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PubMed2004

Defensin deficiency, intestinal microbes, and the clinical phenotypes of Crohn's disease.

Crohn's disease is a chronic, inflammatory disease of the intestinal mucosa. Although intestinal bacteria are implicated in disease pathogenesis, the etiology is still unclear. The main location of disease is the small intestine (ileum) and the colon. Ileal disease has been linked to a mutation in the NOD2 gene. Defensins are antimicrobial peptides and in the ileum, are mainly expressed in Paneth cells, epithelial cells that also express NOD2. In the colon, defensins are expressed by enterocytes or metaplastic Paneth cells. Crohn's disease patients with ileal involvement, compared with controls or Crohn's patients without ileal involvement, have diminished expression of ileal Paneth cell defensins. This decrease is even more pronounced in Crohn's patients displaying a NOD2 mutation. In contrast, Crohn's disease of the colon is characterized by an impaired induction of beta-defensins in enterocytes. The colonic expression of the constitutive beta-defensin 1 is also decreased in the inflamed colonic mucosa, but this decrease is less specific to Crohn's disease, as it can also be found in ulcerative colitis patients. In conclusion, the regional localizations of Crohn's disease, ileal or colonic disease, can be linked to different defensin profiles. Crohn's disease of the ileum is associated with diminished defensin expression in Paneth cells. Crohn's disease of the colon is associated with diminished beta-defensin expression in enterocytes. Thus, it can be speculated that decreased defensin levels lead to a weakened intestinal barrier function to intestinal microbes and might be crucial in the pathophysiology of Crohn's disease.

Crohn Disease↗
PubMed1998

Enteric defensin expression in necrotizing enterocolitis.

Immaturity of local innate defenses has been suggested as a factor involved in the pathophysiology of necrotizing enterocolitis (NEC). The mRNA of enteric human defensins 5 (HD5) and 6 (HD6), antibiotic peptides expressed in Paneth cells of the small intestine, have significantly lower levels of expression in fetal life compared with the term newborn and adult. In the current study, intracellular HD5 was demonstrated by immunohistochemistry at 24 wk of gestation, but at low levels, consistent with findings at the mRNA level. These data suggest that the low level enteric defensin expression, characteristic of normal intestinal development, may contribute to the immaturity of local defense, which predisposes the premature infant to NEC. To test if levels of defensin expression are altered in NEC, specimens from six cases of patients with NEC and five control subjects (four patients with atresia and one with meconium ileus) were analyzed to determine HD5 and HD6 mRNA levels by in situ hybridization. Compared with the control group, the level of enteric defensin expression per Paneth cell assessed by image analysis was increased 3-fold in cases of NEC (p = 0.02, analysis of variance and covariance). In addition, the number of Paneth cells was increased 2-fold in the small intestinal crypts of NEC specimens compared with those of control subjects (p < 0.01, covariance analysis). In healthy tissue, peptide levels within Paneth cells paralleled mRNA levels through development. In tissue from infants with NEC, the steady state level of intracellular peptide was not increased in conjunction with the observed rise in defensin mRNA. A straightforward interpretation of this finding is that HD5 is actively secreted in this setting and the Paneth cells maintain a constant steady state level of intracellular peptide, but the possibility of translational regulation of peptide expression is also consistent with these data. The associations between NEC and enteric defensin expression reported here offer support for future studies to address the role of these endogenous host defense factors in the pathophysiology of this disease.

Adult↗
PubMed1982

Ovarian Sertoli-Leydig cell tumors with heterologous elements. I. Gastrointestinal epithelium and carcinoid: a clinicopathologic analysis of thirty-six cases.

Thirty-six ovarian Sertoli-Leydig cell tumors containing heterologous elements in the form of gastrointestinal-type epithelium are described. Eight of the cases contained microscopic foci of insular carcinoid; in two of these cases of foci of goblet cell carcinoid were also present. The patients' ages ranged from 4-67 years (average, 23 years). The presenting manifestations were androgenic changes (18 cases), abdominal swelling (15 cases), acute abdominal symptoms (one case), menstrual irregularities (one case), and postmenopausal bleeding (two cases). All of the tumors were unilateral. The surface of the tumor was intact in 32 cases, ruptured in three, and adherent to the diaphragm in one. The tumors usually had both cystic and solid components and averaged 15.0 cm in diameter; mucinous fluid was noted on gross examination in 12 cases. On microscopic examination, the gastrointestinal epithelium contained mucinous cells, columnar cells and argentaffin cells; rarely Paneth cells were seen. It was cytologically benign in 29 cases, of borderline malignancy in five and malignant in two. Follow-up information, available for 31 patients, revealed that 29 were alive without evidence of disease from 1-17 years (average, six years) postoperatively. One patient, who had a poorly differentiated tumor, died of recurrent disease six years after the initial operation.

