Eukaryotic expression and functional characterization of PD-1 extracellular domain.
- Author:
Yu-Fei HE
1
;
Gui-Mei ZHANG
;
Xiao-Hong WANG
;
Hui ZHANG
;
Ye YUAN
;
Dong LI
;
Zuo-Hua FENG
Author Information
1. Department of Biochemistry & Molecular Biology, Tongji Medical College, Huazhong University of Science & Technology, Wuhan 430030, China.
- Publication Type:Journal Article
- MeSH:
Animals;
Antigens, CD;
physiology;
Antigens, Surface;
chemistry;
genetics;
physiology;
Apoptosis Regulatory Proteins;
chemistry;
genetics;
physiology;
CTLA-4 Antigen;
Cell Line, Tumor;
Cloning, Molecular;
Flow Cytometry;
Genetic Therapy;
Mice;
Neoplasms;
therapy;
Plasmids;
Programmed Cell Death 1 Receptor;
Protein Structure, Tertiary;
Transfection
- From:
Chinese Journal of Biotechnology
2004;20(5):699-703
- CountryChina
- Language:Chinese
-
Abstract:
The negative signal provided by interactions of costimulatory molecules, programmed death-1 (PD-1) and its ligands, PD-L1 (also B7-H1) and PD-L2 (also B7-DC), is involved in the mechanisms of tumor immune evasion. To block PD-Ls-PD-1 interactions by a soluble receptor of PD-1, we constructed a eukaryotic expression plasmid that expresses extracellular region (aa1-aa167) of murine PD-1 (pPD-1A) and, another version of pPD-1A, pPD-1B, carrying cDNAs encoding for both extracellular region of PD-1 and green fluorescent protein (GFP) reporter gene, which was inserted downstream of PD-1. Experiment of BHK cells transfected with pPD-1B determined that most expression product (sPD-1) in the cells was secreted out. FACS analysis revealed that sPD-1 was specific and bound efficiently to PD-1 ligands. Cytotoxicity assay showed that blocking PD-Ls on either tumor cells or spleen cells by sPD-1 mediated enhanced lysis of H22 cells by Hsp70-H22 peptides complexstimulated spleen cells. The constructed plasmid vector would provide a novel method of tumor gene therapy of blocking PD-Ls-PD-1 interactions by expression of soluble receptor of PD-1 in tumor sites, which could increase the antitumor activity.
