SEA对PML-RARα多肽体外诱导外周血单个核细胞TCRζ链表达的作用  

Effect of SEA on the TCRζ chain expression of T cells stimulated by PML-RARα peptide in vitro

作　　者：高珂[1] 林晨[1] 白雪[1] 陈少华[2] 杨力建[2] 陈思[2] B.N.Selvakumar 李扬秋[2] 

机构地区：[1]暨南大学医学院微生物学与免疫学教研室,广东广州510632 [2]暨南大学医学院血液病研究所,广东广州510632

出　　处：《暨南大学学报（自然科学与医学版）》2008年第4期347-350,共4页Journal of Jinan University(Natural Science & Medicine Edition)

基　　金：广东省科技计划项目(2005B50301016);广东省自然科学基金项目(06025169);广州市科技计划项目(2005Z1-E4015)

摘　　要：目的:了解金黄色葡萄球菌肠毒素A(SEA)联合PML-RARα融合多肽体外诱导正常人外周血T细胞活化TCRζ链基因表达情况。方法:利用T细胞液体培养法分别与正常人外周血淋巴细胞加入PML-RARα融合多肽、SEA和SEA联合PML-RARα多肽诱导培养T细胞,其中SEA刺激包括培养初始或培养第5天加入SEA两组(PS、PSI),并设空白对照组(不加多肽及SEA)。分别收集各组培养20 d后细胞提取mRNA并合成cDNA,采用SYBR G reenⅠ荧光定量PCR和相对定量检测TCRζ链在不同组别T淋巴细胞中的表达情况,以β2微球蛋白基因(β2M)作为内参,根据相对定量公式:2-△△C t分析TCRζ链表达差异。结果:与空白组相比,联合诱导组在培养初始加入SEA及第5天加入SEA的培养T细胞中TCRζ链表达上升,而单独SEA诱导组的TCRζ链表达下降。结论:超抗原SEA联合PML-RARα多肽体外诱导T细胞可使TCRζ链基因表达水平升高,有望为研制急性早幼粒细胞白血病疫苗提供新的切入点。Aim：To measure the expression of signaling transduction factor T lymphocyte-TCRζ chain gene in peripheral blood mononuclear cells which stimulated by PML-RARα fusion peptide or SEA or both of them. To analyze the effect of staphylococcal enterotoxin A（SEA） to strengthen the immunogenicity of PML-RARα to human peripheral T lymphocyte. Methods： Real-Time PCR with SYBR Green Ⅰ technique was used for detecting TCRζ chain expression level in peripheral blood mononuclear ceUs from normal individuals which were induced by PML-RARα peptide for 20 days, respectively add the SEA at the beginning or the fifth day,and a comparison of SEA was set. β2-microglobulin gene （β2M） was used as an endogenous reference. Relative changes in TCRζ chain expression level were used by the 2 ^-△△Ct method between each group and the control. Results： The expression level of TCRζ chain in groups which were added the SEA at the beginning and the fifth day were over expressed, the other groups were decreased. Conclusion： The expression of signaling transduction factor T lymphocyte-TCRζ chain gene in peripheral blood mononuclear ceils, which stimulated by SEA and PML-RARα fusion peptide, could be over expressed. This study could be provided a new insight into the research on vaccine for Acute promyelocytic leukemia.

关 键 词：T细胞受体ζ链 实时定量PCR 金黄色葡萄球菌肠毒素A T细胞受体 PML-RARα融合多肽 

分 类 号：R371.22[医药卫生—病原生物学] R733.74[医药卫生—基础医学]