EB病毒转化细胞模型在IgA肾病研究中的适应性探讨  

The feasibility of EBV-immortalized B cells for study of IgA nephropathy

作　　者：朱厉[1,2,3,4] 翟亚玲[1,2,3,4] 张宏[1,2,3,4] 

机构地区：[1]北京大学第一医院肾内科 [2]北京大学肾脏疾病研究所 [3]卫生部肾脏疾病重点实验室 [4]教育部慢性肾脏病防治重点实验室,北京100034

出　　处：《中国血液净化》2015年第3期173-176,共4页Chinese Journal of Blood Purification

基　　金：国家自然科学基金面上项目(81470945);北京市自然科学基金重点项目(7131016);首都临床特色应用研究(Z141107002514037)

摘　　要：目的越来越多的证据表明Ig A分子异常是Ig A肾病发病始动因素。外周血B淋巴细胞是产生Ig A分子的主要细胞。因从外周血中分离出来的B淋巴细胞数量有限,限制了体外细胞学进一步机制研究的开展。近期有学者发现EB病毒转化可使B淋巴细胞获得永生化的增殖,同时保持Ig A1分子糖基化水平不受影响,因此可作为开展糖基化异常Ig A1分子产生机制研究的体外细胞模型。近期Ig A肾病的一项全基因组关联分析研究提示增生诱导配体（APRIL）是Ig A肾病的发病易感基因。本文旨在探讨EB病毒转化细胞模型是否可用于探讨APRIL参与Ig A肾病发病的机制研究。方法本研究选取3例个体的外周血B淋巴细胞进行EB病毒转化,获得永生化B淋巴细胞后,经APRIL处理后检测培养上清中Ig A的浓度。同时,从1例健康对照中分离获得原代B淋巴细胞,给予APRIL处理并检测其产生Ig A的水平。结果我们的研究发现APRIL可显著上调原代B淋巴细胞产生Ig A的水平（294.3±35.7ng/ml vs.368.2±36.2ng/ml,P=0.043）,但对于EB病毒转化后获得的永生化B淋巴细胞产生的Ig A水平无影响（各剂量浓度1.56-200ng/ml下,P值均大于0.05）。结论本研究发现EBV病毒转化的永生化B淋巴细胞,在APRIL对其影响Ig A产生方面,具有与原代B淋巴细胞不同的生物学效应。因此,当开展APRIL在Ig A肾病发病中的机制探讨时,EB病毒转化后的B淋巴细胞不是合适的体外细胞学模型。Objective Increasing evidences indicated that the initial pathogenic factor for Ig A nephropathy（Ig AN） was the abnormal Ig A molecule. Ig A molecules are produced from peripheral B- lymphocytes.However, the limited availability of B- lymphocytes greatly hindered the studies in depth on mechanisms in Ig AN. Recently, EBV（Epstein-Barr virus）-immortalized B-cells were reported to maintain the characteristic of secreting aberrant glycosylated Ig A1 molecules, and were regarded as a useful cell model for studies on mechanisms leading to aberrant Ig A1 glycosylation in Ig AN. A recent genome- wide association study in Ig AN identified a proliferation-inducing ligand（APRIL） as one of Ig AN susceptible genes. The objective of this study was to investigate the feasibility of EBV-immortalized B-cells for study of APRIL in Ig AN. Methods We established three EBV-immortalized B-cell lines from three individuals with distinct diseases. After treated with different doses of APRIL（1.56~200 ng/ml） for 48 hours, conditioned media from these EBV-immortalized B- cells were collected for detection of Ig A level by ELISA. Meanwhile, primary B- lymphocytes isolated from peripheral blood of a healthy control were treated with 25 ng/ml APRIL, and Ig A in the conditioned media were also detected. Results APRIL significantly promoted the secretion of Ig A in primary Blymphocytes（294.3±35.7 ng/ml vs. 368.2±36.2 ng/ml, P=0.043）, but not in EBV-immortalized B-cells（P value 0.05 was observed in APRIL of 1.56-200 ng/ml）. Conclusion Our study identified the different effect on Ig A secretion induced by APRIL between primary B-lymphocytes and EBV-immortalized B-cells, suggesting that EBV-immortalized B-cells may not be suitable for the study of APRIL on Ig A in Ig AN.

关 键 词：IgA肾病 EB病毒 增生诱导配体 B淋巴细胞 

分 类 号：R692.5[医药卫生—泌尿科学]