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PubMed1986

Immunocytochemical localization of pancreatic carboxylic ester hydrolase in human paneth cells.

The protein-A gold method using specific rabbit sera directed against pure human pancreatic chymotrypsinogen and carboxylic ester hydrolase was applied to locate these (pro)enzymes in human pancreatic acinar cells and intestinal Paneth cells. Quantitative evaluation of the labelling indicated that both (pro)enzymes are present in pancreatic acinar secretory granules. In Paneth cell secretory granules, only carboxylic ester hydrolase was present in significant amounts, although the labelling for this enzyme was less intense than that observed in pancreatic zymogen granules. The results obtained support the view that Paneth cells represent a "diffuse exocrine gland" scattered along the intestine, whose role is either to act as a substitute in the event of a deficient pancreas or to regulate the intestinal flora.

Carboxylesterase↗
PubMed1991

Selective killing of Paneth cells by intravenous administration of dithizone in rats.

Paneth cells are zinc-containing cells widely distributed in Lieberkühn's crypts of intestine in a variety of species. We found that rapid selective killing of Paneth cells took place after the intravenous (i.v.) injection of diphenylthiocarbazone (dithizone), a chelator forming a zinc dithizonate complex, in the rat. As soon as 5 min after the i.v. injection of dithizone, degeneration of Paneth cells occurred. At this stage, zinc dithizonate complexes were observed as purple-red granules in bright field microscopy. Thirty to 60 min later, Paneth cells were detached from the basement membrane and shed into the cryptic lumen. After 6 h, the cell debris in the crypts was no longer seen and the crypts once housing Paneth cells were now occupied by neighbouring crypt base columnar cells. Histochemically demonstrable zinc totally disappeared. After 12-24 h, however, definite Paneth cells began to resume. Histochemical staining for zinc was again positive at the apex of these cells. One week after dithizone administration, the number of Paneth cells increased twice as much as in uninjected control and histochemical staining for zinc was highly positive. After 2 weeks, Paneth cell hyperplasia subsided. X-ray microanalysis revealed that zinc was the most abundant metal in Paneth cells. We concluded that chelation of zinc and formation of zinc-dithizone complexes in Paneth cells' cytoplasm would be responsible for the selective degeneration observed after dithizone administration.

Animals↗
PubMed1990

Ultrastructure of Paneth cells in the intestine of various mammals.

Paneth cells in the following species were observed under an electron microscope: human, rhesus monkey, hare, guinea pig, rat, nude rat, mouse, golden hamster, and insect feeder bat. Secretory granules containing homogeneous electron-dense materials were observed in the Paneth cells of humans, monkeys, hares, guinea pigs, and bats; mouse Paneth-cell granules were bipartite (central core and peripheral halo), and the Paneth cells in rats and golden hamsters had secretory granules showing various electron densities. In humans, monkeys, and bats, immature granules near the Golgi apparatus sometimes showed bipartite substructure. The number and size of secretory granules were also diverse among various animal species. Some lysosome-like bodies were commonly observed in peri- or supranuclear regions, though the size and shape of the bodies differed from cell to cell. In apical cytoplasm, small clear vesicles (100-200 nm diameter) were more-or-less observed in all species examined, and it was especially note that rat Paneth cells contained many clear vesicles. Small dense-cored vesicles (150-200 nm diameter) were rare. It is unlikely that the various ultrastructural features of Paneth cells correlate with the phylogenetical classification.

Adult↗
PubMedDate not supplied

A simple method to demonstrate normal and metaplastic Paneth cells in tissue sections.

Using hematoxylin and eosin (H&E)-stained sections, Paneth cells are identified by the presence of eosinophilic granules. Several other methods have been proposed to stain Paneth cells. Recently, we noticed that Paneth cells were autofluorescent in H&E sections. To assess the number of Paneth cells by this method, 30 surgical specimens from the small bowel and 10 from the colon with IBD were investigated. Consecutive sections were stained with H&E and immunostained with lysozyme (Lz). Paneth cell counting was done in 20 well-oriented, vertically cut crypts per-case using alternatively transmitted light (TL) and incident-light fluorescence (ILF). When H&E and Lz sections were observed with TL, the number of Paneth cells was easily assessed in some crypts, but in crypts having numerous Paneth cells, their exact number was ascertainable. On the other hand, using IFL, the number of H&E-stained Paneth cells-crypt was easily assessed, even in crypts with Paneth cell clustering. Using H&E-ILF, a mean of 3.9 (range 3-8) Paneth cells-pe-crypt was found in the normal small bowel and a mean of 3.3 (range 1-14) in the colon with IBD. ILF provided the adequate visual conditions--i.e. a dark, non-autofluorescent background--for assessing the actual number of normal and metaplastic Paneth cells, even when arranged in tight cell groups. The present method will improve the enumeration of the actual number of normal and metaplastic Paneth cells both in experimental research and in clinical trials.

