本文關(guān)鍵詞：超抗原SEA對(duì)PML-RARα融合多肽誘導(dǎo)TCR Vβ亞家族T細(xì)胞克隆性的影響　出處：《暨南大學(xué)》2007年碩士論文　論文類型：學(xué)位論文

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

【摘要】： 目的：了解PML-RARα、SEA及聯(lián)合應(yīng)用誘導(dǎo)人外周血T淋巴細(xì)胞后，TCR Vβ亞家族的特異性表達(dá)及殺傷性情況，以期尋找能有效增強(qiáng)PML-RARα免疫原性的方法，為APL免疫治療和開(kāi)發(fā)特異性高效價(jià)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(dú)及聯(lián)合誘導(dǎo)人外周血T細(xì)胞增殖，并設(shè)IL-2對(duì)照組和ConA對(duì)照組。利用RT-PCR-基因掃描技術(shù)分析誘導(dǎo)20天后的外周血TCR Vβ亞家族的優(yōu)勢(shì)利用和克隆性增殖的特點(diǎn)；同時(shí)用CCK-8試劑盒檢測(cè)對(duì)白血病細(xì)胞株NB4和K562的細(xì)胞毒性；利用流式細(xì)胞術(shù)檢測(cè)細(xì)胞表面CD3~+細(xì)胞中CD4和CD8分子的表達(dá)情況；并利用ELISA方法檢測(cè)培養(yǎng)液上清中IFN-γ、IL-4、IL-10細(xì)胞因子的分泌水平。 結(jié)果：限制性表達(dá)和克隆性增殖的TCR Vβ亞家族T細(xì)胞均可見(jiàn)于PML-RARα多肽單獨(dú)及聯(lián)合SEA誘導(dǎo)后的外周血T細(xì)胞；而經(jīng)SEA單獨(dú)誘導(dǎo)后的外周血T細(xì)胞僅表現(xiàn)出了對(duì)TCR Vβ亞家族T細(xì)胞的優(yōu)勢(shì)利用。經(jīng)CCK-8檢測(cè)細(xì)胞殺傷活性，顯示誘導(dǎo)后的T細(xì)胞均具有較強(qiáng)的細(xì)胞毒作用，且PML-RARα多肽聯(lián)合SEA誘導(dǎo)的T細(xì)胞比二者單獨(dú)誘導(dǎo)的T細(xì)胞表現(xiàn)出更強(qiáng)烈的細(xì)胞毒作用。流式細(xì)胞術(shù)結(jié)果顯示經(jīng)PML-RARα多肽或聯(lián)合SEA誘導(dǎo)后T細(xì)胞(CD3~+細(xì)胞)，以CD8~+ CTL增殖為主。ELISA結(jié)果顯示，經(jīng)PML-RARα多肽單獨(dú)及聯(lián)合SEA誘導(dǎo)后，上清中Th1型細(xì)胞因子IFN-γ水平明顯升高；Th2型細(xì)胞因子(IL-4，IL-10)水平低下。 結(jié)論：PML-RARα多肽在體外聯(lián)合SEA誘導(dǎo)外周血T細(xì)胞增殖以CD8~+ CTL為主，呈克隆性增殖，，此優(yōu)勢(shì)增殖的克隆性T細(xì)胞具有特異性細(xì)胞毒作用，該細(xì)胞毒作用強(qiáng)于單獨(dú)應(yīng)用PML-RARα多肽或SEA誘導(dǎo)后的T細(xì)胞，故SEA可以協(xié)同PML-RARα多肽增強(qiáng)其對(duì)T細(xì)胞的特異性誘導(dǎo)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(xué)位授予單位】：暨南大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2007
【分類號(hào)】：R392

【參考文獻(xiàn)】

相關(guān)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(duì)白血病細(xì)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(yáng)秋,陳少華,周羽z

本文編號(hào)：1388857