Objective: The incidence of leukocytospermia ranges from 10% to 20%among infertile men. However, the role of leukocytospermia in the pathogenesis of male infertility remains highly controversial. Our study evaluated the relationship between leukocytospermia and standard semen parameters, levels of seminal reactive oxygen species (ROS), and the extent of sperm nuclear DNA damage.
Design: Prospective study examining the relationship between leukocytospermia and semen quality.
Materials/Methods: Semen specimens from 48 men attending the male infertility clinic were examined according to the World Health Organization (WHO) criteria. Patients were classiﬁed into two groups, leukocytospermic (1 x 106 leukocytes/mL) (n = 16) and non-leukocytospermic (≤ 1x106 leukocytes/mL) (n = 32). A group of normal donors (n = 13) served as a control. ROS production was determined by the chemiluminescence assay in the washed cell suspension (containing both sperm and leukocytes).
Results were expressed as x106 counted photons per minute (cpm)/20x106 sperm/mL. Sperm nuclear DNA damage was assessed by sperm chromatin structure assay (SCSA) to determine the percentage of cells outside the main population (COMP at) with abnormal chromatin structure.
Results: Leukocytospermia was associated with an abnormality in one or more of the standard semen parameters in 93% (15/16) of cases. Sperm motility (mean ± SD) was signiﬁcantly lower in the leukocytospermic group (37.9 ± 15.2) compared to the non-leukocytospermic group (47.7 ±15.3%; P < 0.04) and the donor group (66.6 6 10%; P < 0.001). Patients with leukocytospermia had poor sperm morphology (16.9 ± 9.1%) compared to the donor group (36.7± 6.1%; P = 0.0003). Abnormal sperm morphology was observed in 88% of leukocytospermic samples versus 47% of non-leukocytospermic samples. ROS levels [median (25%, 75% interquartile range)] were signiﬁcantly higher in the leukocytospermic group [177.9 (31.5, 430.1)], compared to the non-leukocytospermic group [2.7
(0.5, 12.2)] (P < 0.0001) and the donor group [0.4 (0.1, 2.5)] (P< 0.0001). Signiﬁcantly higher levels of %COMP at (mean 6 SD) were observed in the leukocytospermic group (28.8 6 9.3) versus non-leukocytospermic group (22.6 6 10.8; P < 0.02), and the donors (15.8 6 6.8; P = 0.002). A
COMP at value of $27% was seen in 62% of leukocytospermic samples compared to 25% in non-leukocytospermic samples. A signiﬁcant negative correlation was seen between leukocyte concentration in semen and sperm motility (r = 0.31, P = 0.01) and sperm morphology (r = 0.28, P = 0.03).
Leukocytospermia was positively correlated with high ROS levels (r = 0.7, P <0.0001) and %COMP
at values (r = 0.32, P = 0.01).
Conclusions: Our results show a strong correlation between leukocytospermia and poor semen quality, oxidative stress and increased sperm nuclear DNA damage. COMP at values ≥ 27% have been recently reported to be an indicator of negative pregnancy outcome in assisted reproduction. Therefore, infertile men with leukocytospermia should be counseled concerning the potential negative effects of increased sperm DNA damage in their pregnancy outcome.