Cell Count↗
PubMed2006

Adenomyoma with goblet and Paneth cells of the ileum.

A case of ileal adenomyoma with goblet and Paneth cells is reported. A 75-year-old man died of ruptured hepatocellular carcinoma. As an incidental finding at autopsy, a 9 x 7 x 6 mm(3)-sized nodule was found in the ileal wall. Histologically, the lesion occupied the submucosa and muscularis propria, and consisted of glandular structures of various sizes and interlacing smooth muscle bundles surrounding the glandular elements. Goblet cells and Paneth cells were interspersed in the glandular element. Immunohistochemically, the glandular element was positive for cytokeratin (CK) 7 and negative for CK 20. This is the first reported case of adenomyoma of the gastrointestinal tract that contained Paneth cells. The result of the immunohistochemical staining favored the heterotopic pancreas theory concerning its pathogenesis. The appearance of goblet and Paneth cells might be the result of metaplasia.

Adenomyoma↗
PubMed1987

The morphogenesis of the human Paneth cell. An immunocytochemical ultrastructural study.

In human duodenal mucosa Paneth cells originate away from the base of crypts and migrate towards the base during maturation. The earliest cells in the Paneth cell lineage could be identified by labelling of lysozyme in the Golgi apparatus. Specific labelling for lysozyme was present in the rough endoplasmic reticulum, Golgi apparatus, condensing vacuoles, granules and many lysosomes of mature Paneth cells. The maturation of the Paneth cell is accompanied by an increase in the content of lysozyme in the secretory granules and with senescence lysozyme diffuses into the cytoplasm.

Cell Differentiation↗
PubMed1995

[Overexpressed p53 protein and cell-proliferative activity disappear or remarkably decrease in neoplastic CGA-positive endocrine and Paneth cells in cancer or dysplasia of ulcerative colitis].

We examined over-expression of p53 protein and proliferative activity of neoplastic endocrine and Paneth cells in cancer or dysplasia of ulcerative colitis (UC). The labeling index of p53 or Ki-67 of endocrine cells was counted with double immunostaining of p53 or Ki-67 and chromogranin A (CGA). 540 CGA-positive cells and 65 Paneth cells were all negative for p53. 5.1% of CGA-positive cells and none of 56 Paneth cells were positive for Ki-67 reaction. These observation suggest that even in cancer or dysplasia of UC cell-differentiation to neoplastic endocrine and Paneth cells decrease proliferative activity, and that these types of cell-differentiation suppress over-expression of p53 protein.

Cell Division↗
PubMed2001

Activation of Paneth cell alpha-defensins in mouse small intestine.

Paneth cells in small intestine crypts secrete microbicidal alpha-defensins, termed cryptdins, as components of enteric innate immunity. The bactericidal activity of cryptdins requires proteolytic activation of precursors by matrix metalloproteinase-7 (MMP-7; matrilysin) (Wilson, C. L., Ouellette, A. J., Satchell, D. P., Ayabe, T., Lopez-Boado, Y. S., Stratman, J. L., Hultgren, S. J., Matrisian, L. M., and Parks, W. C. (1999) Science 286, 113-117). Here, we report on the intracellular processing of cryptdin proforms in mouse Paneth cells. Peptide sequencing of MMP-7 digests of purified natural procryptdins identified conserved cleavage sites in the proregion between Ser(43) and Val(44) as well as at the cryptdin peptide N terminus between Ser(58) and Leu(59). Immunostaining co-localized precursor prosegments and mature cryptdin peptides to Paneth cell granules, providing evidence of their secretion. Extensive MMP-7-dependent procryptdin processing occurs in Paneth cells, as shown by Western blot analyses of intestinal crypt proteins and proteins from granule-enriched subcellular fractions. The addition of soluble prosegments to in vitro antimicrobial peptide assays inhibited the bactericidal activities of cryptdin-3 and -4 in trans, suggesting possible cytoprotective effects by prosegments prior to secretion. Levels of activated cryptdins were normal in small bowel of germ-free mice and in sterile implants of fetal mouse small intestine grown subcutaneously. Thus, the initiation of procryptdin processing by MMP-7 does not require direct bacterial exposure, and the basal MMP-7 content of germ-free Paneth cells is sufficient to process and activate alpha-defensin precursors. MMP-7-dependent procryptdin activation in vivo provides mouse Paneth cells with functional peptides for apical secretion into the small intestine lumen.

