[1]王超男,李伟皓,沈宝艳,等.PLA2R 真核表达细胞株的构建及其在膜性肾病诊断中的应用[J].现代检验医学杂志,2020,35(04):1-5,13.[doi:10.3969/j.issn.1671-7414.2020.04.001]
　WANG Chao-nan,LI Wei-hao,SHEN Bao-yan,et al.Establishment of A Stable Cell Line CHO-PLA2R with High Expression of PLA2R and Its Application in the Diagnosis of Membranous Nephropathy[J].Journal of Modern Laboratory Medicine,2020,35(04):1-5,13.[doi:10.3969/j.issn.1671-7414.2020.04.001]
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PLA2R 真核表达细胞株的构建及其在膜性肾病诊断中的应用()
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《现代检验医学杂志》[ISSN:/CN:]

卷:
第35卷
期数:
2020年04期
页码:
1-5,13
栏目:
论　著
出版日期:
2020-09-05

文章信息/Info

Title:
Establishment of A Stable Cell Line CHO-PLA2R with High Expression of PLA2R and Its Application in the Diagnosis of Membranous Nephropathy
文章编号:
1671-7414（2020）04-001-06
作者:
王超男1; 李伟皓2; 沈宝艳3; 李江雪1; 张 佳1; 刘晓梅2; 张菲菲2; 张智萍2; 冯晓燕1; 张贺秋1
（1. 东方海洋（北京）医学研究院，北京 100071； 2. 河北医科大学第二医院特检科，石家庄 050000；3. 蒙城县第一人民医院肾病免疫风湿科，安徽亳州233500）
Author(s):
WANG Chao-nan1;  LI Wei-hao2;  SHEN Bao-yan3;  LI Jiang-xue1;  ZHANG Jia1;  LIU Xiao-mei2;  ZHANG Fei-fei2; ZHANG Zhi-ping2;  FENG Xiao-yan1;  ZHANG He-qiu1
（1. Medical Institute of Oriental Ocean (Beijing) Ltd. , Beijing 100071,China; 2. Special Survey Division, the Second Hospital of Hebei Medical University, Shijiazhuang 050000,China; 3. Department of Nephropathy Immunology,the First Hospital of Mengcheng, Anhui Bozhou 233500,China)
关键词:
膜性肾病; 磷脂酶A2 受体; 细胞株; 质粒; 免疫荧光方法
分类号:
R692；R392.12
DOI:
10.3969/j.issn.1671-7414.2020.04.001
文献标志码:
A
摘要:
目的 构建表达磷脂酶A2受体（phospholipase A2 receptor, PLA2R）的稳定细胞株并对其在膜性肾病（membranousnephropathy，MN）诊断中的应用进行初步评价。方法 人工合成pIRES-PLA2R 真核表达质粒并稳定转染CHO 细胞，筛选后分别进行RT-PCR,Western 免疫印迹和细胞免疫荧光方法（immunofluorescence assay，IFA 法）鉴定。以构建的稳定细胞株建立PLA2R 自身抗体IFA 法并对2018~2019 年收集的97 例肾病患者血清样本进行初步检测。结果 成功构建表达PLA2R 蛋白的稳定细胞株CHO-PLA2R 并以该细胞株为基础建立PLA2R 自身抗体IFA 法。与ELISA 试剂盒检测结果相比，阴性样本符合率为100%（55/55）；阳性样本符合率为76.2%（32/42）；总体符合率为89.7%（87/97）。Kappa 检验具有一定的一致性（P ＜ 0.05）。结论 成功构建表达PLA2R 蛋白的稳定细胞株，并应用该细胞株建立了检测PLA2R 自身抗体的IFA 检测方法。

参考文献/References:

