Department of Microbiology, Panjab University, Chandigarh, India.
PLoS One. 2023 Dec 7;18(12):e0289989. doi: 10.1371/journal.pone.0289989. eCollection 2023.
The present study aims to investigate the antigenic cross reactivity between the receptor from Proteus mirabilis and spermatozoa against a common sperm immobilization factor, SIF, by calorimetric and competitive inhibition studies, and the immunogenicity of this receptor to evoke the formation of antisperm antibodies and their subsequent role in fertility outcome. The sperm binding receptor from Proteus mirabilis (PM-SBR) was extracted from ultrasonicated cell debris by treating it for 12 h at 37°C with 1 M NaCl. After being purified by gel permeation chromatography, its molecular weight as determined by SDS-PAGE was observed to be ≈ 47 kDa. The detrimental impacts of Sperm immobilizing factor (SIF) on spermatozoa viz. motility, viability, and morphology were mitigated when SIF was preincubated with various concentrations of PM-SBR. Using isothermal titration calorimetry, the entropy of the SIF-PM-SBR interaction was found to be -18.31 kJ/mol, whereas the free energy was 28.4 J/mol K. FTIR analysis was used to evaluate the binding interactions between PM-SBR and SIF. In addition, mice that were administered antibodies against PM-SBR were unable to conceive, in contrast to mice that were administered Phosphate buffer saline (PBS) or pre-immunization serum as controls. In light of this, we may conclude that anti-PM-SBR antibodies act as anti-sperm antibodies. Our work found that molecular mimicry between Proteus mirabilis and spermatozoa may cause antisperm immune reactivity. As a result of an immunological response to PM-SBR, infected individuals may produce antibodies against an epitope similar to one found on spermatozoa which helps in developing new strategies for managing autoimmune responses and infertility.
本研究旨在通过量热和竞争性抑制研究,探讨变形菌属受体与精子之间的抗原交叉反应,针对共同的精子固定因子 SIF,以及该受体的免疫原性,以引发抗精子抗体的形成及其随后在生育结局中的作用。从超声处理的细胞碎片中提取变形菌属(PM)的精子结合受体(PM-SBR),并用 1 M NaCl 在 37°C 下孵育 12 小时。通过凝胶渗透色谱法纯化后,通过 SDS-PAGE 测定其分子量约为 47 kDa。当 SIF 与各种浓度的 PM-SBR 预孵育时,SIF 对精子的运动、活力和形态的有害影响得到缓解。使用等温滴定量热法,发现 SIF-PM-SBR 相互作用的熵为-18.31 kJ/mol,而自由能为 28.4 J/mol K。傅里叶变换红外(FTIR)分析用于评估 PM-SBR 和 SIF 之间的结合相互作用。此外,给予 PM-SBR 抗体的小鼠无法受孕,而给予磷酸盐缓冲盐水(PBS)或预免疫血清作为对照的小鼠则可以受孕。鉴于此,我们可以得出结论,抗 PM-SBR 抗体充当抗精子抗体。我们的工作发现变形菌属和精子之间的分子模拟可能导致抗精子免疫反应。由于对 PM-SBR 的免疫反应,感染个体可能会产生针对与精子上相似表位的抗体,这有助于开发管理自身免疫反应和不育症的新策略。