本文關鍵詞：超抗原SEA對PML-RARα融合多肽誘導TCR Vβ亞家族T細胞克隆性的影響　出處：《暨南大學》2007年碩士論文　論文類型：學位論文

  更多相關文章： PML-RARα 金黃色葡萄球菌腸毒素A(SEA) T細胞受體(TCR) 急性早幼粒細胞白血病(APL)

【摘要】： 目的：了解PML-RARα、SEA及聯(lián)合應用誘導人外周血T淋巴細胞后，TCR Vβ亞家族的特異性表達及殺傷性情況，以期尋找能有效增強PML-RARα免疫原性的方法，為APL免疫治療和開發(fā)特異性高效價的抗APL基因疫苗提供可行性資料和前期性研究數(shù)據(jù)。 方法：分別以SEA和PML-RARα多肽單獨及聯(lián)合誘導人外周血T細胞增殖，并設IL-2對照組和ConA對照組。利用RT-PCR-基因掃描技術分析誘導20天后的外周血TCR Vβ亞家族的優(yōu)勢利用和克隆性增殖的特點；同時用CCK-8試劑盒檢測對白血病細胞株NB4和K562的細胞毒性；利用流式細胞術檢測細胞表面CD3~+細胞中CD4和CD8分子的表達情況；并利用ELISA方法檢測培養(yǎng)液上清中IFN-γ、IL-4、IL-10細胞因子的分泌水平。 結果：限制性表達和克隆性增殖的TCR Vβ亞家族T細胞均可見于PML-RARα多肽單獨及聯(lián)合SEA誘導后的外周血T細胞；而經SEA單獨誘導后的外周血T細胞僅表現(xiàn)出了對TCR Vβ亞家族T細胞的優(yōu)勢利用。經CCK-8檢測細胞殺傷活性，顯示誘導后的T細胞均具有較強的細胞毒作用，且PML-RARα多肽聯(lián)合SEA誘導的T細胞比二者單獨誘導的T細胞表現(xiàn)出更強烈的細胞毒作用。流式細胞術結果顯示經PML-RARα多肽或聯(lián)合SEA誘導后T細胞(CD3~+細胞)，以CD8~+ CTL增殖為主。ELISA結果顯示，經PML-RARα多肽單獨及聯(lián)合SEA誘導后，上清中Th1型細胞因子IFN-γ水平明顯升高；Th2型細胞因子(IL-4，IL-10)水平低下。 結論：PML-RARα多肽在體外聯(lián)合SEA誘導外周血T細胞增殖以CD8~+ CTL為主，呈克隆性增殖，，此優(yōu)勢增殖的克隆性T細胞具有特異性細胞毒作用，該細胞毒作用強于單獨應用PML-RARα多肽或SEA誘導后的T細胞，故SEA可以協(xié)同PML-RARα多肽增強其對T細胞的特異性誘導作用。
[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偽, SEA and combined application , in order to find a way to effectively enhance the immunogenicity of PML - RAR偽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偽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 - 緯 , 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偽alone and in combination with SEA . Conclusion : The proliferation of peripheral blood T cells induced by PML - RAR偽in vitro is mainly CD8 ~ + CTL and clonal proliferation . The clonal T cells have specific cytotoxicity against PML - RAR偽polypeptide or SEA - induced T cell , so SEA can enhance its specific induction effect on T cells in conjunction with PML - RAR偽polypeptide .

【學位授予單位】：暨南大學
【學位級別】：碩士
【學位授予年份】：2007
【分類號】：R392

【參考文獻】

相關期刊論文 前10條

1 賀鵬程;張梅;吳迪;徐匯;蔡瑞波;劉亞琳;;IFNγ聯(lián)合全反式維甲酸對白血病細胞株NB4、MR2增殖和分化的影響[J];癌癥;2006年12期

2 楊力建,李揚秋,陳少華,周羽z

本文編號：1388857