Gastroenterology

Gastroenterology

Volume 115, Issue 3, September 1998, Pages 551-563
Gastroenterology

Alimentary Tract
Gluten induces an intestinal cytokine response strongly dominated by interferon gamma in patients with celiac disease,☆☆

https://doi.org/10.1016/S0016-5085(98)70134-9Get rights and content

Abstract

Background & Aims: Celiac disease appears to be a T cell–mediated enteropathy induced by gluten in genetically predisposed individuals. Duodenal biopsy specimens from patients with celiac disease and histologically normal controls were investigated to see if cytokine expression is related to disease activity. Methods: Cytokine messenger RNA (mRNA) expression was determined by quantitative reverse-transcription polymerase chain reaction and in situ expression by immunohistochemistry. Results: In normal controls, mRNA levels were usually below the quantitative limit, even after in vitro gluten stimulation. By contrast, interferon (IFN)-γ mRNA was increased more than 1000-fold in untreated disease. In vitro gluten stimulation of specimens from treated patients (gluten-free diet) increased IFN-γ mRNA to the levels of untreated patients. In addition, increased mRNA levels for interleukin (IL)-2, IL-4, IL-6, and tumor necrosis factor α were found after such stimulation, whereas mRNA for IL-5, IL-10, and IL-12p40 was usually below the quantitative level. Biopsy specimens from untreated patients contained on average 10-fold more lamina propria cells positive for IFN-γ than normal controls, whereas cells containing IL-4 were rare in both subject groups. Conclusions: The results show that mucosal gluten exposure in patients with celiac disease rapidly elicits high levels of IFN-γ expression and lower levels of IL-2, IL-4, IL-6, and tumor necrosis factor α even in the virtual absence of IL-12.

GASTROENTEROLOGY 1998;115:551-563

Section snippets

Patients

The patients enrolled in the cytokine mRNA study were evaluated according to the European Society for Pediatric Gastroenterology and Nutrition criteria for celiac disease of 1990.19 Biopsy specimens from the distal duodenum (n = 1–5 per subject) were obtained during upper gastrointestinal endoscopy (Olympus GIF IT20 endoscope; biopsy forceps, Olympus FB 13K; Tokyo, Japan) from 20 patients with celiac disease (median age, 38 years; range, 6–68 years), 7 with untreated disease and 13 on a

Estimation of cytokine mRNA expression by RT-PCR

Intestinal specimens from histologically normal controls, both unstimulated and stimulated in vitro with gluten, were either negative or showed only low expression of IFN-γ, IL-2, IL-4, IL-5, IL-6, and IL-12p40 mRNA. Moreover, we were unable to detect transcripts for IL-10 and TNF-α (Figure 1).

. Agarose gel electrophoresis for semiquantitative determination of PCR-amplified cytokine mRNA in duodenal specimens from 8 control subjects with normal histology. Specimens from 2 subjects (controls

Discussion

Quantification of mRNA levels by RT-PCR showed that the expression of IFN-γ was remarkably increased in the duodenal mucosa of patients with celiac disease after gluten exposure in vivo or in vitro compared with similarly exposed histologically normal control mucosa. Conversely, only low mucosal levels of IL-2, IL-4, IL-5, IL-6, IL-12p40, and TNF-α mRNA were detected even after gluten stimulation, whereas all specimens showed constitutive expression of TGF-β but were negative for IL-10. In

Acknowledgements

The authors thank Aaste Aursjø, Drude Hansson, and Tone Narvesen for excellent technical assistance; Dr. Ø. Molberg for providing T-cell clones; and Dr. M.F. Kagnoff for the generous gift of plasmids to generate standard mRNA. Mabtech AB is acknowledged for covering the costs of the color plate.

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    Address requests for reprints to: Ellen M. Nilsen, Ph.D., LIIPAT, Rikshospitalet, N-0027 Oslo, Norway. e-mail: [email protected]; fax: (47) 22112261.

    ☆☆

    Supported by the Norwegian Cancer Society, the Research Council of Norway, Anders Jahre's Foundation, the Medinnova Governmental Research Organization, and the Norwegian Celiac Disease Association.

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