【作者】 谢敏；

【导师】 吴汉妮；

【作者基本信息】 华中科技大学 ， 内分泌科， 2006， 硕士

【摘要】 背景自身免疫性疾病的发病机理的研究是当前的一个热点,Graves病作为自身免疫性疾病的一种,其发病原因多与异常的免疫应答有关。CD80、CD86作为机体抗原递呈细胞表面的两个配体,与T淋巴细胞表面的CD28、CTLA-4结合后刺激T淋巴细胞的活化,造成机体自身免疫系统功能紊乱,导致自身免疫性疾病包括Graves病的发生。目的研究Graves病中CD80和CD86基因及蛋白在甲状腺滤泡细胞中的表达,探讨CD80和CD86在Graves病发生中的作用。方法实验一:取Graves病患者和甲状腺癌患者甲状腺切除的组织做体外原代细胞培养,将培养的一部分原代细胞用消化酶消化,然后收集,通过逆转录聚合酶链反应(RT-PCR)法检测手术切除的Graves病患者的甲状腺细胞及甲状腺癌的癌旁正常甲状腺组织细胞中CD80和CD86 mRNA的表达。实验二:Graves病患者和甲状腺癌患者甲状腺切除的组织做体外原代细胞培养,细胞培养期间加或不加刺激因子白介素4、10(IL-4、IL-10)观察体外培养的原代细胞抗原表达的情况,培养10-15天后标记结合FITC的兔抗人的CD80、CD86抗体,观察所培养的原代细胞CD80、CD86抗体的表达情况。结果RT-PCR检测显示,Graves病患者的甲状腺细胞有CD80和CD86 mRNA的表达。用Graves病患者的甲状腺组织培养的原代甲状腺细胞的胞膜显绿色荧光提示存在CD80和CD86的表达。Graves病患者的甲状腺细胞表达CD80和CD86的百分率与细胞培养中加IL-4、IL-10刺激因子对CD80和CD86的百分率两者比较无显著性差异(P >0.05);正常对照组甲状腺细胞CD80和CD86的表达呈弱阳性。结论Graves病患者的甲状腺细胞中存在CD80和CD86的异常表达,该表达不受刺激因子IL-4、IL-10的影响(P >0.05);CD80、CD86的异常表达可能是造成机体免疫功能紊乱从而导致Graves病的发病的重要免疫学因素之一。更多还原

【Abstract】 BackgroundRegarding the immunity disease pathogenesis research is a current hot spot,Graves disease as one of immunity diseases,its pathogenesis is associated with immune response disorder. CD80 and CD86 as two ligands of antigen presenting cell, interreaction with CD28 and CTLA-4, stimulate the T lymphocyte activation,resulting in immunologic system function disorder,causes the immunity disease occurrence.ObjectiveInvestigating the expression of CD80 and CD86 in Graves disease and discussion their roles in pathogenesis.MethodTests one:digestion the culturing thyroid gland primary cell in vitro with digestive enzyme,then collection,through RT-polymerase chain reaction method (RT-PCR) to examine the CD80 and CD86 expression of the mRNA in the surgery collections from Graves disease patients thyroid gland cells and Nearby cancer thyroid gland cells.Test two:Takeing the tissue from the Graves sickness and the thyroid cancer patient who made subtotal thyroidectomy or total thyroidectomy to make in vitro original cell culture,during the cell culture period,adding the stimulating factor interleukin 4 and 10 (IL-4,IL-10) observes the stimulating factors influence to antibody express of primary cell in vitro,after culture for 10-15 day to combine marking FITC rabbit anti- person’s CD80,the CD86 antibody and observation the CD80 and CD86 antibody expression of primary cell in vitro.ResultThe result of RT-PCR examination shows that the thyroid gland cell of Graves disease has the mRNA expression of CD80 and CD86.Representation the green fluorescence in culturing primary cell of thyroid gland tissue of the Graves disease showed that the antibody CD80,CD86 expressed in the thyroid gland cells of Graves disease;in addition there was no significance difference between the thyroid gland cells of Graves disease which expressed CD80,CD86 and the thyroid gland cells of Graves disease adding IL-4,IL-10 stimulating factors which expressed CD80 , CD86(P>0.05) ; after subtotal thyroidectomy or total thyroidectomy,the remaining Thyroid gland tissue hardly expressed antibody CD80,CD86 or weakly positive expression.ConclusionCD80 and CD86 in Graves disease was expressed,and the expression was not effect with the stimulating factors IL-4 and IL-10.This phenomenon shows that the abnormal expression of CD80 and CD86 of antigen presenting cell may cause the organism immune function disorder resulting in the invasion of Graves disease.更多还原