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Author Spotlight: Advancing Reproductive Immunology with a Protocol for the Quantitative Evaluation of Endometrial Immune Cells
Published on: October 13, 2023
Conventional microscopy versus digital image analysis for histopathologic evaluation of immune cells in the
Trine Hallager1, Malene Hviid Saxtorph2, Jens Ole Eriksen1
1Department of Pathology, Zealand University Hospital, Denmark; ReproHealth Research Consortium, Zealand University Hospital, Denmark.
Insights
Standardizing the quantification of uterine immune cells is crucial for diagnosing infertility. This study compared conventional microscopy and digital image analysis (DIA) for immune cell counting, finding significant associations but also method-dependent differences.
Area of Science:
- Reproductive Immunology
- Pathology
Background:
- Diagnosing immunological infertility requires reliable biomarkers.
- Uterine immune cell quantification methods are currently heterogeneous, necessitating standardization.
- Previous research identified the CD56/CD16 ratio as a potential prognostic marker.
Purpose of the Study:
- To compare quantification methods for endometrial immune cells (CD56+, CD16+, CD163+) in women undergoing fertility treatment.
- To evaluate conventional microscopy against digital image analysis (DIA).
- To assess the impact of different field selection methods in DIA on the CD56/CD16 ratio.
Main Methods:
- Immune cell density was assessed using conventional microscopy (semiquantitative scale) and DIA (continuous count: % positive cells, cells/mm²).
- Two DIA field selection methods were compared: systematic random sampling (excluding glands) and analysis of large tissue areas (including glands).
- The CD56/CD16 ratio was analyzed using both DIA field selection approaches.
Main Results:
- A significant association was found between conventional microscopy and DIA for CD56, CD16, and CD163 cell counts (p < 0.001).
- Significant differences in the CD56/CD16 ratio were observed between the two DIA field selection methods (p < 0.001).
- DIA demonstrated a significant association with conventional microscopy results.
Conclusions:
- Both conventional microscopy and DIA can provide comparable results for quantifying endometrial immune cells.
- The method of field selection in DIA significantly impacts the CD56/CD16 ratio.
- Further research correlating these quantification methods with clinical outcomes is needed to establish their utility in diagnosing immunological infertility.
Abstract:
In the search for a reliable biomarker able to diagnose immunological causes of infertility, uterine immune cells have been widely investigated. As a result, heterogeneous methods and cutoff values of what constitutes an aberrant number of immune cells have been reported, and a standardized method for quantification is needed. The objective of this study was to compare methods for quantification of immune cells visualized with immunohistochemistry in the endometrium of women in fertility treatment. Evaluation of the density of CD56+, CD16+ and CD163+ cells by conventional microscopy on a semiquantitative scale (low, medium and high) was compared to a continuous count using digital image analysis (DIA) reported as percentage positive cells out of the total number of stromal cells and number of positive cells per mm2, respectively. We previously reported the CD56/CD16 ratio as a possible prognostic marker, and therefore the ratios of CD56/CD16 were compared using two different methods for selecting fields for counting with DIA: one method using principles of systematic random sampling, where glands were excluded, and one method analyzing large parts of the tissue including glands. A significant association between conventional microscopy and DIA was found when the semiquantitative scale was compared to medians of positive cells in CD56, CD16 and CD163, respectively, p < 0.001. A systematic significant difference in the ratios of CD56/CD16 was found when comparing the two methods for field selection, p < 0.001. To determine the possible use of these methods, more knowledge of the correlation to clinical outcome is warranted.
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