Zvezdova E S, Silaeva Iu Iu, Vagida M S, Mariukhnich E V, Deĭkin A V, Ermolkevich T G, Kadulin S G, Sadchikova E R, Gol'dman I L, Kazanskiĭ D B
Mol Biol (Mosk). 2010 Mar-Apr;44(2):311-22.
Transgenic animal studies has become a key approach for gene function analysis as well as for modeling of different human diseases, including autoimmune diseases caused by activation of T-lymphocyte clones whose TCRs possesses high affinity for syngeneic MHC molecules. In this study we cloned genes, encoding alpha- and beta- chains of autoreactive TCR of hybridoma 7, specific for syngeneic MHC class II molecules A(b). Amplified DNA fragments, containing rearranged genomic DNA of alpha- and beta-chains of hybridoma 7 were cloned into special cassette vectors, containing natural promoter and enhancer elements for direct expression of genes encoding TCR alpha- and beta-chains in T-lymphocytes of transgenic animals. Using this cassette vectors we generated animals in which most of peripheral T-lymphocytes carry alpha-chain, as well as animals with expression of beta-chain transgene of autoreactive TCR. Obtained animals may serve to explain a number of intrathymic selection processing features and T cell maturation as well as to serve as experimental models for development of new approaches to therapy of autoimmune diseases.
转基因动物研究已成为基因功能分析以及不同人类疾病建模的关键方法，包括由T淋巴细胞克隆激活引起的自身免疫性疾病，这些T淋巴细胞的TCR对同基因MHC分子具有高亲和力。在本研究中，我们克隆了编码杂交瘤7自身反应性TCR的α链和β链的基因，该杂交瘤对同基因II类MHC分子A(b)具有特异性。将包含杂交瘤7的α链和β链重排基因组DNA的扩增DNA片段克隆到特殊的盒式载体中，该载体含有天然启动子和增强子元件，用于在转基因动物的T淋巴细胞中直接表达编码TCRα链和β链的基因。使用这种盒式载体，我们培育出了大多数外周T淋巴细胞携带α链的动物，以及表达自身反应性TCRβ链转基因的动物。获得的动物可用于解释胸腺内选择过程的一些特征和T细胞成熟，也可作为开发自身免疫性疾病新治疗方法的实验模型。