Adolescent↗
PubMed1980

The origin of nuclear bodies: a study of the undifferentiated epithelial cells of the equine small intestine.

During an electron and light microscopic study of the equine intestinal epithelium, it was observed that some secretory granules of the undifferentiated crypt epithelium were incorporated into the nucleus during mitosis. A study was made of the chemical nature of the granules, using standard histochemical techniques: PAS-Alcian blue, Deamination-PAS, and Ninhydrin-Schiff reactions. The granules contained a neutral protein-polysaccharide complex with many terminal amino groups, possibly an antibody (IgA). The intranuclear granules underwent coalescence and degeneration during differentiation. The end-product was identical with the nuclear bodies seen in other somatic cells and described in the literature. These nuclear bodies were seen in absorptive cells, goblet cells, and Paneth cells; but were not observed in any entero-endocrine cells. This study shows that the nucleus is capable of isolating and degrading unwanted material, foreign and internally generated, and, further, it gives an explanation for the origin of nuclear bodies, structures that have long been an enigma in cell biology.

Animals↗
PubMed1993

Association between expression of sialosyl-Tn antigen and intestinalization of gastric carcinomas.

We examined immunohistochemically the expression of sialosyl-Tn antigen (STN) in 137 primary gastric carcinomas. In the non-neoplastic mucosa, STN was expressed in all goblet cells, most absorptive cells and Paneth cells of intestinalized tubules, whereas it was not seen in the gastric mucosa proper. We evaluated the association between the mucin-histochemical findings of intramucosal carcinomas and the STN-expression. STN was expressed in 8 of 20 undifferentiated-type carcinomas, all of which were mucin-histochemically of the gastric type. STN was expressed in 25 of 51 differentiated-type carcinomas, among which it was expressed in 3 of 12 gastric-type, 11 of 17 intestinal-type and 11 of 22 mixed-type adenocarcinomas. The extent of STN expression in the intestinal-type adenocarcinomas was significantly higher than in gastric-type adenocarcinomas and undifferentiated-type carcinomas, and that seen in mixed-type adenocarcinomas was significantly higher than in the gastric-type adenocarcinomas. We also examined STN expression in early and advanced carcinomas. As the depth of carcinoma invasion progressed, the extent of STN expression was significantly higher and its incidence was significantly higher. Our findings suggest that STN expression reflects intestinalization of the gastric mucosa, and that a kind of intestinal metaplasia occurs in progressing gastric carcinoma cells.

Adenocarcinoma↗
PubMed1976

Fluorescence histochemistry of peptide hormone-producing cells: observations on the nitroso-naphthol method for the demonstration of tyrosine residues.

Nitroso-naphthol reacts with tyrosine residues of peptides (and probably also proteins) to yield intensely fluorescent condensation products. This reaction forms the basis of a fluorescence histochemical procedure designed to demonstrate cells that are rich in tyrosine-containing peptides or proteins. In models the method was found to be specific for p-hydroxylated phenolic compounds. Fluorescence was induced also following formaldehyde vapour fixation. With the nitroso-naphthol technique the zymogen granules of gastric chief cells, intestinal Paneth cells, pancreatic acinar cells and certain peptide hormone-secreting cells such as the GH cells in the adenohypophysis, the insulin cells of the pancreatic islets and the calcitonin cells of the thyroid gave intense fluorescence with spectral characteristics indistinguishable from those ofthe fluorophores of tyrosine-containing peptides. In addition, a population of endocrine-like cells in the antral and intestinal mucosa of certain mammals displayed fluorescence.

Animals↗
PubMed1992

Prostaglandin E2 localization in the rat ileum.

The application of anti-prostaglandin E2 immunoglobulin to plastic-embedded thin sections of the rat ileum has permitted the localization of prostaglandin E2 in this tissue. In agreement with the published data (Chock & Schmauder-Chock (1988), Schmauder-Chock & Chock (1989)), the results also suggest the presence of an arachidonic acid cascade in the granules of various secretory cells of the gut. Since antibody labelling was found within the secretory granules of connective tissue mast cells, goblet cells, and Paneth cells, the presence of the arachidonic acid cascade in these granules is implied. The appearance of prostaglandin E2 over the non-cellular internal elastic lamina of arterioles suggests that it may have been secreted along with the elastin. The even distribution of prostaglandin throughout the cytoplasm of the erythrocyte is consistent with the concept that this cell scavenges the eicosanoid from the circulation. These data further link the secretory granule to the production of eicosanoids and therefore illustrate the potential sources of prostaglandins in the rat ileum.

