients and controls. The majority of these cells were T-cells, and among them the suppressor/cytotoxic cells were preponderant. In the lamina propria of both untreated and treated patients the numbers of T-cells, helper T-cells, and B-cells were elevated compared to controls. In the patients the number of IgG-containing cells was three times that of the controls; the number of IgE positive cells was also elevated. The numbers of IgA- and IgM-containing cells were not different from that of the controls. The results suggest that in ulcerative colitis the place of primary immunological processes inside the large bowel mucosa is the lamina propria.     

Distribution of cells containing different IgG and IgA subclasses in the colonic mucosa in childhood ulcerative colitis and Crohn disease

The distribution of cells containing various subclasses of IgG and IgA were studied in the rectal and colonic mucosa from 14 paediatric patients with ulcerative colitis and four patients with Crohn's disease, as well as, from rectal biopsy specimens of 10 control subjects using monoclonal antibodies and the peroxidase method. The number of IgG3-containing cells in both the colonic and rectal mucosae of untreated patients with ulcerative colitis was significantly increased compared to controls. In the children with ulcerative colitis the number of IgG4 cells in the colonic mucosa was also significantly increased. In the rectal mucosa of patients with ulcerative colitis the percentage of IgG2 cells was decreased compared to the controls (22% versus 27%). In the great majority of specimens (in 37 out of 40) the number of IgA1 cells was higher than that of IgA2. The number of IgA1 cells in the rectal and colonic specimens of untreated patients with ulcerative colitis was slightly higher than that in the rectal specimens of controls. Authors speculate on the basis of their results that the observed IgG3 response in the patients with ulcerative colitis may be specific to the disease.     

Collagen sprue--a rare form of adult celiac disease]

The authors report the first case of collagenous sprue in Hungary. This condition is characterized by coeliac type small bowel malabsorption, resistant to gluten free diet and other therapeutic efforts, associated with poor prognosis. The diagnosis depends on the histological demonstration of extensive collagenization of the lamina propria in the flat jejunal mucosa. This disease must be kept in mind at differential diagnosis of chronic diarrhoea with progressive malabsorption, especially if it is resistant to gluten withdrawal than conventional coeliac disease.     

Functional status of intestinal t lymphocytes, regulatory mechanisms, and their variations in the course of Crohn disease and ulcerative colitis

We analyzed the phenotype, proliferative responsiveness, cytokine production and apoptosis susceptibility of lamina propria lymphocytes to different activation pathways. Lamina propria lymphocytes is a population enriched of activated lymphocytes showing a "memory" phenotype. As opposite to peripheral blood lymphocytes, lamina propria lymphocytes show proliferative hyporesponsiveness when stimulated via TCR/CD3 pathway while proliferative response to the CD2 activation pathway is relatively preserved. Under the latter activation pathway, cytokine production, especially IL-4 and IFN-gamma, is higher than that observed in peripheral lymphocytes. When compared to controls, lamina propria lymphocytes isolated from inflammatory bowel disease (Crohn's disease and ulcerative colitis) show distinctive variation in the cytokine production. In particular, Crohn's disease is characterized by an increased production of IFN-gamma, while in ulcerative colitis an increased production of IL-5 is observable. Among the different regulatory mechanisms contributing to maintain immunological homeostasis we analyzed the susceptibility to apoptosis of lamina propria lymphocytes. We found that CD2-activation pathway is regulated by Fas-mediated apoptosis, which regulates proliferation and cytokine production. In inflammatory bowel disease this apoptosis is defective thus contributing to the chronic inflammation and cytokine dysregulation.     

Familial association between coeliac disease and ulcerative colitis: preliminary communication.

One hundred and twenty patients who were members of the Nottinghamshire Coeliac Group completed a questionnaire about the occurrence of coeliac disease, ulcerative colitis and Crohn's disease amongst first-degree relatives. Siblings were at a 20-fold risk of developing coeliac disease and a 15-fold risk of developing ulcerative colitis, and significantly increased risks for these two conditions were also seen in other family members. The relatives of patients with coeliac disease are at increased risk not only of developing coeliac disease but also ulcerative colitis. This provides further support for a possible role of a dietary allergen in the development of ulcerative colitis.     

