Monos D S, Czanky E, Ono S J, Radka S F, Kappes D, Strominger J L
Department of Biochemistry and Molecular Biology, Harvard University, Cambridge, MA 02138, USA.
Immunogenetics. 1995;42(3):172-80. doi: 10.1007/BF00191222.
cDNAs coding for the HLA class II DR and DQ alpha and beta chains of the diabetogenic haplotypes DR3 and DR4 were introduced into a mammalian expression vector and transfected into L-cell mouse fibroblasts to produce cells expressing individual human class II molecules. Stable L transfectants were generated expressing each of the DR or DQ isotypes of the cis-encoded alpha and beta chains of the DR3 or DR4 haplotypes, as well as the trans-encoded alpha and beta chains of the DQ molecules of the two haplotypes. However, isotype mismatched combinations (DR alpha/DQ beta or DQ alpha/DR beta) did not result in any stable transfectants. The stable DQ L-cell transfectants obtained, along with homozygous B-cell lines expressing the DQ2 and DQ8 specificities, were tested against a large panel of twentyone anti-HLA class II monoclonal antibodies (mAbs). Their unusual reactivity patterns are described including the failure of most "pan-DQ" mAbs to react with all DQ expressing L-cell transfectants. Interestingly, some mAbs react with certain alpha beta heterodimers expressed on B-LCL but fail to recognize the same heterodimers expressed on the transfectants. This is suggestive of minor structural modifications that class II molecules undergo depending on the cells they are expressed on.
将编码致糖尿病单倍型DR3和DR4的HLA II类DR以及DQα和β链的cDNA引入哺乳动物表达载体,并转染到L细胞小鼠成纤维细胞中,以产生表达个体人类II类分子的细胞。生成了稳定的L转染子,它们表达DR3或DR4单倍型的顺式编码α和β链的每种DR或DQ同种型,以及两种单倍型的DQ分子的反式编码α和β链。然而,同种型错配组合(DRα/DQβ或DQα/DRβ)未产生任何稳定的转染子。对获得的稳定DQ L细胞转染子以及表达DQ2和DQ8特异性的纯合B细胞系,用一大组21种抗HLA II类单克隆抗体(mAb)进行检测。描述了它们不同寻常的反应模式,包括大多数“泛DQ”mAb不能与所有表达DQ的L细胞转染子反应。有趣的是,一些mAb与B-LCL上表达的某些αβ异二聚体反应,但不能识别转染子上表达的相同异二聚体。这表明II类分子根据其表达的细胞会发生微小的结构修饰。