Animals↗
PubMed1979

Organ culture of adult mouse intestine. I. Morphological results after 24 and 48 hours of culture.

Explants of adult mouse intestine have been maintained in organ culture for 24 to 48 hr. The best results have been obtained with a mixture of DMEM-HEPES medium and NCTC-135 enriched with 10% fetal bovine serum. The morphology of the mucosa is well preserved at the light and electron microscopic level: absorbing cells exhibit an increase in secondary lysosomes; goblet cells and Paneth cells remain active; numerous mitoses are observed in the crypts; and vigorous re-epithelization takes place on the margin of the explants.

Animals↗
PubMed2005

Innate defenses of the intestinal epithelial barrier.

The innate immune system plays a crucial role in maintaining the integrity of the intestine and protecting the host against a vast number of potential microbial pathogens from resident and transient gut microflora. Mucosal epithelial cells and Paneth cells produce a variety of antimicrobial peptides (defensins, cathelicidins, crytdinrelated sequence peptides, bactericidal/permeabilityincreasing protein, chemokine CCL20) and bacteriolytic enzymes (lysozyme, group IIA phospholipase A2) that protect mucosal surfaces and crypts containing intestinal stem cells against invading microbes. Many of the intestinal antimicrobial molecules have additional roles of attracting leukocytes, alarming the adaptive immune system or neutralizing proinflammatory bacterial molecules. Dysfunction of components of the innate immune system has recently been implicated in chronic inflammatory bowel diseases such as Crohn's disease and ulcerative colitis, illustrating the pivotal role of innate immunity in maintaining the delicate balance between immune tolerance and immune response in the gut.

Animals↗
PubMed1994

Colonic mucosal replacement by syngeneic small intestinal stem cell transplantation.

A novel method of colonic mucosal replacement by transplantation of disaggregated small intestinal epithelium is described. Thirty-one inbred rats had the ascending colon isolated, and surgical mucosectomy was performed on the "free" loop. Epithelial cell aggregates were isolated from postnatal small intestine using collagenase and dispase digestion, then 20 microL of the cell suspension was "seeded" over the denuded colonic muscle of 25 recipient rats. Six control rats had surgical mucosectomy only. All loops were retrieved after 14 days for histologic examination. Stem cell lineage studies were used with selective staining protocols to identify enterocytes, goblet cells, entero-endocrine cells, and Paneth cells. A neomucosa with typical small bowel morphology including crypts and villi and all four stem cell lineages was regenerated by transplanted cells on the colonic muscle in 19 of 25 (76%) recipients. Control loops showed no epithelial regrowth confirming total mucosectomy. With appropriate stromal support, transplanted small intestinal stem cells have the capacity to re-epithelialize denuded colonic muscle with small bowel neomucosa.

Animals↗
PubMed2006

Orthotopic transplantation of intestinal mucosal organoids in rodents.

BACKGROUND: Orthotopic transplantation of intestinal mucosal organoids that contain putative mucosal stem cells serves as an important step toward implementing intestinal gene therapy and treatment for malabsorption syndromes in animals and humans. We hypothesized that intestinal mucosal organoids can be transplanted along the axis of the small bowel giving rise to a neomucosa expressing proteins of its donor origin. METHODS: Epithelial organoids were harvested from neonatal mice or rat small intestine with the use of a combination of enzymatic digestion with dispase and collagenase, and gravity sedimentation. In adult syngeneic recipients, a 7-cm segment of midjejunum was isolated, leaving its vascular pedicle intact. The remaining proximal and distal segments were anastomosed to restore intestinal continuity. The isolated segments were randomly subjected to surgical or chemical mucosectomy with a chelator solution for 30, 45, or 60 minutes and then compared. Histologic examination was used to confirm the presence of enterocytes, goblet cells, enteroendocrine cells, and Paneth cells in the neomucosal segments. To confirm the presence of ileal bile acid transporter (IBAT) gene message and function, we measured sodium-dependent bile acid uptake and IBAT-messenger RNA. Immunohistochemical examination using anti-IBAT antibodies was performed to demonstrate the expression of IBAT in the neomucosal segments. Experiments were repeated in a murine model transgenic for the green fluorescent protein to verify donor origin of the engrafted mucosa expressing IBAT. RESULTS: The area of peak IBAT function was found to be located in the terminal ileum. Organoid units harvested from this region were capable of generating a small-bowel neoileal mucosa after being seeded into the jejunum. This mucosa was histologically confirmed to differentiate into all 4 intestinal lineages and to express IBAT signal, confirming its donor-derived origin. Optimal engraftment of mucosa expressing the IBAT protein was found in isolated jejunal segments debrided for 45 minutes. Sodium-dependent bile acid uptake was 5-fold higher in the neoileum, compared with the jejunum. IBAT-mRNA levels in the neoileum were 18-100-fold higher than those in the jejunum. Areas of green fluorescent protein-positive mucosa stained positively with anti-IBAT antibody in adjacent sections, suggesting that the regenerated mucosa is from transplanted ileal stem cells. CONCLUSIONS: Orthotopic transplantation of epithelial organoids containing ileal stem cells was used to generate a neoileal mucosa that expressed all 4 intestinal lineages along with a new zone of active bile acid uptake and IBAT expression in a recipient jejunal segment.

