title: Anti-CTGF single-chain variable fragment dimers inhibit human airway smooth muscle (ASM) cell proliferation by down-regulating p-Akt and p-mTOR levels.
abstract: Connective tissue growth factor (CTGF) contributes to airway smooth muscle (ASM) cell hyperplasia in asthma. Humanized single-chain variable fragment antibody (scFv) was well characterized as a CTGF antagonist in the differentiation of fibroblast into myofibroblast and pulmonary fibrosis in our previous studies. To further improve the bioactivity of scFv, we constructed a plasmid to express scFv-linker-matrilin-6×His fusion proteins that could self-assemble into the scFv dimers by disulfide bonds in matrilin under non-reducing conditions. An immunoreactivity assay demonstrated that the scFv dimer could highly bind to CTGF in a concentration-dependent manner. The MTT and EdU assay results revealed that CTGF (≥10 ng/mL) promoted the proliferation of ASM cells, and this effect was inhibited when the cells were treated with anti-CTGF scFv dimer. The western blot analysis results showed that increased phosphorylation of Akt and mTOR induced by CTGF could be suppressed by this scFv dimer. Based on these findings, anti-CTGF scFv dimer may be a potential agent for the prevention of airway remodeling in asthma.
authors.affiliation: Medical School, Southeast University, Nanjing 210009, China.
authors.forename: Wei
authors.initials: W
authors.lastname: Gao
authors.affiliation: Medical School, Southeast University, Nanjing 210009, China.
authors.forename: Liting
authors.initials: L
authors.lastname: Cai
authors.affiliation: Department of Biological engineering, Southeast University, Nanjing 210009, China.
authors.forename: Xudong
authors.initials: X
authors.lastname: Xu
authors.affiliation: Medical School, Southeast University, Nanjing 210009, China.
authors.forename: Juxiang
authors.initials: J
authors.lastname: Fan
authors.affiliation: Medical School, Southeast University, Nanjing 210009, China.
authors.forename: Xiulei
authors.initials: X
authors.lastname: Xue
authors.affiliation: Medical School, Southeast University, Nanjing 210009, China.
authors.forename: Xuejiao
authors.initials: X
authors.lastname: Yan
authors.affiliation: Medical School, Southeast University, Nanjing 210009, China.
authors.forename: Qinrong
authors.initials: Q
authors.lastname: Qu
authors.affiliation: Department of Respiration, Zhongda Hospital, Southeast University, Nanjing 210009, China.
authors.forename: Xihua
authors.initials: X
authors.lastname: Wang
authors.affiliation: Center of Clinical Laboratory Medicine, Zhongda Hospital, Southeast University, Nanjing 210009, China.
authors.forename: Chen
authors.initials: C
authors.lastname: Zhang
authors.affiliation: Center of Clinical Laboratory Medicine, Zhongda Hospital, Southeast University, Nanjing 210009, China.
authors.forename: Guoqiu
authors.initials: G
authors.lastname: Wu
mesh_terms: D056151:Airway Remodeling / Q000187:drug effects / Q000276:immunology; D000818:Animals; D000911:Antibodies, Monoclonal / Q000008:administration & dosage; D001249:Asthma / Q000188:drug therapy* / Q000276:immunology / Q000473:pathology; D002459:Cell Fusion; D002460:Cell Line; D049109:Cell Proliferation / Q000187:drug effects; D055513:Connective Tissue Growth Factor / Q000037:antagonists & inhibitors / Q000276:immunology*; D006801:Humans; D051379:Mice; D009130:Muscle, Smooth / Q000187:drug effects / Q000254:growth & development*; D011993:Recombinant Fusion Proteins; D057127:Single-Chain Antibodies / Q000008:administration & dosage* / Q000276:immunology
chemical_list: D000911:Antibodies, Monoclonal; C526317:CCN2 protein, human; D011993:Recombinant Fusion Proteins; D057127:Single-Chain Antibodies; D055513:Connective Tissue Growth Factor