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A murine cytotoxic T lymphocyte clone from the intestinal mucosa that is antigen specific for proliferation and displays broadly reactive inducible cytotoxic activity.

Thy-1+, Lyt-1-,2+, asialo GM1+ cytotoxic T lymphocyte (CTL) clones have been isolated from the intestinal mucosa of mice primed with alloantigens. Two different types of cytotoxic clones have been obtained. The first type is functionally similar to most splenic and lymph node-derived CTL clones in that they are strictly antigen specific with respect to proliferation and cytolytic activity. The second type of CTL clone has several unique characteristics. Although these clones are also antigen specific with regard to proliferation, they are not cytolytic under standard growth conditions in medium containing 4% rat concanavalin A-induced spleen cell supernatant. After culture for 4 days in the presence of high concentrations of interleukin 2, cells become activated and exhibit broad lytic potential. Moreover, during the activation process, these CTL begin to express a murine T cell surface antigen, CT-1, which is associated with activated cytotoxic cells. The findings reported in this report should now allow us to precisely define, both phenotypically and functionally, specific lymphocyte populations that make up the gut-associated lymphoid tissues. These data also describe a new type of effector CTL that differs from other cytotoxic cells reported to date, because it is antigen dependent for proliferation, but requires signals mediated by lymphokines for lytic activation.

Animals↗

Adult worms of the rodent intestinal nematode, Strongyloides venezuelensis, successfully invade chick intestinal mucosa.

In order to study the mucosal invasion of a rodent intestinal nematode in bird intestine, chicks were infected with the intestinal nematode of rodents, Strongyloides venezuelensis, by subcutaneous larva inoculation and adult worm implantation. No evidence was obtained for larvae reaching the lungs or the intestine after infective larva inoculation. Adult worms implanted in the small intestine invaded the mucosa and remained there at least for 24 h, whereas those implanted in the caecum were trapped by mucus, and did not invade the mucosa. Mucosal invasion of adult worms in the small intestine was confirmed by histological examination. The number of adult worms in the intestinal mucosal tissue dropped rapidly within the first 24 h, which was associated with infiltrating granulocytes around the worms. The present study suggests that S. venezuelensis adult worms are able to invade the intestinal tissue of chicks, which do not belong to the vertebrate class of its normal definitive host, but that they are eliminated rapidly by mucosal defense system of the bird.

Animals↗

The expulsion of Echinostoma trivolvis from C3H mice: differences in glycoconjugates in mouse versus hamster small intestinal mucosa during infection.

Mucosal glycoconjugates were examined in C3H mice and in hamster small intestines infected with Echinostoma trivolvis and in uninfected rodents, using periodic-acid Schiff (PAS) and high-iron diamine-alcian blue (HID-AB) staining and three different fluorescein-conjugated lectins: Triticum vulgaris agglutinin (WGA), Helix pomatia agglutinin (HPA) and Griffonia simplicifolia agglutinin (GSA-II). Lectin-labelling by electron microscopy was also undertaken with WGA and HPA lectin-gold probes. HID-AB stain demonstrated that the most mature goblet cells of the mouse villi contain sulfomucins, whereas those of hamsters contain sialomucins. The expression of lectin-binding sites and the intensity of the lectin binding in the small intestines were changed by echinostome infection. Specific differences in the reaction to mucin glycoproteins were clearly observed between the mouse and hamster intestines infected with E. trivolvis; lectin-binding to hyperplastic goblet cells and crypts in the infected mice increased, while no marked increase in the number of goblet cells and reaction to the glycoconjugates were observed in the infected hamsters. These findings indicate that the expression of terminal N-acetyl-D-galactosamine, sialic acid and N-acetyl-D-glucosamine increased in mucins secreted from hyperplastic goblet cells associated with E. trivolvis infection in mice. No marked increase in these glycoconjugates occurred in hamster infections. These findings reflect clear differences in infectivity of E. trivolvis in C3H mice versus hamsters.

Animals↗

Ability of enteroaggregative Escherichia coli strains to adhere in vitro to human intestinal mucosa.

