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《多發(fā)性硬化抗原特異性CD4^＋CD25^＋調(diào)節(jié)性T細胞的體外擴增及其功能效應.pdf》由會員上傳分享，免費在線閱讀，更多相關內(nèi)容在行業(yè)資料-天天文庫。
1、·基礎論著·多發(fā)性硬化抗原特異性CD4+CD25+調(diào)節(jié)性T細胞的體外擴增及其功能效應姜紅劉廣志楊亭亭何洋于鑫向雅娟高旭光【摘要】目的針對多發(fā)性硬化(MS)患者CD4CD25調(diào)節(jié)性T細胞(Treg)的免疫功能缺陷，嘗試利用體外培養(yǎng)法獲取功能正常的Treg。方法密度梯度離心法獲得MS患者(=5)和健康對照者(=5)外周血單個核細胞(PBMC)，免疫磁珠法分選B細胞和CD4CD25一初始T細胞，前者進一步經(jīng)可溶性CD40配體(sCD40L)刺激活化后作為抗原提呈細胞(APC)，與后者混合體外共培養(yǎng)，并以髓鞘堿性蛋白(M
2、BP)85．。。等刺激后收獲細胞；經(jīng)流式細胞儀分選獲得CD4CD25highCD127Treg。通過增殖分析鑒定所獲Treg的免疫抑制功能。結果磁珠法分選B細胞、初始T細胞的純化率分別達到90％和80．62％。體外誘導擴增所獲的MBP反應性Treg比例為3．5％～6％，MS組與健康對照組Treg比較抑制功能活性無明顯差異。結論經(jīng)此方法可擴增獲得免疫功能正常的Treg細胞，進而為今后開展MS的特異性免疫調(diào)節(jié)治療提供客觀的依據(jù)?！娟P鍵詞】多發(fā)性硬化；T淋巴細胞，調(diào)節(jié)性；體外培養(yǎng)Expansionandfunction
3、analysisofantigen-specificCD4CD25htghregulatoryTcellinpatientswithmul~plesclerosisJiangHong,LiuGuangzhi,YangTingting,HeYang,YuXin,XiangYajuan，GaoXuguang．DepartmentofNeurology,PeMngUniversityPeople'sHospital,Beijing100044,ChinaCorrespondingauthor：GaoXuguang,Em
4、ail：jh1991@sina．com[Abstract]ObjeetiveToobtamnormalregulatoryTcells(Treg)throughinvitrocultureinpatientswithmultiplesclerosis(MS)demonstratingimmunedefectofCD4CD25Treg．MethodsPeripheralbloodmononuclearcells(PBMC)ofMSpatientsandhealthycontrolwasisolatedbydensi
5、tygradientcentrifugation，andCD4CD25一naiveTceUsandaUogeneicBcellswere~thersortedwithimmunomagneticbeads．AllogeneicBcell，afteractivationwithsCD40L，wasco—culturedwithCD4CD25一naiveTcellsinthepresenceofmyelinbasicprotein(MBP)85．99peptide，Rapamycinand4-1BBLfor10day
6、s．CD4CD25~hcD127Tregweresortedwithflowcytometry，anditsimmunosuppressivefunctionwasidentifiedbyproliferationassay．ResultsPurificationrateofCD4CD25一naiveTceilsandallogeneicBcellsthroughirrmmomagneticbeadswere90％and80．62％aswellasforMBP—reactiveTreg3．5％-6％through
7、／nvitroculture．TheTregimmunoactivitiesdemonstratednosignificantdiferencecompared、vitbthatofhealthycontro1．ConclusionTregwithnormalimmunoactivitiescallbeobtainedthroughthiscocultu~methodinvitro，andwilprovidereliableevidenceforflLrtht~"antigen-specificimmunomod
8、ulatorytherapy．[Keywords]Multiplesclerosis；T-lymphocytes，regulatory；／nvitroculture多發(fā)性硬化(multiplesclerosis，MS)是一種調(diào)節(jié)性T細胞(Treg)的效應功能較正常對照下降[1】。發(fā)生于中樞神經(jīng)系統(tǒng)(CNS)的炎性脫髓鞘疾病，多項動物實驗發(fā)現(xiàn)，經(jīng)靜脈注射給予實驗性自身免多見
