Wiener E, Wickramasinghe S N
Department of Haematology, St Mary's Hospital Medical School, London, UK.
Int J Exp Pathol. 1990 Dec;71(6):871-7.
Dimethyl-sulphoxide-induced Friend leukaemia erythroblasts (IFLE) which had been damaged by treatment with inhibitors of protein synthesis (cycloheximide, puromycin) were incubated with normal mouse serum or with doubling dilutions of it. The erythroblasts were subsequently tested for their binding of natural antibodies of all the major immunoglobulin (Ig) isotypes and both light-chain types, using Fc- and light-chain-specific FITC-immunoconjugates, and flow cytometry. After short-term (4-h) exposure of IFLE to puromycin, some binding of IgG but not of IgM or IgA could be demonstrated. By contrast, prolonged (17-h) exposure of IFLE to cycloheximide or puromycin resulted in their reaction with antibodies of all major isotypes (IgM, IgG1, IgG2a, IgG2b, IgG3, IgA) containing kappa-light chains. Under these conditions of damage the percentage of IgM- or IgG-binding IFLE significantly exceeded that of IgA-reactive cells. Moreover, the percentage of damaged IFLE which bound IgG2a or IgG3 was greater than that of cells which bound IgG1 or IgG2b. The percentage of damaged IFLE which bound a certain Ig-isotype did not correlate with the concentration of that Ig-isotype in normal mouse serum. The results suggest that natural antibodies of all major isotypes containing kappa-light chains bind specifically to severely damaged IFLE and could facilitate their interaction with macrophages in a concerted manner.
用蛋白质合成抑制剂(环己酰亚胺、嘌呤霉素)处理后受损的二甲亚砜诱导的弗瑞德白血病成红细胞(IFLE),与正常小鼠血清或其倍比稀释液一起孵育。随后,使用Fc和轻链特异性异硫氰酸荧光素免疫缀合物及流式细胞术,检测成红细胞对所有主要免疫球蛋白(Ig)同种型和两种轻链类型天然抗体的结合情况。在IFLE短期(4小时)暴露于嘌呤霉素后,可证明其与IgG有一定结合,但与IgM或IgA无结合。相比之下,IFLE长期(17小时)暴露于环己酰亚胺或嘌呤霉素会导致它们与所有含有κ轻链的主要同种型抗体(IgM、IgG1、IgG2a、IgG2b、IgG3、IgA)发生反应。在这些损伤条件下,结合IgM或IgG的IFLE百分比显著超过结合IgA的反应性细胞百分比。此外,结合IgG2a或IgG3的受损IFLE百分比高于结合IgG1或IgG2b的细胞百分比。结合某一Ig同种型的受损IFLE百分比与正常小鼠血清中该Ig同种型的浓度无关。结果表明,所有含有κ轻链的主要同种型天然抗体都能特异性结合严重受损的IFLE,并能协同促进它们与巨噬细胞的相互作用。