A collection of 44 enteroaggregative Escherichia coli (EAggEC) strains isolated from infants with diarrhea in India and the United Kingdom were examined for their ability to adhere in vitro to human intestinal mucosa and by electron microscopy for production of putative adherence factors. None of the strains adhered to human duodenal mucosa, and six strains tested did not adhere to ileal mucosa; all 44 strains, however, adhered to human colonic mucosa in localized aggregates. Electron microscopy of infected colonic mucosa indicated fimbrially mediated adhesion of the EAggEC strains. Four morphologically distinct kinds of fimbriae, including a new morphological type of E. coli fimbriae consisting of bundles of fine filaments, were identified among the EAggEC strains; this new type of fimbria was observed in 43 of the 44 EAggEC strains. Forty-three of the 44 EAggEC strains were positive with a DNA probe developed to identify EAggEC, and most of the strains belonged to serotypes unrelated to the other major classes of diarrheic E. coli. These results suggest that EAggEC may be a large-bowel pathogen and colonize the colon by a fimbrially mediated adhesion mechanism.

Bacterial Adhesion↗

Scanning electron microscopic study of the intestinal mucosa of an Indian freshwater adult major carp, Labeo rohita (Hamilton).

Fine anatomical structures of the different regions (viz., anterior, middle and posterior) of the intestinal mucosa in adult Labeo rohita (Hamilton) have been studied by using Scanning Electron Microscope (SEM). Surface mucosal modifications of the anterior and the middle of the intestine not only increase the luminal surface by 2-4 times but serve also the storage function of the ingested food due to the absence of stomach in the fish studied. Fine anatomical structures of the columnar epithelial and goblet cells of the mucosa have been studied in detail. The height and complexity of the microridges on the luminal end of each columnar cells in the various regions vary greatly probably due to the functional variations of the region concerned. Communication between the columnar cells and the sub-mucosal layer through the basement membrane is effected by circular orifices. A pear-shaped apical portion provided with prominent microridges of the goblet cell probably allows the mechanical stress to develop for the accumulation of secretory mucins.

Animals↗

Cellular lesions in intestinal mucosa of gnotobiotic calves experimentally infected with a new unclassified bovine virus (Breda virus).

Four gnotobiotic calves were inoculated intranasally with Breda virus within two hours after cesarean section. Three calves developed diarrhea between 48 and 60 hours after inoculation; one calf was killed at 36 hours. Consistent microscopic findings in the intestinal mucosae of calves with diarrhea were cytopathologic changes in enterocytes of the lower small intestine, large intestine, and dome epithelial cells and an acute inflammatory response with cellular infiltration and subtle changes in capillaries. Virions with average dimensions of 35 x 80 nm were seen in rounded cells that had severe cell swelling, dilatation of the cytocavitary network, and hydropic degeneration. Virions were most often in multiple, large autophagolysosomes. There was a good correlation in detection of virus in cells with electron microscopy and indirect immunofluorescence. No lesions were detectable in two control calves killed five days after inoculation.

Animals↗

Fatty acid synthesis in intestinal mucosa of guinea pig.

1. Acetate-CoA ligase, acetyl-CoA-carbon dioxide ligase and fatty acid synthetase were shown to be present in particle-free fractions of guinea-pig intestinal mucosa. 2. Each of these enzymes was partially purified by ammonium sulphate precipitation from the particle-free supernatant. 3. The incorporation of acetate and citrate into fatty acid was measured. 4. Gas-liquid radiochromatography was used to investigate the pattern of fatty acids synthesized. 5. The rate-limiting step in fatty acid synthesis was shown to be acetyl-CoA-carbon dioxide ligase.

Acetates↗

Elevated concentration of adenosine 3':5'-cyclic monophosphate in intestinal mucosa after treatment with cholera toxin.

A heat-labile factor in cell-free filtrate of a Vibrio cholerae culture induces a marked rise in the wet-weight concentration of adenosine 3':5'-cyclic monophosphate (cyclic AMP) in the intestinal mucosa of the dog. The increase becomes appreciable 1-1.5 hr after intraluminal administration of the filtrate, about the same time as the onset of intestinal secretion in response to a heat-labile enterotoxin in the filtrate. These results are consistent with the hypothesis that cyclic AMP may be an intermediary in the intestinal secretory response to cholera toxin.

Animals↗

Effect of orally administered betel leaf (Piper betle Linn.) on digestive enzymes of pancreas and intestinal mucosa and on bile production in rats.

