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PubMed1994

Early morphological changes in Crohn's disease. Transmission electron-microscopic findings and their interpretation: an overview.

Recent ultrastructural investigations revealed early epithelial lesions in Crohn's disease, while a specific morphological pattern was not identifiable. An increase in plasma cells, lymphocytes, macrophages, mast cells, eosinophilic and neutrophilic granulocytes, as well as focal edema and inflammation of tissue structures was seen in the lamina propria, submucosa and deeper layers. The results are consistent with the frequent discussion about a pathogenetically significant defect of the mucosal 'barrier function', which consists of mechanical, cellular, humoral, immunological and nonimmunological mechanisms, including different histotopographically defined lines of defense of the epithelium and lamina propria. An intact epithelial layer plays an important role as the first line of defense. It is evident that components of the epithelial barrier (absorptive cells, goblet cells, Paneth cells, M cells) show ultrastructural signs of alteration or injury, while the primary agent or event remains unknown. Another pathomechanism would be a preexisting defect in intestinal 'barrier function'. Such a defect would result in an increased uptake of, or an inadequate immune reaction to, ubiquitously occurring antigens/agents (with genetic predisposition). However, no primary defect of epithelial or inflammatory cells has been definitely identified so far. Direct toxic damage of tissue secondarily introducing the inflammatory changes is also possible. Although the morphological alterations in Crohn's disease are not yet clearly understood and exactly interpreted, transmission electron microscopy has been helpful in defining early lesions and has led to further knowledge about the pathogenesis of this disease.

Crohn Disease↗
PubMed1998

Immunohistochemical detection of metallothionein in liver, duodenum and kidney after dietary copper-overload in rats.

Metallothionein (MT) has been used in immunohistochemical techniques to indicate presence and distribution of heavy metals within biological tissues. This study describes a comparison of the pattern of MT-immunostaining in the liver, duodenum and kidney during dietary copper overload in rats. Sixteen male 10-week-old Wistar rats were randomly allocated into groups of four. Two groups were fed a pelleted diet containing 1,500 mg/kg copper and two control groups received a rodent diet containing 10 mg/kg copper. After 6 weeks samples of liver, kidney and duodenum were collected for immunohistochemistry and histology. An indirect immunoperoxidase technique, using monoclonal antibody E9 against horse MT and polyclonal sera against rabbit MT, was employed. Copper-loaded rats had marked MT-immunoreactivity within the nucleus and cytoplasm of many periportal hepatocytes, renal proximal convoluted tubule epithelial cells, intestinal columnar epithelial cells and Paneth cells. Immunohistochemical staining was similar using either mouse anti-MT polyclonal serum, or monoclonal antibody E9. Hepatocytes surrounding inflammatory foci were positive for MT, supporting the proposed role of this protein in free radical scavenging. The presence of MT in the kidney appears to be associated with renal excretion of copper-metallothionein (Cu-MT) in copper-loaded rats. Paneth cells were easily detected using MT-immunostaining. MT may play a part in absorption of copper from intestinal contents and possible storage as Cu-MT in Paneth cells. The function of Paneth cells remains unknown but the presence of marked MT-immunoreactivity in these cells, observed in copper-loaded rats, suggests their involvement in homeostasis and metabolism of copper.

Animals↗
PubMed1988

On the presence of intermediate cells in the small intestine.

In the small intestine, the presence of transitional cells or cells intermediate between Paneth cells and goblet cells has been reported frequently for 100 years. Light microscopy and, more recently, fine structural studies have indicated that secretory granules observed in intermediate cells share some morphologic characteristics with those of granular goblet cells and of Paneth cells. In order to verify if intermediate cells in the jejunum and ileum of the adult mouse have functional similarities with either granular goblet or Paneth cells, we have studied the incorporation of sulfur-35 by radioautography and the localization of lysozyme by immunocytochemistry. After radioautography, goblet cells and, to a lesser extent, granular goblet cells had incorporated sulfur-35, whereas Paneth cells and intermediate cells were completely negative. Immunolocalization of lysozyme was done by using rabbit anti-rat lysozyme and protein A-peroxidase. After demonstration of peroxidase activity, only Paneth cells were stained and intermediate cells were negative. Therefore, intermediate cells do not contain sulfomucin or lysozyme, and they are functionally different from goblet and Paneth cells. Their function remains unknown.

