Abney E R, Keeler K D, Parkhouse R M, Willcox H N
Eur J Immunol. 1976 Jun;6(6):443-50. doi: 10.1002/eji.1830060612.
The memory cells of two antibody-forming cell clones had receptors of the IgM class, even though the clones had been producing IgG1 or IgG2a anti-2,4-dinitrophenyl antibodies for 9-15 months previously (on exposure to antigen). Thus a phenotypic switch in heavy chain constant region evidently occurred after re-exposure of these memory cells to antigen. To show that, we first removed the clonal cells' surface immunoglobins by "capping" and "stripping", with class- or subclass-specific antisera. Then, to assay their remaining receptor activity, the cells were incubated with antigen in vitro, washed and transferred (together with carrier primed cells) to irradiated recipients, and their antibody responses to this in vitro boost were assayed by iselectric focusing. Pretreatment with anti-mu serum, as well as with anti-Fab(kappa), prevented the responses of the IgG1 and IgG2a clones to an in vitro boost, while anti-gamma1 and anti-gamma2a antisera had no effect. An antiserum to the putative mouse IgD also had no effect. The anti-mu serum failed to react with the IgG1 and IgG2A clonal serum antibodies in the test tube. Some other contaminating clones were suppressed completely only by the anti-Fab serum. This result strongly suggests that switching in class commitment may occur during the differentiation of memory cells to antibody producers, and may therefore be antigen-dependent. It also implies that some apparently naive cells with surface IgM may, in reality, be B memory cells.
两个抗体形成细胞克隆的记忆细胞具有IgM类受体,尽管这些克隆在之前9 - 15个月(暴露于抗原后)一直在产生IgG1或IgG2a抗2,4 - 二硝基苯基抗体。因此,这些记忆细胞再次暴露于抗原后,重链恒定区明显发生了表型转换。为了证明这一点,我们首先用类或亚类特异性抗血清通过“封帽”和“剥离”去除克隆细胞的表面免疫球蛋白。然后,为了检测它们剩余的受体活性,将细胞在体外与抗原一起孵育,洗涤后转移(连同载体致敏细胞)到经照射的受体中,并通过等电聚焦检测它们对这种体外刺激的抗体反应。用抗μ血清以及抗Fab(κ)预处理可阻止IgG1和IgG2a克隆对体外刺激的反应,而抗γ1和抗γ2a抗血清则没有效果。针对假定的小鼠IgD的抗血清也没有效果。抗μ血清在试管中未能与IgG1和IgG2A克隆血清抗体发生反应。一些其他污染克隆仅被抗Fab血清完全抑制。这一结果有力地表明,类别转换可能发生在记忆细胞分化为抗体产生细胞的过程中,因此可能是抗原依赖性的。这也意味着一些表面带有IgM的明显幼稚细胞实际上可能是B记忆细胞。