多肽類結(jié)核桿菌抗原激活的γδ t細(xì)胞表達(dá)抗原提呈細(xì)胞表型和功能

《多肽類結(jié)核桿菌抗原激活的γδ t細(xì)胞表達(dá)抗原提呈細(xì)胞表型和功能》由會(huì)員上傳分享，免費(fèi)在線閱讀，更多相關(guān)內(nèi)容在學(xué)術(shù)論文-天天文庫(kù)。

1、多肽類結(jié)核桿菌抗原激活的γδT細(xì)胞表達(dá)抗原提呈細(xì)胞表型和功能【摘要】目的:觀察用多肽類結(jié)核桿菌耐熱抗原(MtbHAg)激活的人外周血γδT細(xì)胞是否具有抗原提呈細(xì)胞的表型和抗原提呈作用。方法:外周血單個(gè)核細(xì)胞(PBMC)經(jīng)貼壁法獲取單核細(xì)胞,加GMCSF,IL4和LPS培養(yǎng)誘導(dǎo)為成熟DC。PBMC用MtbHAg優(yōu)勢(shì)擴(kuò)增γδT細(xì)胞后用流式分選純化γδT(Vδ2+)細(xì)胞。PBMC經(jīng)尼龍毛柱法獲得純化T細(xì)胞,用CFSE標(biāo)記后單獨(dú)加Mtb分泌性抗原(MtbSAg),或加單核細(xì)胞和MtbSAg共同培養(yǎng),或與經(jīng)MtbSAg預(yù)處理的成熟DC或活化γδT細(xì)胞共同培養(yǎng)。第11天時(shí)用流式細(xì)胞

2、術(shù)檢測(cè)CD4+T細(xì)胞的增殖。MtbHAg活化的γδT細(xì)胞與FITC標(biāo)記的MtbSAg孵育不同時(shí)間后用流式細(xì)胞術(shù)和激光共聚焦顯微鏡檢測(cè)攝入情況。結(jié)果:MtbHAg活化的γδT細(xì)胞CD80,CD86和MHCⅡ類分子的表達(dá)顯著增加。MtbSAg預(yù)處理活化的γδT細(xì)胞誘導(dǎo)初始T細(xì)胞增殖數(shù)和CD4+T細(xì)胞增殖指數(shù)的活性與成熟DC相似。MtbHAg活化的γδT細(xì)胞能攝入FITC標(biāo)記的MtbSAg。結(jié)論:用多肽類MtbHAg激活的γδT細(xì)胞具有抗原提呈細(xì)胞的表型,并具有攝入和提呈可溶性抗原給初始T淋巴細(xì)胞的能力?！娟P(guān)鍵詞】結(jié)核分枝桿菌;抗原提呈;T細(xì)胞亞群;流式細(xì)胞術(shù);激光共聚焦顯

3、微鏡13［Abstract］AIM:ToexplorewhetherthehumanperipheralγδTcellsstimulatedbypolypeptideheatresistantantigenfromMycobacteriumtuberculosis(MtbHAg)expressphenotypeandhavethefuctionofantigenpresentation.METHODS:Themonocytesisolatedbyadhesionmethodfromperipheralbloodmononuclearcells(PBMCs)wereculturedw

4、ithGMCSF,IL4andinducedintothematuredendriticcellsbyaddingLPS.PBMCswerestimulatedwithMtbHAgandIL2toexpandpredominantallyγδTcellsthatwerefurtherpurifiedbyflowsorting.ThepureTcellswereisolatedfromPBMCsusingadhesionrevomalandnyloncolumnmethod,labeledwithCFSE,andculturedinalonewithMtbsecretoryan

5、tigen(MtbSAg),inmonocyteswithMtbSAg,orwithMtbSAgprepulseddendriticcellsorMtbHAgactivatedγδTcellsfor11days.TheproliferationoftheCD4+Tcellsweremeasuredbyflowcytometry.TheMtbHAgactivatedγδTcellswereincubatedwithFITClabelledMtbSAgfordifferenttime,andtheingestionofMtbSAgbyγδTcellswasmeasured

6、byflowcytometryandobservedwithlaserconfocalmicroscopye.RESULTS:TheexpressionsofMHCIIandcostimulatorymoleculesCD80andCD86onγδTcellsthatactivatedbyMtbHAgmarketlyincrasedincomparisonwiththatontherestingγδTcells.Theexpandedcellcountsandproliferationindex13ofthepurenaveTcellsthatinducedbyMtbSAg

7、pretreatedactivatedγδTcellsweresimilartothatinducedbymatureDC.Byusingflowcytometerandlaserconfocalmicroscope,FITClabellaedMtbSAgwasingestedbytheactivatedγδTcells.CONCLUSION:γδTcellsstimulatedbypolypeptideMtbHAgexpressthephenotyp