Animals↗
PubMed1989

Cell lineage markers in premalignant and malignant colonic mucosa.

Normal colonic epithelial cells consist of several cell types or lineages that are thought to arise from a common stem cell precursor. Neoplastic transformation may occur at different stages in the differentiation of a colonic stem cell to produce tumors that may retain characteristic cell lineage phenotypes. In this study, immunohistochemical techniques were used to identify cell lineage-related markers in fetal, normal, hyperplastic, adenomatous, and cancerous colonic tissue. These markers consisted of secretory component (columnar cells), a purified mucin antigen (mucous or goblet cells), chromogranin A (enteroendocrine cells), lysozyme (Paneth cells), and carcinoembryonic antigen (panepithelial cell marker). Colonic neoplasms, like normal mucosa, predominantly expressed the markers of columnar and goblet cell lineages. Chromogranin A was expressed in a small population of cells in most normal and fetal colonic crypts. Chromogranin A reactive cells were found in 55% of hyperplastic polyps, 31% of adenomatous polyps, and 33% of carcinomas. Lysozyme reactivity was rare in fetal, normal, and hyperplastic specimens, but was present in 86% of adenomas and 40% of carcinomas. Of 42 primary carcinomas, 9% were "pluripotent" and expressed markers of all four cell lineages. In addition to columnar and goblet cell markers, 7% expressed both enteroendocrine and Paneth cell markers, 17% expressed enteroendocrine cell markers, and 24% expressed Paneth cell markers. Two cases (5%) lacked expression of any of the cell lineage markers. The remainder expressed only columnar and goblet cell markers. The markers used in this study appear to identify the major cell lineages of fetal and normal colonic epithelium and can be used to delineate the altered cell lineage phenotypes in premalignant and malignant colonic mucosa.

Biomarkers, Tumor↗
PubMed1981

The quantification of zinc in the mucosal cells of human small intestine using x-ray microanalysis.

The intracellular zinc concentration of human small intestinal mucosal cells was estimated using X-ray microanalysis. Jejunum and ileum from 11 patients resected during surgery was fixed in 2.5% glutaraldehyde and acetone and the estimations performed with an AEI EMMA 4. The zinc content of undifferentiated stem cells, enterocytes, goblet cells and Paneth cells was determined and all the cell types were found to contain significant amounts of zinc. The highest levels were found in the stem cells and enterocytes with a maximum in the ideal enterocytes in Crohn's disease. In non-inflammatory conditions the ileal enterocytes had a zinc content than the jejunal enterocytes. Stem cells had comparable levels in all tissues analysed. Goblet cells showed a wide variation in zinc levels that may be due to their secretory status. Paneth cells had the lowest zinc levels. The whole Paneth cell zinc and granule zinc were of the same order in all tissues sampled. The cytoplasmic zinc was higher than the granule zinc in all samples of ileum analysed, but was no significant difference in the jejunum. The limitations of the technique are considered and observations made on the role of zinc in intestinal mucosa.

Adult↗
PubMed1996

Paneth cell-rich flat adenoma of the rectum: report of a case.

A patient having familiar adenomatosis polyposis and an ileo-rectal anastomosis developed a flat mucosal lesion in the rectum. A punch biopsy revealed a villous adenoma with high-grade dysplasia. The subsequent surgical specimen indicated that the flat villous adenoma was rich in Paneth cells. Special stains included lysozyme muramidase (to visualize Paneth cells), MIB1 proliferation monoclonal antibody and single and multilabel immunohistochemistry for Paneth cells. Other methods included transmission electron microscopy and quantification with an image quantifier (Program Optilab 2.1) of lysozyme-stained Paneth cells. The subjective evaluation of hematoxylin-eosin-stained preparations demonstrated that the Paneth cells were mainly located in the lower half of the villi. Sections labeled with a specific stain (lysozyme muramidase) revealed more Paneth cells in the villi and electron microscopy showed even more in lysozyme-negative areas. Obviously some migrating dysplastic Paneth cells had retained their characteristic granules on their way towards the tip of the villi. Quantitative studies indicated that the lysozyme muramidase-positive material accounted for 41% of the adenomatous tissue. MIB1 revealed intense cell proliferation at the base of the adenoma and in the entire slopes of the villi. Despite the wide distribution of Paneth cells in intestinal metaplasia of the stomach, in the normal small intestine and in the large bowel with chronic inflammatory diseases, it is surprising that tumors arising in Paneth cells are extremely rare. The causes of the apparent natural resistance of Paneth cells to tumor development deserve to be investigated. This is the first case of Paneth cell-rich flat adenoma of the rectum in the literature.

