Preparation and Evaluation of the Fully Humanized Monoclonal Antibody GD-mAb Against Respiratory Syncytial Virus
Preparation and Evaluation of the Fully Humanized Monoclonal Antibody GD-mAb Against Respiratory Syncytial Virus
复制标题
呼吸道合胞病毒全人源化单克隆抗体GD-mAb的制备及评价
DOI：
10.3389/fcimb.2019.00275
复制
发表时间：
2019-07
影响因子：
5.7
通讯作者：
Chen Tingtao
中科院分区：
文献类型：
--
作者：
Tian Puyuan;Wang Yuqing;Liu Hui;Yang Yulu;Wu Xiaoli;Wei Hua;Chen Tingtao
呼吸道合胞病毒 (RSV) 是婴儿、幼儿和免疫功能低下成人肺部和支气管炎症的主要原因，但控制 RSV 的治疗选择有限。在本研究中，使用噬菌体展示文库淘选技术筛选了针对 RSV 的单链抗体（GD-scFv），并根据编码 Ig VH 和 Ig VL 的序列从 GD-scFv 开发了全长单克隆抗体（GD-mAb）。在体外和小鼠体内评估了 GD-mAb 的抗 RSV 潜力。我们的结果表明，GD-scFv (4.25 ± 2 nM) 和 GD-mAb (3.13 ± 0.89 nM) 均表现出对 GD 蛋白的高结合能力和强结合特异性。通过空斑减少中和试验，GD-mAb 有效中和 RSV，并以浓度依赖性方式减少空斑数量。在小鼠中，GD-mAb 降低了小鼠肺部的肺指数并降低了肺病毒滴度 (p < 0.05)。抗体治疗降低了 TLR4/NF-κB、MAPK 和 PI3K/Akt 通路中的磷酸化蛋白水平 (p < 0.05)，并与小鼠肺和血清中促炎因子 TNF-α、IL-1β 和 IL-6 的缺失相关 (p < 0.05)。总之，这些数据表明GD-mAb可能是治疗RSV感染的有效治疗剂。重要性 目前，只有少数治疗方案可用于控制人类呼吸道 RSV。在本研究中，我们课题组开发了全长单克隆抗体（GD-mAb），并报告了与RSV表面糖蛋白G的高结合特异性。此外，GD-mAb在体外有效中和RSV，并显着降低了感染小鼠肺部的肺指数和肺病毒滴度，这表明GD-mAb可能作为RSV感染的有效抗病毒剂。
Respiratory syncytial virus (RSV) is the major cause of pulmonary and bronchial inflammation in infants, young children, and immunocompromised adults, but therapeutic options to control RSV are limited. In the present study a single chain antibody against RSV (GD-scFv) was screened using phage display library panning technology and a full-length monoclonal antibody (GD-mAb) was developed from GD-scFv based on the sequence encoding Ig VH and Ig VL. The anti-RSV potential of GD-mAb was evaluated in vitro and in mice. Our results indicated that both GD-scFv (4.25 ± 2 nM) and GD-mAb (3.13 ± 0.89 nM) showed high binding capability and strong binding specificity to GD protein. GD-mAb effectively neutralized RSV and reduced the plaque number in a concentration-dependent manner through a plaque reduction neutralization assay. In mice, GD-mAb lowered the lung index and reduced the lung virus titer in the mouse lung (p < 0.05). Antibody treatment reduced the phosphorylated protein level in pathways of TLR4/NF-κB, MAPKs, and PI3K/Akt (p < 0.05) and correlated with an absence of pro-inflammatory factors TNF-α, IL-1β, and IL-6 in the mouse lung and serum (p < 0.05). In summary, these data suggest that GD-mAb may be an effective therapeutic agent for the treatment of RSV infections. Importance Currently, only a few therapeutic options are available to control respiratory RSV in humans. In this study, our group developed a full-length monoclonal antibody (GD-mAb) and reported a high binding specificity of the RSV surface glycoproteins G. Moreover, GD-mAb effectively neutralized RSV in vitro, and significantly lowered the lung index and reduced the lung virus titer in an infected mouse lung, which suggests that GD-mAb may serve as an effective antiviral agent for RSV infection.
登录
查看更多内容
影响因子：
6.2
作者：
G. Lewis
通讯作者：
G. Lewis
影响因子：
2.3
作者：
C. Nüsslein-Volhard;E. Wieschaus;B. Edward;Lewis
通讯作者：
C. Nüsslein-Volhard;E. Wieschaus;B. Edward;Lewis
影响因子：
5.4
作者：
Haynes, LM;Tonkin, J;Tripp, RA
通讯作者：
Tripp, RA
影响因子：
2.2
作者：
R. Tripp
通讯作者：
R. Tripp
影响因子：
158.5
作者：
DeVincenzo, John P.;Whitley, Richard J.;Chien, Jason W.
通讯作者：
Chien, Jason W.