三种试剂对红细胞表面CD38抗原去除效果的研究

Study for the Removal of CD38 Antigen on the Surface of Red Blood Cells by Three Different Reagents

目的比较0.2 mol/L二硫苏糖醇(DTT)、0.2 mol/L二巯基乙醇(2-ME)及1%菠萝蛋白酶对红细胞表面CD38抗原去除的效果。方法将2%~5%红细胞悬液与0.2 mol/L DTT、0.2 mol/L 2-ME及1%菠萝蛋白酶按照体积比1∶4分别混匀,37℃孵育30 min洗涤后制备试验用红细胞悬液,以未经处理的红细胞悬液作为阳性对照。上述试验用红细胞悬液与含有达雷妥尤单抗(抗-CD38)的患者血浆进行间接抗人球蛋白试验(IAT)。结果经0.2 mol/L DTT、0.2 mol/L 2-ME及1%菠萝蛋白酶处理过的红细胞与患者血浆进行IAT,前二者处理后结果为阴性,后者显微镜下观察为弱阳性。阳性对照结果为1+。进行IAT期间发现处理过的红细胞均发生不同程度的溶血现象,最严重的为1%菠萝蛋白酶处理过的红细胞。结论上述三种试剂均能去除红细胞表面CD38抗原,但0.2 mol/L DTT、0.2 mol/L 2-ME去除效果优于1%菠萝蛋白酶。

Objective To compare the effects of 0.2 mol/L dithiothreitol(DTT),0.2 mol/L 2-mercaptoethanol(2-ME)and 1%bromelin on the removal of CD38 antigen on the surface of red blood cells(RBCs).Methods 2%~5%RBCs suspension was mixed with 0.2 mol/L DTT,0.2 mol/L 2-ME,and 1%bromelin in a volume ratio of 1∶4 respectively,and then incubated at 37℃for 30 minutes.Untreated RBCs were used as positive control.Indirect anti-human globulin test(IAT)was performed with the RBCs suspension and the plasma of patients with Daratumumab(Anti-CD38).Results The results showed that no agglutination was found in DTT and 2-ME treated RBCs,that weak agglutination was observed under microscope in bromelin treated RBCs,and that 1+agglutination was in positive control.It was found that hemolysis occurred in all treated RBCs during the IAT procedure,and that bromelin treated RBCs were shown most serious hemolysis.Conclusion All three reagents can remove CD38 antigen on the surface of RBCs,and the removal efficiency of 0.2 mol/L DTT and 0.2 mol/L 2-ME is better than that of 1%bromelin.