Expression of interleukin-4, interferon-gamma and lymphocyte surface markers in small intestinal mucosa of adult patients with cereal allergy

INTRODUCTION: The mechanism for adverse reactions to foods in the gastrointestinal tract are poorly understood. Previous studies of other atopic diseases and animal models suggest that lymphocytes and cytokines may be implicated in the pathogenesis of food allergy. AIM: The authors investigated the expression of interleukin-4, interferon-gamma and other lymphocyte markers of patients with cereal allergy (wheat, rye, oats) and of controls. PATIENTS/METHOD: Expressions of cytokines and lymphocyte markers on duodenal mucosa of nine patients (mean age 38.3 years, range 18-50 years, 8 women and one man) and nine controls (mean age 36 years, range 24-54 years, 6 women, 3 men) by means of immunohistochemistry were investigated. RESULTS: The mucosal structure on every biopsy specimens was normal. Despite the normal structure the expression of Ki-67 intranuclear proliferation marker was higher in patients with cereal allergy. Expression of interleukin-4 was markedly elevated in the food allergy group, however, interferon-gamma density showed no inter-group difference. The densities of CD4 (1251 vs. 1053 cells/mm2) and HLA-DR positive cells (1227 vs. 1064 cells/mm2) in the lamina propria of cereal allergy group were significantly elevated when compared with controls (P = 0.05 and P = 0.04, respectively). The densities of CD3, CD8, TCR alpha/beta and gamma/delta, HLA-DP, IgA, IgA1, IgA2-containing cells did not differ in the two groups studied. CONCLUSIONS: The authors results suggest that, despite the normal mucosal structure, the increased expression of CD4 and HLA-DR positive cells show a sign of inflammation in duodenal biopsies of patients with cereal allergy. Moreover, increased density of IL-4 may suggest its role in the pathogenesis of cereal hypersensitivity.     

Changes in the proliferation and apoptosis of colonic epithelial cells in correlation with histologic activity of ulcerative colitis

BACKGROUND: The imbalance of epithelial cell kinetics in ulcerative colitis can lead to several gastrointestinal disorders such as ulcers or neoplasias. The changes of epithelial cell kinetics caused by the inflammatory process have not been fully and uniformly described. Aim of the study was to compare the rate of epithelial cell apoptosis and proliferation within colonic crypts considering the histological activity of ulcerative colitis. MATERIALS AND METHODS: Formalin fixed paraffin embedded biopsy specimens from mild, moderate, and severe active inflammation of ulcerative colitis and from normal colonic tissue were observed. Number of cases (apoptosis/proliferation): normal: 8/10; mild UC: 8/10; moderate: 8/8; severe: 12/8. For detecting apoptosis the TUNEL-POD-DAB method, and for proliferation the PCNA (PC-10)-Biotin-Streptavidin-AEC method were used, constrained with haematoxylin. Using light microscope, at least 500 crypt epithelial cells were encountered axially in whole, well-orientated colonic crypts per each specimen. RESULTS: The number of TUNEL positive epithelial cells were significantly higher in moderate and severe active inflammation compared to each other and to normal (p < 0.0001). Between mild inflammation and normal significant alteration was not found. The number of PCNA positive cells were significantly higher in each groups of inflammation compared to normal or each other. CONCLUSION: These results demonstrate that there is a strong correlation between the histological activity of ulcerative colitis and elevated crypt epithelial cell turnover, although the alterations in proliferation develop in the earlier stages of inflammation.     

Experimental amoebiasis in the guinea-pig: sequential histological abnormalities following intracaecal injection of Entamoeba histolytica

The histological abnormalities of amoebic colitis have been well described but their evolution over a period of time has not been clearly examined. To study this, three- to four-week-old guinea-pigs were inoculated intracaecally with 80,000 to 100,000 Entamoeba histolytica trophozoites, left untreated and killed at 3, 5, 8, 11, 14 and 21 days, when the caecum was removed and the tissue sectioned and stained with H & E and PAS. Although there was considerable variation in the type of histological abnormality seen on different days of the experiment, there was a definite pattern of evolution. Initially there was diffuse infiltration of the lamina propria by lymphomononuclear cells in the presence of an intact surface epithelial lining. This was followed firstly by superficial and then by deep ulceration of the mucosa. Mucosal invasion by the trophozoites was seen only at this stage and was accompanied by tissue necrosis; the most severe abnormalities occurred on day 11. The final development was regenerative activity which was first noted on day 11, and became more prominent subsequently. The presence of cellular infiltrate in the lamina propria in the absence of trophozoites and damage to the surface epithelium, suggests that these abnormalities are not the direct effect of amoebae. It is suggested that the initial damage is caused by an enterotoxin; the trophozoites enter the mucosa only when there is a break in the surface epithelium. Once within the tissues, trophozoites aggravate the damage by their ability to phagocytose and to release cytotoxic enzymes.     

