伴侣素包含T复合蛋白1ε亚基(CCT5)是与人γδT细胞相关的自身抗原.pdfVIP

伴侣素包含T复合蛋白1ε亚基（CCT5）是与人γδT细胞相

关的自身抗原

陈慧;滕达;游红琴;高运安;张建民;何维

【摘要】ObjectiveToexaminetherelationshipbetweenCCT5,γδTcell

andautoimmunediseases.MethodsRecombinantCCT5proteinwas

cloned,expressedandpurifiedinE.coli.ThreepeptidesofCCT5protein

wereusedtoprepareforanti-CCT5monoclonalantibodies.PurifiedCCT5

proteinwasusedtoexpandγδTcellsfrompe-ripheralbloodmononuclear

cells(PBMC).PlasmalevelofCCT5inhealthydonors,patientswith

rheumatoidarthritis(RA)andsystemiclupuserythematosus(SLE)were

detectedbyELISAassays.ThecorrelationanalysisbetweenplasmaCCT5

concentrationandthepercentageofdifferentsubtypesofγδTcells

measuredbyflowcytometrywasmade.ResultsTheCCT5genewas

amplifiedbyPCRandthelengthofthetargetfragmentwas1750bp.The

expressed65kuCCT5proteinwaspurifiedandvalidatedbySDS-

PAGE.Twopairedmonoclonalanti-CCT5antibodieswerenbsp;screened

todetectCCT5proteininplasma.ImmobilizedrecombinantCCT5protein

wasabletoinducespecificsig-nificantamplificationofperipheralγδT

cells.Correlationanalysisof10healthydonorsindicatedsignificantcorre-

lationbetweentheplasmaCCT5concentrationandtheproportionofVγ9

andVδ2γδTcells.TheplasmaCCT5concentrationsignificantlydecreased

inautoimmunediseasespatients,includingRAandSLE.Conclusions

ThesedatasuggestthatCCT5couldbeanovelVγ9δ2γδTcell-related

factorinautoimmunediseases,whichdeepentheunderstandingofVγ9δ2

γδTcellfunctioninautoimmunediseases.%目的：探讨CCT5与γδT细胞及

自身免疫病的关系。方法PCR扩增人CCT5的基因，表达并纯化CCT5重组蛋白；

用3条CCT5蛋白的肽段免疫小鼠，制备并筛选抗CCT5的单克隆抗体；固相化

CCT5重组蛋白，体外扩增人外周血单个核细胞中的γδT细胞；用制备的单抗通

过ELISA检测正常人、类风湿性关节炎（RA）及系统性红斑狼疮（SLE）患者血

浆中CCT5蛋白的含量；用流式细胞技术分析不同亚型γδT细胞的比例，并与血

浆CCT5含量做相关性分析。结果PCR扩增获得CCT5基因，目的片段长度为

1750bp；经SDS分析可见重组CCT5蛋白分子质量约为65ku，并获得

了纯化。通过筛选，获得两株抗CCT5单克隆抗体对