Human monoclonal anti‑TLR4 antibody negatively regulates lipopolysaccharide‑induced inflammatory responses in mouse macrophages.
Human monoclonal anti‑TLR4 antibody negatively regulates lipopolysaccharide‑induced inflammatory responses in mouse macrophages.
复制标题
人单克隆抗 TLR4 抗体负向调节脂多糖诱导的小鼠巨噬细胞炎症反应
DOI：
10.3892/mmr.2020.11500
复制
发表时间：
2020-11
影响因子：
3.4
通讯作者：
Zhu J
中科院分区：
文献类型：
--
作者：
Wang Y;Gong D;Yao C;Zheng F;Zhou T;Cao Q;Zhu X;Wang M;Zhu J
先前的研究表明，Toll样受体4（TLR4）介导的促炎信号通路的激活在急性炎症，败血症和慢性炎症性疾病中起着重要作用，TLR4显着有助于脂肪性疾病（LPS）诱导的免疫反应。这是对TLR4途径的调节是专门针对这些病理的重要策略。人类抗TLR4 IgG2抗体通过从噬菌体播放文库中筛选人类TLR4 Fab，并通过抗体工程将其与重链的人IgG2的恒定区域整合在一起。此外，用人类抗Tlr4 IgG2处理小鼠衍生的巨噬细胞，并在体外诱导转录定量PCR和蛋白质印迹用于确定细胞因子的mRNA表达水平和信号传导途径的磷酸化水平，发现人类抗TLR4 IgG2与TLR4结合的高度亲和力为8.713×10-10 m，和与抗-TLR4 IgG2的预孵育抑制了LPS诱导的肿瘤坏死因子-α，干扰素-β和白介素6 mRNA表达水平的产生小鼠腹膜巨噬细胞也证明了人类抗TLR4 IgG2通过减少NF-κB的磷酸化，促丝裂原激活蛋白激酶和干扰素调节因子3信号传导途径，抑制了LPS诱导的TLR4信号。 IgG2保护小鼠免受LPS挑战的攻击，生存率为40％，并且显着增加了Cecal连接和穿刺模型中的生存时间。因此，据推测，人类抗TLR4 IgG2对败血症相关损伤起受保护的作用，并且可能适用于治疗与感染相关的免疫功能障碍。
Previous studies have revealed that activation of the Toll-like receptor 4 (TLR4)-mediated proinflammatory signaling pathway plays an important role in acute inflammation, sepsis and chronic inflammatory disorders. Moreover, TLR4 significantly contributes to lipopolysaccharide (LPS)-induced immune response. Thus, modulation of the TLR4 pathway is an important strategy to specifically target these pathologies. The aim of the present study was to develop a complete human anti-TLR4 IgG2 antibody by screening human TLR4 Fab from a phage-display library and integrating it with constant regions of the heavy chain of human IgG2 via antibody engineering. ELISA, a BLItz system and fluorescence-activated cell sorting were used to assess its affinity. Furthermore, mouse-derived peritoneal macrophages were treated with human anti-TLR4 IgG2 and induced with LPS in vitro. Reverse transcription-quantitative PCR and western blotting were used to determine mRNA expression levels of cytokines and phosphorylation levels of signaling pathways, respectively. It was found that human anti-TLR4 IgG2 bound to TLR4 with a high affinity of 8.713×10−10 M, and that preincubation with anti-TLR4 IgG2 inhibited the LPS-induced production of tumor necrosis factor-α, interferon-β and interleukin-6 mRNA expression levels in mouse peritoneal macrophages. It was also demonstrated that human anti-TLR4 IgG2 inhibited LPS-induced TLR4 signaling by reducing the phosphorylation of the NF-κB, mitogen-activated protein kinase and interferon regulatory factor 3 signaling pathways. In addition, human anti-TLR4 IgG2 protected mice from LPS challenge with a survival rate of 40% and also significantly increased the survival time in the cecal ligation and puncture model. Therefore, it was speculated that human anti-TLR4 IgG2 plays a protective role against sepsis-associated injury and is potentially applicable for the treatment of infection-associated immune dysfunction.
登录
查看更多内容
影响因子：
5.3
作者：
Kaplon, Helene;Reichert, Janice M.
通讯作者：
Reichert, Janice M.
影响因子：
3.7
作者：
Neal MD;Jia H;Eyer B;Good M;Guerriero CJ;Sodhi CP;Afrazi A;Prindle T Jr;Ma C;Branca M;Ozolek J;Brodsky JL;Wipf P;Hackam DJ
通讯作者：
Hackam DJ
影响因子：
32.4
作者：
Eckert, Jana K.;Kim, Young J.;Schumann, Ralf R.
通讯作者：
Schumann, Ralf R.
影响因子：
2.6
作者：
Liao, YJ;Zhao, P;Cao, B
通讯作者：
Cao, B
影响因子：
3.7
作者：
Andresen L;Theodorou K;Grünewald S;Czech-Zechmeister B;Könnecke B;Lühder F;Trendelenburg G
通讯作者：
Trendelenburg G