Definition of chronic male genital tract inflammation and its impact on male infertility is still a matter of debate. In particular, DNA integrity has been reported to be disturbed in subfertile men. Thus, the aim of this study was to investigate an association of DNA integrity to altered standard semen parameters as well as inflammatory parameters such as peroxidase-positive cells, macrophages and seminal interleukin-6 concentration. Macrophages were detected by CD18/HLA-Dr staining, and DNA integrity was analysed by acridine orange staining using flow cytometry. Interleukin-6 was detected by ELISA. Normal DNA integrity showed a significant correlation to sperm number and progressive motility. Moreover, a significant inverse correlation of DNA integrity to Interleukin-6 and macrophages could be demonstrated. Further on, seminal interleukin-6 also significantly correlated to macrophages. No association has been observed between the number of peroxidase-positive cells and normal DNA integrity. As disturbed DNA integrity has been reported to negatively influence spermatozoon-egg interaction and even fertilisation rates following ICSI, and as early miscarriages have been associated with sperm DNA damage, it should be screened very carefully for male genital tract inflammations in couples undergoing infertility treatment. Measuring Interleukin-6 seems superior to assessment of the number of leucocytes alone and additional assessment of DNA integrity into the diagnostic work-up should be considered.
Silent chronic inflammation of genital tract (CIGT) is considered as a major contributing factor to male fertility disorders. Within CIGT, inflammatory cytokines such as TNF-alpha might induce spermatozoal apoptosis, which in turn has been shown to have a negative influence on the sperm-oocyte penetration capacity. Thus, the aim of this study was to investigate spermatozoal apoptosis in patients with fertility disorders and signs of CIGT. Apoptosis of spermatozoa was determined by expression of annexin V using flow cytometry. Apoptotic spermatozoa were further discriminated from necrotic spermatozoa by 7-amino-actinomycin (7AAD). Ejaculates of patients showing signs of CIGT (P-CIGT) such as high polymorphonuclear (PMN) elastase were compared to control ejaculates of patients lacking signs of CIGT (CON). Thereby we detected a significant correlation between PMN elastase and TNF-alpha in the seminal plasma and apoptotic spermatozoa. Furthermore, we demonstrated a significantly higher percentage of apoptotic spermatozoa in the ejaculate of P-CIGT compared to CON. Interestingly, a significantly higher percentage of apoptotic spermatozoa was detected in the swim-up fraction demonstrating motility of apoptotic spermatozoa. This is in line with the missing correlation of apoptotic spermatozoa and motility. In conclusion, patients with signs of CIGT display high numbers of apoptotic spermatozoa with progressive motility, which might have an impact on reproductive techniques such as intrauterine insemination or in-vitro fertilisation.
Increased numbers of mast cells (MCs) in the testis have been associated with testicular dysfunction, where accumulation of MCs occurs. Furthermore, it has been reported that MCs might affect sperm function as it has been demonstrated that MC-derived tryptase in the seminal fluid might reduce sperm motility. Although MCs have been detected in rat epididymis, only little is known about the presence of MCs in human seminal plasma. Thus, we analysed MC numbers in the ejaculate of men during routine semen analysis of male patients suspected for infertility (n = 100). MCs were detected by c-kit (CD117) expression using flow cytometry. Thereby, we detected significant numbers of MCs in the ejaculate of most patients (559 +/- 525 MCs ml(-1), mean +/- SD). However, we could neither detect a correlation with respect to MCs and sperm count, motility or morphology nor to the seminal inflammatory markers like polymorphonuclear elastase. Nevertheless, a significant correlation of MCs to spermatozoa-bound IgA (r = 0.5; P = 0.03; n = 21) was observed. It is concluded that significant numbers of MCs can be detected in the human ejaculate without necessarily influencing sperm function. A potential role of MCs in seminal plasma as well as the association between MCs and IgA on spermatozoa remains to be elucidated.
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