Adenoma↗
PubMed2003

Candidate markers for stem and early progenitor cells, Musashi-1 and Hes1, are expressed in crypt base columnar cells of mouse small intestine.

Musashi-1, a neural RNA-binding protein, is important for maintaining neural stem cells. Both Musashi-1 and Hes1, a transcriptional factor regulated by Musashi-1, are expressed in the small intestine. Here we show that Musashi-1 is present in a few epithelial cells just above the Paneth cells in the small intestinal crypt, the putative position of stem cells, whereas Hes1 is expressed in lower crypt cells just above the Paneth cells, including Musashi-1-positive cells. Musashi-1 and Hes1 were not expressed in Paneth cells. Notably, Musashi-1 and Hes1 were coexpressed in the crypt base columnar cells located between the Paneth cells. These findings suggest that not only the cells just above Paneth cells but also the crypt base columnar cells between the Paneth cells have stem cell characteristics.

Animals↗
PubMed2005

NOD2 mutation and mice: no Crohn's disease but many lessons to learn.

Many patients with ileal Crohn's disease, a chronic intestinal inflammation, carry mutations in the gene encoding NOD2 (CARD15), but the mechanistic details of how this mutation leads to disease are not fully understood. NOD2 is expressed in antigen-presenting cells and Paneth cells, which are secretory epithelial cells of the small intestine. Two complementary studies using genetically engineered murine models help to explain the association of NOD2 malfunction and mucosal disease. One study observes a dysregulation of proinflammatory responses, suggesting that the most common NOD2 mutation in humans results in a gain of function. The other study determined that NOD2-null mutations impair the Paneth-cell antimicrobial response, which is consistent with recent findings in humans. Together, these studies fuel optimism that new therapeutic directions might emerge to better treat this severe mucosal disease.

Animals↗
PubMed1992

Paneth cell-like change and small cell carcinoma of the prostate. Two divergent forms of prostatic neuroendocrine differentiation.

Paneth cell-like change of the prostate refers to collections of prostatic cells with eosinophilic cytoplasmic granules that bear a striking histological resemblance to normal intestinal Paneth cells. Paneth cell-like change in malignant prostatic epithelium usually represent neuroendocrine differentiation, with neuroendocrine granules confirmed by immunohistochemical and ultrastructural studies. We report the histopathological, immunohistochemical, and electron microscopic findings in a mixed adenocarcinoma with Paneth cell-like change and small cell undifferentiated carcinoma. This case illustrates two divergent forms of neuroendocrine differentiation occurring in a single prostatic neoplasm. The spectrum of neuroendocrine differentiation in the prostate should be expanded to include tumors with Paneth cell-like change in addition to carcinoid tumors and small cell undifferentiated carcinoma. These three distinct forms of prostatic neuroendocrine neoplasia appear to correlate with three size ranges of neuroendocrine granules seen by electron microscopy.

Adenocarcinoma↗
PubMed1976

Precancerous changes in the stomach.