The influence of two varieties of betel leaf (Piper betle Linn.) namely, the pungent Mysore and non-pungent Ambadi, was examined on digestive enzymes of pancreas and intestinal mucosa and on bile secretion in experimental rats. The betel leaves were administered orally at two doses which were either comparable to human consumption level or 5 times this. The results indicated that while these betel leaves do not influence bile secretion and composition, they have a significant stimulatory influence on pancreatic lipase activity. Besides, the Ambadi variety of betel leaf has a positive stimulatory influence on intestinal digestive enzymes, especially lipase, amylase and disaccharidases. A slight lowering in the activity of these intestinal enzymes was seen when Mysore variety of betel leaf was administered, and this variety also had a negative effect on pancreatic amylase. Further, both the betel leaf varieties have shown decreasing influence on pancreatic trypsin and chymotrypsin activities.

Administration, Oral↗

Improved method for the recovery of Ascarus suum larvae from pig intestinal mucosa.

In the course of a comprehensive study on the population biology of Ascaris suum, it became necessary to determine quantitatively the migration of the larvae (L2) through the pig intestine. Because no satisfactory methods for recovering these larvae from the intestinal mucosa were apparently available in the literature, we undertook the development of such a procedure. Direct Baermannization of the intestine proved inadequate, so a series of studies was undertaken to find a method to solubilize the mucosa and free the larvae. Both pepsin digestion and mucus reduction with N-acetyl cysteine were evaluated. The highest recovery rates were obtained with short-term pepsin digestion (45-90 min), followed by a specific settling and washing procedure. A third alternative, migration of larvae out of agar-gel, was also tested. Although the recovery efficiency was low (10% of pepsin digestion), the larvae recovered were clean, with good motility. The agar-gel procedure could be useful when L2 larvae are needed for other purposes, such as antigen preparation or cultivation.

Acetylcysteine↗

[Alkaline phosphatase in the intestinal mucosa of children with the malabsorption syndrome].

110 children suffering from malabsorption underwent several biopsies of the gut to confirm coeliac disease (CD) following the ESPGAN criteria. We studied the values for alkaline phosphatase (AP) in the intestinal mucosa after gluten challenge. In 42 patients the after challenge biopsy was normal, thus excluding coeliac disease. In 68 children the mucosa was severely damaged confirming CD. In all biopsy specimens lactase, invertase, maltase and alkaline phosphatase were measured. We found a good correlation between PA values and severity of mucosal damage, showing that measurement of PA in the mucosa is helpful in assessing the degree of mucosal atrophy in children suffering from malabsorption.

Alkaline Phosphatase↗

[Activation of nuclear factor-kappaB and effects of anti-inflammatory treatment thereon in intestinal mucosa of patients with ulcerative colitis].

OBJECTIVE: To investigate the activation and expression of nuclear factor-kappaB (NF-kappaB) and effects of anti-inflammatory treatment on NF-kappaB in the intestinal mucosa of patients with ulcerative colitis (UC). METHODS: Ten pieces of colon mucosal biopsy specimens were obtained from 31 cases with UC, 17 of which received sulphasalazine (SASP) or SASP plus glucocorticoid and 14 of which received no medication. Samples of normal mucosa around the lesion taken from 11 patients with colon cancer were used as controls. NF-kappaB DNA binding activity was evaluated by electrophoretic mobility shift assay. NF-kappaB p65 expression was determined by Western blot analysis and immunohistochemical staining with a NF-kappaB p65 antibody. The type of cells containing activated NF-kappaBp65 was identified by double immunofluorescence confocal laser scanning microscopy. RESULTS: The expression of NF-kappaB p65 and NF-kappaB DNA binding activity were significantly higher in patients with UC than in the control (P < 0.05), and were correlated with the degree of inflammation. The NF-kappaB expression was significantly stronger in the nuclei than in the cytoplasm in patients with UC without pharmacotherapy. The NF-kappaB expression in nuclei was significantly stronger in the group without pharmacotherapy than in the group with pharmacotherapy (P < 0.05). Only a few NF-kappaB p65 positive cells were seen in the controls. NF-kappaBp65 expression was found in all major subsets of mononuclear cells, including macrophages, B lymphocytes, T lymphocytes, and cryptal epithelial cells. CONCLUSION: The increased activation of NF-kappaB and increased expression of NF-kappaB may be involved in the pathogenesis of UC. Glucocorticoids and SASP strongly inhibited NF-kappaB activation and expression. The inhibition of NF-kappaB activation may be a central part of the anti-inflammatory action of glucocorticoids and SASP, which might represent an important pharmacological mechanism in treatment of patients with UC. NF-kappaB will be an important target for cytokine-based therapy of UC.

