文献类型：期刊文献

英文题名：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

作者：Zhu, Yan-Ping[1];Yue, Feng[1];He, Yong[1];Li, Peng[1];Yang, Yuan[1,2];Han, Yu-Ting[1,2];Zhang, Yan-Fang[1];Sun, Guo-Peng[1];Guo, Dong-Guang[1];Yin, Mei[2];Wang, Xuan-Nian[1,2]

第一作者：朱艳平

通讯作者：Wang, XN[1];Wang, XN[2]

机构：[1]Xinxiang Univ, Sch Life Sci & Technol, Biotechnol Res Ctr, Xinxiang 453003, Henan Province, Peoples R China;[2]Henan Inst Sci & Technol, Coll Vet & Anim Sci, Xinxiang 453003, Henan Province, Peoples R China

第一机构：新乡学院生命科学技术学院

通讯机构：[1]corresponding author), Xinxiang Univ, Sch Life Sci & Technol, Biotechnol Res Ctr, Xinxiang 453003, Henan Province, Peoples R China;[2]corresponding author), Henan Inst Sci & Technol, Coll Vet & Anim Sci, Xinxiang 453003, Henan Province, Peoples R China.|[1107115]新乡学院生命科学技术学院;[11071]新乡学院;

年份：2017

卷号：81

期号：2

起止页码：147-154

外文期刊名：CANADIAN JOURNAL OF VETERINARY RESEARCH-REVUE CANADIENNE DE RECHERCHE VETERINAIRE

收录：;Scopus(收录号：2-s2.0-85017141281);WOS:【SCI-EXPANDED(收录号:WOS:000402154700009)】；

基金：This work was supported by grants (nos. 31272539 and 31201877) from the National Natural Science Foundation of China.

语种：英文

摘要：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-ExPD-1 and His-ExPD-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-ExPD-1-FITC and His-ExPD-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.