Assessment of Intraepithelial Lymphocytes Count in Potential Celiac Disease
Roberta Mandile1,2, Mariantonia Maglio1, Antonella Marano1
1European Laboratory for the Investigation of Food Induced Diseases (ELFID) University of Naples Federico II, Naples, Italy.
Insights
Intraepithelial lymphocytes (IELs) counts for celiac disease diagnosis show low agreement between H&E staining and IHC methods. This impacts Marsh grading, especially in potential celiac disease patients.
Area of Science:
- Gastroenterology
- Histopathology
- Immunology
Background:
- Intraepithelial lymphocytes (IELs) are crucial for diagnosing celiac disease (CD).
- IEL counts can be assessed via H&E staining of paraffin sections or IHC on frozen sections.
- Standardized comparison of these methods for Marsh grading is needed.
Purpose of the Study:
- To evaluate the concordance in Marsh grading between H&E and IHC methods for IEL counting.
- To assess agreement in different patient groups: normal, potential celiac disease (PCD), and controls (CTR).
Main Methods:
- Comparison of Marsh grading based on IEL counts from H&E and IHC anti-CD3 staining.
- Analysis of 280 normal, 210 PCD, and 70 CTR patient sections.
- Statistical analysis using Cohen Kappa to measure concordance.
Main Results:
- Overall concordance was 66.8% (Kappa=0.33), with lower agreement in PCD patients (63%, Kappa=0.26) compared to controls (77%, Kappa=0.54).
- Marsh-1 classification showed particularly low concordance (48.6% overall, 45.6% in PCD).
- Differences in concordance between groups were statistically significant (p<0.05).
Conclusions:
- There is low concordance in IEL counts and Marsh grading between H&E and IHC methods, especially in potential celiac disease patients.
- The choice of technique significantly affects Marsh grading, necessitating caution in clinical interpretation.
- Further standardization or clarification is required for accurate celiac disease diagnosis based on IEL counts.
Abstract:
Intraepithelial lymphocytes (IELs) count, central for coeliac disease (CD) diagnosis, can be performed either directly on hematoxylin and eosin (H&E)-stained paraffined sections or on optimal-cutting-temperature-compound (OCT)-embedded frozen sections stained by immunohistochemistry (IHC) with anti-CD3. We evaluated the concordance in Marsh grading between these two techniques on a large sample of sections. A total of 280 patients with a normal intestinal architecture, 210 potential celiac disease (PCD) patients, and 70 controls (CTR) were included. At the H&E histological evaluation, 136/280 were classified as Marsh-0 (showing < 25 IELs/100 enterocytes) and 144 Marsh-1, while at the IHC evaluation, 191 were classified as Marsh-0 (showing ≤ 34 CD3+/mm of epithelium) and 89 Marsh-1. The overall concordance was 66.8% (48.6% Marsh-1 and 86% Marsh-0) with a Cohen Kappa value of 0.33. In the PCD group, the overall concordance was 63% (45.6% Marsh-1 and 84% Marsh-0) with a Cohen Kappa value of 0.26, while in the CTR group it was 77% (60% Marsh-1, 90% Marsh-0) with a Cohen Kappa value of 0.54. Differences between the two groups were statistically significant (p < 0.05). In conclusion, the concordance of IELs counts between histological and IHC evaluation is low (Kappa Cohen 0.54) in no-CD and even more in PCD patients (0.26). Caution must be paid when classifying a patient as Marsh-0 or Marsh-1 according to the technique used.