Adult↗

Mannan and oligomers of N-acetylglucosamine protect intestinal mucosa of celiac patients with active disease from in vitro toxicity of gliadin peptides.

Wheat flour and other cereals toxic for celiac patients contain an alcohol-soluble protein fraction that, under experimental conditions simulating in vivo protein digestion, yields peptides that agglutinate undifferentiated K 562(S) cells. In contrast, cereals well tolerated in celiac disease (i.e., rice and maize) do not. Furthermore, purified A-gliadin peptides that damage in vitro-cultured flat celiac mucosa are powerful agglutinins for K 562(S) cells, whereas A-gliadin peptides that do not show any adverse in vitro effect on celiac intestine lack agglutinating activity. Mannan, acetylglucosamine, and its oligomers (N,N'-diacetylchitobiose and N,N',N"-triacetylchitotriose) were able to prevent and reverse cell agglutination induced by peptides from all the toxic cereals. Moreover, mannan and N,N',N"-triacetylchitotriose exhibited a protective effect on intestinal mucosa specimens of patients with active celiac disease cultured with wheat protein-derived peptides. These data are consistent with the hypothesis that the agglutinating and toxic peptides are bound by carbohydrates.

Acetylglucosamine↗

Lactic acid bacteria (LAB) bind to human B- or H-antigens expressed on intestinal mucosa.

Twenty lactobacilli isolated from human feces were studied for binding to the human blood type B-antigen [Galalpha1-3 (Fucalpha1-2) Gal-] and H-antigen (Fucalpha1-2Gal-] expressed sugar chains in human intestinal mucosa. We found two strains, L. gasseri OLL2755 and L. gasseri OLL2877 that firmly adhere to human B-antigen, and we found L. gasseri OLL2827 bound to the H-antigen.

Bacterial Adhesion↗

The mononuclear cells of human mesenteric blood, intestinal mucosa and mesenteric lymph nodes: compartmentalization of NK cells.

The proportions of T cell subsets and Leu 7+ cells and the spontaneous cell-mediated cytotoxicity (SCMC) of isolated mononuclear cells have been determined across the mesenteric vascular bed and along the intestinal mucosal-mesenteric lymph node (MLN) axis in patients undergoing abdominal surgery. Whereas the proportion of T4+ and T8+ cells were similar in simultaneously taken PVB and mesenteric venous blood (MVB), the proportion of Leu 7+ cells was higher in MVB in 16 of 17 studies (15.4 +/- 6.8%, 10.8 +/- 5.1%). Additional studies showed that the proportions of lymphocyte subsets in peripheral arterial blood are the same as those in PVB. Thus, an enrichment of Leu 7+ cells occurs across the mesenteric vascular bed. Isolated intestinal and MLN mononuclear cells contained similarly high proportions of T4+ and T8+ cells as in PVB but Leu 7+ cells made up a minority subpopulation in intestinal (1.3 +/- 0.8%) and MLN mononuclear cells (1.0 +/- 0.9%). The SCMC of intestinal and MLN mononuclear cells was low and paralleled the proportion of Leu 7+ cells. Despite the higher proportions of Leu 7+ cells in MVB, the SCMC was less than that of PVB in eight patients with inflamed intestine and not significantly different from PVB in seven patients with normal intestines. These paradoxical findings were at least in part due to inhibitory factors in mesenteric plasma. In conclusion, NK cells appear to be largely confined within the vascular system and the enrichment of Leu 7+ cells across the mesenteric vascular bed suggests that this compartmentalization may be due to differences in the traffic of lymphocyte subpopulations through the intestinal mucosa and MLN.

Cell Movement↗

An Escherichia coli strain that causes diarrhea by invasion of the small intestinal mucosa and induces monosaccharide intolerance.

E. coli can induced diarrhea either by enterotoxin production or by invasion of the colonic mucosa. Here we report a 2/12 year old infant with caute diarrhea induced by E. coli strain, isolated from the jejunal fluid, that had no enterotoxigenic activity but invaded the small intestinal mucosa and induced severe morphological alterations. Total villous atrophy and monosaccharide intolerance occured. After 51 days of hospitalization there was a partial recovery of the small intestinal morphology and the patient could also tolerate disaccharides again.

Acute Disease↗