Xiang Yougui, Garrard William T
Department of Molecular Biology University of Texas, Southwestern Medical Center, Dallas, TX 75390, USA.
J Immunol. 2008 May 15;180(10):6725-32. doi: 10.4049/jimmunol.180.10.6725.
The mouse Igkappa locus has three known transcriptional enhancers: the matrix association region/intronic enhancer, the 3' enhancer (E3'), and the further downstream enhancer (Ed). Previous studies have shown that both matrix association region/intronic and E3' enhancers are required for maximal gene rearrangement of the locus, and that E3' is also required for maximal expression and somatic hypermutation (SHM). To functionally elucidate Ed in vivo, we generated knockout mice with a targeted germline deletion of Ed. Ed deleted homozygous mice (Ed-/-) have moderately reduced numbers of Igkappa expressing B cells and correspondingly increased numbers of Iglambda expressing B cells in spleen. Ed-/- mice also have decreased Igkappa mRNA expression in resting and T cell-dependent activated splenic B cells and reduced Igkappa chains in sera. However, our analysis indicates that Igkappa gene rearrangement is normal in Ed-/- mice. In addition, our results show that Ed-/- mice exhibit reduced SHM in the Igkappa gene J-C intronic region in germinal center B cells from Peyer's patches. We conclude that Ed positively regulates Igkappa gene expression and SHM, but not gene rearrangement.
小鼠免疫球蛋白κ轻链基因座有三个已知的转录增强子:基质附着区域/内含子增强子、3'增强子(E3')和更下游的增强子(Ed)。先前的研究表明,基质附着区域/内含子增强子和E3'增强子对于该基因座的最大基因重排都是必需的,并且E3'对于最大表达和体细胞超突变(SHM)也是必需的。为了在体内从功能上阐明Ed,我们构建了靶向种系缺失Ed的基因敲除小鼠。Ed缺失的纯合小鼠(Ed-/-)脾脏中表达免疫球蛋白κ轻链的B细胞数量适度减少,相应地表达免疫球蛋白λ轻链的B细胞数量增加。Ed-/-小鼠在静息和T细胞依赖性活化的脾脏B细胞中免疫球蛋白κ轻链mRNA表达也降低,血清中免疫球蛋白κ轻链减少。然而,我们的分析表明,Ed-/-小鼠中免疫球蛋白κ轻链基因重排是正常的。此外,我们的结果表明,Ed-/-小鼠派尔集合淋巴结生发中心B细胞免疫球蛋白κ轻链基因J-C内含子区域的体细胞超突变减少。我们得出结论,Ed正向调节免疫球蛋白κ轻链基因表达和体细胞超突变,但不调节基因重排。