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为了在体外研究HPA-1抗原的多态性,同时建立一种更简便灵敏的检测抗HPA-1同种(异型)抗体的方法,将含有HPA-1a和HPA-1b的DNA质粒分别转染CHO细胞(中国仓鼠卵巢细胞)并获得稳定表达β_3HPA-1a和β_3HPA-1b的细胞株。流式细胞分析结果表明,CHO细胞结合抗HPAPA-1a同种(异型)抗体的能力与细胞表面β_3整合素表达量直接相关。同时,针对HPA-1a表位的单克隆抗体SZ21仅能抑制CHOβ_3HPA-1a细胞的粘附,对 CHOβ_3HPA-1b细胞却没有影响。抗HPA-1a参照血清能阻断CHOβ_3HPA-1a细胞的粘附,但对CHOβ_3HPA-1b细胞却影响甚微。结果表明,CHO细胞模型可以成功地用来表达并研究HPA-1a的多态性及其抗原特性,同时可以作为用于临床检测抗HPA-1a同种(异型)抗体的一种替代方法。
In order to study the polymorphism of HPA-1 antigen in vitro and to establish a more simple and sensitive method for detecting the anti-HPA-1 allotype antibody, the DNA plasmids containing HPA-1a and HPA-1b were transfected separately CHO cells (Chinese hamster ovary cells) and obtain cell lines stably expressing β_3HPA-1a and β_3HPA-1b. Flow cytometry analysis showed that the ability of CHO cells to bind anti-HPAPA-1a allotype antibody was directly related to the cell surface β 3 integrin expression. Meanwhile, the monoclonal antibody SZ21 directed against the HPA-1a epitope only inhibited the adhesion of CHOβ_3HPA-1a cells and had no effect on CHOβ_3HPA-1b cells. Anti-HPA-1a reference serum blocked the adhesion of CHOβ_3HPA-1a cells, but had little effect on CHOβ_3HPA-1b cells. The results show that the CHO cell model can be successfully used to express and study the polymorphism of HPA-1a and its antigenic properties, and can be used as an alternative method for clinical detection of anti-HPA-1a allotypic antibodies.