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国家自然科学基金(30671954)

作品数:3 被引量:17H指数:1
相关作者:沈关心张金元廖雯君周玮吴雄文更多>>
相关机构:华中科技大学解放军第四五五医院湖北大学更多>>
发文基金:国家自然科学基金上海市自然科学基金武汉市青年科技晨光计划更多>>
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小鼠PDL1-Red2融合蛋白真核表达载体的构建、表达及其效应
2008年
目的:小鼠跨膜型PD-L1与红色荧光蛋白(RFP)融合基因真核表达载体的构建、表达与鉴定,及其效应初步研究。方法:应用基因工程技术构建重组载体,脂质体转染NIT细胞株,以流式细胞术(FCM)和倒置相差荧光显微镜检测融合蛋白的表达;采用混合淋巴细胞培养,以FCM测定脾细胞的增殖反应。结果:融合基因在转染的真核细胞中获得表达,并呈膜分布,转染PD-L1/pDsRed2-N1的细胞对淋巴细胞增殖反应有一定的抑制作用,表明PD-L1/pDsRed2-N1融合蛋白中分子标签未影响PD-L1的结构及其生物学活性。结论:PD-L1与DsRed2融合基因载体构建成功,表达的融合蛋白具有生物学活性。
廖雯君孙晶朱慧芬张金元沈关心
关键词:PD-L1红色荧光蛋白融合基因
IL-10 Gene Modified Dendritic Cells Inhibit T Helper Type 1-Mediated Alloimmune Responses and Promote Immunological Tolerance in Diabetes被引量:17
2008年
Dendritic cells (DCs) have the potency to regulate the outcome of autoimmunity through the modulation of immune responses. The induction of antigen specific tolerance is critical for prevention and treatment of allograft rejection. In the present study,we transfected IL-10 gene into DCs and investigated their effect on inhibition of lymphocyte activity in vitro and induction of immune tolerance on islet allograft in mice. An IDDM C57BL/6 mouse model was induced by streptozotocin. The islet cells isolated from the BALB/c mice were transplanted into the kidney capules of the model mice followed by injection of IL-10 modified DCs (mDCs). The results showed that mDCs could significantly inhibit T lymphocyte proliferation mediated by allotype cells and induce its apoptosis,whereas,unmodified DCs (umDCs) could promote the murine lymphocyte proliferation markedly. The injection of mDCs could prolong the survival of allotype islet transplanted IDDM mice. The average plasma glucose (PG) level in mDCs treated mice returned to normal within 3 days and lasted for about 2 weeks. The rejection response in control mice occurred for 5 days after transplantation. The level of IFN-γ was lower while IL-4 was higher in mDCs treated mice than that in umDCs treated mice,which indicated that Th1/Th2 deviation occurred. Our studies suggest that IL-10 gene modified DCs can induce the immune tolerance to islet graft and prolong survival of the recipients by the inhibiting of T cell proliferation in allotype mice.
Huifen ZhuWenhong QiuPing LeiWei ZhouXue WenFengrong HeLi LiHong DaiGuanxin ShenFeili Gong
关键词:抵抗力
可溶性mPDL1-hIgGFc表达载体构建、表达及对诱导细胞增殖与凋亡的影响
2008年
目的可溶性mPDL1-hIgGFc(mouse PDL1-human IgG Fc)表达载体的构建与表达,及其对诱导细胞增殖与凋亡的研究。方法以含有mPDL1基因的载体pmPDL1为模板,采用PCR的方法获得mPDL1胞外段基因,定向连接于含有hIgGFc基因片段的载体phIgGFc,构建表达载体pmPDL1-hIgGFc;采用脂质体将重组子转染CHO细胞,转染后的CHO命名为CHOp;ELISA和Western blot法检测目的蛋白的表达,流式细胞术检测CHOp培养上清对混合淋巴细胞培养(MLC)的影响。结果测序结果和酶切鉴定显示构建的表达载体pmPDL1-hIgGFc完全正确;ELISA和Western blot法检测CHOp培养上清中有目的蛋白表达;流式细胞术检测表明CHOp培养上清可体外抑制小鼠MLC,并诱导活化T细胞凋亡,其效应呈剂量依赖性。结论成功构建mPDL1-hIgGFc表达载体;mPDL1-hIgGFc在体外可抑制MLC和诱导活化T细胞凋亡,具有负性调节T细胞的功能。
杨敬廖雯君王国华何峰蓉朱惠芬戴红周玮吴雄文张金元沈关心
关键词:混合淋巴细胞培养增殖凋亡
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