多重细胞因子检测试剂的开发及其在骨髓瘤中的应用  

Development of multiplex cytokine detection reagents and its application in myeloma

作　　者：彭火英 张之尧 郑向君[4] 韦鹏[4] 胡迪 陈文明[3] 于晓波[2] PENG Huoying;ZHANG Zhiyao;ZHENG Xiangjun;WEI Peng;HU Di;CHEN Wenming;YU Xiaobo(School of Basic Medicine,Qingdao University,Qingdao 266071,China;National Center for Protein Sciences-Beijing(PHOENIX Center),State Key Laboratory of Medical Proteomics,Beijing Proteome Research Center,Beijing 102206,China;Beijing Chao-Yang Hospital,Capital Medical University,Beijing 100020,China;Beijing Sunbio Biotech Co.,Ltd.,Beijing 100176,China;Proteomics Era Medical Co.,Ltd.(Beijing),Beijing 102206,China)

机构地区：[1]青岛大学基础医学院,青岛266071 [2]国家蛋白质科学中心-北京(凤凰中心),医学蛋白质组全国重点实验室,北京蛋白质组研究中心,北京102206 [3]首都医科大学附属北京朝阳医院,北京100020 [4]北京沙东生物技术有限公司,北京100176 [5]旦生(北京)医学科技有限责任公司,北京102206

出　　处：《中国免疫学杂志》2024年第9期1944-1950,共7页Chinese Journal of Immunology

基　　金：国家科技部项目(2021YFA1301604);国家重点基础研究计划项目(2018ZX09733003);蛋白质组学国家重点实验室开放课题项目(SKLP-O202007);北京市自然科学基金面上项目(M21003);广东省科技厅重点领域研发计划项目(2020B1111100006)。

摘　　要：目的:开发多重细胞因子检测试剂,分析复发难治性多发性骨髓瘤(RRMM)的细胞因子、血管生成、骨重塑蛋白表达水平变化。方法:采用多重微球流式荧光免疫技术建立多重细胞因子定量检测试剂,并应用于55例RRMM患者和22例健康对照血清样本的测定,分析患者细胞因子、血管生成、骨重塑蛋白表达水平及其与临床特征的相关性。结果:本研究成功开发了十重细胞因子免疫分析检测试剂,平均灵敏度为7.1 pg/ml,平均回收率为97.4%,平均批内变异系数为4.8%,平均批间变异系数为9.0%。此外,RRMM样本结果发现IL-2、IL-17、DKK1、RANKL、OPG水平与IgG单克隆蛋白水平呈正相关,TIMP1与IgG、IgA单克隆蛋白水平呈正相关。结论:本研究开发出10种具有较高灵敏度和特异性的细胞因子检测试剂,并发现IL-2、IL-17、DKK1、RANKL、OPG、TIMP1在RRMM中具有追踪疾病进程的潜在价值。建立的多重细胞因子试剂开发流程对今后拓展蛋白标志物多重检测试剂的研发和应用具有重要参考价值。Objective:To develop multiplex cytokine detection reagents to analyze expression levels of cytokines,angiogenesis and bone remodeling proteins in relapse/refractory multiple myeloma(RRMM).Methods:Multiplex bead-based immunoassay by flow cytometry was used to develop quantitative detection reagents of multiplex cytokines,which were applied to detect serum samples from 55 RRMM patients and 22 healthy controls.Expression levels of cytokines,angiogenesis,and bone remodeling proteins in patients,and their correlation with clinical characteristics were analyzed.Results:Detection reagents of 10-plex cytokine immunoassay were successfully developed in this study,with average sensitivity of 7.1 pg/ml,average recovery rate of 97.4%,average intra-assay CV of 4.8%,and average inter-assay CV of 9.0%.In addition,results of RRMM samples found that levels of IL-2,IL-17,DKK1,RANKL and OPG were positively correlated with the level of IgG monoclonal protein,and TIMP1 was positively correlated with levels of IgG and IgA monoclonal protein.Conclusion:In this study,ten kinds of cytokine detection reagents with high sensitivity and specificity are developed,and we found that IL-2,IL-17,DKK1,RANKL,OPG and TIMP1 have potential value in tracking disease progression in RRMM.The established development process of multiplex cytokine reagents has important reference significance for expanding the development and application of multiplex detection reagents for protein markers in the future.

关 键 词：多重微球流式荧光免疫技术 复发难治性多发性骨髓瘤 细胞因子 血管生成 骨重塑 

分 类 号：R392-33[医药卫生—免疫学]