去除雷达木干扰输血相容性检测的方法及输血疗效评估对比

Removing the interference of daratumumab on transfusion compatibility testing and transfusion efficacy comparison

目的探讨DARA-Fab片段处理经抗CD38单克隆抗体雷达木(daratumumab,DARA)治疗后的多发性骨髓瘤(multiple myeloma,MM)患者输血相容性检测方法的可行性,并通过对比二硫苏糖醇(Dithiothreitol,DTT)方法评估其输血疗效。方法将DARA使用PierceFab制备试剂盒制备成DARA-Fab片段后,确认不同体积(5、10、15、30μL)DARA-Fab片段对于抗筛细胞和抗体鉴定细胞的中和效果;DARA-Fab片段和明确相应抗原的抗筛细胞与对应的单抗试剂为试验组,以添加同体积的生理盐水为对照组孵育后离心;各选取20名经DARA治疗后的MM患者,分别使用DARA-Fab封闭RBC表面抗原和与DTT破坏红细胞表面抗原的方法来进行输血相容性检测,对其输血前后实验室指标进行统计分析,并比较2种配血方法。结果15、30μL DARA-Fab片段与抗筛细胞与混有DARA的血清孵育离心后结果为阴性,5、10μL DARA-Fab结果为阳性,15μL DARA-Fab处理抗体鉴定细胞(2、3、4、5、7、9、11)后为阴性,抗体鉴定细胞(1、6、8、10、12)经30μL DARA-Fab片段处理后,结果为阴性;添加DARA-Fab的实验组对MNS系统、Duffy、Kidd、Kell、Lewis、Rh血型系统和对照组结果一致;DARA-Fab片段这20名患者输注RBC后Hb(hemoglobin,Hb)(73.90±1.90)(g/L)比输血前(63.60±1.58)明显提高,统计学差异明显(P<0.01);患者总胆红素(total bilirubin,TBil)(μmol/L)(16.25±3.54 vs 17.87±3.57)、直接胆红素(direct bilirubin,DBIL)(μmol/L)(6.31±2.32 vs 7.10±2.80)和间接胆红素(Indirect Bilirubin,I-Bil)(9.94±1.38 vs 10.77±1.22)输注前后无明显差异,无统计学差异(P>0.05)。DTT处理方法与DARA-Fab片段封闭方法的输血前后Hb差值(10.75±1.04 vs 10.30±0.98)、TBil差值(3.31±1.47 vs 3.31±0.55)、DBIL差值(2.76±1.24 vs 2.60±0.83)和I-Bil差值(1.97±0.40 vs 2.82±0.53)无明显差异,无统计学差异(P>0.05)。结论DARA-Fab片段可通过封闭RBC表面CD38抗原,来去除DARA对红细胞输血相容性检测的干扰,这种封闭方法对于红细胞临床常见血型系统的抗原无影响,且通过这种配血方法实验室指标输血疗效明显,与DTT方法的输血疗效无差异。

Objective To explore the feasibility of blood transfusion compatibility testing for multiple myeloma(MM)patients treated with anti-CD38 monoclonal antibody daratumumab(DARA)after DARA-Fab fragment blocking,and to evaluate the transfusion efficacy by comparing with dithiothreitol(DTT)method.Methods After DARA was prepared into DARA-Fab fragments using PierceFab preparation kit,the neutralization effects of different volumes(5,10,15,30μL)on screening cells and panel cells were confirmed.DARA-Fab fragments and screening cells with specific antigens and corresponding monoclonal antibody reagents were used as the experimental group and the control group with the same volume of saline for incubating and centrifugin.Twenty MM patients treated with DARA were selected for cross-matching with DARA-Fab and DTT respectively,and the laboratory indexes before and after transfusion were statistically analyzed,and the two blood matching methods were compared.Results After incubating and centrifuging,the results of DARA-Fab fragments(15,30μL)with screening cells and serum mixed with DARA were negative,while those of DARA-Fab(5,10μL)were positive.15μL DARA-Fab treated antibody identification cells(2,3,4,5,7,9,11)were negative,antibody identification cells(1,6,8,10,12)were negative after 30μL DARA-Fab fragments treatment;the results of MNS,Duffy,Kidd,Kell,Lewis,Rh blood group system of the experimental group were consistent with those of the control group;the hemoglobin(Hb)(g/L)of 20 patients after infusion of RBC(73.90±1.90)was significantly higher than that before transfusion(63.60±1.58),P<0.01.There was no significant difference in total bilirubin(TBil)(μmol/L)(16.25±3.54 vs 17.87±3.57),direct bilirubin(DBIL)(μmol/L)(6.31±2.32 vs 7.10±2.80)and indirect bilirubin(I-Bil)(9.94±1.38 vs 10.77±1.22)before and after infusion(P>0.05).And no statistical difference was noticed in Hb(10.75±1.04 vs 10.30±0.98),TBil(3.31±1.47 vs 3.31±0.55),DBIL(2.76±1.24 vs 2.60±0.83),and I-Bil(1.97±0.40 vs 2.82±0.53)between the DTT treatment method and the DARA Fab fragment treatment before and after transfusion(P>0.05).Conclusion DARA-Fab can remove the interference of RBC on cross matching by blocking CD38 antigen.This method has no effect on the antigens of common RBC blood group systems,and shows significant blood transfusion efficacy as that of DTT method.