超抗原SEA对PML-RARα融合多肽诱导TCR Vβ亚家族T细胞克隆性的影响

发布时间：2018-01-06 17:24

本文关键词：超抗原SEA对PML-RARα融合多肽诱导TCR Vβ亚家族T细胞克隆性的影响　出处：《暨南大学》2007年硕士论文　论文类型：学位论文

【摘要】： 目的：了解PML-RARα、SEA及联合应用诱导人外周血T淋巴细胞后，TCR Vβ亚家族的特异性表达及杀伤性情况，以期寻找能有效增强PML-RARα免疫原性的方法，为APL免疫治疗和开发特异性高效价的抗APL基因疫苗提供可行性资料和前期性研究数据。方法：分别以SEA和PML-RARα多肽单独及联合诱导人外周血T细胞增殖，并设IL-2对照组和ConA对照组。利用RT-PCR-基因扫描技术分析诱导20天后的外周血TCR Vβ亚家族的优势利用和克隆性增殖的特点；同时用CCK-8试剂盒检测对白血病细胞株NB4和K562的细胞毒性；利用流式细胞术检测细胞表面CD3~+细胞中CD4和CD8分子的表达情况；并利用ELISA方法检测培养液上清中IFN-γ、IL-4、IL-10细胞因子的分泌水平。结果：限制性表达和克隆性增殖的TCR Vβ亚家族T细胞均可见于PML-RARα多肽单独及联合SEA诱导后的外周血T细胞；而经SEA单独诱导后的外周血T细胞仅表现出了对TCR Vβ亚家族T细胞的优势利用。经CCK-8检测细胞杀伤活性，显示诱导后的T细胞均具有较强的细胞毒作用，且PML-RARα多肽联合SEA诱导的T细胞比二者单独诱导的T细胞表现出更强烈的细胞毒作用。流式细胞术结果显示经PML-RARα多肽或联合SEA诱导后T细胞(CD3~+细胞)，以CD8~+ CTL增殖为主。ELISA结果显示，经PML-RARα多肽单独及联合SEA诱导后，上清中Th1型细胞因子IFN-γ水平明显升高；Th2型细胞因子(IL-4，IL-10)水平低下。结论：PML-RARα多肽在体外联合SEA诱导外周血T细胞增殖以CD8~+ CTL为主，呈克隆性增殖，，此优势增殖的克隆性T细胞具有特异性细胞毒作用，该细胞毒作用强于单独应用PML-RARα多肽或SEA诱导后的T细胞，故SEA可以协同PML-RARα多肽增强其对T细胞的特异性诱导作用。
[Abstract]:Objective : To investigate the specific expression and anti - traumatic condition of TCR V尾 subfamily after human peripheral T lymphocytes induced by PML - RAR伪, SEA and combined application , in order to find a way to effectively enhance the immunogenicity of PML - RAR伪and provide feasibility and preliminary research data for APL immunotherapy and development of specific high - titer anti - APL gene vaccine . Methods : The proliferation of peripheral blood T cells was induced by SEA and PML - RAR伪respectively and IL - 2 control group and Con A control group were established . The cytotoxicity of TCR V尾 subfamily in peripheral blood was analyzed by RT - PCR - gene scanning technique . The expression of CD4 and CD8 was detected by flow cytometry . The levels of IFN - 纬 , IL - 4 and IL - 10 cytokines in the supernatant of the culture medium were detected by flow cytometry . Results : TCR V尾 subfamily T cells in both restriction expression and clonal proliferation were found in peripheral blood T cells induced by PML - RAR伪alone and in combination with SEA . Conclusion : The proliferation of peripheral blood T cells induced by PML - RAR伪in vitro is mainly CD8 ~ + CTL and clonal proliferation . The clonal T cells have specific cytotoxicity against PML - RAR伪polypeptide or SEA - induced T cell , so SEA can enhance its specific induction effect on T cells in conjunction with PML - RAR伪polypeptide .

【学位授予单位】：暨南大学
【学位级别】：硕士
【学位授予年份】：2007
【分类号】：R392

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