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Updated: Jul 7, 2026
Isolating Lymphocytes from the Mouse Small Intestinal Immune System
Published on: February 28, 2018
The normal range of duodenal intraepithelial lymphocytes
Siavosh Nasseri-Moghaddam1, Azadeh Mofid, Mehdi Nouraie
1Digestive Disease Research Center, Shariati Hospital, Tehran University of Medical Sciences, Tehran, Iran.
Insights
The normal upper limit for duodenal intraepithelial lymphocytes is 35/100EC for immunohistochemistry and 34/100EC for H&E staining. Counts above these thresholds may indicate increased lymphocytes, crucial for celiac disease diagnosis.
Area of Science:
- Gastroenterology
- Histopathology
- Immunology
Background:
- Increased duodenal intraepithelial lymphocytes (IELs) are a hallmark of celiac disease.
- Existing studies indicate a broad normal range for IELs (10-40 IELs/100 epithelial cells).
- The normal range of IELs in the distal duodenum requires precise definition.
Purpose of the Study:
- To establish the normal range of intraepithelial lymphocytes in the distal duodenum.
- To compare diagnostic accuracy between H&E and immunohistochemistry (IHC) for IEL assessment.
- To determine the upper limit of normal for IEL counts in distal duodenal biopsies.
Main Methods:
- Collected distal duodenal biopsies from individuals undergoing endoscopy for non-intestinal reasons.
- Stained biopsies using Hematoxylin-Eosin (H&E) and immunohistochemistry (CD45).
- Assessed at least 1000 cells per biopsy, calculating the villous height to crypt depth ratio.
Main Results:
- The upper limit of normal for IELs was determined as 35/100EC for IHC and 34/100EC for H&E staining.
- Counts between 36-39 (IHC) and 35-37 (H&E) are considered borderline.
- H&E staining demonstrated excellent agreement with IHC, suggesting its adequacy for clinical use.
Conclusions:
- Normal IEL counts are below 35/100EC (IHC) and 34/100EC (H&E).
- Elevated IEL counts (above 39/100EC for IHC, 37/100EC for H&E) suggest pathology.
- H&E staining is a viable and cost-effective method for assessing IELs in routine diagnostics. Consideration of regional variations and temporal changes is important for accurate interpretation.
Background:
Increased duodenal intraepithelial lymphocytes is a key histological finding in celiac disease. Available studies suggest a wide normal range (10 - 40 intraepithelial lymphocytes /100 epithelial cells, EC). We assessed the normal range of distal duodenal intraepithelial lymphocytes.
Methods:
Biopsies from the normal-looking distal duodenum from individuals referring for endoscopy for reasons other than intestinal pathologies were stained with hematoxylin-eosin and immuno-histochemistry (immunohistochemistry, CD45). At least 1000 cells from the tip and body of the villi were assessed. Villous height to crypt depth ratio was calculated.
Results:
Fifty individuals were enrolled. Four were excluded (inadequate biopsies). Mean intraepithelial lymphocyte count was similar in villous tip and body. In the whole villous, upper limit of normal (mean+2SD) was 35 intraepithelial lymphocytes/100EC (95% CI: 31 - 39) in immunohistochemistry and 34 intraepithelial lymphocytes/100EC (95%CI: 29 - 37) in H&E staining. The villous height to crypt depth ratio was 3.9. Hematoxylin-eosin and immunohistochemistry had excellent agreement.
Conclusion:
Intraepithelial lymphocyte counts less than 35/100EC in IHC and 34/100EC in hematoxylin-eosin staining can be considered normal. Counts between 36-39 (immunohistochemistry) and 35 - 37 (hematoxylin-eosin) are borderline and more than 39 (immunohistochemistry) and 37 (hematoxylin-eosin) are increased. The hematoxylin-eosin staining method seems adequate for clinical purposes. Regional differences in the normal upper limit for intraepithelial lymphocytes as well as changes over time should be considered when interpreting duodenal biopsies.
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