应用改良流式法检测猕猴和食蟹猴体内血型抗体水平的分布情况

Application of flow cytometry to detect ABO blood group antibody levels in rhesus monkeys and cynomolgus monkeys

绝大部分灵长类动物存在与人类相似的ABO血型系统,该研究采用改良流式法(flow cytometry method,FCM)检测猕猴及食蟹猴血清中血型抗体水平的分布情况。以流式细胞术为基础,使用商品化人源红细胞为靶细胞,并通过加入特异性荧光标记的抗人IgM或IgG二抗,对收集的实验用猕猴及食蟹猴的血清样本进行检测,以人类健康受试者的血清样本为对照,比较两者血型抗体水平的差异。结果显示:预先用人O型浓缩红细胞吸附猴血清中所含种属间非特异性抗体后,FCM法能够准确检测其血型抗体水平及分型,并且发现猴血清中天然血型抗体的水平明显低于健康人(P<0.05)。由此得出:通过预处理清除非特异性抗体的干扰后,FCM法同样适用于灵长类动物血清中血型抗体的检测,也为构建灵长类动物模拟人ABO血型不合器官移植模型提供了技术保障和实验数据。

Similar to humans,most of non-human primates also contain the ABO blood group system.In this study,we sought to evaluate the ABO antibody（Ab） levels in monkeys using a modified flow cytometry method（FCM）.The standard commercial human A or B-red blood cells（RBCs） were used as target cells.The binding of target cells and anti-A or B blood group Ab in sera of rhesus or cynomolgus monkeys was detected by flow cytometry after adding secondary specific fluorescence-labeled anti-human IgG or IgM Ab.Human healthy blood donor sera were used as controls.The results revealed that,using clear monkey sera,which were pre-absorbed on normal human type O RBCs to remove non-specific anti-human Abs,the modified FCM gave an accurate detection of ABO Ab levels in monkeys.When compared with the results of human sera,the distribution of ABO Ab levels in monkey sera were significantly lower（P0.05）.We concluded that the modified FCM could be used for the detection of monkey ABO Ab levels.The technique and data will be very valuable for the future establishment of ABO-incompatible organ transplant models in non-human primates,which could improve clinical applications.