人源化基因修饰猪红细胞与人血清免疫相容性的体外研究  

In vitro study of immunocompatibility of humanized genetically modified pig erythrocytes with human serum

作　　者：陈蕾佳 崔梦一 宋翔宇 王恺 贾志博 杨鎏璞 董阳辉 左浩辰 杜嘉祥 潘登科 许文静 任洪波[4] 赵亚群 彭江 Chen Leijia;Cui Mengyi;Song Xiangyu;Wang Kai;Jia Zhibo;Yang Liupu;Dong Yanghui;Zuo Haochen;Du Jiaxiang;Pan Dengke;Xu Wenjing;Ren Hongbo;Zhao Yaqun;Peng Jiang(Graduate School of Hebei North University,Zhangjiakou 075132,China)

机构地区：[1]河北北方学院研究生院,河北张家口075132 [2]中国人民解放军总医院第一医学中心神经外科医学部 [3]中国人民解放军总医院第四医学中心骨科医学部研究所 [4]邯郸市中心医院神经外科 [5]成都中科奥格生物科技有限公司

出　　处：《器官移植》2024年第3期415-421,共7页Organ Transplantation

基　　金：国家重点研发计划(2019YFA0110704);解放军总医院青年自主创新科学基金项目(22QNFC014)。

摘　　要：目的 探讨野生型(WT)、四基因修饰(TKO/hCD55)和六基因修饰(TKO/hCD55/hCD46/hTBM)猪红细胞与人血清的免疫相容性和免疫差异。方法 收集20名不同血型志愿者的血液,将WT、TKO/hCD55和TKO/hCD55/hCD46/hTBM猪红细胞、ABO-相容(ABO-C)及ABO-不相容(ABO-I)人红细胞分别暴露于不同血型人血清中,检测血凝集、抗原抗体结合(IgG、IgM)水平和补体依赖细胞毒性,评估2种基因修饰猪红细胞与人血清的免疫相容性。结果 ABO-C组未出现明显凝集;WT组和ABO-I组凝集水平高于TKO/hCD55组和TKO/hCD55/hCD46/hTBM组(均为P<0.001)。WT组猪红细胞裂解水平高于ABO-C组、TKO/hCD55组和TKO/hCD55/hCD46/hTBM组;ABO-I组猪红细胞裂解水平高于TKO/hCD55组、TKO/hCD55/hCD46/hTBM组(均为P<0.01)。TKO/hCD55组猪红细胞IgM和IgG结合水平均低于WT组和ABO-I组;TKO/hCD55/hCD46/hTBM组猪红细胞IgG和IgM结合水平低于WT组,IgG低于ABO-I组(均为P<0.05)。结论基因修饰猪红细胞的免疫相容性优于野生型猪,并接近于ABO-C,人源化猪红细胞在血资源奇缺时可考虑作为血液来源。Objective To investigate the differences and the immunocompatibility of wild-type(WT),four-gene modified(TKO/hCD55)and six-gene modified(TKO/hCD55/hCD46/hTBM)pig erythrocytes with human serum.Methods The blood samples were collected from 20 volunteers with different blood groups.WT,TKO/hCD55,TKO/hCD55/hCD46/hTBM pig erythrocytes,ABO-compatible(ABO-C)and ABO-incompatible(ABO-I)human erythrocytes were exposed to human serum of different blood groups,respectively.The blood agglutination and antigenantibody binding levels(IgG,IgM)and complement-dependent cytotoxicity were detected.The immunocompatibility of two types of genetically modified pig erythrocytes with human serum was evaluated.Results No significant blood agglutination was observed in the ABO-C group.The blood agglutination levels in the WT and ABO-I groups were higher than those in the TKO/hCD55 and TKO/hCD55/hCD46/hTBM groups(all P<0.001).The level of erythrocyte lysis in the WT group was higher than those in the ABO-C,TKO/hCD55 and TKO/hCD55/hCD46/hTBM groups.The level of erythrocyte lysis in the ABO-I group was higher than those in the TKO/hCD55 and TKO/hCD55/hCD46/hTBM groups(both P<0.01).The pig erythrocyte binding level with IgM and IgG in the TKO/hCD55 group was lower than those in the WT and ABO-I groups.The pig erythrocyte binding level with IgG and IgM in the TKO/hCD55/hCD46/hTBM group was lower than that in the WT group and pig erythrocyte binding level with IgG was lower than that in the ABO-I group(all P<0.05).Conclusions The immunocompatibility of genetically modified pig erythrocytes is better than that of wild-type pigs and close to that of ABO-C pigs.Humanized pig erythrocytes may be considered as a blood source when blood sources are extremely scarce.

关 键 词：输血 异种移植 基因修饰猪 异种抗原 红细胞 抗原抗体结合 凝集 补体依赖细胞毒性 

分 类 号：R457.1[医药卫生—治疗学] R392[医药卫生—临床医学]