Antibody production by single, hapten-specific B lymphocytes: an antigen-driven cloning system free of filler or accessory cells.
Antibody production by single, hapten-specific B lymphocytes: an antigen-driven cloning system free of filler or accessory cells.
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由单个半抗原特异性 B 淋巴细胞产生抗体：一种抗原驱动的克隆系统，不含填充细胞或辅助细胞。
DOI：
10.1073/pnas.78.12.7702
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发表时间：
1981
影响因子：
11.1
通讯作者：
Nossal,GJ
中科院分区：
文献类型：
--
作者：
Vaux,DL;Pike,BL;Nossal,GJ
CBA小鼠脾细胞在荧光素（FLU）-明胶薄层上进行半抗原亲和分级分离。该程序产生了97%具有不同FU结合亲和力的B细胞。将1至30个细胞置于10微升微量培养物中，不含任何填充细胞或辅助细胞。当单独使用时，偶联到FLU的T-非依赖性抗原聚合鞭毛蛋白（FLU-POL）在刺激这些细胞克隆增殖或抗体产生方面是无效的。富含白细胞介素的未纯化制剂也不能刺激细胞。当特异性抗原，而不是无关的半抗原-POL，与白细胞介素结合，刺激克隆增殖，大多数克隆产生抗FLU抗体形成细胞。抗体形成克隆的频率仅略低于使用抗原加填充细胞的系统中的频率。在没有添加白细胞介素的情况下，有丝分裂原大肠杆菌脂多糖加硫酸葡聚糖诱导等效的抗体产生。然而，克隆增殖的频率较高。添加白细胞介素并不能帮助这些促分裂原驱动的反应。这种抗原依赖性克隆系统，不含填充细胞和辅助细胞，应该允许更精确地分析抗原和白细胞介素在抗体形成克隆形成的生理学中的各自作用。
CBA mouse spleen cells were subjected to hapten affinity fractionation on thin layers of fluorescein (FLU)-gelatin. This procedure yields 97% B cells with varying FLU-binding avidities. One to 30 cells were placed in 10-microliter microcultures without any filler or accessory cells. the T-independent antigen polymerized flagellin coupled to FLU (FLU-POL) was ineffective in stimulating these cells to clonal proliferation or antibody production when used alone. Unpurified preparations rich in interleukins also failed to stimulate the cells. When specific antigen, but not in irrelevant hapten-POL, was combined with the interleukins, clonal proliferation was stimulated and most clones produced anti-FLU antibody-forming cells. The frequency of antibody-forming clones was only slightly lower than that in a system using antigen plus filler cells. In the absence of added interleukins, the mitogens Escherichia coli lipopolysaccharide plus dextran sulfate induced equivalent antibody production. However, a higher frequency of clonal proliferation was noted. Added interleukins did not aid these mitogen-driven responses. Such an antigen-dependent cloning system, free of filler and accessory cells, should permit more precise analysis of the respective roles of antigens and interleukins in the physiology of antibody-forming clone formation.