Tim-3抗体的生物学活性及应用研究

发布时间：2018-02-27 12:56

本文关键词： Tim- 抗体单克隆 LD U细胞炎性因子脓毒症　出处：《军事医学》2014年08期 　论文类型：期刊论文

【摘要】：目的制备抗人Tim-3单抗并评价其生物学活性,为干预Tim-3表达失调引发的疾病提供实验基础。方法制备重组人Tim-3蛋白,常规方法免疫BALB/c小鼠,筛选出阳性克隆,并对其识别、中和活性进行体内外验证,探讨其作为免疫干预手段的可行性。结果 1获得一株能够分泌具有较好结合活性的抗人Tim-3单克隆抗体细胞株(克隆号L3D),其分泌的免疫球蛋白亚型为IgG2a;2流式实验结果表明,该抗体能够结合人U937细胞上的Tim-3分子,且与小鼠RAW 264.7细胞上Tim-3有一定的交叉结合活性;3该抗体能够阻断Gal-9分子诱导的人THP1细胞凋亡,具有良好的体外中和活性;4体内注射L3D,显示该抗体能够显著加重脓毒血症模型小鼠病情,提高炎性因子的表达水平,显示出其良好的体内中和活性。结论成功制备一株分泌抗人Tim-3抗体的细胞株,其良好的结合及中和活性为基于Tim-3表达异常相关疾病的干预提供了实验基础。
[Abstract]:Objective to prepare a monoclonal antibody against human Tim-3 and evaluate its biological activity. Methods Recombinant human Tim-3 protein was prepared and immunized BALB/c mice by routine method, and positive clones were screened and recognized. Neutralization activity was verified in vitro and in vivo. Results 1 A monoclonal antibody cell line with good binding activity against human Tim-3 (clone L3DX) was obtained and its immunoglobulin subtype was IgG2aF2. The antibody can bind to the Tim-3 molecule in human U937 cells, and has a certain cross-binding activity with Tim-3 on mouse RAW 264.7 cells. The antibody can block the apoptosis of human THP1 cells induced by Gal-9 molecule. In vivo injection of L3D with a good neutralization activity in vitro showed that the antibody could significantly aggravate the condition of sepsis model mice and increase the expression of inflammatory factors. Conclusion A cell line secreting anti-human Tim-3 antibody was successfully prepared and its good binding and neutralizing activity provided experimental basis for intervention based on abnormal expression of Tim-3.
【作者单位】：军事医学科学院基础医学研究所免疫学研究室;解放军总医院第一附属医院呼吸科;河南大学医学院免疫学教研室;
【基金】：国家973计划资助项目(2013CB530506) 国家自然科学基金资助项目(81072475,81172800,81272320)
【分类号】：R392

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