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Counting Intraepithelial Lymphocytes: A Comparison Between Routine Staining and CD3 Immunohistochemistry
Rosalin Cooper1, Neil James Papworth2, Chloe Harris2
1John Radcliffe Hospital, Oxford, UK.
Insights
Comparing intraepithelial lymphocyte (IEL) counts in duodenal biopsies using hematoxylin and eosin (H&E) and CD3 immunohistochemistry (IHC) revealed significant differences. CD3 IHC yielded higher counts, indicating these methods are not interchangeable for IEL assessment.
Area of Science:
- Gastroenterology
- Histopathology
- Immunohistochemistry
Background:
- Intraepithelial lymphocytes (IELs) are crucial for duodenal biopsy assessment.
- Hematoxylin and eosin (H&E) and CD3 immunohistochemistry (IHC) are used for IEL counting.
- Comparability of IEL counts between H&E and CD3 IHC is not well-established.
Purpose of the Study:
- To compare IEL counts in duodenal biopsies using H&E stains versus CD3 IHC.
- To evaluate interobserver variability in IEL counting with both methods.
Main Methods:
- Thirty-five paired H&E and CD3 stained duodenal biopsy sections were analyzed.
- Six pathologists counted IELs per 100 enterocytes.
- Counts were categorized into normal, mildly raised, and markedly raised groups.
Main Results:
- CD3 IHC demonstrated significantly higher IEL counts compared to H&E.
- Four cases with normal H&E counts showed elevated counts with CD3 IHC.
- Moderate interobserver agreement was observed for both H&E and CD3 methods.
- CD3 IHC did not significantly improve interobserver variability.
Conclusions:
- IEL counts obtained by CD3 IHC and H&E staining are not directly comparable.
- The distinct results suggest these methods should not be considered equivalent for IEL quantification.
- Clinical interpretation of IEL counts may require method-specific reference ranges.
Abstract:
Counting intraepithelial lymphocytes (IELs) is a key part of the assessment of duodenal biopsies. Immunohistochemistry (IHC) for CD3 can aid identification of lymphocytes in this context, but it is not evident that counts on hematoxylin and eosin (H&E) and CD3 are comparable. This study aimed to compare the IEL counts in duodenal biopsies using H&E stains and CD3 IHC, and to examine the interobserver variability. Thirty-five paired H&E and CD3 sections were reviewed by 6 pathologists who counted the number of IELs per 100 enterocytes. The counts were categorized into groups: normal (<25 lymphocytes), mildly raised (25-40 lymphocytes), and markedly raised (>40 lymphocytes). CD3 IHC was associated with significantly higher IEL counts than H&E. Four cases with normal H&E counts had raised counts with CD3. There was moderate agreement between observers for both H&E and CD3. Lack of concordance between CD3 and H&E IEL counts suggests that counts derived from the 2 methods may not be comparable to each other and should not be considered equivalent. There was no significant improvement in interobserver variability with CD3 IHC.
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