[1] CATTRAN D. Management of membranous nephropathy[J]. Nephrology, 2001, 5(4): 209-213.
[2] DU BUF-VEREIJKEN P W G, BRANTEN A J W,WETZELS J FM. Idiopathic membranous nephropathy:outline and rationale of a treatment strategy[J].American Journal of Kidney Diseases,2005,46(6):1012-1029.
[3] SEGAL P E,CHOI M J. Recent advances andprognosis in idiopathic membranous nephropathy [J].Advances in Chronic Kidney Disease,2012,19(2): 114-119.
[4] BECH A P, HOFSTRA J M,BRENCHLEY P E,et al.Association of anti-PLA2R antibodies with outcomesafter immunosuppressive therapy in idiopathicmembranous nephropathy[J]. Clinical Journal of theAmerican Society of Nephrology : CJASN,2014,9(8):1386-1392.
[5] SAITO T, IWANO M, MATSUMOTO K, et al.Significance of combined cyclosporine-prednisolonetherapy and cyclosporine blood concentrationmonitoring for idiopathic membranous nephropathywith steroid-resistant nephrotic syndrome:arandomized controlled multicenter trial[J]. Clinical andExperimental Nephrology, 2014, 18(5): 787-794.
[6] 李正东，张任．血清学PLA2R 抗体和THSD7A 抗体检测辅助诊断特发性膜性肾病及对评估疗效和预后的价值研究[J]. 中国煤炭工业医学杂志，2019，22（2）: 149-157． LI Zhengdong, ZHANG Ren. Value of serumPLA2R antibody and THSD7A antibody in efficacyassessment and prognosis of idiopathic membranousnephropathy[J]. Chinese Journal of Coal IndustryMedicine，2019, 22(2): 149-157.
[7] BECH L H, BONEGIO R B, GERARD L, et al. M-Typephospholipase a2 receptor as target antigen in idiopathicmembranous nephropathy[J]. New England Journal ofMedicine, 2009, 361(1): 11-21.
[8] HOFSTRA J M,DEBIEC H,SHORT C D, et al.Antiphospholipase A2 receptor antibody titer andsubclass in idiopathic membranous nephropathy[J].Journal of the American Society of Nephrology :JASN,2012,23(10): 1735-1743.
[9] LI Weihao, GUO Yaping, ZHANG Zhiping, et al.Comparison of 2 anti-PLA2R immunoassays for thediagnosis of primary membranous nephropathy[J].Laboratory Medicine,2018,46（4）: 316-322.
[10] TOTH-MANIKOWSKI S M, BECK L H . PLA2R- andTHSD7A-associated primary membranous nephropathy[M]// Glomerulonephritis. 2016：301-331.
[11] 陈瑞颖，鲁鉴达，谢琼虹，等．成人磷脂酶A2 受体相关特发性膜性肾病的自然病程和治疗反应的影响因素[J]．中华肾脏病杂志,2019,35（1）:1-8. CHEN Ruiying, LU Jianda, XIE Qionghong, et al.The clinical outcome and risk factors of phospholipaseA2 receptor-associated idiopathic membranousnephropathy in adults[J]. Chinese Journal ofNephrology, 2019, 35(1):1-8.
[12] GRANATA F,PETRAROLI A,BOILARD E , etal. Activation of cytokine production by secretedphospholipase A2 in human lung macrophagesexpressing the M-type receptor[J]. Journal ofImmunology ,2005,174(1): 464-474.
[13] SILLIMAN C C, MOORE E E, ZZLLEN G, et al.Presence of the M-type sPLA(2)receptor on neutrophilsand its role in elastase release and adhesion[J].American Journal of Physiology. Cell Physiology,2002, 283(4): C1102-C1113.
[14] HOXHA E , THIELE I, ZAHNER G, et al.Phospholipase A2 receptor autoantibodies and clinicaloutcome in patients with primary membranousnephropathy[J]. Journal of the American Society ofNephrology, 2014,25(6): 1357-1366.
[15] YEO M K,KIM Y H,CHOI DE,et al. The usefulnessof phospholipase A2 receptor and IgG4 detection indifferentiation primary membranous nephropathy fromsecondary membranous nephropathy in renal biopsy[J]. Applied Immunohistochemistry & MolecularMorphology, 2018,26(8):591-598.
[16] 唐琦，邬碧波，李林，等．特发性膜性肾病血清抗磷脂酶A2 受体抗体与免疫治疗关系的研究[J]．药学服务与研究，2019,19（3）:180-184． TANG Qi, WU Bibo, LI Lin, et al. Relationshipbetween serum anti-phospholipase A2 receptorantibodies and immunotherapy of the patients withidiopathic membranous nephropathy[J]. PharmaceuticalCare and Research, 2019,19(3):180-184.
[17] 伍小香，文思，朱雪婧，等．肾组织磷脂酶A2 受体和血清抗磷脂酶A2 受体抗体在膜性肾病中的诊断价值[J]. 中南大学学报（医学版），2017,42（4）:395-399． WU Xiaoxiang, WEN Si, ZHU Xuejing, et al.Diagnostic value of renal phospholipase A2 receptorand serum anti-phospholipase A2 receptor antibody inmembranous nephropathy [J]. Journal of Central SouthUniversity(Medical Science) ,2017,42（4）:395-399．[18] DAI Huanzi,ZHANG Huhai,HE Yani. Diagnosticaccuracy of PLA2R autoantibodies and glomerularstaining for the differentiation of idiopathic andsecondary membranous nephropathy: an updated metaanalysis[J]. Scientific Reports,2015,5(1):8803.
[19] RADICE A, PIERUZZI F, TREZZI B, et al. Diagnosticspecificity of autoantibodies to M-type phospholipaseA2 receptor in differentiating idiopathic membranousnephropathy from secondary forms and otherglomerular disease[J]. Journal of Nephrology, 2018,31(2): 271-278.
[20] AKIYAMA S, AKIYAMA M, IMAI E, et al. Prevalenceof anti-phospholipase A2 receptor antibodies inJapanese patients with membranous nephropathy[J].Clin Exp Nephroll, 2015, 19(4): 653-660.

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备注/Memo

备注/Memo:
收稿日期：2020-04-17修回日期：2020-05-06基金项目：河北省卫生计生委，河北省2015 年度医学科学研究重点课题计划（20150673）。作者简介：王超男（1990-），女，硕士研究生，研究方向：诊断试剂研发，E-mail：vanchywang@qq.com。李伟皓（1975-），男，硕士研究生，副主任检验师，研究方向：免疫学检验，E-mail：13722992006@139.com，共同第一作者。通讯作者：冯晓燕（1973-），女，博士，研究员，研究方向：生物标志物筛选及应用，E-mail：xyfeng2002@126.com。
更新日期/Last Update: 2020-10-08