Amino Acid Sequence↗
PubMed1975

The Paneth cell in the adenoma of familial polyposis coli.

It is said that the Paneth cells are found in the large intestine in a pathological state such as ulcerative colitis or adenoma. We examined the Paneth cells in the adenomas of familial polyposis coli. Nine cases including one case of Gardner's syndrome comprised the material for the examination of the Paneth cells because the caecum was available for the examination. The remaining one case had no Paneth cells. In two cases, the Paneth cells were found among the adenomas in the areas beyond the caecum and the proximal part of the colon ascendens. In one remarkable case, the Paneth cells were found in 43% of the adenomas in the caecum. Seven cases were carcinomas but no Paneth cells were found in or near the carcinoma. In the control cases, which were taken from the resected colon with a disease other than familial polyposis coli, the Paneth cells were found confined to the caecum. We concluded that the distribution of the Paneth cell-bearing adenomas reflects the distribution of the Paneth cells in the normal mucosa of the large intestine and that the Paneth cells in the adenoma may have differentiated in the adenoma.

Adenoma↗
PubMed1976

Immunocytochemical identification and localization of immunoglobulin A within Paneth cells of the rat small intestine.

Light microscopic immunocytochemistry was used to identify Paneth cells by their lysozyme content and to detect immunoglobulin antigens within a subpopulation of these cells. Antisera specific for the heavy chains of rat or human immunoglobulin A and for immunoglobulin light chain antigens produced specific staining of rat Paneth cells. The distribution of immunoglobulin staining varied between adjacent Paneth cells in the same crypt and between Paneth cells in adjacent crypts, as well as between Paneth cell populations of different animals. No staining of rat Paneth cells was detected using antisera specific for the heavy chain of immunoglobulins G or M. The specific staining of Paneth cells for immunoglobulin A and light chain antigens was blocked by absorption of each antiserum with its respective purified antigen. Absorption of these antisera with purified rat lysozyme did not affect staining and thereby eliminated the possibility of immunologic cross-reactivity between lysozyme and immunoglobulin antigens. It is suggested, in light of current concepts of Paneth cell function, that the immunoglobulin staining of Paneth cells may reflect their ability to phagocytize immunoglobulin A-coated microorganisms or immune complexes containing immunoglobulin A.

Animals↗
PubMed1994

Paneth cells of African giant rats (Cricetomys gambianus).

The ultrastructure of Paneth cells of African giant rats (Cricetomys gambianus), which were captured on the savanna in western Africa, was studied. The Paneth cells of Cricetomys were clustered at the bottom of crypts of the small intestine, but not of the colon. In the normal state, Paneth cells had a few secretory granules showing high electron density. Small clear vesicles which are a characteristic in laboratory albino rats were not conspicuous. Vacuolated Paneth cells and secreted materials from the Paneth cells were frequently found. This suggests that the secretion of Paneth cells of Cricetomys is active in the natural state. No phagocytotic figures were observed. After atropine sulfate treatment, secretory granules increased in size and number, whereas the electron density decreased, similar to that of goblet cell granules. However, the granules were not stained by alcian blue or by PAS. Inhibition of secretory stimuli by atropine can alter the intracellular processing of secretory substances in Paneth cells.

Animals↗
PubMed1996

Expression of CD15 in normal and metaplastic Paneth cells of the digestive tract.

