Nossal G J, Lewis H
J Exp Med. 1972 Jun 1;135(6):1416-22. doi: 10.1084/jem.135.6.1416.
Spleen cells from CBA mice that had been primarily or secondarily immunized with sheep red blood cells were reacted at 0 degrees C with a (125)I-labeled polyvalent rabbit anti-mouse globulin reagent. After suitable washing, the cells were placed in a plaque-revealing monolayer and warmed to 37 degrees C. Plaques appeared within 10-20 min. Single plaque-forming cells (PFC) were taken from the middle of plaques, were washed by micromanipulation, and were singly dried on glass slides. The amount of attached antireceptor was assessed by quantitative radioautography. Great variation in "receptor density" was encountered among the 258 single cells studied. However, early, immature PFC in both primary and secondary responses had statistically significantly more receptors than late, mature PFC. On any given day point, no difference was found between IgM- and IgG-forming cells. The results were consistent with the view that cells still able to be driven to further proliferation by antigen retain receptors, and conversely that cells, as they mature, lose both receptors and ability to be influenced by antigen.
用绵羊红细胞进行初次或二次免疫的CBA小鼠的脾细胞,在0℃下与(125)I标记的多价兔抗小鼠球蛋白试剂反应。适当洗涤后,将细胞置于斑块显影单层中并加热至37℃。10 - 20分钟内出现斑块。从斑块中间取出单个斑块形成细胞(PFC),通过显微操作洗涤,并单独干燥在载玻片上。通过定量放射自显影评估附着的抗受体量。在所研究的258个单细胞中,“受体密度”存在很大差异。然而,在初次和二次反应中,早期、未成熟的PFC在统计学上比晚期、成熟的PFC具有明显更多的受体。在任何给定的时间点,形成IgM和IgG的细胞之间没有差异。结果与以下观点一致:仍能被抗原驱动进一步增殖的细胞保留受体,相反,细胞在成熟时会失去受体以及受抗原影响的能力。