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Updated: Jun 16, 2026

The Examination of Peroxidase-Positive Leukocytes in Semen
Published on: January 19, 2024
Macrophage activity in semen is significantly correlated with sperm quality in infertile men
1Research Centre for Reproductive Health, Discipline of Obstetrics and Gynaecology, School of Paediatrics and Reproductive Health, University of Adelaide, Adelaide, SA, Australia. kelton.tremellen@adelaide.edu.au
Abstract:
The presence of leucocytes within semen has the potential to impair sperm function. Neutrophils and macrophages make up 95% of seminal leucocytes, with both having the ability to damage sperm via the generation of reactive oxygen species, proteases and the induction of apoptosis. Existing cytological techniques for quantifying leucocyte activity within semen (peroxidase, CD45) are less than ideal as they merely count the number of leucocytes, rather than assess their activity. Seminal plasma elastase effectively determines neutrophil activity, yet gives no insight into macrophage activity. Neopterin, a molecule released from activated macrophages, may be a useful marker for macrophage activity in the male reproductive tract. To examine this possibility a total of 63 asymptomatic subjects with male factor infertility and 11 fertile controls provided semen samples for measurement of various inflammatory markers. We were able to confirm for the first time that seminal plasma does indeed contain neopterin and that the levels of this macrophage activity marker are threefold higher in infertile than fertile men. Furthermore, seminal plasma neopterin concentration was significantly correlated with sperm oxidative stress, DNA fragmentation (TUNEL) and apoptosis (Annexin V), making it a useful marker of sperm quality. By contrast, seminal plasma elastase showed no correlation with any marker of sperm quality.
Insights
Seminal neopterin, a marker for macrophage activity, is threefold higher in infertile men. Elevated neopterin levels correlate with impaired sperm quality, indicating its potential as a diagnostic tool for male infertility.
Area of Science:
- Reproductive Biology
- Immunology
- Urology
Background:
- Leukocytes in semen can impair sperm function through reactive oxygen species, proteases, and apoptosis.
- Current methods like peroxidase and CD45 only count leukocytes, not their activity.
- Seminal plasma elastase measures neutrophil activity but not macrophage activity.
Purpose of the Study:
- To investigate neopterin as a marker for macrophage activity in semen.
- To assess the correlation between seminal neopterin levels and sperm quality in infertile men.
Main Methods:
- Collected semen samples from 63 infertile men and 11 fertile controls.
- Measured seminal plasma neopterin and elastase levels.
- Assessed sperm oxidative stress, DNA fragmentation (TUNEL), and apoptosis (Annexin V).
Main Results:
- Neopterin was detected in seminal plasma for the first time.
- Seminal neopterin levels were threefold higher in infertile men compared to fertile controls.
- Neopterin concentration correlated significantly with sperm oxidative stress, DNA fragmentation, and apoptosis.
- Seminal plasma elastase showed no correlation with sperm quality markers.
Conclusions:
- Seminal neopterin is a reliable marker of macrophage activity in the male reproductive tract.
- Neopterin levels reflect sperm quality and may serve as a diagnostic indicator for male infertility.
- Neopterin offers a more comprehensive assessment of seminal inflammation than elastase.
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