AIMS: To substantiate that incubation with monoclonal antibody CD15 (C3D-1) elicits a distinctive immunoreaction in normal small intestinal Paneth cells, normal and metaplastic Paneth cells along the digestive tract were assessed to determine whether they are also immunoreactive to CD15. METHODS: Paneth cells in paraffin wax embedded specimens of normal small intestine, appendix and proximal ascending colon, and from cases of chronic gastritis and ulcerative colitis were investigated immunohistochemically for lysozyme and CD15 antigen expression by means of the avidin-biotin peroxidase complex method. RESULTS: CD15 antibody reacted with a high proportion of both normal and metaplastic Paneth cells. Paneth cell immunoreactivity to CD15, however, was less intense and less extensive than to antilysozyme antibody, though the latter also stained many other cell types and was more commonly associated with nonspecific background staining. CONCLUSIONS: CD15 seems to be a valuable adjuvant for the study of Paneth cells in the normal and diseased digestive tract. Furthermore, as CD15 has been shown to be involved in activation of phagocytes, its expression in Paneth cells reinforces their proposed role as antimicrobial agents and regulators of the intestinal flora.

Appendix↗
PubMed2004

Paneth cell carcinoma of the ampulla of Vater.

We describe a Paneth cell carcinoma arising within the ampulla of Vater in a 64-year-old man. The phenotype of virtually all neoplastic cells was consistent with that of Paneth cells, based on routine morphology and their strong positive immunostaining for lysozyme. Additional widespread positive immunostaining for carcinoembryonic antigen and CA 19.9 supports a totipotential cell as the origin of such neoplastic cells. This case, therefore, represents a true Paneth cell carcinoma, as opposed to inclusion of occasional neoplastic Paneth cells into a poorly differentiated adenocarcinoma. This pattern of differentiation is rare, and predictions regarding its ultimate biological behavior and malignant potential must be guarded.

Adenocarcinoma↗
PubMed2005

In vivo killing of Giardia trophozoites harbouring bacterial endosymbionts by intestinal Paneth cells: an ultrastructural study.

To date Paneth cells have not previously been reported to kill Giardia trophozoites and other protozoa in vivo. Here we report the first evidence for in vivo killing of Giardia trophozoites by intestinal Paneth cells. Transmission electron microscopic (TEM) examination of duodenal specimens taken from naturally infected mice revealed that only Giardia trophozoites harbouring peripheral bacterial endosymbionts were destroyed and lysed in the vicinity of the activated Paneth cells. Additionally, intestinal epithelium was more affected by Giardia harbouring bacterial endosymbionts than Giardia with no endosymbionts. Our findings imply that the bacterial endosymbionts within Giardia trophozoites have a role in both host protective and pathological mechanisms, probably through altering the trophozoite antigencity. These observations might shed light on the diversity in infectivity and host specificity of Giardia species.

Animals↗
PubMed2006

Paneth cell alpha-defensin synthesis and function.

Endogenous antimicrobial peptides (AMPs) mediate innate immunity in every species in which they have been investigated. Cathelicidins and defensins are the two major AMP families in mammals, and they are abundant components of phagocytic leukocytes and are released by epithelial cells at mucosal surfaces. In the small intestine, Paneth cells at the base of the crypts of Lieberkühn secrete alpha-defensins and additional AMPs at high levels in response to cholinergic stimulation and when exposed to bacterial antigens. Paneth cell alpha-defensins evolved to function in the extracellular environment with broad-spectrum antimicrobial activities, and they constitute the majority of bactericidal peptide activity secreted by Paneth cells. The release of Paneth cell products into the crypt lumen is inferred to protect mitotically active crypt cells from colonization by potential pathogens and confers protection from enteric infection, as is evident from the immunity of mice expressing a human Paneth cell alpha-defensin transgene to oral infection by Salmonella enterica serovar Typhimurium. alpha-Defensins in Paneth cell secretions also may interact with bacteria in the intestinal lumen above the crypt-villus boundary and influence the composition of the enteric microbial flora. Mutations that cause defects in the activation, secretion, dissolution, and bactericidal effects of Paneth cell AMPs may alter crypt innate immunity and contribute to immunopathology.

Amino Acid Sequence↗
PubMed1982

Light microscopic morphometric analysis of rat ileal mucosa: II. Component quantitation of Paneth cells.

