Kwok W W, Kovats S, Thurtle P, Nepom G T
Virginia Mason Research Center, Seattle, WA 98101.
J Immunol. 1993 Mar 15;150(6):2263-72.
HLA-DQ class II molecules are complex heterodimers in which polymorphic alpha- and beta-polypeptides form noncovalent associations leading to cell surface expression. Using gene transfer with retroviral vectors, we demonstrate that surface expression of heterodimers among various DQ alpha and -beta polypeptides is not universal. Two types of allele-specific constraints were found: DQ beta chains that are associated with the DQw1 specificities do not form stable cell surface heterodimers with DQ alpha chains encoded on haplotypes that are associated with DQw2, DQw3, or DQw4. Similarly, surface expression of heterodimers composed of DQw2- or DQw3-associated DQ beta chains and DQw1 DQ alpha chains were not detected. The formation of mixed isotype heterodimers also showed allelic preference: DR alpha/DQw1 beta dimers were detected on the cell surface, but DR alpha/DQw3 beta dimers were not. Failure to express class II heterodimers on the cell surface was apparently due to either inefficient assembly of the alpha- and beta-chains or retention of the mismatched dimers in an intracellular compartment. DQ beta cDNA encoding DQw1- and DQw3-associated beta-polypeptides were modified to create chimeric expression vectors in which portions of the DQ beta polypeptides were exchanged between different alleles. The resulting chimeric DQ beta polypeptides were tested for their ability to form stable heterodimers with different DQ alpha polypeptides. A cluster of polymorphic amino acid residues encoded by the 3' end of the DQB1 second exon, corresponding to residues 60 to 91 in the DQ beta polypeptide, were found to be critical determinants constraining stable cell surface heterodimer formation among preferential alpha- and beta-combinations.
HLA - DQ II类分子是复杂的异二聚体,其中多态性的α和β多肽形成非共价结合,从而导致细胞表面表达。通过逆转录病毒载体进行基因转移,我们证明了不同DQα和β多肽之间异二聚体的表面表达并非普遍存在。发现了两种类型的等位基因特异性限制:与DQw1特异性相关的DQβ链不会与与DQw2、DQw3或DQw4相关的单倍型上编码的DQα链形成稳定的细胞表面异二聚体。同样,未检测到由与DQw2或DQw3相关的DQβ链和DQw1 DQα链组成的异二聚体的表面表达。混合同种型异二聚体的形成也表现出等位基因偏好:细胞表面检测到DRα/DQw1β二聚体,但未检测到DRα/DQw3β二聚体。未能在细胞表面表达II类异二聚体显然是由于α链和β链组装效率低下,或者是错配的二聚体滞留在细胞内区室中。编码与DQw1和DQw3相关的β多肽的DQβ cDNA被修饰以创建嵌合表达载体,其中DQβ多肽的部分在不同等位基因之间进行了交换。测试了所得嵌合DQβ多肽与不同DQα多肽形成稳定异二聚体的能力。发现由DQB1第二外显子3'端编码的一组多态性氨基酸残基,对应于DQβ多肽中的60至91位残基,是限制优先α和β组合之间稳定细胞表面异二聚体形成的关键决定因素。