Total parenteral nutrition alters molecular and cellular indices of intestinal inflammation in neonatal piglets

BACKGROUND: The adverse effects of TPN on systemic immunity are well-documented; however, the impact of IV feeding on neonatal intestinal immunity is unknown. METHODS: A piglet TPN model was used to compare immune cell composition within the intestinal epithelium and lamina propria of parenterally and orally fed piglets. RESULTS: Small intestinal weight of piglets maintained intravenously was reduced 50% after 7 days. Intestinal atrophy in piglets fed parenterally was evidenced by decreased width of intestinal villi and colon cuffs and reduced intestinal crypt depth. The numbers of CD4+ and CD8+ T lymphocytes were threefold greater within the lamina propria of jejunal and ileal villi of piglets supported intravenously. Inverse correlations were observed between villus height or width and T-lymphocyte numbers (r = -.80; p < .05). Major histocompatibility complex class II mRNA expression, an indicator of localized inflammation, was increased in the ileum and colon of piglets receiving parenteral nutrition. Goblet cell numbers were two-fold greater in jejunal and ileal villi, and mast cells were more abundant in the colon of piglets fed parenterally. Furthermore, jejunal T-lymphocyte numbers were correlated with goblet cell numbers (r = .80; p = .01). CONCLUSIONS: These data identify molecular and cellular indices of intestinal inflammation that are responsive to IV feeding in neonates and provide a novel framework to investigate mechanisms underlying gut atrophy during TPN.     

白细胞介素8在溃疡性结肠炎患者肠黏膜中的表达

目的探讨白细胞介素8(IL-8)在溃疡性结肠炎(UC)发病机制中的作用。方法 48例UC肠黏膜标本取自行结肠镜检查的患者,按病理组织学对炎症进行分级,Ⅲ、Ⅳ级30例,I、Ⅱ级18例;对照组为30名健康成人。应用半定量RT-PCR检测对照组和UC组肠黏膜IL-8 mRNA表达水平,免疫组织化学方法检测IL-8蛋白的表达。结果与对照组相比,UC肠黏膜IL-8 mRNA和其蛋白过度表达(P0.05),IL-8 mRNA表达与疾病严重程度成正相关。IL-8蛋白表达位于UC肠黏膜固有层单核细胞,病理分级Ⅲ、Ⅳ级肠黏膜IL-8蛋白表达较病理分级I、Ⅱ级组明显增加(P0.01)。结论 UC黏膜组织中IL-8表达水平明显上调,其表达水平与病情轻重和炎症严重程度呈正相关。     

Therapeutic effect of antisecretory factor-rich egg yolk on the late phases of 2,4,6-trinitrobenzenesulphonic acid colitis in mice

Antisecretory factor (AF) is expressed in all tissues of mammals, inhibits intestinal hypersecretion and has anti-inflammatory properties as well. Endogenous AF synthesis may be stimulated by feeding hydrothermally processed cereals. Alternatively, freeze-dried egg yolk can be used as a source of exogenous AF. Several reports have suggested that AF from freeze-dried egg yolk may be useful in inflammatory bowel disease. We assessed the effect of freeze-dried, AF-rich egg yolk intake on 2,4,6-trinitrobenzenesulphonic acid (TNBS) colitis. Balb/c mice were randomised to receive (1) AF in sterile drinking-water (4?g/l, n 38) and (2) sterile drinking-water alone (vehicle, n 38) from TNBS or saline administration onwards. Different subsets of mice were killed at weeks 1-3 after TNBS or saline administration. Macroscopic and microscopic damage was assessed in colonic specimens. Eicosanoid and cytokine production was evaluated in supernatants of 24?h-incubated colonic explants. Myeloperoxidase activity was measured in frozen colonic samples, while apoptosis was assessed in paraffined samples by the in situ oligoligation method. AF-treated mice showed a milder colonic damage compared with the vehicle group, which became statistically significant at week 3. This was accompanied by decreased IL-2, IL-1 and leukotriene B4 production at weeks 2 and 3, as well as increased interferon-γ at week 1, in AF-treated mice compared with vehicle-treated mice. AF-treated mice had significantly increased counts of apoptotic cells in the lamina propria at weeks 1 and 2 post-TNBS. In conclusion, the administration of AF-rich egg yolk has a therapeutic effect in the late phases of TNBS colitis in Balb/c mice.     

