中国血液净化

中国血液净化 ›› 2015, Vol. 14 ›› Issue (03): 173-176.doi: 10.3969/j.issn.1671-4091.2015.03.00

• 基础研究 • 上一篇    下一篇

EB病毒转化细胞模型在IgA肾病研究中的适应性探讨

朱厉,翟亚玲,张宏   

  1. 北京大学第一医院肾内科
  • 收稿日期:2014-11-14 修回日期:2015-01-04 出版日期:2015-03-12 发布日期:2015-04-20
  • 通讯作者: 张宏 hongzh@bjmu.edu.cn E-mail:hongzh@bjmu.edu.cn
  • 基金资助:
    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)

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

  • Received:2014-11-14 Revised:2015-01-04 Online:2015-03-12 Published:2015-04-20

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摘要： 【摘要】目的 越来越多的证据表明IgA分子异常是IgA肾病发病始动因素。外周血B淋巴细胞是产生IgA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IgA1分子糖基化水平不受影响，因此可作为开展糖基化异常IgA1分子产生机制研究的体外细胞模型。近期IgA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IgA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IgA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IgA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IgA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IgA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IgA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IgA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IgA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。

关键词: IgA肾病, EB病毒, 增生诱导配体, B淋巴细胞

Abstract: 【Abstract】Objective Increasing evidences indicated that the initial pathogenic factor for IgA nephropathy (IgAN) was the abnormal IgA molecule. IgA molecules are produced from peripheral B-lymphocytes. However, the limited availability of B-lymphocytes greatly hindered the studies in depth on mechanisms in IgAN. Recently, EBV (Epstein-Barr virus)-immortalized B-cells were reported to maintain the characteristic of secreting aberrant glycosylated IgA1 molecules, and were regarded as a useful cell model for studies on mechanisms leading to aberrant IgA1 glycosylation in IgAN. A recent genome-wide association study in IgAN identified a proliferation-inducing ligand (APRIL) as one of IgAN susceptible genes. The objective of this study was to investigate the feasibility of EBV-immortalized B-cells for study of APRIL in IgAN. 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 IgA level by ELISA. Meanwhile, primary B-lymphocytes isolated from peripheral blood of a healthy control were treated with 25 ng/ml APRIL, and IgA in the conditioned media were also detected. Results APRIL significantly promoted the secretion of IgA in primary B-lymphocytes (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 IgA 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 IgA in IgAN.

Key words: IgA nephropathy, Epstein–Barr virus, a proliferation-inducing ligand, B lymphocyte