Rao N, Udani M, Nelson J, Reid M E, Telen M J
Department of Medicine, Duke University Medical Center, Durham, North Carolina, USA.
Transfusion. 1995 Jun;35(6):459-64. doi: 10.1046/j.1537-2995.1995.35695288762.x.
The high-frequency Hy and Gya antigens have been shown to reside on the same protein. Gy(a-) Hy-negative red cells are also Do(a-b-). A mouse monoclonal antibody, 5B10, was produced with specificity related to the human Gregory, Holley, and Dombrock blood group antigens.
The antibody reacted in direct hemagglutination assays, and its specificity was investigated by radioimmunoassay, inhibition assay, and Western blotting.
The 5B10 antibody failed to bind to abnormal paroxysmal nocturnal hemoglobinuria red cells and human erythroleukemia cell line K562, but it was weakly reactive with HEL cells. Red cells, but not other circulating hematopoietic cells, express the 5B10 antigen. The 5B10 antibody had a specificity similar but not identical to that of Gya. Gy(a-) Hy-negative red cells reacted extremely weakly with 5B10 antibody, but Gy(a-) Hy-negative red cells treated with a variety of proteases bound 5B10 antibody strongly. This suggests that these cells express a variant form of the protein recognized by 5B10.
Identification of a monoclonal antibody to this glycosylphosphatidylinositol-linked protein opens a new avenue for investigation of the biochemistry, genetics, and function of the glycosylphosphatidylinositol-linked protein that bears the Gya, Hy, and Do antigens.
高频Hy和Gya抗原已被证明存在于同一蛋白质上。Gy(a-)Hy阴性红细胞也是Do(a-b-)。制备了一种与人类格雷戈里、霍利和唐布罗克血型抗原相关的小鼠单克隆抗体5B10。
该抗体在直接血凝试验中发生反应，并通过放射免疫测定、抑制试验和蛋白质印迹法研究其特异性。
5B10抗体未能与异常阵发性夜间血红蛋白尿红细胞和人类红白血病细胞系K562结合，但与HEL细胞有弱反应。红细胞而非其他循环造血细胞表达5B10抗原。5B10抗体的特异性与Gya相似但不完全相同。Gy(a-)Hy阴性红细胞与5B10抗体反应极弱，但用多种蛋白酶处理的Gy(a-)Hy阴性红细胞能强烈结合5B10抗体。这表明这些细胞表达5B10识别的蛋白质的变异形式。
鉴定出针对这种糖基磷脂酰肌醇连接蛋白的单克隆抗体，为研究携带Gya、Hy和Do抗原的糖基磷脂酰肌醇连接蛋白的生物化学、遗传学和功能开辟了一条新途径。
