Yang S Y, Morishima Y, Collins N H, Alton T, Pollack M S, Yunis E J, Dupont B
Immunogenetics. 1984;19(3):217-31. doi: 10.1007/BF00364765.
A new mouse monoclonal antibody (MoAb) 4E, which detects an epitope shared by HLA-B locus antigens, together with the MoAb W6/32, detecting a common HLA, B, C, determinant, and the MoAb4B, detecting HLA-A2 and A28, were used to isolate HLA-A and -B antigens in sequential immunoprecipitation. The HLA antigens obtained from metabolically labeled cell extracts of B-lymphoblastoid cell lines or from phytohemagglutinin (PHA)-activated peripheral blood lymphocytes were compared by one-dimensional isoelectric focusing (1D-IEF). The IEF banding patterns obtained with native HLA antigens segregated in a family with HLA. Neuraminidase treatment of isolated antigens reduced the number of bands to one or two, simplifying the analysis of characteristic patterns. Thus, we have cataloged IEF banding patterns for the majority of the serologically recognized HLA-A and -B allotypes obtained from 57 unrelated American Caucasians. While no HLA-A locus or HLA-B locus specific banding patterns were observed, the HLA-A antigens had, in general, slightly higher pI values than the HLA-B antigens. HLA-C antigens could not be detected in this assay system. The polymorphism detected by IEF banding patterns was as extensive as the serologically detected polymorphism identified by classical HLA serology. Moreover, variants for some HLA allotypes could be detected by this method. In addition to previously recognized A2 variants, new variants were identified for HLA-A1, A26, and Bw44. Each A1 and Bw44 variant was associated with particular haplotypes. The HLA-A2 antigens occurred on 43 HLA haplotypes in the unrelated Caucasian population. Only one of each A2 variants was identified in this population.
一种新的小鼠单克隆抗体（MoAb）4E，可检测HLA - B位点抗原共有的一个表位，与检测常见HLA - B、C决定簇的MoAb W6/32以及检测HLA - A2和A28的MoAb4B一起，用于在连续免疫沉淀中分离HLA - A和 - B抗原。通过一维等电聚焦（1D - IEF）比较了从B淋巴母细胞系的代谢标记细胞提取物或从植物血凝素（PHA）激活的外周血淋巴细胞中获得的HLA抗原。用天然HLA抗原获得的IEF条带模式在一个与HLA相关的家族中分离。对分离抗原进行神经氨酸酶处理可使条带数量减少至一两条，从而简化了特征模式的分析。因此，我们已经编目了从57名无关的美国白种人获得的大多数血清学识别的HLA - A和 - B同种异型的IEF条带模式。虽然未观察到HLA - A位点或HLA - B位点特异性条带模式，但HLA - A抗原的等电点值总体上略高于HLA - B抗原。在该检测系统中无法检测到HLA - C抗原。IEF条带模式检测到的多态性与经典HLA血清学检测到的血清学多态性一样广泛。此外，通过该方法可以检测到一些HLA同种异型的变体。除了先前识别的A2变体之外，还鉴定出了HLA - A1、A26和Bw44的新变体。每个A1和Bw44变体都与特定的单倍型相关。在无关的白种人群体中，HLA - A2抗原出现在43种HLA单倍型上。在该群体中每种A2变体仅鉴定出一种。