Animals↗
PubMed1990

Mucosal ultrastructure of continent cecal reservoir for urine and its ileal nipple valve 2-9 years after construction.

Biopsy specimens from continent cecal reservoir for urine and from its ileal nipple valve were studied with electron microscopy after two to nine years of function in ten patients. In the colonic mucosa from the reservoir there was shortening of microvilli, in some cases with random orientation and numerical reduction-changes unrelated to the time from reservoir construction. Filamentous core rootlets were also randomly oriented and numerically diminished. Glycocalyceal bodies were present in most cases. Mucosal edema and reduced numbers of goblet cells were found in six cases and increased amount of collagen in two. In the ileal nipple valve mucosa there were no microvillous changes, but metaplastic formation of glycocalyceal bodies was interpreted as adaptation to physiologic conditions comparable with those in the reservoir's colonic mucosa. Collagen increase was found in two of the nipple valves. Neurogenic processes, enterochromaffin cells and Paneth cells were always well preserved in normal amounts in the cecal as well as the ileal mucosa.

Biopsy↗
PubMed1979

Loss of differentiation in intestinal metaplasia in cancerous stomachs. A comparative morphologic study.

229 stomachs resected for duodenal and gastric ulcer and carcinoma were examined with special regard to the morphological and histochemical pattern of intestinal metaplasia (IM). The results of qualitative and semiquantitative studies were analysed statistically. Whereas duodenal and gastric ulcer cases are best discriminated by the presence or absence of IM, the strongest discriminating factor between carcinoma and gastric ulcer is the content of goblet cells in metaplastic crypts. Metaplastic crypts lined exclusively with goblet cells producing sulfated acid glycoproteins could be identified in more than one third of the cancer cases. The increase in goblet cells coincides with a loss of the more differentiated cells in the metaplastic glands, such as enterocytes, APUD cells, or Paneth cells. This "enterocoli metaplasia" seems to be specific for cancer bearing mucosa and occurs more often in cancer of intestinal type; it may represent a form of a derepressive dedifferentiation. The significance of enterocoli metaplasia as a premalignant lesion remains to be elucidated.

Adult↗
PubMed1980

Morphometric studies of normal and inflamed duodenal mucosa.

Corresponding and differing parameters in normal and inflamed duodenal mucosa are compared in a morphometric, histologic study. Results of this study show that the density of cellular infiltration into the lamina propria is decisive for the diagnosis of duodenitis, in particular the composition of plasma cells and lymphocytes. This diagnosis is supported by dilatation of Brunner's glands, cellular infiltration of the muscularis mucosae, and by gastric metaplasia of the duodenal epithelium. The length and width of the villi and crypts, the number of goblet cells and Paneth cells, the mitotic rate, and hemorrhage and hyperemia are inappropriate differential diagnosticc criteria.

Anthropometry↗
PubMed1988

Entero-endocrine cell differentiation in carcinomas of the gallbladder and mucinous cystadenocarcinomas of the pancreas.

Forty two carcinomas of the gallbladder and 25 mucinous cystadenocarcinomas of the pancreas were analyzed using silver stains and immunohistochemical techniques. Fourteen (33.3%) gallbladder carcinomas had argyrophil and argentaffin cells and 17 (40%) contained endocrine cells as shown by immunoperoxidase stains. The gallbladder tumors that had the largest number of endocrine cells were the well differentiated adenocarcinomas with colonic features. The most common endocrine cell in these tumors was the serotonin-containing (EC) cell followed by somatostatin-containing cells and cells that reacted to pancreatic polypeptide and gastrin. Intestinal metaplasia with pseudopyloric gland hyperplasia was present in the gallbladder mucosa adjacent to 11 carcinomas and had an endocrine cell population similar to that of the tumors. Endocrine cells were demonstrated in 18 (70%) of the 25 mucinous cystadenocarcinomas of the pancreas by the immunoperoxidase method although only 9 had argyrophil and argentaffin cells. The population of endocrine cells in these mucinous pancreatic tumors was similar to that found in gallbladder carcinomas. Endocrine cells were more numerous in areas with colonic-type glands, goblet cells and Paneth cells. The secretory products of the endocrine cells in these gallbladder and pancreatic tumors did not give rise to systemic endocrine manifestations. The presence of endocrine cells in these tumors can be explained on the basis of intestinal differentiation.