Nonoxynol-9 causes rapid exfoliation of sheets of rectal epithelium

Nonoxynol-9 (N-9) containing spermicides and other N-9 containing products are commonly used as lubricants during rectal intercourse. We have previously demonstrated that rectal application of N-9 products in mice can cause exfoliation of epithelial cells, increasing the probability of infection by HSV-2. To determine if N-9-containing products would have a similar effect on the rectal epithelium in humans, the application of K-Y Plus and ForPlay, both over-the-counter (OTC) N-9 products, were compared to the application of two formulations, carrageenan and methyl cellulose, that do not contain N-9. The effects of each formulation were evaluated in 4 human participants. Light and electron microscope examination of rectal lavage specimens collected 15 min post application of N-9 products revealed the presence of sheets of epithelium. Each sheet contained hundreds of epithelial cells that included columnar and goblet cells, varieties of cells typical of rectal epithelial morphology. Sheets of epithelium were not observed in rectal lavage specimens collected 8 to 12 hr post N-9 product use or in either of the timed lavages involving non-N-9 containing formulations. In addition, no sheets of epithelial cells were observed in the baseline lavage specimens. We conclude that the rectal use of N-9-containing products causes a rapid exfoliation of extensive areas of the rectal epithelium. Exfoliation of the epithelium is no longer observed at 8 hr. It is reasonable to assume that the loss of the protective epithelium would render a person more at risk for infection by HIV and other sexually transmitted pathogens. We, therefore, caution against the use of N-9-containing products during rectal intercourse.     

Influences of Short‐Term Fasting and Carbohydrate Supplementation on Gut Immunity and Mucosal Morphology in Mice

Background: Preoperative carbohydrate (CHO) supplementation has been recommended in enhanced recovery after surgery protocols. However, the effects of CHO supplementation on gut and systemic immunity are not well understood. Methods: Mice (n = 60) were randomized to 1 of the following 5 groups: control (ad lib feeding), 12‐hour fasting without CHO administration (fasting), and 12 hours of fasting with CHO administration at 2, 4, and 8 hours before sacrifice. Then, lymphocytes were isolated from gut‐associated lymphoid tissue, that is, Peyer's patches, the intraepithelial space, and the lamina propria of the small intestine. These lymphocyte numbers and phenotypes were evaluated. IgA levels in respiratory and small‐intestinal washings were determined by ELISA. Morphology, proliferation, and apoptosis of the intestinal epithelium were also evaluated histologically. Results: Although there were no significant differences in IgA levels among the 5 groups, fasting decreased intraepithelial and lamina propria, but not Peyer's patches lymphocyte numbers. CHO at 2 hours prevented lymphocyte loss in intraepithelial, whereas CHO at 4 hours reversed lamina propria lymphocytes numbers. Percentages of lymphocyte phenotypes were similar in each site among the 5 groups. Fasting caused villous atrophy; however, CHO at 2 hours restored villous structure along with maintenance of epithelial cell proliferation rate. Conclusions: Only 12 hours of fasting causes marked gut‐associated lymphoid tissue cell loss along with gut atrophy. However, CHO at 2 hours preserves gut immunity and morphology not completely but moderately.     

直肠癌微血管密度与临床病理因素相关性研究

目的本研究回顾性评价直肠癌微血管密度(MVD)与临床病理因素之间的关系。方法采用免疫组化法对87例手术切除直肠癌标本的癌灶中心运用单克隆鼠抗人CD34抗体进行血管标记和染色,然后与临床病理因素分析。结果不同临床病理因素癌灶中心MVD计数表达不同,有无淋巴结转移的MVD计数分别为98.78±26.15、80.12±18.40(P<0.05);浸润深肌层以内及浆膜层及以外分别为83.39±19.70、89.00±28.21(P>0.05);Dukes分期A+B与C+D分别为82.47±18.40、96.50±21.36(P<0.05)。结论直肠癌不同病理因素中MVD表达不同,随着肿瘤分期的进展、淋巴结转移,微血管密度增加。MVD可以作为直肠癌分期、预测直肠癌转移及预后的重要指标。     

Issues related to gluten-free diet in coeliac disease

PURPOSE OF REVIEW: In the last few years, knowledge about coeliac disease has significantly improved, resulting in a better understanding of disease pathogenesis, diagnosis and therapy. This review describes the latest progress in research concerning treatment with gluten-free diet in patients with coeliac disease. RECENT FINDINGS: Gluten-free diet is generally admitted as effective therapy in symptomatic patients, but a life-long dietary treatment in some challenging cases such as 'silent' and 'latent' patients is under discussion. Tolerance to gluten may be acquired later in life, but, as latency may be transient, a strict follow-up is necessary in these patients. The composition of gluten-free diet needs a better definition; latest evidence demonstrates that oats are tolerated by most patients with coeliac disease. Finally, the amount of gluten permitted in gluten-free products is still a matter of debate; significant progress has been made in the sensitivity of techniques for gluten detection, but the daily amount of gluten that can be safely consumed is not yet defined. SUMMARY: Gluten-free diet remains the cornerstone of therapy of coeliac disease. More studies addressing the need of gluten-free diet for cases of 'potential' coeliac disease are necessary, as well as studies linking the best available analytical detection of gluten to the clinical threshold of tolerance.     

