人类全基因组编码蛋白质芯片技术筛选胆道闭锁自身免疫靶抗原  

Identification of antoantigens in biliary atresia using human proteome microarrays

作　　者：邓蕾 庞舒尹 杨丽媛[1] 代玉梅[1] 刘云锋[1] 张锐忠[3] 刘海英[1] DENG Lei;PANG Shuyin;YANG Liyuan;DAI Yumei;LIU Yunfeng;ZHANG Ruizhong;LIU Haiying(Department of Clinical Laboratory;Pediatric Surgery Laboratory,Guaagzhou Women and Children＇ s Medical Center Affiliated to Guangzhou Medical University,Guangzhou 510623,Guangdong;Graduate School of Guangzhou Medical University,Guangzhou 511456,Guangdong,China)

机构地区：[1]广州医科大学附属广州市妇女儿童医疗中心检验科,广州510623 [2]广州医科大学研究生院,广州511436 [3]广州医科大学附属广州市妇女儿童医疗中心小儿外科实验室,广州510623

出　　处：《临床检验杂志》2018年第10期732-737,共6页Chinese Journal of Clinical Laboratory Science

基　　金：广东省科技厅社会发展领域项目(2014A020212520);广州市卫生局西医类重点项目(201102A212023);广州市妇女儿童医疗中心内部基金(YIP-2018-013)

摘　　要：目的筛选胆道闭锁(biliary atresia,BA)自身免疫相关靶抗原,建立疾病差异性抗原谱,寻找疾病诊断及发病机制研究线索。方法以20例BA患儿、5例疾病对照组患儿(包括婴儿肝炎综合征2例、胆总管囊肿3例)和5例正常儿童对照组的血清为探针,采用全基因组蛋白质芯片技术进行检测,并进行生物学聚类分析和GO功能分析。结果通过比较BA组与对照组,共筛选出存在明显差异的免疫响应蛋白281个,其中Ig G响应105个,Ig M响应176个,Ig G和Ig M同时响应的26个;依据差异倍数≥2、表达稳定的标准得出49个高响应蛋白。GO功能分析发现差异抗原主要涉及调控细胞发育、增殖、凋亡及胆道形成。其中CENP6、UBQLN3、PDCD2、SPEG和RBPJ蛋白在BA组中阳性率显著高于正常对照组(100%vs 0%,P<0.05)。结论成功建立BA相关自身抗原差异谱,为BA的早期诊断、发病机制的阐明提供新的科学依据,但仍需更为深入的鉴定和更多临床样本的验证。Objective To screen the autoantigens in the patients with biliary atresia（ BA）,establish its differential antigen spectra,and provide the clues for its diagnosis and pathogenic mechanism researches. Methods Sera from 20 BA children patients,5 disease controls（ 2 with infant hepatitis syndrome and 3 with choledochocyst） and 5 healthy controls were detected by the human proteome microarrays,and then the cluster analysis and GO functional analysis were performed. Results By comparing the BA group and controls,a total of 281 candidate autoantigens,including 105 reacted with Ig G and 176 reacted with Ig M,were picked out,and there were 26 candidate autoantigens reacted with Ig G and Ig M simultaneously. Based on the criteria of fold change ≥2 and stable expression,49 BArelated candidate autoantigens were identified. GO functional analysis showed that differential autoantigens were mainly involved in the regulation of cell development,proliferation,apoptosis and the formation of biliary tract. The positive rates of CENP6,UBQLN3,PDCD2,SPEG and RBPJ proteins in BA patients were significantly higher than that in the control（ 100% vs 0%,P〈0.05）. Conclusion The BA-related candidate autoantigen spectra is established successfully,which provides a new clue for the early diagnosis of BA and the elucidation of its pathogenesis,but further validation by larger clinical samples is necessary.

关 键 词：胆道闭锁 自身抗原 自身抗体 蛋白质芯片 

分 类 号：R446.6[医药卫生—诊断学]