Eukaryotic expression and functional characterization of PD-1 extracellular domain / 生物工程学报
Chinese Journal of Biotechnology
;
(12): 699-703, 2004.
Article
in Chinese
| WPRIM
| ID: wpr-249951
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.
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Physiology
/
Plasmids
/
Therapeutics
/
Transfection
/
Genetic Therapy
/
Antigens, CD
/
Chemistry
/
Cloning, Molecular
/
Protein Structure, Tertiary
/
Cell Line, Tumor
Limits:
Animals
Language:
Chinese
Journal:
Chinese Journal of Biotechnology
Year:
2004
Type:
Article
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