P–099 MYD88 dependent pathway through TLR 1,2 and 6 activation play a role in interaction of high DNA fragmented human sperm with fallopian tube epithelial cells

Abstract

Abstract Study question How does sperm with high DFI disrupt the female reproductive system? Summary answer Sperm with damaged DNA could disrupt the female reproductive system by production of high inflammatory factors through activation of TLRs and MyD88-dependent pathway . What is known already Previous studies have also shown that the interaction between sperm and fallopian tube is a complex process and activates a number of immune mechanisms. Many TLRs, including TLR1, TLR2, and TLR6 are expressed in fallopian tube epithelial cells. TLRs have an important role in the immune interaction between fallopian tube and sperm. They are also involved in sperm capacitation, fertilization and pregnancy. Examining this interaction may lead us to valuable data. Study design, size, duration In this study, 10 Recurrent Implantation failure couples with high DFI ( >30% by SCSA evaluation) and 10 healthy donors men with low DFI (<30%) were considered as high DFI group and low DFI group respectively. After fresh semen preparation, sperm were co-cultured with human fallopian tube epithelial cell line( OE-E6/E7) for 24 hours. Participants/materials, setting, methods RNA were extracted from the cell line and PCR array for human innate and adaptive immune responses was performed by RT2 profiler PCR array. Main results and the role of chance Analysis of PCR array data showed that the expression of TLR–1, TLR–2, TLR–6, MYD88, TIRAP, IRAKS,TRAF6, MAPKS, NF-KB, G-CSF, GM-CSF, CXCL8, CXCL10, CCL2, IL–6, IL–1, TNFa in high DFI group was significantly higher than control group. All these factors are involved in the MyD88-dependent pathway. Our research suggests that the MyD88-dependent pathway through TLR1,2 and 6 activation may be the main inflammatory pathway activated by sperm with high DFI. The released DAMPs through damaged sperms were recognized by TLRs on the epithelial cells. Following the TLR1,2 and 6 signaling, and MYD88 activation, epithelial cells were producing the inflammatory cytokines which result in the neutrophils infiltration. In the lumen of the fallopian tube, the neutrophils were activated via TLR1/TLR2, complexes that lead to the neutrophils activation, phagocytosis, NET formation, and apoptosis. Limitations, reasons for caution Studying with a larger sample size and examining the final factors at the protein level will be better to detect the effect of sperm DNA fragmentation on fallopian tube. Wider implications of the findings: MyD88-dependent pathway could be one of the mechanisms that involved in interaction of high DNA fragmented sperm with female reproductive tract. Trial registration number Not applicable