α-1,2-L-岩藻糖苷酶处理人红细胞H抗原的实验研究

Study of removing H antigen on erythrocyte by α-1,2-L-fucosidase

目的通过对α-1,2-L-岩藻糖苷酶处理人红细胞H抗原的研究,明确岩藻糖苷酶对H抗原处理的效果及对红细胞的影响,确保经酶处理的红细胞可以成为指导H抗原自身抗体检测的试剂红细胞,为输血安全性提供进一步的保障。方法用20 000 U/mL的α-1,2-L-岩藻糖苷酶处理人红细胞,然后用单克隆抗-H在血清学及流式细胞术方面检测H抗原的酶处理效果,用其它相关的单克隆抗体检测岩藻糖苷酶处理后红细胞相关抗原的变化,在显微镜下观察红细胞的形态变化以及用流式细胞术检测经酶处理红细胞膜电位的变化。结果经α-1,2-L-岩藻糖苷酶处理的红细胞H抗原被清除,红细胞的形态、膜电位及红细胞上其它相关抗原与处理前无明显变化。结论α-1,2-L-岩藻糖苷酶处理的红细胞可以作为检测H抗原自身抗体的试剂红细胞,为孟买血型及类孟买血型人的输血提供进一步的保障。

Objective To observe the effects of enzymatic treatment with a-1,2-L-fucosidase on H antigen on erythro- cyte and to study the influence of erythrocyte, guide the detection of anti-H auto-antibody and make sure the safety of blood transfusion. Methods Erythrocyte was treated by 20 000 units/mL ct-1,2-L-fucosidase and the detection of H antigen was evaluated using monoclonal anti-H by serology experiments and flow cytometry （FCM）. The main relative blood group anti- gens were tested using the other relative monoclonal antibodies. The shape of erythl＂ocytes was observed by microscope. The absolute membrane potential of erythrocytes was determinated by flow cytometry （ FCM ）. Results The H antigen was cleaved completely by the or-1,2-L-fucosidase. The shape of enzyme-converted erythrocyte hadn＇t changed. And after the en zymatic treatment, the main relative blood group antigens hadn＇t changed either. Conclusion Human RBCs were treated with a-1,2-L-fucosidase could be detection of anti-H auto-antibody and make sure the safety of blood transfusion for bombay and para-hombay.