中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2011年
11期
1866-1868
,共3页
陈丽红%黄爱民%刘景丰%邱明链
陳麗紅%黃愛民%劉景豐%邱明鏈
진려홍%황애민%류경봉%구명련
树突状细胞%免疫耐受%转化生长因子－β1%基因转染
樹突狀細胞%免疫耐受%轉化生長因子－β1%基因轉染
수돌상세포%면역내수%전화생장인자－β1%기인전염
Dendritic cells%Immue tolerance%TGF-β1%Gene transfection
目的 探讨转化生长因子(TGF) -β1基因转染后树突状细胞(DC)细胞表型和免疫生物学功能的变化.方法 利用大鼠骨髓细胞诱导、培养不成熟树突状细胞(imDC),以脂质体介导的pIRES2-EGFP-hTGF-β1转染imDC细胞,通过酶联免疫吸附试验(ELISA)和Western blot法检测转染后各组imDC培养上清TGF-β1蛋白表达；通过流式细胞法、免疫细胞化学法及混合淋巴细胞反应检测各组imDC表面分子的分子表型和免疫功能变化.结果 TGF-β1基因转染的imDC上清中均检测相应蛋白的高表达,TGF-β1基因转染组TGF-β1蛋白表达(252.75±11.31) μg/L均高于mDC组(11.19±2.29) μg/L、imDC组(35.18±6.17) μg/L、空载体组(33.67±4.61)μg／L；基因转染组的表面分子CD86、CD80及MHCⅡ表达明显低于空载体组和无任何转染的imDC组；基因转染组的imDC对T的增殖研究结果显示有显著抑制作用,低于空载体和无任何转染的imDC组.结论 TGF-β1基因成功导人imDC,基因改造的imDC不但能维持不成熟状态同时分泌免疫耐受抑制蛋白,致免疫耐受作用显著增强.
目的 探討轉化生長因子(TGF) -β1基因轉染後樹突狀細胞(DC)細胞錶型和免疫生物學功能的變化.方法 利用大鼠骨髓細胞誘導、培養不成熟樹突狀細胞(imDC),以脂質體介導的pIRES2-EGFP-hTGF-β1轉染imDC細胞,通過酶聯免疫吸附試驗(ELISA)和Western blot法檢測轉染後各組imDC培養上清TGF-β1蛋白錶達；通過流式細胞法、免疫細胞化學法及混閤淋巴細胞反應檢測各組imDC錶麵分子的分子錶型和免疫功能變化.結果 TGF-β1基因轉染的imDC上清中均檢測相應蛋白的高錶達,TGF-β1基因轉染組TGF-β1蛋白錶達(252.75±11.31) μg/L均高于mDC組(11.19±2.29) μg/L、imDC組(35.18±6.17) μg/L、空載體組(33.67±4.61)μg／L；基因轉染組的錶麵分子CD86、CD80及MHCⅡ錶達明顯低于空載體組和無任何轉染的imDC組；基因轉染組的imDC對T的增殖研究結果顯示有顯著抑製作用,低于空載體和無任何轉染的imDC組.結論 TGF-β1基因成功導人imDC,基因改造的imDC不但能維持不成熟狀態同時分泌免疫耐受抑製蛋白,緻免疫耐受作用顯著增彊.
목적 탐토전화생장인자(TGF) -β1기인전염후수돌상세포(DC)세포표형화면역생물학공능적변화.방법 이용대서골수세포유도、배양불성숙수돌상세포(imDC),이지질체개도적pIRES2-EGFP-hTGF-β1전염imDC세포,통과매련면역흡부시험(ELISA)화Western blot법검측전염후각조imDC배양상청TGF-β1단백표체；통과류식세포법、면역세포화학법급혼합림파세포반응검측각조imDC표면분자적분자표형화면역공능변화.결과 TGF-β1기인전염적imDC상청중균검측상응단백적고표체,TGF-β1기인전염조TGF-β1단백표체(252.75±11.31) μg/L균고우mDC조(11.19±2.29) μg/L、imDC조(35.18±6.17) μg/L、공재체조(33.67±4.61)μg／L；기인전염조적표면분자CD86、CD80급MHCⅡ표체명현저우공재체조화무임하전염적imDC조；기인전염조적imDC대T적증식연구결과현시유현저억제작용,저우공재체화무임하전염적imDC조.결론 TGF-β1기인성공도인imDC,기인개조적imDC불단능유지불성숙상태동시분비면역내수억제단백,치면역내수작용현저증강.
Objective To assess the biological functions of transforming growth factor-β1 (TGF-β1 ) in rat bone marrow-derived dendritic cells (DCs).Methods In the in vitro study,rat bone marrowderived imature dendritic cells (imDCs) and mature DCS (mDCs) were obtained after primary culture of bone marrow cells in RPMI 1640 medium with different cytokines.We used imDCs transfected with plasmid vectors encoding TGF-β1,imDCs transfected with plasmid vectors encoding enhanced GFP ( eGFP),untransfected imDCs,and un-transfected mDCs.The latter three kinds of imDCS were used as controls.CD86,CD80,and MHC class Ⅱ were compared by using flow cytometry and immuocytochemistry,and T lymphocyte proliferation was observed under mixed lymphocyte reaction.The biological functions of TGF-β1were assessed in rat bone marrow-derived DCs between two groups.Results The expression of TGF-β1 in imDCs transfected with TGF-β1 genes was (252.75 ± 11.31 ) μg/L,which was higher than in imDC group (35.18 ±6.17) μg/L,vector only imDC group (33.67 ±4.61 ) μg/L and mDC group ( 11.19 ±2.29)μg/L.The expression of MHC class Ⅱ,CD80 and CD86 in imDCs transfected with TGF-β1 genes was lower than the other three groups.These results suggest that the expression of MHC class Ⅱ,CD80 and CD86 can be significantly down-regulated,especially in imDCs transfected with TGF-β1.Strong allergenic T lymphocytes proliferation was induced in mDCs,and imDCs transfected with TGF-β1gene showed lower capability in stimulating T lymphocyte proliferation than the other groups.Conclusion The present study demonstrated that TGF-β1 gene can be successfully transferred to imDCs through plasmid vectors.The use of plasmid vectors encoding TGF-β1 gene can inhibit multiple immunological functions of imDCs and enhance their tolerogenicity.
