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重组人白细胞介素21的制备及免疫学活性分析

Preparation of recombinant human interleukin 21 and its immunologic activity
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摘要 目的:纯化制备rhIL-21,并分析其免疫学活性。方法:在含重组质粒pGEX4T-2/IL-21的大肠杆菌DH5α中,经IPTG诱导表达出GST-融合蛋白,蛋白经过一系列的纯化,SDS-PAGE电泳鉴定蛋白纯度。用MTT法研究rhIL-21对T细胞增殖、NK细胞杀伤活性的影响。结果:表达出了可溶性融合蛋白,纯化后的rhIL-21纯度达95%。rhIL-21能促进T细胞增殖,能增强NK细胞杀伤活性。结论:成功制备了rhIL-21,其在体外具有一定的免疫学活性,为其大量制备和体内的生物学活性研究奠定了基础。 Objective：To prepare and purify recombinant human interleukin 21（rhIL-21）,and to study its immunologicd activity in vitro.Methods：The E.coli DH5α containing recombinant plasmid pGEX4T-2/IL-21 was induced to express protein by IPTG.Supernatants of lysate were purified by affinity chromatography,thrombin digestion and cation chromatography sequentially.The purified products were analyzed by SDS-PAGE.The effect of rhIL-21 on proliferation of T lymphocyte and cytotoxicity of NK cells was studied by using MTT method.Results：The target protein was soluble in supernatants of the germ culture.Purity of rhIL-21 was 95% after the procedures for purification.rhIL-21 augmented proliferation of T lymphocytes and cytotoxicity of NK cells in vitro.Conclusion：rhIL-21 was successfully prepared in small scale and purified.It laid a foundation of study on biological activity in vivo,the procedure of pilot and large-scale preparations of rhIL-21.
作者 刘明军 孙秀芳 钱冬萌 王斌
机构地区 青岛大学医学院附属医院临床免疫检验科 青岛大学医学院医药生物技术重点实验室
出处 《中国免疫学杂志》 CAS CSCD 北大核心 2009年第1期72-75,共4页 Chinese Journal of Immunology
基金 山东省自然科学基金(E99C02) 青岛市科技局计划项目(04-2-J2-95)资助
关键词 人白细胞介素21 大肠杆菌 表达 纯化 活性 Human interleukin 21 Escherichia coli Expression Purification Activity
分类号 Q78 [生物学—分子生物学]
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参考文献8

  • 1Parrish-Novak J, Dillon S R, Nelson A et al. Interleukin 21 and its receptor are involved in NK cell expansion and regulation of lymphocyte function [J]. Nature,2000;408:57-63.
  • 2Kasaian M T,Whitters M J,Carter L et al.IL-21 limits NKcell responses and promotes antigen-specific T cellactivation: a mediator of the transition from innate to adaptive immunity [J]. Immunity,2002; 16 (4) :559-569.
  • 3付强,钱冬萌,闫志勇,侯伟,赵百慧,王斌.IL-21编码基因的克隆及其在大肠杆菌中表达[J].青岛大学医学院学报,2003,39(1):15-17. 被引量：6
  • 4黎阳,黄绍良,魏菁,吴燕峰,周敦华.不同细胞因子培养体系对CIK、NK及NKT细胞等诱导产率的影响[J].中山大学学报（医学科学版）,2003,24(4):363-367. 被引量：5
  • 5Asano R, Kudo T, Makabe K et al. Antitumor activity of interleukin-21 prepared by novel refolding procedure from inclusion bodies expressed in Escherichia coli [J]. FEBS Lett,2002;528 (123):70-76.
  • 6Mari Strengell, Sampsa Matikainen, Jukka Siren et al. IL-21 in synergy with IL-15 or IL-18 enhances IFN-γ production in human NK and T ceils [J] .J Immunology,2003; 170(11):5464-5469.
  • 7Adrianna Moroz, Cheryl Eppolito, Li Qingsheng et al. IL-21 enhances and sustains CD8^+ T cell responses to achieve durable tumor immunity: comparative evaluation of IL-2, IL-15, and IL-21 [ J ]. J Immunology, 2004; 173 : 900-909.
  • 8Fotmtoulakis M, Takaes B. Design of protein purification path-ways:application to the protege of Haemophilus influenza using heparin chromatography [J].Protein Expr Purif, 1998;14 (2):113-116.

二级参考文献19

  • 1Jean D,Sara L,Cliona 0 F,et al.Selective expansion and partial activation of human NK cells and NK reeepfor-positive T cells by IL-2 and IL-15[J].J Immunol.2001,167(6):3129.
  • 2Lu P H.Negrin R S.A novel population of expanded human CD3+CD56+cells derived from T cells with potent in vivo antitumor activity in mice with severe combined immunodeficiency[J]. J Immunol, 1994,153(4):1687.
  • 3Zoll B,Lefterova P,Csipai M,et al.Generation of cytokine—induced killer cells using exogenous interleukin-2,-7 or-12[J].Cancer Immunol Immunother,1998,47(4):221.
  • 4Martino R, Badell I, Brunet S, et al. Second bone marrow transplantation for leukemia in untreated relapse[J]. Bone Marrow Transplant, 1994,14(4): 589.
  • 5Lee P T, Benlagha K, Teyton L, et al. Distinct functional lineages of human V(alpha) 24 natural kiLler T cells[J]. J Exp Med, 2002,195(5): 637.
  • 6Rocha V,Cornish J,Sievers EL,et al.Comparison of outcomes of unrelated bone marrow and umbilical cord blood transplants in children with acute leukemia[J].Blood,2001,97(10):2962.
  • 7Stremmel C,Exley M,Balk S,et al.Characterization of the phenotype and function of CD8(+),alpha／beta(+)NKT cells from tumor-bearlng mice that show a natural killer cell activity and lyse multiple tumor targets[J].Eur J Immunol,2001,31(9):2818.
  • 8Van der V H,Nishi N,Koezuka Y.et al.Potent expansion of human natural killer T cells using alpha-galactosylcerdmide(KRN7000)-loaded monocytederived dendritic cells,cultured in the presence of IL-7 and IL-15[J] Immunol Methods,2001,247(1-2):61.
  • 9Habib T,Senadheera S,Weinberg K,et al.The common gamma chain (gamma C) is a required signaling component of the IL-21 receptor and supports IL-21-induced cell proliferation via JAK3[].Biochemistry.2002
  • 10Asano R,Kudo T,Makabe K,et al.Antitumor activity of interleukin-21 prepared by novel refolding procedure from inclusion bodies expressed in Escherichia coli[].FEBS Letters.2002

共引文献9

中国免疫学杂志
2009年 第1期

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