An optimized Factor H-Fc fusion protein against multidrug-resistant Neisseria gonorrhoeae.
An optimized Factor H-Fc fusion protein against multidrug-resistant Neisseria gonorrhoeae.
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DOI：
10.3389/fimmu.2022.975676
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发表时间：
2022
影响因子：
7.3
通讯作者：
中科院分区：
文献类型：
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迫切需要针对多重耐药淋病奈瑟菌全球威胁的新疗法。淋球菌通过结合因子 H (FH)（旁路途径的关键抑制剂）逃避补体杀伤。 FH 由 20 个短一致重复 (SCR) 域组成，组织为单链。淋球菌通过结构域 6 和 7，以及 C 端结构域 18 至 20 结合 FH。此前，我们发现，包含（从 N 端到 C 端）FH 结构域 18-20（结构域 19 中包含点突变以防止宿主细胞裂解）与人 IgG1 Fc（称为 FH*/Fc1）融合的嵌合蛋白以补体依赖性方式杀死淋球菌，并减少持续时间和细菌负荷在淋病小鼠阴道定植模型中。考虑到淋病奈瑟菌结合 FH 结构域 18-20 位于天然 FH 的 C 端，我们推断将 Fc N 端定位至 FH* (Fc1/FH*) 将提高结合和杀菌活性。尽管两种分子与淋球菌的结合相似，但 Fc1/FH* 的 IC50（补体依赖性杀菌测定中 50% 杀灭所需的浓度）比 FH*/Fc1 低 5 倍。为了进一步增强补体激活，我们将 Fc1/FH* 中的人 IgG1 Fc 替换为来自人 IgG3（最有效的补体激活 IgG 亚类）的 Fc，以获得 Fc3/FH*。与 Fc1/FH* 相比，Fc3/FH* 的杀菌活性进一步提高约 2.3 倍。 Fc3/FH* 以补体依赖性方式杀死（定义为 <50% 存活率）45/45 (100%) 表达 PorB1B 的不同淋球菌，但仅杀死 2/15 表达 PorB1A 的分离株。 Fc3/FH* 结合减少导致针对 PorB1A 菌株的活性有限。当每天阴道内施用 5 µg 剂量时，Fc3/FH* 对小鼠阴道定植模型中所有四种测试的 PorB1B 淋球菌菌株均有效。此外，当冻干或喷雾干燥后重构时，Fc3/FH* 保留了杀菌活性，这表明配制到阴道环中的可行性。总之，Fc3/FH* 代表了一种有前途的针对多重耐药淋球菌的预防性免疫疗法。
Novel therapeutics against the global threat of multidrug-resistant Neisseria gonorrhoeae are urgently needed. Gonococci evade killing by complement by binding factor H (FH), a key inhibitor of the alternative pathway. FH comprises 20 short consensus repeat (SCR) domains organized as a single chain. Gonococci bind FH through domains 6 and 7, and C-terminal domains 18 through 20. Previously, we showed that a chimeric protein comprising (from the N- to C-terminus) FH domains 18-20 (containing a point mutation in domain 19 to prevent lysis of host cells) fused to human IgG1 Fc (called FH*/Fc1) killed gonococci in a complement-dependent manner and reduced the duration and bacterial burden in the mouse vaginal colonization model of gonorrhea. Considering the N. gonorrhoeae-binding FH domains 18-20 are C-terminal in native FH, we reasoned that positioning Fc N-terminal to FH* (Fc1/FH*) would improve binding and bactericidal activity. Although both molecules bound gonococci similarly, Fc1/FH* displayed a 5-fold lower IC50 (the concentration required for 50% killing in complement-dependent bactericidal assays) than FH*/Fc1. To further increase complement activation, we replaced human IgG1 Fc in Fc1/FH* with Fc from human IgG3, the most potent complement-activating IgG subclass, to obtain Fc3/FH*. Bactericidal activity was further increased ~2.3-fold in Fc3/FH* compared to Fc1/FH*. Fc3/FH* killed (defined by <50% survival) 45/45 (100%) diverse PorB1B-expessing gonococci, but only 2/15 PorB1A-expressing isolates, in a complement-dependent manner. Decreased Fc3/FH* binding accounted for the limited activity against PorB1A strains. Fc3/FH* was efficacious against all four tested PorB1B gonococcal strains in the mouse vaginal colonization model when administered at a dose of 5 µg intravaginally, daily. Furthermore, Fc3/FH* retained bactericidal activity when reconstituted following lyophilization or spray-drying, suggesting feasibility for formulation into intravaginal rings. In conclusion, Fc3/FH* represents a promising prophylactic immunotherapeutic against multidrug-resistant gonococci.
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DOI：
10.4049/jimmunol.1102746
发表时间：
2012-04-01
期刊：
Journal of immunology (Baltimore, Md. : 1950)
影响因子：
--
作者：
Gulati S;Agarwal S;Vasudhev S;Rice PA;Ram S
通讯作者：
Ram S
DOI：
10.5731/pdajpst.2015.01003
发表时间：
2015-01-01
影响因子：
--
作者：
Gikanga, Benson;Turok, Robert;Maa, Yuh-Fun
通讯作者：
Maa, Yuh-Fun
影响因子：
6.4
作者：
Cohen, CR;Sinei, S;Kreiss, J
通讯作者：
Kreiss, J
影响因子：
36.8
作者：
FIGUEROA, JE;DENSEN, P
通讯作者：
DENSEN, P
影响因子：
6.4
作者：
Gulati, Sunita;Pennington, Michael W.;Rice, Peter A.
通讯作者：
Rice, Peter A.