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Biomedical subjects

P D Howdle

Publications and source records attributed to P D Howdle.

At least 73 records · Page 4Linked to original sources

Jejunal plasma cells and in vitro immunoglobulin production in adult coeliac disease.

IgA, IgE, IgG and IgM plasma cells in small bowel mucosal biopsies from 15 controls, 16 untreated and 14 treated coeliac patients and five patients with selective serum IgA deficiency (four of whom also had coeliac disease) were quantified using an indirect immunoperoxidase technique. The IgA, IgG and IgM plasma cell counts were significantly increased in the untreated coeliac patients. The cell counts were intermediate in the treated coeliac group. These changes were in parallel to production in vitro of IgA and sIgA, IgG, and IgM by cultured mucosal biopsies from the same patients. The IgA deficient patients had very few mucosal IgA cells but elevated IgG and IgM plasma cell numbers; again these changes were reflected in the production in vitro of immunoglobulins. IgE plasma cell counts were very low in all patients and there were no differences between patient groups. The changes in cell counts and mucosal immunoglobulin production were not reflected in serum IgA, IgM and IgG concentrations but serum secretory IgA was significantly elevated in the untreated coeliac patients compared with controls, with the treated coeliac patients being intermediate. The raised mucosal plasma cell counts reflect the local mucosal production of immunoglobulin but not the immunoglobulin concentrations of serum, emphasising the importance of studying the immune function of the gut itself in coeliac disease rather than immunological abnormalities in serum.

Adolescent↗

Organ culture of fetal rat small intestine for testing gluten toxicity: a reappraisal.

Jejunal segments from fetal rats of 18 days gestation were maintained in an organ culture system for up to 72 h. During this period, villi developed within the intestinal lumen and the epithelium changed from stratified to simple columnar. Peristaltic activity was observed during in-vitro culture. Alkaline phosphatase specific activity of the bowel segments fell after 48 hours culture, compared with pre-culture values (P less than 0.05), but that of alpha-glucosidase increased. The addition of Frazer's gluten fraction III to the culture medium caused slowing in the rate of morphological maturation of the jejunal explants, but there was no additional effect on enzyme specific activities compared with segments cultured in gluten-free medium. The place of organ culture of fetal rat intestine as an animal model for testing cereal toxicity in the study of coeliac disease remains unclear.

Animals↗

Immunoglobulin production by coeliac biopsies in organ culture.

The production of immunoglobulins by jejunal mucosa during organ culture has been studied. In 18 untreated coeliac patients the amounts of IgA, secretory IgA and IgM in the culture medium were higher than in those from 17 normal controls. The results in 15 treated coeliac patients did not differ from the control subjects. Mucosal biopsies from patients with serum IgA deficiency produced very little IgA, but large amounts of IgG and IgM. The addition of neither Frazer's gluten fraction III, nor alpha-gliadin to the culture system stimulated any additional increased secretion of immunoglobulins from the untreated coeliac patients. Production of IgA and IgM by the small bowel mucosa continues during organ culture, but that of IgG appears insignificant.

Adolescent↗

Secretion of gliadin antibody by coeliac jejunal mucosal biopsies cultured in vitro.

The role of gliadin antibodies in coeliac disease is unknown. We have investigated this by maintaining jejunal biopsies from control subjects, treated and untreated coeliac patients in organ culture for 24 h. The tissue culture supernatants were assayed by enzyme-linked immunosorbent assay (ELISA) for total IgG, IgM and IgA, anti-gliadin and casein antibodies. Jejunal biopsies from untreated coeliac patients secreted more total IgM and IgA but not IgG, than biopsies from control subjects: values for treated coeliac patients were similar to those from control subjects. There was increased secretion of IgG and IgM gliadin antibody, but not casein antibody, by jejunal biopsies from untreated coeliac patients compared with those from control subjects.

Antibody Formation↗

The distribution of gluten-sensitive lymphocytes in coeliac patients--is it related to dietary gluten?

Cell-mediated immunity to gluten within the peripheral blood and jejunal mucosa of the same coeliac patients was measured simultaneously using migration inhibition tests. There was an inverse correlation in migration indices indicating that as cell-mediated immunity became more detectable in the peripheral blood, it was less so in the jejunal mucosa and vice versa. There was virtually no relationship, however, between this correlation and the gluten content of the diet in these coeliac patients.

