Evaluation of T-Bet immunostaining in the counting of intraepithelial lymphocytes in celiac disease
Keywords:T-bet, Immunostaining, Gluten Sensitive Enteropathy, Distribution pattern, Marsh grading, Immunohistochemistry
Objective: In this study, we aimed to evaluate CD3, T-bet and GATA3 staining of intraepithelial lymphocytes (IELs) in comparison to the routine H&E stains in celiac disease.
Methods: a total of 50 patients, in whom celiac disease was diagnosed based on a combination of clinicopathological features, were enrolled in the study. Duodenal biopsied tissues were processed routinely into formalin fixed paraffin embedded (FFPE) blocks. Sections were prepared and stained with H&E and each of CD3, T-bet and GATA3. A number of histological criteria were measured to calculate the Marsh score. The results were analyzed using the IBM SPSS analytic software.
Results: a positive correlation was found between the count of T-bet stained cells and the increase of intraepithelial lymphocytes (P-value = 0.001). In addition, low count of GATA3 stained cells was seen in almost all cases. The count of GATA3 stained cells was not affected by the increase in IELs count.
Conclusions: the majority of increased IELs were stained with T-bet. Whereas in normal IELs count is less than half the IELs were stained with T-bet. This would indicate that T-bet immunostaining is a potential alternative to H&E and/or CD3 based counting of IELs.
2. Lerner A. Serological diagnosis of celiac diseaseâ€“moving beyond the tip of the iceberg. Int J Celiac Dis. 2014;2:64-6.
3. Leon F. Flow cytometry of intestinal intraepithelial lymphocytes in celiac disease. Journal of Immunological Methods. 2011;363(2):177-86.
4. Mustalahti K, Catassi C, Reunanen A, Fabiani E, Heier M, McMillan S, et al. The prevalence of celiac disease in Europe: results of a centralized, international mass screening project. Annals of medicine. 2010;42(8):587-95.
5. Barada K, Bitar A, Mokadem MA-R, Hashash JG, Green P. Celiac disease in Middle Eastern and North African countries: A new burden? World Journal of Gastroenterology : WJG. 2010;16(12):1449-57.
6. Barada K, Daya HA, Rostami K, Catassi C. Celiac disease in the developing world. Gastrointestinal Endoscopy Clinics. 2012;22(4):773-96.
7. Al-Hussaini A, Troncone R, Khormi M, AlTuraiki M, Alkhamis W, Alrajhi M, et al. Mass Screening for Celiac Disease Among School-aged Children: Toward Exploring Celiac Iceberg in Saudi Arabia. Journal of Pediatric Gastroenterology and Nutrition. 2017;65(6):646-51.
8. Cappello M, Morreale GC, Licata A. Elderly Onset Celiac Disease: A Narrative Review. Clinical Medicine Insights: Gastroenterology. 2016;9:CGast.S38454.
9. GonzÃ¡lez DA, de Armas LG, RodrÃguez IM, Almeida AA, GarcÃa MG, Gannar F, et al. Strategies to improve the efficiency of celiac disease diagnosis in the laboratory. Journal of Immunological Methods. 2017;449:62-7.
10. Reinton N, Helgheim A, Shegarfi H, Moghaddam A. A one-step real-time PCR assay for detection of DQA1âŽ05, DQB1âŽ02 and DQB1âŽ0302 to aid diagnosis of celiac disease. Journal of Immunological Methods. 2006;316(1):125-32.
11. Green PH, Cellier C. Celiac disease. New England Journal of Medicine. 2007;357(17):1731-43.
12. Agardh D, Lee H-S, Kurppa K, Simell V, Aronsson CA, JÃ¶rneus O, et al. Clinical features of celiac disease: a prospective birth cohort. Pediatrics. 2015;135(4):627-34.
13. Fei Bao, Peter H.R. Green, Govind Bhagat. An Update on Celiac Disease Histopathology and the Road Ahead. Archives of pathology & laboratory medicine. 2012;136(7):735-45.
14. Volta U, Villanacci V. Celiac disease: diagnostic criteria in progress. Cellular And Molecular Immunology. 2011;8:96.
15. Ludvigsson JF, Leffler DA, Bai JC, Biagi F, Fasano A, Green PH, et al. The Oslo definitions for coeliac disease and related terms. Gut. 2013;62(1):43-52.
16. Walker MM, Murray JA. An update in the diagnosis of coeliac disease. Histopathology. 2011;59(2):166-79.
17. Charlesworth RPG, Andronicos NM, Scott DR, McFarlane JR, Agnew LL. Can the sensitivity of the histopathological diagnosis of coeliac disease be increased and can treatment progression be monitored using mathematical modelling of histological sections? â€“ A pilot study. Advances in Medical Sciences. 2017;62(1):136-42.
18. Bai JC, Fried M, Corazza GR, Schuppan D, Farthing M, Catassi C, et al. World Gastroenterology Organisation global guidelines on celiac disease. Journal of clinical gastroenterology. 2013;47(2):121-6.
19. Kurppa K, Ashorn M, Iltanen S, Koskinen LLE, Saavalainen P, Koskinen O, et al. Celiac Disease without Villous Atrophy in Children: A Prospective Study. The Journal of Pediatrics. 2010;157(3):373-80.e1.
