Voluntary control of fertility is of paramount importance to the modern society. But since the contraceptive methods available for women have their limitations such as urinary tract infections, allergies, cervical erosion and discomfort, a desperate need exists to develop safe methods. Vaginal contraceptives may be the answer to this problem, as these are the oldest ways of fertility regulation, practiced over the centuries. With minimal systemic involvement, these are also the safest. Natural substances blocking or impairing the sperm motility offer as valuable non-cytotoxic vaginal contraceptives. Antimicrobial peptides (AMPs) isolated from plants, animals and microorganisms are known to possess sperm immobilizing and spermicidal properties. Following this, in the quest for alternative means, we have cloned, over expressed and purified the recombinant sperm agglutinating factor (SAF) from Staphylococcus warneri, isolated from the cervix of a woman with unexplained infertility. Genomic library of Staphylococcus warneri was generated in Escherichia coli using pSMART vector and screened for sperm agglutinating factor (SAF). The insert in sperm agglutinating transformant was sequenced and was found to express ribonucleotide-diphosphate reductase-α sub unit. The ORF was sub-cloned in pET28a vector, expressed and purified. The effect of rSAF on motility, viability, morphology, Mg++-dependent ATPase activity and acrosome status of human sperms was analyzed in vitro and contraceptive efficacy was evaluated in vivo in female BALB/c mice. The 80 kDa rSAF showed complete sperm agglutination, inhibited its Mg2+-ATPase activity, caused premature sperm acrosomal loss in vitro and mimicked the pattern in vivo showing 100% contraception in BALB/c mice resulting in prevention of pregnancy. The FITC labeled SAF was found to bind the entire surface of spermatozoa. Vaginal application and oral administration of rSAF to mice for 14 successive days did not demonstrate any significant change in vaginal cell morphology, organ weight and tissue histology of reproductive and non-reproductive organs and had no negative impact in the dermal and penile irritation tests. The Sperm Agglutinating Factor from Staphylococcus warneri, natural microflora of human cervix, showed extensive potential to be employed as a safe vaginal contraceptive.

中文翻译：

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从莫名其妙的不育症妇女的子宫颈中分离出的华纳葡萄球菌精子凝集因子的避孕功效

自愿控制生育对现代社会至关重要。但是由于妇女可用的避孕方法有其局限性，例如尿路感染，过敏，宫颈糜烂和不适，因此迫切需要开发安全的方法。阴道避孕药可能是解决这个问题的方法，因为这是数百年来实行的最古老的生育控制方法。只需很少的系统干预，它们也是最安全的。阻止或损害精子活力的天然物质可作为有价值的非细胞毒性阴道避孕药。已知从植物，动物和微生物中分离出的抗菌肽（AMPs）具有固定精子和杀精子的特性。此后，在寻求替代方法的过程中，我们克隆了，从无法解释的不育症妇女子宫颈中分离出的华氏葡萄球菌的重组精子凝集因子（SAF）过度表达和纯化。使用pSMART载体在大肠杆菌中产生了金黄色葡萄球菌的基因组文库，并筛选了精子凝集因子（SAF）。对精子凝集转化体中的插入片段进行测序，发现其表达核糖核苷酸-二磷酸还原酶-α亚单位。将ORF亚克隆到pET28a载体中，表达并纯化。体外分析了rSAF对人精子的活力，活力，形态，Mg ++依赖性ATPase活性和顶体状态的影响，并在雌性BALB / c小鼠体内评估了避孕效果。80 kDa rSAF显示出完全的精子凝集，抑制了其Mg2 + -ATPase活性，引起体外精子顶体早丢失，并模仿了BALB / c小鼠体内100％避孕的体内模式，从而防止了怀孕。发现FITC标记的SAF结合了精子的整个表面。连续14天向小鼠阴道施用和口服rSAF并未显示出生殖和非生殖器官的阴道细胞形态，器官重量和组织组织学变化没有任何显着变化，并且对皮肤和阴茎刺激性试验没有负面影响。来自人类子宫颈天然菌群的金黄色葡萄球菌的精子凝集因子显示出广泛的潜力，可以用作安全的阴道避孕药。发现FITC标记的SAF结合了精子的整个表面。连续14天向小鼠阴道施用和口服rSAF并未显示出生殖和非生殖器官的阴道细胞形态，器官重量和组织组织学变化没有任何显着变化，并且对皮肤和阴茎刺激性试验没有负面影响。来自人类子宫颈天然菌群的金黄色葡萄球菌的精子凝集因子显示出广泛的潜力，可以用作安全的阴道避孕药。发现FITC标记的SAF结合了精子的整个表面。连续14天向小鼠阴道施用和口服rSAF并未显示出生殖和非生殖器官的阴道细胞形态，器官重量和组织组织学变化没有任何显着变化，并且对皮肤和阴茎刺激性试验没有负面影响。来自人类子宫颈天然菌群的金黄色葡萄球菌的精子凝集因子显示出广泛的潜力，可以用作安全的阴道避孕药。生殖器官和非生殖器官的器官重量和组织组织学，对皮肤和阴茎刺激试验没有负面影响。来自人类子宫颈天然菌群的金黄色葡萄球菌的精子凝集因子显示出广泛的潜力，可以用作安全的阴道避孕药。生殖器官和非生殖器官的器官重量和组织组织学，对皮肤和阴茎刺激试验没有负面影响。来自人类子宫颈天然菌群的金黄色葡萄球菌的精子凝集因子显示出广泛的潜力，可以用作安全的阴道避孕药。