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
Dans Chinois
| 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.
Texte intégral:
Disponible
Indice:
WPRIM (Pacifique occidental)
Sujet Principal:
Physiologie
/
Plasmides
/
Thérapeutique
/
Transfection
/
Thérapie génétique
/
Antigènes CD
/
Chimie
/
Clonage moléculaire
/
Structure tertiaire des protéines
/
Lignée cellulaire tumorale
Limites du sujet:
Animaux
langue:
Chinois
Texte intégral:
Chinese Journal of Biotechnology
Année:
2004
Type:
Article
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