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Anti-CTGF single-chain variable fragment dimers inhibit human airway smooth muscle (ASM) cell proliferation by down-regulating p-Akt and p-mTOR levels
- Source :
- PLoS ONE, Vol 9, Iss 12, p e113980 (2014), PLoS ONE
- Publication Year :
- 2014
- Publisher :
- Public Library of Science (PLoS), 2014.
-
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.
- Subjects :
- Pulmonology
Recombinant Fusion Proteins
medicine.medical_treatment
Science
chemical and pharmacologic phenomena
Biology
Cell Line
Biomaterials
Cell Fusion
Mice
Medicine and Health Sciences
medicine
Animals
Humans
MTT assay
Fibroblast
Protein kinase B
PI3K/AKT/mTOR pathway
Cell Proliferation
Multidisciplinary
integumentary system
Cell growth
Growth factor
Connective Tissue Growth Factor
Biology and Life Sciences
Antibodies, Monoclonal
Muscle, Smooth
respiratory system
Molecular biology
Asthma
CTGF
medicine.anatomical_structure
Airway Remodeling
Medicine
Myofibroblast
Research Article
Biotechnology
Single-Chain Antibodies
Subjects
Details
- Language :
- English
- ISSN :
- 19326203
- Volume :
- 9
- Issue :
- 12
- Database :
- OpenAIRE
- Journal :
- PLoS ONE
- Accession number :
- edsair.doi.dedup.....cca963d2ec86558d1f243c22dc44b684