Prokaryotic expression of the extracellular domain of porcine programmed death 1 (PD-1) and its ligand PD-L1 and identification of the binding with peripheral blood mononuclear cells in vitro .

Autor:	Zhu YP; Biotechnology Research Center, School of Life Science and Technology, Xinxiang University, Xinxiang, Henan Province 453003, China (Zhu, Yue, He, Li, Yang, Han, Zhang, Sun, Guo, Wang); College of Veterinary and Animal Sciences, Henan Institute of Science and Technology, Xinxiang, Henan Province 453003, China (Yang, Han, Yin, Wang)., Yue F; Biotechnology Research Center, School of Life Science and Technology, Xinxiang University, Xinxiang, Henan Province 453003, China (Zhu, Yue, He, Li, Yang, Han, Zhang, Sun, Guo, Wang); College of Veterinary and Animal Sciences, Henan Institute of Science and Technology, Xinxiang, Henan Province 453003, China (Yang, Han, Yin, Wang)., He Y; Biotechnology Research Center, School of Life Science and Technology, Xinxiang University, Xinxiang, Henan Province 453003, China (Zhu, Yue, He, Li, Yang, Han, Zhang, Sun, Guo, Wang); College of Veterinary and Animal Sciences, Henan Institute of Science and Technology, Xinxiang, Henan Province 453003, China (Yang, Han, Yin, Wang)., Li P; Biotechnology Research Center, School of Life Science and Technology, Xinxiang University, Xinxiang, Henan Province 453003, China (Zhu, Yue, He, Li, Yang, Han, Zhang, Sun, Guo, Wang); College of Veterinary and Animal Sciences, Henan Institute of Science and Technology, Xinxiang, Henan Province 453003, China (Yang, Han, Yin, Wang)., Yang Y; Biotechnology Research Center, School of Life Science and Technology, Xinxiang University, Xinxiang, Henan Province 453003, China (Zhu, Yue, He, Li, Yang, Han, Zhang, Sun, Guo, Wang); College of Veterinary and Animal Sciences, Henan Institute of Science and Technology, Xinxiang, Henan Province 453003, China (Yang, Han, Yin, Wang)., Han YT; Biotechnology Research Center, School of Life Science and Technology, Xinxiang University, Xinxiang, Henan Province 453003, China (Zhu, Yue, He, Li, Yang, Han, Zhang, Sun, Guo, Wang); College of Veterinary and Animal Sciences, Henan Institute of Science and Technology, Xinxiang, Henan Province 453003, China (Yang, Han, Yin, Wang)., Zhang YF; Biotechnology Research Center, School of Life Science and Technology, Xinxiang University, Xinxiang, Henan Province 453003, China (Zhu, Yue, He, Li, Yang, Han, Zhang, Sun, Guo, Wang); College of Veterinary and Animal Sciences, Henan Institute of Science and Technology, Xinxiang, Henan Province 453003, China (Yang, Han, Yin, Wang)., Sun GP; Biotechnology Research Center, School of Life Science and Technology, Xinxiang University, Xinxiang, Henan Province 453003, China (Zhu, Yue, He, Li, Yang, Han, Zhang, Sun, Guo, Wang); College of Veterinary and Animal Sciences, Henan Institute of Science and Technology, Xinxiang, Henan Province 453003, China (Yang, Han, Yin, Wang)., Guo DG; Biotechnology Research Center, School of Life Science and Technology, Xinxiang University, Xinxiang, Henan Province 453003, China (Zhu, Yue, He, Li, Yang, Han, Zhang, Sun, Guo, Wang); College of Veterinary and Animal Sciences, Henan Institute of Science and Technology, Xinxiang, Henan Province 453003, China (Yang, Han, Yin, Wang)., Yin M; Biotechnology Research Center, School of Life Science and Technology, Xinxiang University, Xinxiang, Henan Province 453003, China (Zhu, Yue, He, Li, Yang, Han, Zhang, Sun, Guo, Wang); College of Veterinary and Animal Sciences, Henan Institute of Science and Technology, Xinxiang, Henan Province 453003, China (Yang, Han, Yin, Wang)., Wang XN; Biotechnology Research Center, School of Life Science and Technology, Xinxiang University, Xinxiang, Henan Province 453003, China (Zhu, Yue, He, Li, Yang, Han, Zhang, Sun, Guo, Wang); College of Veterinary and Animal Sciences, Henan Institute of Science and Technology, Xinxiang, Henan Province 453003, China (Yang, Han, Yin, Wang).
Jazyk:	angličtina
Zdroj:	Canadian journal of veterinary research = Revue canadienne de recherche veterinaire [Can J Vet Res] 2017 Apr; Vol. 81 (2), pp. 147-154.
Abstrakt:	Programmed cell death protein 1 (PD-1), a costimulatory molecule of the CD28 family, has 2 ligands, PD-L1 and PD-L2. Our previous studies showed that the expression of PD-1 and PD-L1 is up-regulated during viral infection in pigs. Extensive studies have shown that blockade of the PD-1/PD-L1 pathways by anti-PD-L1 antibody or soluble PD-1 restores exhausted T-cells in humans and mice. In the present study the extracellular domains of PD-1 and PD-L1 were used to evaluate the binding of PD-1 and PD-L1 with peripheral blood mononuclear cells (PBMCs). We amplified the cDNA encoding the extracellular domains of PD-1 and PD-L1 to construct recombinant expression plasmids and obtain soluble recombinant proteins, which were then labeled with fluorescein isothiocyanate (FITC). The His- Ex PD-1 and His- Ex PD-L1 recombinant proteins were expressed in the form of inclusion bodies with a relative molecular weight of 33.0 and 45.0 kDa, respectively. We then prepared polyclonal antibodies against the proteins with a multi-antiserum titer of 1:102 400. Binding of the proteins with PBMCs was evaluated by flow cytometry. The fluorescence signals of His- Ex PD-1-FITC and His- Ex PD-L1-FITC were greater than those for the FITC control. These results suggest that the soluble recombinant proteins may be used to prepare monoclonal antibodies to block the PD-1/PD-L1 pathway.
Databáze:	MEDLINE
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