Cell Transformation, Neoplastic↗
PubMed1989

Dysplasia of the gallbladder. Its histogenesis and correlation to gallbladder adenocarcinoma.

A total of 50 cases of gallbladder dysplasia found adjacent to carcinoma were examined histologically and immunohistochemically for metaplastic changes in order to elucidate the characteristics of the dysplasia of the gallbladder. The incidence of metaplastic changes in the dysplastic mucosa such as the occurrence of endocrine cells, lysozyme-immunoreactive cells, goblet cells, and Paneth cells were 58%, 74%, 32%, and 22%, respectively. Based on the presence or absence of metaplastic changes, these 50 cases of dysplasia were divided into 40 cases (80%) of metaplastic type dysplasia showing at least one marker of metaplasia and 10 cases (20%) of non-metaplastic type dysplasia showing no metaplasia. On the other hand, these 50 cases of carcinoma were divided into 10 cases of non-metaplastic type carcinoma and 40 cases of metaplastic type carcinoma based on the presence or absence of metaplasia in the tumor tissue. The incidence of metaplastic changes in the dysplastic lesions was compared between the dysplasia adjacent to non-metaplastic type carcinoma and that adjacent to metaplastic type carcinoma. The incidence of metaplasia in the dysplastic mucosa adjacent to metaplastic type carcinoma was higher than that adjacent to non-metaplastic type carcinoma with a statistically significant difference. Moreover, the presence or absence of metaplastic changes was also examined in the surrounding dysplasia and non-tumorous mucosa in each case. Most cases of metaplastic type carcinoma were surrounded by dysplasia with metaplastic changes and by metaplastic epithelium, whereas most cases of non-metaplastic type carcinoma were surrounded by dysplasia without metaplasia.(ABSTRACT TRUNCATED AT 250 WORDS)

Adenocarcinoma↗
PubMed1998

The role of enterocytes in gut dysfunction.

Stem cells in the intestinal epithelium give rise to enterocytes, goblet cells, enteroendocrine cells, and Paneth cells. Each of these cell lines plays a role in cytoprotection of the intestinal mucosa. In particular, it has been demonstrated that mature enterocytes can act as antigen presenting cells. Parenteral and enteral nutrition are used to nourish critically ill patients. However, these regimens are unfortunately associated with gut atrophy. Glutamine, the preferred intestinal nutrient, reverses this gut atrophy and plays a key role in maintaining the barrier function of the gut. Specific nutrients (putrescine, spermidine, spermine) have been used to modulate intestinal adaption. In addition, ornithine has been shown to act as a regulator of intestinal adaption. In this review, we discuss the relationship between the biology of enterocytes and failure of the gut barrier.

Animals↗
PubMed1997

Down-regulation of CD95 (Fas/Apo-1) in the epithelia of adenovirus-infected appendices.

AIMS: Adenoviral inclusions are commonly seen in appendices from infants with intussusception. They are associated with focal epithelial budding and less frequently with epithelial shedding. These morphological changes could depend on the opposing effects of adenoviral gene products on CD95-mediated apoptosis. METHODS AND RESULTS: Appendices from intussusceptions with viral inclusions (n = 4) and normal appendices (n = 10) were studied by immunochemistry with anti-adenovirus, anti-CD95 and anti-HLA-DR antibodies. Apoptosis was studied by the TUNEL method. The mucosa of normal appendices contained no adenoviral protein. CD95 was present in all epithelial cells except Paneth cells. HLA-DR was absent in epithelial cells and apoptosis was seen only in germinal centres and in a few surface epithelial cells. The epithelium of appendices from intussusceptions contained nuclear inclusions labelled with anti-adenovirus antibody, always found in the epithelial buds. The epithelial CD95 pattern was drastically altered in adenovirus-infected appendices. CD95 was absent from the budding foci. In these foci, HLA-DR was overexpressed. There was also increased epithelial apoptosis in areas remote from those lacking CD95 antigen. CONCLUSIONS: The appearance of epithelial budding or shedding in appendices from intussusception could be due to focal in situ differences in the expression of adenoviral genes.

Adenoviridae Infections↗