Intestinal metaplasia is often associated with human gastric carcinoma. Intestinalization seems to be a typical example of abnormal differentiation and is possibly a precancerous state. For investigation of intestinal metaplasia, a method for visualizing disaccharidases using Tes-Tape was developed; this method was applied to many specimens of stomach surgically removed for the treatment of gastric carcinoma. More than 130 specimens of human stomach were investigated. Intestinalization was classified into types I and II intestinal metaplasia. In type I intestinal metaplasia, sucrase, maltase, trehalase, alkaline phosphatase, goblet cells, and Paneth cells were present; while the type II intestinal metaplasia, sucrase and maltase were present but alkaline phosphatase and trehalase were absent. In type II, goblet cells were present but not Paneth cells. The histochemical technique for sucrase was newly devised. Some of the villi with goblet cells in the area of intestinalization in the stomach were not stained by sucrase activity, although most of the villi were stained. The presence of a third type of metaplasia was suggested. Purified sucrases obtained from the intestine and one case of type I intestinal metaplasia showed blood group reactivity due to the oligosaccharide side chain. However, purified sucrases obtained from two cases of type II intestinal metaplasia were negative in blood group reactivity. A close relation between distribution of alpha-fetoprotein and carcinoembryonic antigen in gastric carcinoma and that in surrounding intestinal metaplasia is discussed.

Carcinoembryonic Antigen↗
PubMed1982

Intestinal metaplasia of the stomach as a precancerous stage.

Intestinal metaplasia is defined as the appearance of intestinal epithelium in the stomach. Intestinal metaplasia is frequently found in populations with a high incidence of gastric cancer. Macroscopic demonstration of sucrase and trehalase with Tes-tape in many resected stomachs yielded new information for understanding the nature of intestinal metaplasia. Intestinal metaplasia can be classified into two types, complete and incomplete. The former is associated with the presence of sucrase, trehalase, leucine aminopeptidase, alkaline phosphatase, goblet cells and Paneth cells, and the latter with that of sucrase, leucine aminopeptidase and goblet cells, but not trehalase or Paneth cells. Goblet cells in the complete type of intestinal metaplasia contain sialomucin, as does the small intestine, while those in the incomplete type contain sulphomucin and sialomucin, as does the large intestine. Well-differentiated adenocarcinoma is closely related to intestinal metaplasia, especially the incomplete type. Atypical epithelium of intestinal metaplasia has been proposed as a more proximate stage of gastric cancer. Intestinal metaplasia can be diagnosed by staining with dye under endoscopic observation. A reduced level of pepsinogen I in the blood reflects the presence of severe intestinal metaplasia, which is understood to be a sign of high risk of gastric cancer. Intestinal metaplasia is supposed to be produced by components of food. Mutagens/carcinogens such as N-methyl-N'-nitro-soguanidine and N-propyl-N'-nitro-N-nitrosoguanidine can produce intestinal metaplasia in the glandular stomach of rats and gastric cancers. The formation of intestinal metaplasia precedes the appearance of adenocarcinoma in the glandular stomach. Intestinal metaplasia, which is a kind of host reaction to environmental agents, may result either from genetic change - change in DNA structure - or from epigenetic change - change in the differentiation mechanism. Preventive measures could be developed to suppress the development of intestinal metaplasia and to suppress the process of conversion of metaplastic cells to cancer cells.

Cell Differentiation↗
PubMed2002

Parameters of intestinal inflammation in mice given graded infections of the nematode Trichinella spiralis.

Four parameters of the intestinal inflammatory response (numbers of mucosal mast cells (MMC) and Paneth cells, villus:crypt ratios and mitotic figures) were measured in mice exposed to varying doses of infective larvae of Trichinella spiralis. The aim of the experiments was to determine whether generation of these components of inflammation required a threshold level of infection and whether, once triggered, inflammation became pan-mucosal. Near maximal MMC and Paneth cell responses were elicited even with infections as low as 35 larvae; changes in villus:crypt ratios and in mitotic indices also occurred at this level of infection, but were progressively greater with increasing levels of infection. In all infected mice, including those infected with 35 larvae, MMC and Paneth cell responses extended over most of the small intestine. These data are interpreted as showing: (i) that the intestinal mucosa is highly responsive to T. spiralis infection; (ii) that once triggered, components of the inflammatory response are amplified by T cell-dependent mechanisms, becoming pan-mucosal; and (iii) that MMC and Paneth cell responses, which require cell division and differentiation, become maximal at a lower infection threshold than changes in the villus:crypt ratio or in mitotic indices, which directly reflect increased rates of division in crypt cells.

Animals↗
PubMed1979

[Particular ultrastructural aspects of human Paneth's cells].