20. Hudacko R, Kathy Zhou X, Yantiss RK. Immunohistochemical stains for CD3 and CD8 do not improve detection of gluten-sensitive enteropathy in duodenal biopsies. Modern pathology : an official journal of the United States and Canadian Academy of Pathology, Inc. 2013;26(9):1241-5.
21. Abadie V, Discepolo V, Jabri B. Intraepithelial lymphocytes in celiac disease immunopathology. Seminars in immunopathology. 2012;34(4):551-66.
22. Corazza G, Villanacci V. Coeliac disease. Journal of clinical pathology. 2005;58(6):573-4.
23. Pellegrino S, Villanacci V, Sansotta N, ScarfÃ¬ R, Bassotti G, Vieni G, et al. Redefining the intraepithelial lymphocytes threshold to diagnose gluten sensitivity in patients with architecturally normal duodenal histology. Alimentary pharmacology & therapeutics. 2011;33(6):697-706.
24. Siriweera EH, Qi Z, Yong JL. Validity of intraepithelial lymphocyte count in the diagnosis of celiac disease: A histopathological study. Int J Celiac Dis. 2015;3:156-8.
25. Kabiraj A, Gupta J, Khaitan T, Bhattacharya PT. PRINCIPLE AND TECHNIQUES OF IMMUNOHISTOCHEMISTRY â€“ A REVIEW2015. 5204-10 p.
26. Cheroutre H, Madakamutil L. Acquired and natural memory T cells join forces at the mucosal front line. Nature Reviews Immunology. 2004;4(4):290.
27. Neutra MR, Mantis NJ, Kraehenbuhl J-P. Collaboration of epithelial cells with organized mucosal lymphoid tissues. Nature immunology. 2001;2(11):1004.
28. Chang F, Mahadeva U, Deere H. Pathological and clinical significance of increased intraepithelial lymphocytes (IELs) in small bowel mucosa. Apmis. 2005;113(6):385-99.
29. McAllister CS, Kagnoff MF, editors. The immunopathogenesis of celiac disease reveals possible therapies beyond the gluten-free diet. Seminars in immunopathology; 2012: Springer.
30. Harris KM, Fasano A, Mann DL. Monocytes differentiated with IL-15 support Th17 and Th1 responses to wheat gliadin: Implications for celiac disease. Clinical Immunology. 2010;135(3):430-9.
31. Di Sabatino A, Corazza GR. Coeliac disease. The Lancet. 2009;373(9673):1480-93.
32. Rajendran A, Sivapathasundharam B. Shafer's Textbook of Oral Pathology: Elsevier Health Sciences; 2014.
33. JÃ¶hrens K, Anagnostopoulos I, Stein H. Tâ€bet expression patterns in coeliac disease, cryptic and overt enteropathyâ€type Tâ€cell lymphoma. Histopathology. 2005;47(4):368-74.
34. Salvati V, MacDonald T, Bajaj-Elliott M, Borrelli M, Staiano A, Auricchio S, et al. Interleukin 18 and associated markers of T helper cell type 1 activity in coeliac disease. Gut. 2002;50(2):186-90.
35. Stanford Medicine. Celiac Disease 2018. Available from: http://surgpathcriteria.stanford.edu/gi/celiac-disease/marsh.html.
36. Villanacci V, Ceppa P, Tavani E, Vindigni C, Volta U. Coeliac disease: The histology report. Digestive and Liver Disease. 2011;43:S385-S95.
37. Ludvigsson JF, Bai JC, Biagi F, Card TR, Ciacci C, Ciclitira PJ, et al. Diagnosis and management of adult coeliac disease: guidelines from the British Society of Gastroenterology. Gut. 2014:gutjnl-2013-306578.
38. Mubarak A, Wolters VM, Houwen RHJ, ten Kate FJW. Immunohistochemical CD3 staining detects additional patients with celiac disease. World Journal of Gastroenterology : WJG. 2015;21(24):7553-7.
39. Veress B, FranzÃ©n L, Bodin L, Borch K. Duodenal intraepithelial lymphocyteâ€count revisited. Scandinavian journal of gastroenterology. 2004;39(2):138-44.
40. Ensari A. Gluten-sensitive enteropathy (celiac disease): controversies in diagnosis and classification. Archives of pathology & laboratory medicine. 2010;134(6):826-36.
41. Rostami K, Marsh MN, Johnson MW, Mohaghegh H, Heal C, Holmes G, et al. ROC-king onwards: intraepithelial lymphocyte counts, distribution & role in coeliac disease mucosal interpretation. Gut. 2017:gutjnl-2017-314297.
42. Holtmann MH, Neurath MF. T helper cell polarisation in coeliac disease: any (T-)bet ? Gut. 2004;53(8):1065-7.
43. JÃ¶hrens K, GrÃ¼nbaum M, Anagnostopoulos I. Differences in the T-bet and GATA-3 expression patterns between lymphocytic colitis and coeliac disease. Virchows Archiv. 2010;457(4):451-6.
44. Omran TZ, Ahmed MM, Metib NJ. Assessment of GATA3 expression in duodenal biopsies of celiac disease suspected and diagnosed patients. Iraq Medical Journal. 2018(3):72-4%V 2.