Elevated CD4+, CD8+, and Pro-inflammatory Cytokine Levels and H19 Hypomethylation are Associated with Reduced Sperm Motility in Oligoasthenoteratozoospermia: A Case-control Study

Introduction: Oligoasthenoteratozoospermia (OAT) is a frequent cause of male reproductive issues, but its etiology remains unclear. This study investigated the associations between immune markers, epigenetic alterations, and sperm parameters in men with OAT compared to healthy individuals. Methods: A case-control study included 40 OAT patients and 40 healthy, fertile men as a control group. Flow cytometric analysis was performed to enumerate specific T lymphocyte populations (CD4+ and CD8+). To assess key inflammatory markers, including TNF-α and IL-6, ELISA technique was used. Gene methylation of H19 was evaluated using bisulfite sequencing. Sperm parameters were analyzed according to World Health Organization (WHO) guidelines. Results: Findings indicated significantly elevated levels of helper and cytotoxic T lymphocytes, along with increased concentrations of pro-inflammatory mediators in OAT patients relative to healthy controls. These immune markers exhibited strong inverse relationships with key sperm quality parameters. Statistical analysis using the area under the curve (AUC) showed high differential power of these biomarkers in distinguishing OAT patients from controls. As well, significantly reduced H19 gene methylation in OAT patients was observed relative to controls. Subgroup analyses revealed associations between these immune markers and various socio-demographic and clinical factors. Discussion: The observed elevations in immune markers and decreased H19 methylation in OAT patients suggest probable roles for immune dysfunction and epigenetic alterations in OAT pathogenesis. The strong correlations between these markers and sperm parameters support their usefulness in clinical practice. The subgroup analyses draw attention to the complex relationship between socio-environmental factors and male fertility. Conclusion: This investigation demonstrates significant alterations in T lymphocyte subsets and inflammatory cytokine levels, alongside H19 gene hypomethylation, in men with OAT. These immune and epigenetic changes strongly correlate with impaired sperm parameters. Collectively, these findings indicate the potential of these markers as diagnostic indicators for male infertility assessment.