Our research is about the ultrastructural aspects of Paneth cells obtained from human jejunal mucosa. The cells appear columnar in shape and show a roundish nucleus; their cytoplasm is characterized by a great amount of R.E.R. and by typical large granules in sopranuclear position. Our results confirm those previously described by other authors about the morphology of Paneth cells. In our specimens we have found moreover a variable number of particular cytoplasmic structures; these may show a fibrillar or granular matrix and are always surrounded by a membrane.

Cell Nucleus↗
PubMedDate not supplied

Cloning of intestinal phospholipase A2 from intestinal epithelial RNA by differential display PCR.

Differential display polymerase chain reaction (DD-PCR) is a powerful technique for comparing gene expression between cell types, or between stages of development or differentiation. Differentially expressed genes may be cloned and analysed further. Here we extend the use of DD-PCR to analyse differences in gene expression between two complex epithelia: that of the small intestine and of the large intestine. The aim of this study was to identify genes expressed preferentially in Paneth cells. Paneth cells are secretory epithelial cells putatively involved in host defense and regulation of crypt cell proliferation and are found at the base of the small intestinal crypts adjacent to the stem cell zone. Of 34 clones that were analysed, partial sequencing identified two clones related to known Paneth cell products: a homologue of secretory phospholipase A2 (clone B1) and a homologue of a neutrophil defensin (clone C5). B1 was strongly expressed in Paneth cells, as demonstrated by in-situ hybridization. B1 was also expressed at a lower level in the large intestinal epithelium. A full length B1 cDNA clone was isolated and sequenced, and shown to be highly homologous to type II secretory phospholipase A2 genes, and almost identical to the enhancing factor gene and the putative gene for the MOM-1 locus. B1 expression is limited to the intestinal tract, and we propose that it be designated intestinal phospholipase A2, or i-PLA2. The method we describe is well suited to the rapid identification of genes expressed exclusively or predominantly in Paneth cells.

Animals↗
PubMed1997

Reliability of rectal biopsy in distinguishing between chronic inflammatory bowel disease and acute self-limiting colitis.

AIMS: In order to assess the validity of previously proposed criteria for the differentiation of chronic inflammatory bowel disease (CIBD) from acute self-limiting colitis (ASLC) all rectal biopsies were reported by a single histopathologist with a long-term gastrointestinal interest over a 4.5-year period. METHODS AND RESULTS: The presence or absence of distorted crypt architecture, increased lymphocytes and plasma cells, villous mucosal architecture, granulomata, basal lymphoid aggregates, basal giant cells and Paneth cell metaplasia was noted for each biopsy. The definite presence of any of the above features, with the exception of intramucosal granulomata, was regarded as indicative of CIBD. Eighteen months later all available case notes were examined and the presenting clinical symptoms and working clinical diagnosis extracted. The final diagnosis, histopathological diagnosis and the presence or absence of any of the above histopathological features were correlated and the positive predictive value of each histopathological feature was calculated. A correct diagnosis of either CIBD or ASLC was made in 80 of 84 and 29 of 31 cases, respectively. CONCLUSIONS: Villous mucosal architecture and Paneth cell metaplasia were found to be specific features of CIBD. Distorted crypt architecture, basal lymphoid aggregates and plasma cell infiltration of the lamina propria were also useful features but strict definition of these features is required and discussed. Intramucosal epithelioid granulomas were identified in eight cases of CIBD and four cases of ASLC. In association with ruptured crypts intramucosal granulomas are not specific features of Crohn's colitis.

Acute Disease↗
PubMed1994

Generation of neomucosa in vivo by transplantation of dissociated rat postnatal small intestinal epithelium.

