全人单克隆抗核抗体的制备和鉴定  

Construction and identification of full human monoclonal antinuclear autoantibody

作　　者：吴静[1] 李洋[2] 韩秋媛 于鑫[4] 考文萍[1] 宋武琦[1] 翟爱霞[1] 李玉军[1] 李爱梅[1] 张庆猛[1] 潘超[1] 张凤民[1] Wu J Kao WP Song WQ Zhai AX Li Y J Li AM Zhang QM Pan C Zhang FM Li Y Han QY Yu X(Department of Microbiology,Harbin Medical University,the Heilongjiang Key Laboratory of Immunity and Infec- tion ,Pathogenic Biology, Wu Lien- Teh Institute, Harbin 150081, China Department of Radiotherapy, the Third Affiliated Hospital of Harbin Medical University, Harbin 150081, China Department of Critical Care Medicine, the Second Affiliated Hospital of Harbin Medical University,Harbin 150081, China Department of Medical Laboratory Medi- cine,Affiliated Zhongshan Hospital of Dalian University, Dalian 116001, China)

机构地区：[1]哈尔滨医科大学微生物学教研室,黑龙江省感染与免疫重点实验室,黑龙江省感染与免疫科技创新团队,伍连德研究所,150081 [2]哈尔滨医科大学附属第三医院放疗科,150081 [3]哈尔滨医科大学附属第二医院重症医学科,150086 [4]大连大学附属中山医院医学检验部,116001

出　　处：《国际免疫学杂志》2017年第1期11-15,共5页International Journal of Immunology

基　　金：基金项目：黑龙江省教育厅科技研究项目（12541492）

摘　　要：目的本研究旨在从系统性红斑狼疮（systemmiclupuserythematosus，SLE）患者外周血淋巴细胞中获得自身抗体，并从中鉴定筛选出抗核抗体。方法利用人B细胞杂交瘤技术制备SLE患者来源的全人单克隆自身抗体，采用酶联免疫吸附实验（enzyme-linkedimmunosorbenkassay，ELISA）方法测定阳性杂交瘤细胞上清抗体浓度，并以HEp-2细胞为底物，采用间接免疫荧光法鉴定筛选出抗核抗体。结果共获得48株稳定分泌IgG型全人单克隆自身抗体的杂交瘤细胞株，其中12种细胞上清抗体浓度大于1500ng／mL，占25％，通过免疫荧光鉴定从中筛选出3种抗核抗体，荧光核型分别为周边型、斑点型和核仁型。结论全人单克隆自身抗体的制备及抗核抗体的筛选鉴定，为开发新型单克隆抗体药物以及研究SLE的致病机制奠定了良好的实验基础。Objective This study is aimed to obtain autoantibodies from the peripheral blood lympho- cytes of systemic lupus erythematosus （SLE） patients, and to screen the antinuclear antibody by indirect immu- nofluorescence identification. Methods The technology of human B lymphocyte hybridoma was used in the present study for the preparation of the full human autoantibody derived from SLE patients. ELISA was per- formed to detect the concentration of the monoclonal antibodies in the positive hybridoma cell culture superna- tant. Indirect assay was performed to identify the antint^clear antibody by using HEp-2 cells as a basal substrate. Results Forty eight strains of full human monoclonal autoantibody secreting hybridoma cells strains were prepared,and the concentration in twelve strains is higher than 1 500 ng/mL with a ratio of 25%. Three types of antinuclear antibody were identified and screened from the full human monoclonal autoanti-body library, and the immunofluorescence pattern was peripheral, speckled, and nuclear, respectively. Conclu- sion The preparation of full human monoclonal autoantibody and identification of antinuclear antibody could provide a experimental basis for the development of new monoclonal antibody drugs and for the study of researc- hing the pathogenesis of SLE.

关 键 词：系统性红斑狼疮 自身抗体 抗核抗体 人B细胞杂交瘤技术 

分 类 号：R392-33[医药卫生—免疫学]