Systemic-oral immunization with 56 kDa molecule of Giardia lamblia affords protection in experimental mice.

Prior systemic-oral immunization of inbred mice with Giardia lamblia surface-associated antigen of molecular mass 56 kDa not only significantly blocked colonization but also resulted in elimination of G. lamblia trophozoites by 9-11 days following challenge. The colonization and multiplication of the trophozoites in unprotected animals were accompanied by a pronounced influx of suppressor T cells in intraepithelial or lamina propria of the small intestine and a significant decline in IgA-bearing plasma cells in the lamina propria. An induction of helper/inducer T cells in the intraepithelial and lamina propria and significant enhancement of IgA and IgG-bearing cells in the lamina propria of the small gut resulted in a decline, and eventual elimination, of the trophozoites from the gut. The completion of the immunization of animals with 56 kDa G. lamblia antigen resulted in: significant enhancement of helper/inducer T lymphocytes with no effect on suppressor T cells in the intraepithelial and lamina propria of the small gut; significant enhancement of IgA- and IgG-bearing plasma cells in lamina propria; and significant elevation of antibodies to 56 kDa G. lamblia antigen in the systemic circulation. The stimulation of such effector mechanisms in 56 kDa-immunized animals appears to result in failure of the trophozoites to get established, prevention of multiplication and earlier elimination from the gut. The data suggest that the 56 kDa molecule of G. lamblia immunoregulates the giardial infection.     

MPO与TGF-β1在溃疡性结肠炎中的表达

目的探讨髓过氧化物酶（MPO）和转化生长因子β1（TGF—β1）在溃疡性结肠炎中的表达及意义。方法通过免疫组织化学染色法（两步法）检测37例溃疡性结肠炎,10例克罗恩肠病及20例正常结肠组织中MPO和TGF-β1的表达。结果MPO和TGF—β1阳性表达在溃疡性结肠炎组织中（89．2％,86．5％）均显著高于克罗恩肠病（55．0％,35．0％）和正常结肠组织（10．0％,20．0％）,差异均有统计学意义（P〈0．05）。MPO和TGF—β1在溃疡性结肠炎组织中的表达有相关性（r=0．51,P〈0．05）。结论MPO和TGF—β1参与溃疡性结肠炎的发病,检测MPO和TGF-β1对研究溃疡性结肠炎的发病机制和判断溃疡性结肠炎的病情具有一定的参考价值。     

Surgical management of inflammatory bowel diseases

The development of the medical management of the inflammatory bowel disease (IBD-ulcerative colitis, Crohn's disease) has reduced the number of the acute surgical interventions. Beyond the medical treatment the surgical management, the operative modalities, the pre and postoperative care has gone through a lot of changes. They review the different types of surgical alternatives and debating on the advantages and disadvantages, they put emphasis on the different surgical solutions of the Crohn's disease and that of the ulcerative colitis. Among the applied surgical alternatives to the ulcerative colitis they discuss the traditional proctocolectomy with end-ileostomy, the Kock-reservoir (as continent stoma), as well as the restorative proctocolectomy which is sufficient to preserve the anal continence. Principles of the surgery of the Crohn's disease are discussed according to the localisation of the inflammatory process (small bowel, colonic, rectal, anal channel). Because of the predisposition of relapses and the necessity of the successive surgical therapy, the extensive resections should be avoided.     

Current concepts on celiac disease physiopathology

Celiac sprue (CS) is defined as a chronic small bowel malabsorption disorder caused by ingestion of gluten, affecting those genetically predisposed individuals. It is characterized by intestinal villi atrophy, increased number of intraepithelial lymphocytes and extense inflammatory infiltrate in the intestinal lamina propria. The role of gluten as responsible for the intestinal damage seen in CS patients is clear, however, the physiopathological mechanisms involved are still unknown. Several factors and theories have been proposed: 1) Genetic predisposition, based on the association to mendelian factors as well to the presence of particular major histocompatibility complex (MHC) haplotypes in CS patients; 2) Immunological factors, that consider the derangements that occur in the immune response of CS patients, and 3) Gliadin partial deamination by the tissular transglutaminase (tTG). In an effort to explain all these complex mechanisms, recently, all these theories have been unified, yielding one complex physiopathogenic mechanism that we tray to explain in the present review.