A novel method to study the generation of rat small intestinal mucosa, by transplantation of disaggregated postnatal rat small intestinal epithelium is described. Cellular aggregates, comprised of epithelium with attached proliferative cells and closely associated stromal tissue, were isolated from postnatal rat small intestine by enzymatic digestion, then grafted immediately to the subcutaneous plane of adult recipients. On graft retrieval after 14 days, 39% of cellular transplants to nude mice, and 84% of cellular transplants to inbred rats had developed into small intestine-like structures. These structures were comprised of a circumferential layer of epithelium surrounding a central mucin filled lumen. This neomucosal layer exhibited well formed crypts and villi, and contained all epithelial stem cell lineages i.e. absorptive enterocytes, goblet cells, Paneth's cells and entero-endocrine cells. Proliferative activity within this neomucosa was confined to crypt regions as in normal postnatal small intestine. Developmental maturation within the regenerated neomucosa was demonstrated by organotypic morphogenesis, i.e. formation of mature crypts and villi, and progressive cytodifferentiation with increased numbers of goblet cells, entero-endocrine cells and Paneth's cells. Altered patterns of brush border enzyme expression further confirmed a temporal progression of development within neomucosal enterocytes. It is concluded that after "extensive" mucosal disaggregation, postnatal small intestinal epithelial progenitor cells retain the capacity for organotypic regeneration of neomucosa when transplanted to ectopic sites in adult recipients. These small aggregates of epithelium and stroma are capable of generating the topographical signals necessary for the three dimensional regeneration of this tissue. Furthermore, the multipotent generative potential of the stem cells within these cellular aggregates is maintained with production of all progeny.

Animals↗
PubMed2006

In vitro culture of embryonic mouse intestinal epithelium: cell differentiation and introduction of reporter genes.

BACKGROUND: Study of the normal development of the intestinal epithelium has been hampered by a lack of suitable model systems, in particular ones that enable the introduction of exogenous genes. Production of such a system would advance our understanding of normal epithelial development and help to shed light on the pathogenesis of intestinal neoplasia. The criteria for a reliable culture system include the ability to perform real time observations and manipulations in vitro, the preparation of wholemounts for immunostaining and the potential for introducing genes. RESULTS: The new culture system involves growing mouse embryo intestinal explants on fibronectin-coated coverslips in basal Eagle's medium+20% fetal bovine serum. Initially the cultures maintain expression of the intestinal transcription factor Cdx2 together with columnar epithelial (cytokeratin 8) and mesenchymal (smooth muscle actin) markers. Over a few days of culture, differentiation markers appear characteristic of absorptive epithelium (sucrase-isomaltase), goblet cells (Periodic Acid Schiff positive), enteroendocrine cells (chromogranin A) and Paneth cells (lysozyme). Three different approaches were tested to express genes in the developing cultures: transfection, electroporation and adenoviral infection. All could introduce genes into the mesenchyme, but only to a small extent into the epithelium. However the efficiency of adenovirus infection can be greatly improved by a limited enzyme digestion, which makes accessible the lateral faces of cells bearing the Coxsackie and Adenovirus Receptor. This enables reliable delivery of genes into epithelial cells. CONCLUSION: We describe a new in vitro culture system for the small intestine of the mouse embryo that recapitulates its normal development. The system both provides a model for studying normal development of the intestinal epithelium and also allows for the manipulation of gene expression. The explants can be cultured for up to two weeks, they form the full repertoire of intestinal epithelial cell types (enterocytes, goblet cells, Paneth cells and enteroendocrine cells) and the method for gene introduction into the epithelium is efficient and reliable.

Adenoviridae↗
PubMed1987

Argyrophilic adenocarcinoma of the prostate with Paneth cell-like granules.

A prostatic adenocarcinoma with argyrophilia and many Paneth cell-like granules in a 91-year-old man is reported. The initial symptom was pollakisuria, and the laboratory data showed no significant abnormality except for moderate anemia. Through radiologic examinations a right hydronephrosis and metastatic cancer of the lumbar vertebra were suspected. Because prostatic cancer was suspected, a needle biopsy of the prostate was performed. Routine histologic examinations revealed a moderately differentiated adenocarcinoma of the prostate. By hematoxylin and eosin stain, eosinophilic large granules like those of Paneth cells were found in many cancer cells. In addition, argyrophilic cancer cells were seen by Grimelius' stain. Immunoperoxidase staining elucidated a definite reactivity for lysozyme in the granules like Paneth cells. Ultrastructurally, the granules of Paneth cell-like cancer cells were electron-dense spherical bodies with a medium diameter of 540 nm. Prostatic adenocarcinoma with argyrophilia is rare, and the argyrophilic adenocarcinoma with Paneth cell-like granules is very rare in the prostate.

Adenocarcinoma↗