摘要
目的探讨FAM234A是否为一种新的非Gal异种移植抗原。方法用α-1,3-半乳糖基转移酶基因敲除(GTKO)小型猪的原代猪主动脉内皮细胞(PAEC)免疫食蟹猴。将免疫的猴血清与PAEC孵育,其中经免疫产生的猴抗猪细胞抗体能够识别并结合PAEC表面的未知抗原。采用流式细胞术测定PAEC表面结合的猴抗猪细胞抗体的平均荧光强度(MFI),用以表示PAEC表面抗原的含量。利用慢病毒介导的短发夹核糖核酸(shRNA)敲减PAEC中的FAM234A基因,检测该细胞结合的猴抗猪细胞抗体含量是否减少,以确定该基因是否为潜在的移植抗原。结果 PAEC中的非Gal抗原能够在猴体内引发大量的猴抗猪细胞抗体。在GTKO小型猪的PAEC中用shRNA敲减FAM234A基因后,PAEC结合的猴IgG抗体减少。结论 FAM234A是一种新的非Gal异种移植抗原。
Objective To investigate whether FAM234A is a novel non-Gal antigen in xenotransplantation.Methods Cynomolgus monkeys were immunized with primary porcine aorta endothelial cells(PAEC)fromα-1,3-galactosyltransferase gene-knockout(GTKO)minipigs.The serum samples of immunized monkeys were incubated with PAEC.After immunization,the monkey anti-pig cell antibodies were able to recognize and bind with the unknown antigens on the PAEC surface.Flow cytometry was adopted to determine the mean fluorescence intensity(MFI)of monkey anti-pig cell antibodies binding with the PAEC surface to characterize the content of PAEC surface antigen.Lentivirus-mediated short hairpin ribonucleic acid(shRNA)was utilized to knock down the FAM234A gene in PAEC to detect whether the content of the cell-bound monkey anti-pig cell antibody was reduced and determine if this gene was a potential transplant antigen.Results The non-Gal antigen in PAEC could trigger a large quantity of monkey anti-pig cell antibodies in monkeys.After knock down of the FAM234A gene with shRNA in the PAEC of the GTKO miniature pigs,the monkey IgG antibody binding with PAEC was declined.Conclusions FAM234A is a novel non-Gal antigen in xenotransplantation.
作者
谢崇伟
戴文杰
张军方
蔡志明
牟丽莎
Xie Chongwei;Dai Wenjie;Zhang Junfang;Cai Zhiming;Mou Lisha(Zhongshan School of Medicine,Sun Yat-sen University,Guangzhou 510275,China)
出处
《器官移植》
CAS
CSCD
北大核心
2018年第5期354-359,共6页
Organ Transplantation
基金
国家重点研发计划(2017YFC1103704)
深圳市医疗卫生三名工程(SZSM201412020)
深圳市医学高水平学科建设(2016031638)
深圳市科技计划(JCJY20160229204849975
GJHZ20170314171357556)
深圳市卫生计生系统科研项目(SZXJ2017021)
