Eisen A F, Atkinson M J, Celler J W, Paige C J, Wu G E
Department of Immunology, University of Toronto, Ontario, Canada.
Int Immunol. 1991 May;3(5):477-84. doi: 10.1093/intimm/3.5.477.
The VDJ recombination potential of a number of Abelson murine leukemic virus transformed fetal liver cell lines derived from (C57BL/6 x BALB/c) F1 mice was measured. The specific developmental stage of each line was determined using Southern blot analysis to ascertain their rearrangement status at the immunoglobulin heavy chain locus (DJ/DJ, VDJ/DJ or VDJ/VDJ). It was observed that DNA from DJ/DJ lines gave many more 'subhaploid' bands hybridizing with JH than did DNA from VDJ/VDJ lines. While the lack of appropriate substrate (VDJ/VDJ lines have exhausted the normal IgH substrate) contributes to the decrease in 'subhaploid bands', this result indicates that the rate of ongoing immunoglobulin heavy chain gene rearrangement was higher in the DJ/DJ lines. However, when the lines were examined using the assay developed by Hesse et al. (4) to measure VDJ recombinase activity, it was found that although all lines had recombinase activity, the DJ/DJ lines had four times more VDJ recombinase activity than did the VDJ/VDJ lines.
对源自(C57BL/6×BALB/c)F1小鼠的多种阿贝尔逊鼠白血病病毒转化的胎肝细胞系的VDJ重组潜能进行了测定。使用Southern印迹分析确定每个细胞系的特定发育阶段,以确定它们在免疫球蛋白重链基因座处的重排状态(DJ/DJ、VDJ/DJ或VDJ/VDJ)。观察到,与VDJ/VDJ细胞系的DNA相比,DJ/DJ细胞系的DNA与JH杂交产生的“亚单倍体”条带更多。虽然缺乏合适的底物(VDJ/VDJ细胞系已耗尽正常的IgH底物)导致“亚单倍体条带”减少,但这一结果表明DJ/DJ细胞系中正在进行的免疫球蛋白重链基因重排速率更高。然而,当使用Hesse等人(4)开发的测定方法检测这些细胞系以测量VDJ重组酶活性时,发现虽然所有细胞系都有重组酶活性,但DJ/DJ细胞系的VDJ重组酶活性是VDJ/VDJ细胞系的四倍。