Francés Rubén, Tumang Joseph R, Kaku Hiroaki, Gurdak Sean M, Rothstein Thomas L
Department of Medicine, Boston University School of Medicine, Boston, MA 02118, USA.
Mol Immunol. 2006 May;43(13):2124-9. doi: 10.1016/j.molimm.2005.12.011. Epub 2006 Feb 17.
B-1 cells constitute a unique B cell subset that differs phenotypically, biochemically, and functionally from the predominant population of conventional B-2 cells. Functional differences include constitutive secretion of natural immunoglobulin and failure of BCR signaling to initiate proliferation. The origin of these differences remains uncertain. We hypothesized that unbiased analysis of differences in protein expression between highly pure populations of B-1 and B-2 cells might provide information not readily available through other means. To pursue this, we undertook 2D gel analysis of B-1 and B-2 cells combined with mass spectrometry. We identified the smooth muscle protein, transgelin 2, in peritoneal (but not splenic) B-1 cells and did not find it in splenic B-2 cells; these results were confirmed by Western blot analysis, which showed a more than 60-fold difference in transgelin 2 expression between peritoneal B-1 and splenic B-2 cells. In contrast, levels of transgelin 2 RNA differed to a much lesser extent (3-fold) in the two B cell populations, so transgelin 2 is an example of a molecule whose subset-specific expression is more readily detected by proteomic than transcriptomic analyses. Finally, transgelin 2 protein expression was induced in splenic B-2 cells; thus, transgelin 2 joins a number of other inducible molecules that are constitutively expressed by peritoneal B-1 but not splenic B-2 cells. Although the role of transgelin 2 in B-1 cell function remains uncertain, identification of this molecule demonstrates the value of examining protein expression in this B cell subset.
B-1细胞构成了一个独特的B细胞亚群,在表型、生化和功能上与主要的传统B-2细胞群体不同。功能差异包括天然免疫球蛋白的组成性分泌以及BCR信号传导无法启动增殖。这些差异的起源仍不确定。我们假设,对高度纯化的B-1和B-2细胞群体之间蛋白质表达差异进行无偏分析,可能会提供通过其他方式不易获得的信息。为了实现这一点,我们对B-1和B-2细胞进行了二维凝胶分析并结合质谱分析。我们在腹膜(而非脾脏)B-1细胞中鉴定出平滑肌蛋白转胶蛋白2,而在脾脏B-2细胞中未发现;蛋白质印迹分析证实了这些结果,该分析显示腹膜B-1细胞和脾脏B-2细胞之间转胶蛋白2的表达差异超过60倍。相比之下,两种B细胞群体中转胶蛋白2 RNA的水平差异要小得多(3倍),因此转胶蛋白2是一个例子,说明通过蛋白质组学比转录组学分析更容易检测到其亚群特异性表达的分子。最后,转胶蛋白2蛋白表达在脾脏B-2细胞中被诱导;因此,转胶蛋白2加入了许多其他可诱导分子的行列,这些分子由腹膜B-1细胞组成性表达,但脾脏B-2细胞不表达。尽管转胶蛋白2在B-1细胞功能中的作用仍不确定,但该分子的鉴定证明了在这个B细胞亚群中研究蛋白质表达的价值。