Adult↗

The microenvironment of coeliac disease: T cell phenotypes and expression of the T2 'T blast' antigen by small bowel lymphocytes.

Peroral jejunal biopsies were studied by double-label immunofluorescence on cryostat tissue sections from control patients of normal histology and patients with coeliac disease. A panel of monoclonal antibodies was used to identify T cells and T cell subsets, together with antibody markers of cell stimulation (the T2 40 kdalton T blast specificity), proliferation (T9 and Ki67) and activation (HLA-DR and Ig receptors). In normal mucosa, expression of T2 was predominantly found in the T8+ intraepithelial cytotoxic/suppressor population. In coeliac disease there was little alteration of T4:T8 ratios per se, but a much higher percentage of T8-T4+ helper/inducer cells expressed the T2 antigen (approx. 30% compared to approx. 10%), manifest as an accretion of T2+T8- cells in the epithelium and subjacent stroma. Additionally, T cell lymphoid aggregates were observed in the lamina propria, consisting of greater than 90% T4+ cells, of which the majority (60-80%) were also T2+. The increase of stimulated helper cells correlated well with the pathology of coeliac disease, being most marked in untreated cases of maximal tissue damage, and least in well treated cases with restoration of normal morphology. By contrast, an increased tendency for the T8+ cells to co-express T1 in untreated coeliac disease was not observed in treated disease, even in cases where failure to adhere to a strict gluten free diet had resulted in little histological improvement. There was little evidence of T cell activation or proliferation. The observation that T helper cells undergo migration and stimulation suggests an imbalance in immunoregulation which causes a breakdown of normal immune tolerance to dietary gluten, and therefore may be a crucial mechanism of tissue damage in coeliac disease.

Adolescent↗

Routine colonoscopy service.

We report our experience of 50 paediatric colonoscopies in a relatively unselected group of patients. Collaboration between an experienced colonoscopist and a paediatrician has provided a smooth, routine service and the examination has proved extremely useful for the diagnosis and management of children with colonic symptoms.

Child↗

Leucocyte migration inhibition test in coeliac disease - a reappraisal.

Results of the direct leucocyte migration inhibition (LMI) test using gluten fraction III as antigen were unaffected by incorporation of puromycin into the culture medium at concentrations shown to prevent lymphokine mediated inhibition. Results of the LMI test performed with purified polymorphs were similar to and correlated with results of the standard LMI test using mixed leucocytes in both coeliacs and controls. The addition of purified T lymphocytes did not increase migration inhibition. Normal leucocytes incubated with serum from coeliac patients and washed showed marked migration inhibition when incubated with gluten fraction III. This sensitisation of normal leucocytes was prevented by preincubation with aggregated human IgG. These results suggest that leucocyte migration inhibition by gluten in coeliac disease is not due to lymphokine production by sensitised lymphocytes but is caused by cytophilic antibody.

Adolescent↗

Low fibronectin in portal hypertension.

We have measured plasma fibronectin in 28 patients with well-compensated chronic liver disease, 4 patients with non-cirrhotic portal hypertension and 6 patients who have undergone shunt surgery for the relief of portal hypertension and splenectomy. 17 patients with portal hypertension had significantly lower levels of fibronectin (207 +/- 54; mean +/- SD) compared with 15 patients without portal hypertension (315 +/- 75, p less than 0.001) and the 6 patients who had undergone shunting and splenectomy (330 +/- 66, p less than 0.001). We suggest that in patients with portal hypertension and splenomegaly, low levels of fibronectin might result from increased consumption by the enlarged spleen.

Fibronectins↗

Jejunal biopsy and lymphocyte co-culture in coeliac disease.

Jejunal biopsies from 13 patients with treated coeliac disease were maintained in organ culture for 24 h with and without the addition of gluten fraction III, in each case with and without autologous peripheral blood mononuclear cells. Biopsies cultured with gluten alone or with lymphocytes alone showed no significant reduction in mean enterocyte height compared with those cultured in control medium, but those cultured with both gluten and lymphocytes had significantly lower enterocyte height than control biopsies, than those cultured with gluten alone, and than those cultured with lymphocytes alone. These results demonstrate that the combination of gluten and lymphocytes is more toxic to treated coeliac mucosa in vitro than either one alone and support the suggestion that immune reactions to gluten may be important in the pathogenesis of the mucosal lesion of coeliac disease.

