机构：[1]Department of Neurosurgery, The Second Affiliated Hospital of Soochow University, Suzhou 215004, China[2]Department of Neurosurgery, Wuxi Third People’s Hospital, Wuxi 214041, China[3]Department of Neurosurgery, The First Affiliated Hospital of Soochow University, Suzhou 215006, China
The objective of this study is to investigate the negative immunomodulatory capacity of human amniotic mesenchymal cells (AMSCs) and their possible intrinsic mechanism, by which we can confirm that they modulate microglial activation of central nervous system from multiple perspectives at the molecular level. The identification of the immune phenotype of AMSCs and microglial cells was executed by immunohistochemical methods and flow cytometry. Meanwhile, the influence and mechanism of amniotic mesenchymal cells in vitro on proliferation, cell cycle, and cytokine release of activated microglia (MI) would be detected by ELISA, beta-liquid scintillation counting method, and flow cytometry. Human amnion mesenchymal cells highly expressed negative co-stimulatory molecules PD-L1, while its ligand PD1 was expressed with high level by activated MI. When adding the PD-L1mAb to the mixed culture system composed of AMSCs and activated MI, the proliferation inhibitory effect and the cycle-blocking effect produced by the former on the latter would be partially reversed; at the same time, the impact of the latter cytokine secretion would be adjusted. As a conclusion, AMSCs play inhibitory effects on microglial activation, proliferation, and immune effects partially through the PD-L1-PD1 signaling pathways.
基金：
RPMI1640 (Gibco BRL, Gaithersburg, MD, USA); 10 % FBS (Gibco BRL); trypsin (Sigma, St. Louis, MO, USA); mouse anti-human monoclonal antibodies of PD-1, CD40, CD80, CD83, and CD86 (eBioscience, San Diego, CA); mouse anti-human PD-L1mAb (Stem Cell Research Institute of Suzhou University, Suzhou, China) LPS (Sigma); penicillin and streptomycin (Gibco BRL); PI (Sigma); TNF-α ELISA kits (R&D, Minneapolis, MN, USA) NO ELISA kits (R&D); the rabbit anti-mouse PE fluorescent secondary antibodies (Shanghai Yeli Biotechnology Co. Ltd., Shanghai, China); 3H-TdR (the Atomic Energy Research Institute of Chinese Academy of Sciences, Beijing, China); mouse anti-human antibody MAC-1 (eBio-science) (San Diego, CA, USA); mouse anti-human OX40L monoclonal antibodies (Chemicon International, Single Oak Drive Temecula, CA); polylysine (Sigma); antibody CD68 (DAKO, Copenhagen, Denmark); mouse anti-human monoclonal antibody GFAP (Glial fibrillary acidic protein) IgG (Santa Cruz, City of Santa Cruz, CA, USA); the sheep anti-mouse IgG secondary antibody combined with FITC (Biosource, United BioSource Corporation, Chevy Chase, MD, USA); tar purple (Shanghai Kaishi Technology Company, Shanghai, China); and the monoclonal antibody vimentin IgG (Santa Cruz) were used.
第一作者机构：[1]Department of Neurosurgery, The Second Affiliated Hospital of Soochow University, Suzhou 215004, China[2]Department of Neurosurgery, Wuxi Third People’s Hospital, Wuxi 214041, China
通讯作者：
通讯机构：[1]Department of Neurosurgery, The Second Affiliated Hospital of Soochow University, Suzhou 215004, China
推荐引用方式(GB/T 7714)：
Weijiang Wu,Qing Lan,Hua Lu,et al.Human Amnion Mesenchymal Cells Negative Co-stimulatory Molecules PD-L1 Expression and Its Capacity of Modulating Microglial Activation of CNS[J].CELL BIOCHEMISTRY AND BIOPHYSICS.2014,69(1):35-45.doi:10.1007/s12013-013-9763-9.
APA：
Weijiang Wu,Qing Lan,Hua Lu,Jie Xu,Aihua Zhu...&Guozhen Hui.(2014).Human Amnion Mesenchymal Cells Negative Co-stimulatory Molecules PD-L1 Expression and Its Capacity of Modulating Microglial Activation of CNS.CELL BIOCHEMISTRY AND BIOPHYSICS,69,(1)
MLA：
Weijiang Wu,et al."Human Amnion Mesenchymal Cells Negative Co-stimulatory Molecules PD-L1 Expression and Its Capacity of Modulating Microglial Activation of CNS".CELL BIOCHEMISTRY AND BIOPHYSICS 69..1(2014):35-45
