| foxp3转染的CD4~+ CD25~- T细胞治疗急性肺损伤的研究 |
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| 引用本文： | 张方,施毅,李子玲,胡波,宋勇. foxp3转染的CD4~+ CD25~- T细胞治疗急性肺损伤的研究[J]. 中国组织化学与细胞化学杂志, 2011, 20(1): 17-22. DOI: 10.3870/zgzzhx.2011.01.004 |
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| 作者姓名： | 张方 施毅 李子玲 胡波 宋勇 |
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| 作者单位： | 1. 南京军区南京总医院呼吸内科
2. 南京军区联勤部卫生部,南京,210002 |
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| 基金项目： | 中国博士后科学基金资助(20070411050); 江苏省博士后科研资助计划(0701023B); 南京军区面上课题(07M074) |
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| 摘 要： | 目的通过基因转染技术将foxp3基因导入CD4+CD25-T淋巴细胞,生成CD4+CD25-Foxp3+T细胞,通过输入急性肺损伤小鼠模型体内,促进炎症消退,为体外定向操纵调节T细胞生成和细胞过继治疗急性肺损伤奠定基础。方法采用RT-PCR法从小鼠胸腺来源cDNA文库中扩增获得foxp3 cDNA,亚克隆至构建pIRES2-EGFP质粒中,构建并筛选出重组质粒pmFoxp3-IRES2-EGFP;采用免疫磁珠法分离CD4+CD25-T淋巴细胞,穿孔法转染Foxp3基因至CD4+CD25-T淋巴细胞,流式细胞仪检测CD4+和EGFP共表达的细胞比例;将foxp3基因转染的小鼠CD4+CD25-T细胞通过尾静脉输注急性小鼠体内,通过ELISA法检测小鼠肺泡灌洗液中TNF-α以及IL-1β细胞因子的表达,通过肺组织病理研究肺部炎症的消退状况。结果成功构建双基因共表达重组质粒pmFoxp3-IRES2-EGFP;免疫磁珠法较高纯度分离CD4+CD25+、CD4+CD25-T淋巴细胞,电穿孔法成功将重组质粒pmFoxp3-IRES2-EGFP转染致CD4+CD25-T细胞,CD4+和EGFP共表达的细胞比例为35.5%;过继Foxp3基因转染的小鼠CD4+CD25-T细胞以及Treg均能抑制急性肺损伤小鼠肺泡灌洗液中TNF-α以及IL-1β炎性细胞因子的表达,促进炎症的消退。结论转染Foxp3基因的CD4+CD25-细胞同Treg(CD4+CD25-)细胞一样可以通过细胞过继治疗急性肺损伤。
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| 关 键 词： | 急性肺损伤 Foxp3 基因治疗 Treg |
Therapeutic study on the actue lung injury by adoptive transfer of gene foxp3-transfected CD4+ CD25- T lymphocytes |
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| Zhang Fang,Shi Yi,Li Ziling,Hu Bo,Song Yong. Therapeutic study on the actue lung injury by adoptive transfer of gene foxp3-transfected CD4+ CD25- T lymphocytes[J]. Chinese Journal of Histochemistry and Cytochemistry, 2011, 20(1): 17-22. DOI: 10.3870/zgzzhx.2011.01.004 |
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| Authors: | Zhang Fang Shi Yi Li Ziling Hu Bo Song Yong |
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| Affiliation: | Zhang Fang,Shi Yi,Li Ziling,Hu Bo1,Song Yong(Department of Respiratory Diseases,Nanjing Genaral Hospital of Nanjing Military Command,1 Joint Logistics Department of the Nanjing Military Region,Nanjing 210002,China) |
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| Abstract: | Objective To transfer the foxp3 gene into CD25-CD4+ T lymphocytes to induce the production of CD25-CD4+ Foxp3+ T lymphocytes,then infuse it to the mouse model of acute lung injury to promote the subsidence of inflammation.It is a novel way to treat acute lung injury by guiding T regulatory lymphocytes in vitro.Methods The foxp3 cDNA was amplified from Balb/c mouse monocyte suspension by RT-PCR,and subcloned into pIRES2-EGFP plasmids to construct the recombinant plasmids pmFoxp3-IRES2-EGFP.CD4+ CD25-T lympho... |
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| Keywords: | Acute lung injury Foxp3 Gene therapy Treg |
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