Adult↗

Cell-mediated immunity to gluten within the small intestinal mucosa in coeliac disease.

Jejunal biopsies from controls and coeliac patients were maintained in organ culture in the presence of gluten fraction III. The culture media were assayed for evidence of lymphokine activity in a migration inhibition test using normal peripheral blood leucocytes. Significant inhibition of migration was produced by media from untreated coeliac patients compared with controls (P less than 0.005) or treated coeliac patients (P less than 0.001), indicating the production of a leucocyte migration inhibition factor (LIF) by untreated coeliac mucosa in response to gluten fraction III. The degree of inhibition correlated with the preculture interepithelial lymphocyte count in the coeliac biopsies (P less than 0.02). In six coeliac patients studied when on a normal diet and on a gluten-free diet, LIF was produced while on a normal diet, but not while on a gluten-free diet. These results suggest that a local cell-mediated immune reaction to gluten is present in the mucosa of patients with untreated coeliac disease but that this is reversed by treatment with a gluten-free diet.

Adult↗

Cell-mediated immunity to dietary antigens in coeliac disease.

By means of the leukocyte migration inhibition test a significant depression of migration index (indicating increased immunity) was found in 10 untreated coeliac patients compared with 24 control subjects with the dietary antigens bovine serum albumin (BSA) and egg white but not with milk. The degree of immunity was similar to that obtained with gluten fraction III as antigen. Treatment with a gluten-free diet led to a decrease in immunity to egg white and BSA, but immunity to gluten fraction III was increased in the early months of treatment.

Adolescent↗

In vitro diagnosis of coeliac disease: an assessment.

Jejunal biopsies from controls and coeliac patients were maintained in organ culture for up to 48 hours. The in vitro effect of gluten fraction III during the period of culture was assessed by measurement of the activity of the brush border enzymes alkaline phosphatase and alpha-glucosidase. Mucosa from controls and treated and untreated coeliacs behaved similarly and no reproducible in vitro effect of gluten was demonstrated. These results cast doubt on the in vitro diagnosis of coeliac disease by monitoring brush border enzyme activity.

Alkaline Phosphatase↗

Gluten sensitivity of small intestinal mucosa in vitro: quantitative assessment of histologic change.

In view of the importance to the patient of demonstrating mucosal sensitivity to gluten we have quantitated morphologic change during organ culture of small bowel mucosa by measuring enterocyte height. In 27 normal controls the mean preculture enterocyte height was 28.3 micrometer +/- 1.8 (+/- 1 SD), decreasing to 25.6 micrometer +/- 1.9 and 26.1 micrometer +/- 2.0 after 24 h culture in the absence and presence of gluten, respectively. Both these decreases were significant (p < 0.001), but there was no difference between culture with or without gluten. In 9 abnormal controls, the mean preculture enterocyte height was 27.0 micrometer +/- 2.7, decreasing significantly (p < 0.05) to 24.5 micrometer +/- 1.1 after 24 h culture without gluten, and to 25.4 micrometer +/- 2.3 with gluten (NS). In 17 untreated celiac patients, the mean preculture enterocyte height was 19.6 micrometer +/- 1.9 increasing significantly to 23.8 micrometer +/- 2.0 (p < 0.001) after 24 h culture without gluten but decreasing to 18.7 micrometer +/- 3.1 after 24 h culture with gluten (NS). There was a significant difference between culture with or without gluten (p < 0.001). In 21 treated celiac patients, the mean preculture enterocyte height was 28.1 micrometer +/- 1.9 decreasing to 25.4 micrometer +/- 2.0 and 24.2 +/- 1.9 in the absence and presence of gluten, respectively. Both these decreases were significant (p < 0.001), but more importantly there was a significant decrease (p < 0.001) after culture with gluten compared with culture without gluten. These data indicate that it is possible to quantify morphologic change during organ culture and to demonstrate gluten-sensitivity not only in untreated, but also in treated celiac mucosa. These studies have important implications as regards the diagnosis and further investigation of the etiology of celiac disease.

Caseins↗