CAJ | 학술논문

目的:探讨在异戊烯焦磷酸(IPP)作用下,健康人外周血和新生儿脐带血中γδT细胞的增殖和亚型变化,以期获得足够的具有不同特征的γδT细胞用于后续的实验研究.方法:流式细胞术分析经IPP诱导的单个核细胞表面分子的表达状态,评估其γδT细胞的比例、亚型和表型格局.结果:健康人外周血和新生儿脐带血中γδT细胞数量虽占较低比例,但两者的γδT细胞存在显著异质性.健康人外周血γδT细胞主要以Vγ9Vδ2 TCR亚群为主;初始分选的外周血单个核细胞中Vγ9Vδ2T细胞主要为中央记忆型(CD27~+CD45RA~-)和效应记忆型(CD27~-CD45RA~-);IPP诱导后,γδT细胞显著扩增,VγVδ2 T细胞亚群转变为以效应记忆型为主,并高表达HLA-DR和B7分子.而新生儿脐带血来源的γδT细胞却呈现出较为复杂的亚群异质性,其Vγ9Vδ2 T细胞主要为CD27~+CD45RA~+的幼稚型细胞;经IPP诱导14天后,γδT细胞获得扩增(比例增高)且Vγ9Vδ2 T细胞趋向中央记忆型和效应记忆型分化,但仍以幼稚型为主.结论:健康人外周血和新生儿脐带血中γδT细胞在数量、亚群诸方面存在差异;IPP具有诱导和扩增外周血γδT细胞的牛物学作用,而脐带血来源的γδT细胞还需相关细胞因子的协同作用,才能呈现出其分化成熟为可用于理论研究和临床实验的效应记忆型γδT细胞的潜能.关于其具有的免疫调节和效应功能将另文报道.
목적:탐토재이무희초린산(IPP)작용하,건강인외주혈화신생인제대혈중γδT세포적증식화아형변화,이기획득족구적구유불동특정적γδT세포용우후속적실험연구.방법:류식세포술분석경IPP유도적단개핵세포표면분자적표체상태,평고기γδT세포적비례、아형화표형격국.결과:건강인외주혈화신생인제대혈중γδT세포수량수점교저비례,단량자적γδT세포존재현저이질성.건강인외주혈γδT세포주요이Vγ9Vδ2 TCR아군위주;초시분선적외주혈단개핵세포중Vγ9Vδ2T세포주요위중앙기억형(CD27~+CD45RA~-)화효응기억형(CD27~-CD45RA~-);IPP유도후,γδT세포현저확증,VγVδ2 T세포아군전변위이효응기억형위주,병고표체HLA-DR화B7분자.이신생인제대혈래원적γδT세포각정현출교위복잡적아군이질성,기Vγ9Vδ2 T세포주요위CD27~+CD45RA~+적유치형세포;경IPP유도14천후,γδT세포획득확증(비례증고)차Vγ9Vδ2 T세포추향중앙기억형화효응기억형분화,단잉이유치형위주.결론:건강인외주혈화신생인제대혈중γδT세포재수량、아군제방면존재차이;IPP구유유도화확증외주혈γδT세포적우물학작용,이제대혈래원적γδT세포환수상관세포인자적협동작용,재능정현출기분화성숙위가용우이론연구화림상실험적효응기억형γδT세포적잠능.관우기구유적면역조절화효응공능장령문보도.
Objective: To investigate the variation of γδ T cells from healthy human peripheral blood(PB)and neonatus cord blood (CB)in proliferation and subtypes with isopentenyl pyrophosphate(IPP), and to acquire enough γδ T cells possessing distinct characteristics for experimental study.Methods: Mononuclear-cells from peripheral blood and cord blood induced by IPP were stained separately with monoclonal antibodies,which were fluorescein-labeled,and then used for assaying the expressing condition of surfaco molecules,as well as to evaluate the variation of γδ T cells on the percentage, subtypes and pbenotypes by FCM.Results:γδ T cells only account for a small proportion in both PB and CB.However,there was a significant difference in the heterogeneity between peripheral blood and cord blood γδ T cells.Vγ9Vδ2 T cells were dominant in peripheral blood γδ T cells.Most of Vγ9Vδ2 T cells in fresh isolated PBMC were central memory-type(CD27~+ CD45RA~-)and effector memory-type(CD27~-CD45RA~-)with IPP, PB γδ T cells proliferated strongly;The effector memory-typo(CD27~-CD45RA~-)turned into the main subtype in all Vγ9Vδ2 T cells,and HLA-DR and B7 molecules were highly expressed on the populations.But the cord blood γδ T cells showed rather complex subgroup heterogeneity,and Vγ9Vδ2 T cells were almost na(i)ve-type(CD27~+ CD45RA~+); Though γδ T cells were expanded(the percent of γδ T cells was increased),and Vγ9Vδ2 T cells were differentiated towards central memory-type and effector memory-type on day 14 with IPP,most of γδ T celLs still remained in the phase of na(i)ve-type(CD27~+ CD45RA~+).Conclusion:Tbere lies great differences of γδ T cells in quantity and subtypes between healthy person peripheral blood and neonatus cord blood.Peripheral blood γδ T cells can be activated and proliferated with IPP, while cord blood γδ T cells have the potential to deferentiate into director memory-type which can be used for experimental and clinical study with the synergy of corresponding cytokines.The immuno-regulation and effector function will be reported in other papers.