title: The Fab Fragment of a Humanized Anti-Toll Like Receptor 4 (TLR4) Monoclonal Antibody Reduces the Lipopolysaccharide Response via TLR4 in Mouse Macrophage.
abstract: Lipopolysaccharides (LPS) can induce acute inflammation, sepsis, or chronic inflammatory disorders through the Toll receptor 4 (TLR4) signaling pathway. The TLR4/MD2 (myeloid differentiation protein 2) complex plays a major role in the immune response to LPS. However, there is not a good method to suppress the immune response induced by LPS via this complex in macrophages. In this article, we aimed to evaluate the effects of humanized anti-TLR4 monoclonal antibodies on LPS-induced responses in mouse macrophages. The peritoneal macrophages of mice were incubated with anti-TLR4 monoclonal antibodies and stimulated with LPS. The expression levels of cytokines were analyzed by quantitative polymerase chain reaction and enzyme-linked immunosorbent assays. Additionally, activation of various signaling pathways was evaluated by Western blotting. The results showed that the humanized anti-TLR4 monoclonal antibody blocked the inflammatory cytokines expression at both the mRNA and protein level. We also found that the Fab fragment significantly inhibited the nuclear factor kappaB signaling pathway by reducing the phosphorylation of the inhibitor of kappaBalpha and decreasing the translocation of p65, resulting in the suppression of p38, extracellular signal-regulated kinase 1/2, c-Jun N-terminal kinase 1/2, and IFN-β regulatory factor 3 phosphorylation. Therefore, our study showed that this humanized anti-TLR4 monoclonal antibody could effectively protect against LPS-induced responses by blocking the TLR4 signaling pathway in mouse peritoneal macrophages.
authors.affiliation: Department of Infectious Disease, Anhui Medical University Affiliated with Bayi Clinical College, Hefei 230000, China. cbg1021@163.com.
authors.forename: Binggang
authors.initials: B
authors.lastname: Cai
authors.affiliation: Department of Infectious Disease, Anhui Medical University Affiliated with Bayi Clinical College, Hefei 230000, China. maorongwang@126.com.|Institute of Liver Disease, Nanjing Jingdu Hospital, Nanjing 210002, China. maorongwang@126.com.
authors.forename: Maorong
authors.initials: M
authors.lastname: Wang
authors.affiliation: Department of Microbiology, Huadong Medical Institute of Biotechnology, Nanjing 210002, China. xh_zhu@163.com.
authors.forename: Xuhui
authors.initials: X
authors.lastname: Zhu
authors.affiliation: Institute of Liver Disease, Nanjing Jingdu Hospital, Nanjing 210002, China. xujingjerry@126.com.
authors.forename: Jing
authors.initials: J
authors.lastname: Xu
authors.affiliation: Department of Pathology, Key Laboratory of Antibody Technique of the Ministry of Health, NJMU, Nanjing 210029, China. zwk1987617@163.com.
authors.forename: Wenkai
authors.initials: W
authors.lastname: Zheng
authors.affiliation: Department of Pathology, Key Laboratory of Antibody Technique of the Ministry of Health, NJMU, Nanjing 210029, China. rachel_zhang2005@163.com.
authors.forename: Yiqing
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Department of Microbiology, Huadong Medical Institute of Biotechnology, Nanjing 210002, China. zhengf82@gmail.com.
authors.forename: Feng
authors.initials: F
authors.lastname: Zheng
authors.affiliation: Department of Pathology, Key Laboratory of Antibody Technique of the Ministry of Health, NJMU, Nanjing 210029, China. fengzhenqing@njmu.edu.cn.
authors.forename: Zhenqing
authors.initials: Z
authors.lastname: Feng
authors.affiliation: Department of Microbiology, Huadong Medical Institute of Biotechnology, Nanjing 210002, China. zhujin1968@njmu.edu.cn.|Department of Pathology, Key Laboratory of Antibody Technique of the Ministry of Health, NJMU, Nanjing 210029, China. zhujin1968@njmu.edu.cn.
authors.forename: Jin
authors.initials: J
authors.lastname: Zhu
mesh_terms: D000818:Animals; D061067:Antibodies, Monoclonal, Humanized / Q000276:immunology*; D002478:Cells, Cultured; D007140:Immunoglobulin Fab Fragments / Q000276:immunology*; D050838:Interferon Regulatory Factor-3 / Q000378:metabolism; D048031:JNK Mitogen-Activated Protein Kinases / Q000378:metabolism; D008070:Lipopolysaccharides / Q000276:immunology; D017737:Macrophages, Peritoneal / Q000276:immunology*; D051379:Mice; D008810:Mice, Inbred C57BL; D016328:NF-kappa B / Q000378:metabolism; D015398:Signal Transduction; D051197:Toll-Like Receptor 4 / Q000276:immunology*
chemical_list: D061067:Antibodies, Monoclonal, Humanized; D007140:Immunoglobulin Fab Fragments; D050838:Interferon Regulatory Factor-3; D008070:Lipopolysaccharides; D016328:NF-kappa B; C493487:Tlr4 protein, mouse; D051197:Toll-Like Receptor 4; D048031:JNK Mitogen-Activated Protein Kinases
keywords: Fab fragment; Toll-like receptor 4 signaling; humanized anti-Toll like receptor 4 antibody; lipopolysaccharide