Licence Steve, Persson Christine, Mundt Cornelia, Mårtensson Inga-Lill
Developmental Immunology, The Babraham Institute, Cambridge, GB.
Eur J Immunol. 2003 Apr;33(4):1117-26. doi: 10.1002/eji.200323702.
In adult mice, the VpreB genes are expressed in bone marrow progenitor (pro-) and precursor (pre-) B cells. As part of the pre-B cell receptor, the proteins are crucial for the proliferation of these cells and consequently normal B lymphocyte development. Using cell lines, we identified a lineage- and developmental-stage-specific VpreB1 enhancer. Here, we analyze its specificity in vivo by generating transgenic mice in which expression of a reporter gene (human CD122) is regulated by the VpreB1 enhancer in the context of its own promoter. All transgenic lines expressed the reporter gene in the bone marrow in a copy number-independent manner, whereas expression levels were integration site-dependent. While the enhancer is not tissue specific, within the B cell lineage the expression pattern of human CD122 mimicked that of endogenous VpreB1. Thus, low levels were detected in pro-B cells, high levels in pre-BI and slightly lower levels in pre-BII cells; no expression was detected in immature/mature B cells. Furthermore, when in vitro cultured transgenic pre-B cells differentiated into immature B cells there was concomitant down-regulation of human CD122 and endogenous VpreB1. Thus the VpreB1 enhancer is sufficient to ensure developmental stage-specific expression of a reporter gene in B lymphocytes in vivo.
在成年小鼠中,VpreB基因在骨髓祖B细胞和前B细胞中表达。作为前B细胞受体的一部分,这些蛋白质对于这些细胞的增殖以及正常B淋巴细胞的发育至关重要。利用细胞系,我们鉴定出一种谱系和发育阶段特异性的VpreB1增强子。在此,我们通过构建转基因小鼠来分析其在体内的特异性,在这些转基因小鼠中,报告基因(人CD122)的表达在其自身启动子的背景下受VpreB1增强子调控。所有转基因系均以拷贝数无关的方式在骨髓中表达报告基因,而表达水平则依赖于整合位点。虽然该增强子不是组织特异性的,但在B细胞谱系中,人CD122的表达模式模仿了内源性VpreB1的表达模式。因此,在祖B细胞中检测到低水平表达,在前B I细胞中高水平表达,在前B II细胞中表达水平略低;在未成熟/成熟B细胞中未检测到表达。此外,当体外培养的转基因前B细胞分化为未成熟B细胞时,人CD122和内源性VpreB1会伴随下调。因此,VpreB1增强子足以确保报告基因在体内B淋巴细胞中进行发育阶段特异性表达。