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Updated: May 14, 2026

Multiplexed Fluorescent Immunohistochemical Staining of Four Endometrial Immune Cell Types in Recurrent Miscarriage
Published on: August 4, 2021
Inter-observer and intra-observer variability in immunohistochemical detection of endometrial stromal plasmacytes in
Kotaro Kitaya1, Tadahiro Yasuo
1Department of Anatomy and Cell Science, Kansai Medical University, Osaka 570-8506;
Insights
Immunohistochemistry using CD138 is a reliable method for diagnosing chronic endometritis (CE). This technique shows high accuracy and reduced bias compared to traditional methods, even with varying observer experience.
Area of Science:
- Reproductive Medicine
- Pathology
- Immunohistochemistry
Background:
- Chronic endometritis (CE) is an endometrial inflammation often overlooked due to subtle symptoms and complex diagnosis.
- Current diagnostic methods for CE, including immunohistochemistry for CD138, require further validation regarding accuracy and potential biases.
Purpose of the Study:
- To assess the inter- and intra-observer variability in detecting endometrial stromal plasmacytes using CD138 immunohistochemistry for chronic endometritis diagnosis.
- To compare the reliability of CD138 immunohistochemistry with conventional histopathology in diagnosing CE.
Main Methods:
- Evaluation of 80 CE and 20 non-pathological endometrial samples by experienced and inexperienced observers.
- Counting stromal plasmacytes in both hematoxylin-stained and CD138-immunostained preparations.
- Assessing inter- and intra-observer agreement using linear weighted kappa coefficients.
Main Results:
- CD138 immunohistochemistry demonstrated superior inter- and intra-observer agreement compared to conventional histopathology for CE diagnosis.
- Agreement was consistently high across observers regardless of experience level and between repeated evaluations.
- No significant variability was found in paired sample comparisons, validating the method's consistency.
Conclusions:
- Immunohistochemistry for CD138 is a validated, accurate, and less biased diagnostic tool for chronic endometritis.
- The method offers improved reliability in CE diagnosis, benefiting both experienced and inexperienced pathologists.
Abstract:
Chronic endometritis (CE) is an unusual endometrial inflammation characterized by stromal plasmacyte infiltration. CE is easily missed due to its subtle symptoms and demanding histopathological examinations. Although the immunohistochemistry for the plasmacyte marker CD138 has facilitated the detection of endometrial stromal plasmacytes, the accuracy and biases of this method for CE diagnosis remain poorly understood. The aim of this study was to investigate the inter- and intra-observer variability in the immunohistochemical detection of stromal plasmacytes in the human endometrium. A total of 80 CE and 20 non-pathological archival hematoxylin-stained endometrial preparations with or without immunostaining for CD138 were evaluated independently by two experienced observers and two inexperienced observers. Endometrial stromal plasmacytes in unit areas were counted in the hematoxylin-stained and CD138-immunostained preparations. Each preparation was subdivided into 11 categories by every five plasmacyte counts. The second evaluation was performed four weeks after the first evaluation. The immunohistochemical detection method was superior to conventional histopathological evaluation in both the inter- and intra-observer agreement, irrespective of the experience level of the observers. The linear weighted κ coefficient for intra-observer agreement was higher in the experienced observers than in the inexperienced observers. The inter-observer agreement among the four observers by the immunohistochemical detection method was similarly good between the first and second evaluation. There was no significant inter- or intra-observer variability in the paired comparison of the individual samples. These findings validate the use of immunohistochemistry for CD138 as an accurate and less biased diagnostic tool for CE.
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