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人类免疫缺陷病毒(HIV)感染那些细胞表面表达 CD4分子的 T 辅助细胞,CD4是 HIV 的细胞受体,CD4分子介导细胞表面和病毒衣壳糖蛋白 gp120之间的相互作用。本文作者用重组 DNA 方法构建了两个编码 CD4分子的嵌合基因,其中 HT4-Y1编码 CD4的整个细胞外区域,HT4-X6编码免疫球蛋白样和第二 CD4区域,两个嵌合蛋白的 C 末端均由免疫球蛋白 K 轻链稳定区组成。将这两个嵌合基因导入骨髓瘤细胞以分泌相应的两个嵌合蛋白质,用Western 印迹法测定的分子量分别为60KDa(HT4-Y1)和30KDa(HT4-X6),用抗 CD4抗体进一步对上清进行免疫沉淀确定:这些分泌的蛋白质均保持原有构象。可溶性 CD4蛋白质能与纯化的 HIV 衣壳蛋白 gp120结合,有趣的是:小鼠 CD4类似物 L3T4即使用与 CD4相同的稀释法,配成较 CD4高100倍的浓度,
Human immunodeficiency virus (HIV) infects T-helper cells that express CD4 molecules on the cell surface. CD4 is the cellular receptor for HIV and CD4 molecules mediate the interaction between the cell surface and the gp120 viral capsid protein. The authors constructed two chimeric genes encoding CD4 molecules using recombinant DNA methods, in which HT4-Y1 encodes the entire extracellular region of CD4, HT4-X6 encodes immunoglobulin-like and second CD4 regions, and two chimeric proteins The C-terminus consists of immunoglobulin K light chain stabilizing regions. The two chimeric genes were introduced into myeloma cells to secrete the corresponding two chimeric proteins with molecular weights of 60 kDa (HT4-Y1) and 30 kDa (HT4-X6), respectively, as determined by Western blotting and further with anti-CD4 antibodies Supernatants were subjected to immunoprecipitation to confirm that the secreted proteins retained their original conformation. The soluble CD4 protein binds to the purified HIV capsid protein gp120. Interestingly, the mouse CD4 analogue L3T4 is formulated at a 100-fold higher concentration than CD4 using the same dilution method as CD4,