Sato Kota, Shikano Sojin, Xia Guohong, Takao Joe, Chung Jin-Sung, Cruz Ponciano D, Xie Xiao-Song, Ariizumi Kiyoshi
Department of Dermatology, The University of Texas Southwestern Medical Center, Dallas Veterans Affairs Medical Center, 5323 Harry Hines Building, Dallas, TX 75390-9069, USA.
Mol Immunol. 2006 Mar;43(9):1443-53. doi: 10.1016/j.molimm.2005.07.035. Epub 2005 Sep 6.
Dendritic cells (DC) are far more potent to activate T cells than other antigen presenting cells (e.g., macrophages) and distributed to many organs where DC develop to functionally and phenotypically distinctive subsets. To isolate DC-differentially expressed genes, we used a subtractive cDNA cloning (XS52 DC minus J774 macrophages), resulting in the identification of d2 isoform of vacuolar (V) H+-ATPase subunit d. Unlike the ubiquitously expressed isoform (d1), d2 mRNA manifested expression restricted to particular subsets of DC (e.g., skin- and bone marrow-derived DC) among leukocytes and encoded two transcripts (1.6 and 3.0 kb) that differed in the length of the 3'-untranslated region. The d2 protein displayed association with membranes and the localization in lysosomes and antigen-containing endosomes. Interestingly, XS52 DC expressed seven-fold higher V-ATPase proton-pump activity than J774 macrophages and distinguished from the macrophage by high levels of isoforms a1 and a2 expression among V-ATPase subunits. These results indicated that d2 is a new marker for DC and it may, co-operatively with subunit a isoforms, regulate V-ATPase activity.
树突状细胞(DC)比其他抗原呈递细胞(如巨噬细胞)更有效地激活T细胞,并且分布于许多器官,在这些器官中DC发育为功能和表型独特的亚群。为了分离DC差异表达基因,我们采用了消减cDNA克隆技术(XS52 DC减去J774巨噬细胞),从而鉴定出液泡(V)H + -ATP酶亚基d的d2亚型。与普遍表达的亚型(d1)不同,d2 mRNA在白细胞中仅在特定的DC亚群(如皮肤和骨髓来源的DC)中表达,并且编码两种转录本(1.6和3.0 kb),它们在3'非翻译区的长度上有所不同。d2蛋白与膜相关,定位于溶酶体和含抗原的内体中。有趣的是,XS52 DC的V-ATP酶质子泵活性比J774巨噬细胞高7倍,并且在V-ATP酶亚基中,a1和a2亚型的表达水平高于巨噬细胞。这些结果表明d2是DC的一个新标志物,它可能与a亚型亚基协同调节V-ATP酶活性。