Georgoutsou-Spyridonos Maria, Ricklin Daniel, Pratsinis Haris, Perivolioti Eustathia, Pirmettis Ioannis, Garcia Brandon L, Geisbrecht Brian V, Foukas Periklis G, Lambris John D, Mastellos Dimitrios C, Sfyroera Georgia
Department of Biodiagnostic Sciences and Technologies, I/NRASTES, National Center for Scientific Research "Demokritos," 15310 Athens, Greece;
Department of Pathology and Laboratory Medicine, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104;
J Immunol. 2015 Oct 15;195(8):3946-58. doi: 10.4049/jimmunol.1500966. Epub 2015 Sep 4.
Staphylococcus aureus can cause a broad range of potentially fatal inflammatory complications (e.g., sepsis and endocarditis). Its emerging antibiotic resistance and formidable immune evasion arsenal have emphasized the need for more effective antimicrobial approaches. Complement is an innate immune sensor that rapidly responds to bacterial infection eliciting C3-mediated opsonophagocytic and immunomodulatory responses. Extracellular fibrinogen-binding protein (Efb) is a key immune evasion protein of S. aureus that intercepts complement at the level of C3. To date, Efb has not been explored as a target for mAb-based antimicrobial therapeutics. In this study, we have isolated donor-derived anti-Efb IgGs that attenuate S. aureus survival through enhanced neutrophil killing. A phage library screen yielded mini-Abs that selectively inhibit the interaction of Efb with C3 partly by disrupting contacts essential for complex formation. Surface plasmon resonance-based kinetic analysis enabled the selection of mini-Abs with favorable Efb-binding profiles as therapeutic leads. Mini-Ab-mediated blockade of Efb attenuated S. aureus survival in a whole blood model of bacteremia. This neutralizing effect was associated with enhanced neutrophil-mediated killing of S. aureus, increased C5a release, and modulation of IL-6 secretion. Finally, these mini-Abs afforded protection from S. aureus-induced bacteremia in a murine renal abscess model, attenuating bacterial inflammation in kidneys. Overall, these findings are anticipated to pave the way toward novel Ab-based therapeutics for S. aureus-related diseases.
金黄色葡萄球菌可引发一系列潜在致命的炎症并发症(如败血症和心内膜炎)。其新出现的抗生素耐药性以及强大的免疫逃逸机制凸显了采用更有效抗菌方法的必要性。补体是一种先天性免疫传感器,能对细菌感染迅速做出反应,引发C3介导的调理吞噬作用和免疫调节反应。细胞外纤维蛋白原结合蛋白(Efb)是金黄色葡萄球菌的一种关键免疫逃逸蛋白,可在C3水平拦截补体。迄今为止,Efb尚未被探索作为基于单克隆抗体的抗菌治疗靶点。在本研究中,我们分离出了供体来源的抗Efb IgG,其通过增强中性粒细胞杀伤作用来减弱金黄色葡萄球菌的存活。噬菌体文库筛选产生了微型抗体,其通过破坏复合物形成所必需的接触部分选择性抑制Efb与C3的相互作用。基于表面等离子体共振的动力学分析使得能够选择具有良好Efb结合谱的微型抗体作为治疗先导物。微型抗体介导的Efb阻断在菌血症全血模型中减弱了金黄色葡萄球菌的存活。这种中和作用与增强中性粒细胞介导的金黄色葡萄球菌杀伤、增加C5a释放以及调节IL-6分泌有关。最后,这些微型抗体在小鼠肾脓肿模型中提供了针对金黄色葡萄球菌诱导的菌血症的保护作用,减轻了肾脏中的细菌炎症。总体而言,这些发现有望为金黄色葡萄球菌相关疾病的新型基于抗体的治疗方法铺平道路。