ccr7基因转染对小鼠未成熟树突状细胞成熟及迁移功能的影响

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1、CCR7基因转染对小鼠未成熟树突状细胞成熟及迁移功能的影响辛海明，彭毅志，郭浩，袁志强（第三军医大学西南医院全军烧伤研究所，创伤、烧伤与复合伤国家重点实验室，重庆400038）摘要：目的研究趋化因子受体-7(chemokinereceptor-7，CCR7)重组腺病毒体外转染对小鼠未成熟树突状细胞（immaturedendriticcells，imDCs）成熟和迁移功能的影响。方法小鼠骨髓经梯度离心及rmIL-4、rmGM-CSF诱导培养未成熟树突状细胞；CCR7重组腺病毒经增强离心法转染未成熟树突状细胞，观察C

2、CR7重组腺病毒对未成熟树突状细胞的形态、细胞表型及趋化功能的影响。结果扫描电镜显示CCR7腺病毒转染后细胞树枝状突起较未成熟DC增多，即轻微地诱使imDCs向成熟转化，但IL-10可抑制成熟；流式结果表明转染后细胞表型符合典型imDCs的特征；WesternBlotblot证实CCR7重组腺病毒转染imDCs后可增强CCR7的表达；体外趋化实验显示转染后imDCs体外迁移百分比由4.2%增加到36.1%。结论CCR7重组腺病毒转染imDCs后，其仍保留未成熟特性，CCR7蛋白表达增强，体外迁移能力明显增强。关键

3、词：CCR7；未成熟树突状细胞；重组腺病毒；免疫耐受中图法分类号：文献标识码：Transfectionofccr7recombinantadenovirusenhancesTheinfluenceofmatureizationandmigrateionfunctioninofmurineimmaturedendriticcellswhichtransfectedwithccr7recombinantadenovirusinvitroXINHai-ming,PENGYi-zhi,GUOHao,YUANZhi-qia

4、ng(StateKeyLaboratoryofTrauma，,BurnsandCombinedInjury，,InstituteofBurns，,SouthwestHospital，,ThirdMilitaryMedicalUniversity,，Chongqing400038,，China)Abstract：ObjectiveToinvestigatetheinfluenceofmatureizationandmigrateionfunctioninofmurineimmaturedendriticcells(

5、DCs)whichaftertransfectdionwithchemokinereceptor-7(ccr7)recombinantadenovirusinvitro.MethodsBonemarrowcellswereisolatedfromhealthBalb/cmiceandthenthemononuclearcellswerescreenedwithgradientcentrifugation.Immaturedendriticcellsweregeneratedinducedinthepresence

6、ofrecombinantmurineinterleukin-4andgranulocytemacrophagecolony-stimulatingfactorfrommurinebonemarrowbygradientcentrifugationmethod;.Recombinantadenovirusvectorencodingfull-lengthmurineccr7cDNAwastransfectedintotheimmaturedendriticcellsbycentrifugalforcemethod

7、,.Thenthemorphology、,cellphenotype,CCR7expressionandmigrationintheimmaturedendriticcellswereobservedafterthetransfectedionwithccr7recombinantadenovirusbyscanningelectronmicroscopy,flowcytometry,WesternblotanalysisandTranswellcellmigrationassay.ResultsTherecom

8、binantadenoviruscanslightlyinducedimDCstotransformedintomatureDCswhichwasobservedbyscanningelectronmicroscopye,butIL-10caninhibitedittheprocess;FlowcytometryindicatedTthecellphenotypeoffl