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Original research
Diagnosis of coeliac disease by flow cytometry of intraepithelial lymphocytes: a new ‘gold’ standard?
  1. Kaninika Basu1,
  2. Hannah Creasey2,
  3. Nina Bruggemann1,
  4. Jennifer Stevens2,
  5. David Bloxham2,
  6. Jeremy Mark Woodward1
  1. 1Gastroenterology and Clinical Nutrition, Cambridge University Hospitals NHS Foundation Trust, Cambridge, UK
  2. 2Haematology Oncology Diagnostic Service, Cambridge University Hospitals NHS Foundation Trust, Cambridge, UK
  1. Correspondence to Dr Jeremy Mark Woodward, Gastroenterology and Clinical Nutrition, Cambridge University Hospitals NHS Foundation Trust, Cambridge CB2 1TN, UK; jeremy.woodward{at}addenbrookes.nhs.uk

Abstract

Objective The analysis of intraepithelial lymphocytes (IELs) by flow cytometry of duodenal biopsies—the ‘IEL’ lymphogram—has been proposed as a diagnostic test for coeliac disease. However, its clinical applicability has been limited due to variability in methods and definitions. This study set out to define useful parameters for the application of the IEL lymphogram to the diagnosis of coeliac disease.

Design Flow cytometry was performed on 117 sets of duodenal biopsies in 107 adult patients with active coeliac disease, long-term coeliac disease on a gluten free diet and a control group. The initial 95 samples were used for hypothesis generation for the subsequent samples comprising 12 patients with coeliac disease and 10 controls.

Results Rather than using single linear cut-offs for CD3 and T-cell receptor γδ (TCRγδ)+ve IELs, a discriminant function was identified as %CD3+ve IELs+2x(%TCRγδ+IELs)>100. This differentiated coeliac disease from control biopsies in the hypothesis generating group. These results were replicated in the validation group and found to be independent of histology in patients on long-term gluten free diet up to 12 years (combined sensitivity, 98.5%; specificity, 97.7%).

Conclusions Flow cytometric analysis of IELs is a highly sensitive and specific adjunct to serology and histological examination for the diagnosis of coeliac disease, even in individuals with coeliac disease following a gluten free diet who exhibit normal duodenal histology.

  • coeliac disease
  • immunology
  • gluten sensitive enteropathy
  • gluten
  • gluten free diet

Data availability statement

Data are available upon reasonable request. Deidentified participant data available from the author on reasonable request.

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Data availability statement

Data are available upon reasonable request. Deidentified participant data available from the author on reasonable request.

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Footnotes

  • Contributors KB contributed to the writing and laboratory work. HC, NB, JS and DB carried out the cell separation and flow cytometry and contributed intellectual input. JMW conceived the project, carried out the gastroscopies and duodenal biopsies and contributed to the writing.

  • Funding The authors have not declared a specific grant for this research from any funding agency in the public, commercial or not-for-profit sectors.

  • Competing interests None declared.

  • Provenance and peer review Not commissioned; externally peer reviewed.

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