Carra G, Accolla R S
J Exp Med. 1987 Jan 1;165(1):47-63. doi: 10.1084/jem.165.1.47.
Structural analysis by two-dimensional peptide maps (2D-PM) of the human Ia molecular pool expressed on the cell surface of two distinct lymphoblastoid cell line, LG-2 and Raji, revealed the existence of a novel MHC class II molecular heterodimer that differs at the level of both alpha and beta subunits from the previously described DP, DQ, and DR antigens. These differences were also seen at the level of two-dimensional electrophoresis (2D-PAGE) of biosynthetically labeled intact molecules, although to a lesser extent, due to the intrinsic limitations of this technique in resolving fine structural differences. We have designated this new class II antigen as the fourth Ia subset. The fourth Ia subset seems to represent a small proportion of the human Ia pool. Comparative analysis by 2D-PM of the two cell lines showed the presence of structural variations in the alpha chains of the fourth Ia subset, suggesting the existence of polymorphism for these subunits. Cell surface iodination did not show appreciable labeling of the fourth subset beta chain in LG-2 cells, and this prevented analysis of the structural polymorphism of this subunit. Furthermore, for the first time, we have shown that DP alpha chains display distinct peptide maps in LG-2 and Raji cells, thus suggesting the presence of structural polymorphism for these Ia subunits also. The DQ1 alpha and beta allelic products present in LG-2 cells (DQ homozygous) did not show appreciable structural variation when compared with the homologous allelic products present in Raji cells (DQ heterozygous). Finally, we have confirmed the absence of polymorphism for the DR alpha subunits. By 2D-PM, relatively low structural variation was instead found for the highly polymorphic DR beta subunits expressed in the two cell lines, suggesting that cell surface iodination preferentially labels constant domains of DR beta chains.
通过二维肽图(2D-PM)对在两种不同的淋巴母细胞系LG-2和Raji细胞表面表达的人Ia分子库进行结构分析,发现了一种新的MHC II类分子异二聚体,其α和β亚基水平均与先前描述的DP、DQ和DR抗原不同。在生物合成标记的完整分子的二维电泳(2D-PAGE)水平上也观察到了这些差异,不过由于该技术在解析精细结构差异方面的固有局限性,差异程度较小。我们将这种新的II类抗原指定为第四个Ia亚群。第四个Ia亚群似乎只占人Ia库的一小部分。通过2D-PM对这两种细胞系进行的比较分析显示,第四个Ia亚群的α链存在结构变异,表明这些亚基存在多态性。细胞表面碘化未显示LG-2细胞中第四个亚群β链有明显标记,这妨碍了对该亚基结构多态性的分析。此外,我们首次表明,DP α链在LG-2和Raji细胞中显示出不同的肽图,因此也表明这些Ia亚基存在结构多态性。与Raji细胞(DQ杂合)中存在的同源等位基因产物相比,LG-2细胞(DQ纯合)中存在的DQ1 α和β等位基因产物未显示出明显的结构变异。最后,我们证实了DR α亚基不存在多态性。通过2D-PM,在这两种细胞系中表达的高度多态的DR β亚基反而发现结构变异相对较低,这表明细胞表面碘化优先标记DR β链的恒定区。