表达CD20scFv-IgGFc-CD28-ζ及CD20scFv-IgGFc T淋巴细胞的建立及其对B细胞淋巴瘤靶向杀伤作用的比较  

Establishment of T-lymphocytes that express CD20scFv-IgGFc-CD28-ζ and CD20scFv-IgGFc and their killing activity of B-lymphoma cells

作　　者：谭映霞[1] 俞康[2] 胡永仙[2] 章圣辉[1] 高申孟[1] 吴建波[1] 

机构地区：[1]温州医学院附属第一医院医学科学研究所,浙江温州325011 [2]温州医学院附属第一医院血液内科,浙江温州325011

出　　处：《中国病理生理杂志》2008年第7期1302-1307,共6页Chinese Journal of Pathophysiology

基　　金：浙江省自然科学基金资助项目(No.302781;No.206344)

摘　　要：目的:研究CD20scFv嵌合T淋巴细胞靶向杀伤Daudi细胞时杀伤的效果和T细胞活化情况。方法:将两种质粒转染至PA317细胞中,用转染成功的PA317上清液感染外周血T淋巴细胞后经800mg/L的G418筛选1周后去杀伤Daudi、K562细胞,分别在不同时点用流式细胞仪检测Daudi细胞AnnexinⅤ的阳性率,用ELISA检测细胞因子IL-2、IFNγ。结果:Daudi细胞AnnexinⅤ的阳性率在24h内两实验组与K562组相比明显增高,而实验组之间没有显著差异。72h时CD20scFv-IgGFc-CD28-ζ组比CD20scFv-IgGFc组IL-2(1509.00ng/L比220.54ng/L)和IFNγ(912.16ng/L比251.42ng/L)的分泌量有显著增高。结论:①两种CD20scFv特异的T细胞在引起Daudi细胞早期凋亡无显著差异,说明在CD20scFv的靶向杀伤过程中CD28-ζ基因可能不主导Daudi细胞的早期凋亡。②CD20scFv-IgGFc-CD28-ζ组IL-2、IFNγ增幅更加明显,说明CD3ζ和CD28嵌合的T细胞可以自身活化而不受MHC的限制,从而增强了T细胞的活化和杀伤等功能。AIM： To investigate the target killing effect of T lymphocytes with chimeric CD2OscFv gene on Daudi cells and the activation of T lymphocytes. METHODS： Two kinds of plasmids were transfected into retrovirus - packed PA317 cell lines. The supernatant was collected from successfully transfected PA317 culture and was used to infect peripheral blood T lymphocytes. After one - week screening with G418, the cells were used to kill Daudi and K562 cells. The positive rates of Annexin V in Daudi cells were measured at different times points respectively by flow cytometry. Meanwhile, the level of IL - 2 and IFN -γ were determined by ELISA. RESULTS ： The Annexin V positive rate was significant higher in Daudi cells compared to control K562 cell lines at 24 h. No difference of AnnexinV in Daudi cells was ob- served in CD20 modification T lymphocyte groups. The secretions of IL - 2 and IFN - γ in CD2OscFv - CD80 - lgGFc - CD28 -ζ gene modified T cells co - cultured with Daudi cells were dramatically higher than that in CD20scFv - IgGFc group at 72 h. CONCLUSION ： ①The two kinds of genetic modified specific T cells have no significant difference in inducing early apeptosis of Daudi cells. CD28 -ζ can＇t affect Daudi cell early apeptosis at the CD20scFv target killing. ② The increase in the secretions of IL - 2 and IFN-γ is more obvious in CD20scFv - IgGFc - CD28 - ζ group, indicating that the self - activation takes place in CD3ζ and CD28 modified T cells without MHC restriction and then increases the activation and killing function of T cells.

关 键 词：淋巴瘤 免疫疗法 抗体 CD20 

分 类 号：R733[医药卫生—肿瘤]