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同种抗原激活的T淋巴母细胞表面独特型的部分特性分析

Partial characterization of cell surface idiotypes on alloantigen-activated T lymphoblasts.

作者信息

Binz H, Frischkencht H, Mercolli C, Wigzell H

出版信息

Scand J Immunol. 1978;7(6):481-5. doi: 10.1111/j.1365-3083.1978.tb00481.x.

Abstract

T lymphoblasts of specificity Lewis anti-DA, Lewis anti-BN, BN anti-DA and L.BN anti-DA were purified on Ficoll-Paque from mixed leucocyte cultures on day 5. Blasts were radiolabelled with iodine-125 by the lactoperoxidase method and lysed by the aid of Nonidet P40. Idiotypic molecules were puried from the lysates by the use of anti-idiotypic antiserum of specificity anti-(Lewis anti-DA) and Staphylococcus aureus bearing protein-A. In this way, molecules with a molecular weight of 150,000 and 75,000 daltons could be isolated from Lewis anti-DA and L.BN anti-DA T lymphoblasts but no significant radioactivity was brought down by the same procedure from Lewis Lewis anti-BN or BN anti-DA T lymphoblasts. No additional molecules were detected on the lower molecular weight regions where light chains of Ig type as well as conventional products of the MHC genes would appear. The 150,000 dalton molecules are composed of two single polypeptide chains with a molecular weight of around 75,000 daltons. These data are in complete agreement with earlier results on antigen-binding, idiotypic receptors obtained from normal rat T lymphocytes.

摘要

在第5天,从混合白细胞培养物中通过Ficoll-Paque纯化出具有Lewis抗DA、Lewis抗BN、BN抗DA和L.BN抗DA特异性的T淋巴母细胞。通过乳过氧化物酶法用碘-125对母细胞进行放射性标记,并借助Nonidet P40进行裂解。通过使用特异性为抗-(Lewis抗DA)的抗独特型抗血清和带有蛋白A的金黄色葡萄球菌,从裂解物中纯化独特型分子。通过这种方式,可从Lewis抗DA和L.BN抗DA T淋巴母细胞中分离出分子量为150,000和75,000道尔顿的分子,但通过相同程序从Lewis Lewis抗BN或BN抗DA T淋巴母细胞中未获得明显的放射性。在免疫球蛋白类型轻链以及MHC基因常规产物可能出现的较低分子量区域未检测到其他分子。150,000道尔顿的分子由两条分子量约为75,000道尔顿的单条多肽链组成。这些数据与早期从正常大鼠T淋巴细胞获得的关于抗原结合、独特型受体的结果完全一致。

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