摘要
IL-1β可介导炎症病理,阻断IL-1活性具有重要抗炎治疗作用。本研究通过构建猪IL-1β原核表达质粒,进行大肠杆菌表达和蛋白纯化,制备重组蛋白。采用细胞杂交瘤技术制备获得4株稳定分泌IL-1β单克隆抗体杂交瘤细胞株(1E2、2H3、5H3及7E7);4株单抗腹水ELISA效价高达1∶1024000;5H3单抗识别的抗原表位为149 DLKREVVFCM 158;1E2、2H3和7E7单抗所识别的抗原表位为202 KRYPKRD 208。脂多糖(LPS)小鼠炎症模型抗炎试验结果显示,相对LPS对照组,IL-1β单抗处理组小鼠血清NO、TNF-α、IL-6和IL-1β含量及临床症状评分显著降低,表明4株单抗均有较好抗炎效果,其中1E2单抗抗炎作用最优,为猪IL-1β阻断药物研究奠定了重要基础。
IL-1βmediates inflammatory pathology,and blocking IL-1 activity has important anti-inflammatory effects.In this study,recombinant protein was prepared by constructing porcine IL-1βprokaryotic expression plasmid,expression by Escherichia coli and protein purification.Four hybridoma cell lines(1E2,2H3,5H3 and 7E7)with stable IL-1βmonoclonal antibody secretion were prepared by cell hybridoma technique.The ELISA titer of the 4 mAbs in ascites were up to 1:1024000.The epitope of 5H3 mAb was 149 DLKREVVFCM 158.The epitope recognized by 1E2,2H3 and 7E7 mAb was 202 KRYPKRD 208.The anti-inflammatory test results of lipopolysaccharide(LPS)mice inflammation model showed that compared with LPS control group,serum NO,TNF-α,IL-6 and IL-1βcontents and clinical symptom score of mice treated with IL-1βmonoclonal antibody were significantly reduced,indicating that all the four strains of monoclonal antibody had better anti-inflammatory effect,and 1E2 monoclonal antibody had the best anti-inflammatory effect.It laid an important foundation for the study of porcine IL-1βblocking drugs.
作者
李璠
孙海凤
孙萌
高雁怩
孙杨杨
张路捷
白娟
姜平
LI Fan;SUN Haifeng;SUN Meng;GAO Yanxiao;SUN Yangyang;ZHANG Lujie;BAI Juan;JIANG Ping(Key Laboratory of Animal Bacteriology of Ministry of Agriculture and Rural Affairs,Nanjing Agricultural University,Nanjing 210095,China)
出处
《畜牧兽医学报》
北大核心
2025年第2期890-899,共10页
ACTA VETERINARIA ET ZOOTECHNICA SINICA
基金
国家自然科学基金项目(32230103)
国家生猪产业技术体系专项(CARS-35)
江苏省自主创新研发基金项目(CX(22)1011)。