A quantitative light microscopic morphometric analysis of lysozyme- and IgA-containing Paneth cells within the ileal mucosa of physiologically manipulated and control (sham operation) animals was performed. The experimental groups of rats included animals raised in a gnotobiotic environment (microbial reduction) and animals with ileal self-filling blind (microbial proliferation) and Thiry-Vella (intestinal discontinuity) loops. The unlabeled antibody enzyme immunohistochemical localization technique was employed for the identification of intracellular lysozyme and IgA. Component quantitation was performed by use of a micrometer component quantitator. Marked Paneth cell hyperplasia was noted in association with gnotobiosis and with the Thiry-Vella fistula. This observation quantitatively confirms previous subjective impressions of increased Paneth cell differentiation in association with those physiologic states. Since the neurovascular component of the Thiry-Vella fistula is intact, the normal intraluminal succus entericus would appear to be involved in modulation of Paneth cell differentiation. The recognition of Paneth cell hyperplasia in association with the Thiry-Vella fistula suggests that this may be a useful experimental model for an evaluation of the life cycle and functional characteristics of this cell population. The results also revealed that no significant change in the volume percentage of Paneth cells and a decreased volume percentage of Paneth cells containing IgA occurred in association with the self-filling blind loop. A decreased volume percentage of IgA-containing immunocytes in association with the blind loop has previously been reported. The data are most consistent with the interpretation that the Paneth cell and immunocyte response to antigenic stimulation are interrelated and that the Paneth cell population has a restricted latitude of response to microbial proliferation.

Animals↗
PubMed2000

Secretion of microbicidal alpha-defensins by intestinal Paneth cells in response to bacteria.

Paneth cells in mouse small intestinal crypts secrete granules rich in microbicidal peptides when exposed to bacteria or bacterial antigens. The dose-dependent secretion occurs within minutes and alpha-defensins, or cryptdins, account for 70% of the released bactericidal peptide activity. Gram-negative bacteria, Gram-positive bacteria, lipopolysaccharide, lipoteichoic acid, lipid A and muramyl dipeptide elicit cryptdin secretion. Live fungi and protozoa, however, do not stimulate degranulation. Thus intestinal Paneth cells contribute to innate immunity by sensing bacteria and bacterial antigens, and discharge microbicidal peptides at effective concentrations accordingly.

Animals↗
PubMed1998

Activation of cAMP-dependent C1- currents in guinea-pig paneth cells without relevant evidence for CFTR expression.

1. To determine whether Paneth cells exhibit functional expression of cAMP-activated Cl- currents and molecular expression of the cystic fibrosis transmembrane conductance regulator (CFTR), we applied whole-cell patch clamp and single-cell mRNA analysis by reverse transcription (RT) followed by polymerase chain reaction (PCR) amplification to single Paneth cells in crypts isolated from the guinea-pig small intestine. 2. Prominent activation of Cl- currents was consistently observed after stimulation with dibutyryl cAMP and forskolin or with vasoactive intestinal polypeptide (VIP). The cAMP-activated Cl- current was inhibited by removal of intracellular ATP or administration of an inhibitor of protein kinase A. 3. Many of the biophysical and pharmacological properties of the currents were phenotypically similar to those of the CFTR Cl- channel, such as the ohmic current-voltage relationship, the anion selectivity with a Type III sequence (Br- > Cl- > I- >> F- >= gluconate-), I--induced blockage, insensitivity to a stilbene-derivative Cl- channel blocker, and sensitivity to a carboxylate analogue Cl- channel blocker. The sensitivity of the current to glibenclamide was, however, much weaker than that reported for the CFTR Cl- channel current. In contrast to the time independence of CFTR currents, the inward component of the Paneth cell Cl- currents exhibited inactivation kinetics. 4. Expression of CFTR mRNA could not be detected by RT-PCR analysis in almost all single Paneth cells, although its expression was consistently detected at the whole-crypt level. The presence of a small number of CFTR-expressing epithelial cells, which were scattered both in villi and crypts but not at the crypt base where Paneth cells were located, was demonstrated by immunocytochemistry. 5. Taken together, it appears that guinea-pig Paneth cells functionally express cAMP-activated Cl- conductance without relevant evidence for molecular expression of CFTR. Functional expression of VIP receptors in the Paneth cells was also demonstrated.

Animals↗