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[1]陶雷,黎江,朱海莉,等.PD 1抗体表达载体的构建以及其功能的探究[J].浙江理工大学学报,2014,31-32(自科6):697-701.
　TAO Lei,LI Jiang,ZHU Hai li,et al.Research on Construction of Expression Vector ofPD 1 Antibody and Its Function[J].Journal of Zhejiang Sci-Tech University,2014,31-32(自科6):697-701.
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PD 1抗体表达载体的构建以及其功能的探究()
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浙江理工大学学报[ISSN:1673-3851/CN:33-1338/TS]

卷:
第31-32卷
期数:
2014年自科6期
页码:
697-701
栏目:
(自科)生物与生命科学
出版日期:
2014-11-10

文章信息/Info

Title:
Research on Construction of Expression Vector ofPD 1 Antibody and Its Function
文章编号:
1673-3851 (2014) 06-0697-05
作者:
陶雷;  黎江;  朱海莉;  钱其军
1. 浙江理工大学生命科学学院新元医学与生物技术研究所, 杭州 310018;2. 第二军医大学东方肝胆外科医院病毒基因治疗实验室, 上海 200438
Author(s):
TAO Lei;  LI Jiang;  ZHU Haili;  QIAN Qijun
1. Xinyuan Institute of Medicine and Biotechnology, School of Life Science, Zhejiang SciTechUniversity, Hangzhou 310018, China; 2. Laboratory of Viral and Gene Therapy, EasternHepatobiliary Surgical Hospital, Second Military Medical University, Shanghai 200438, China
关键词:
CTL;  PD 1抗体;  抗肿瘤;  免疫细胞治疗;  免疫抑制
分类号:
Q255
文献标志码:
A
摘要:
通过构建含PD 1抗体基因的重组腺病毒表达载体Ad35 anti PD 1,表达的PD 1抗体以探究其对细胞毒性T淋巴细胞(cytotoxic T lymphocyte, CTL)抗肿瘤功能的影响。用ELISA实验检测Ad35 anti  PD 1在293细胞中PD 1抗体的表达量;利用Protein G亲和层析法纯化PD 1抗体;通过体外杀伤实验研究PD 1抗体阻断PD 1信号通路的CTL杀伤肝癌细胞能力的变化。显示成功构建重组腺病毒载体Ad35 anti PD 1,滴度为1×10 10  pfu/mL;ELISA实验检测在72 h时293细胞上清中PD 1抗体的表达量约在600 ng/mL;Westernblotting实验证明PD 1抗体包含正确的重链(50 kD)和轻链(25 kD);ELISA实验检测从100 mL细胞上清中得到1 mL浓度在40 μg/mL的PD 1抗体;体外杀伤实验证明CTL在添加PD 1抗体的环境中杀伤肝癌细胞的能力显著增强。

参考文献/References:

［1］ Okazaki T, Honjo T. PD 1 and PD 1 ligands: from discovery to clinical application［J］. International Immunology, 2007, 19(7): 813-824.
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［7］ Paley M A, Kroy D C, Odorizzi P M, et al. Progenitor and terminal subsets of CD8+ T cells cooperate to contain chronic viral infection［J］. Science, 2012, 338(6111): 1220-1225.
［8］ Yan Zhang S H, Dejun Gong, Yanghua Qin, et al. Programmed death 1 upregμLation is correlated with dysfunction of tumorinfiltrating CD81 T lymphocytes in human nonsmall cell lung cancer［J］. Cellular & Molecular Immunology, 2010, 7: 389-395.
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备注/Memo

备注/Memo:
收稿日期： 2014-01-08
基金项目： 国家自然科学基金(81071850)
作者简介： 陶雷(1988-),男,安徽池州人,硕士研究生,主要从事肿瘤的免疫细胞治疗研究
通信作者： 钱其军,E-mail:qianqi@163.com
更新日期/Last Update: 2014-11-19