靶向IL-12 p40和TNF-α的双功能抗体可抑制小鼠银屑病发生

发布时间：2018-11-05 12:29

【摘要】：目的构建具有能同时阻断白细胞介素12(IL-12)/IL-23 p40与肿瘤坏死因子α(TNF-α)的双功能抗体,并对其生物学功能进行鉴定,观察其对小鼠银屑病的阻断效果。方法根据已有的阿达木单抗(adalimumab)和优特克单抗(ustekinumab)蛋白序列,设计、构建并制备了全新的双功能抗体Bi AU003、Bi AU022和Bi AU023。采用ELISA检测抗原抗体结合能力;用TNF-α和双功能抗体处理人脐静脉内皮细胞(HUVEC),加入FITC标记的内皮白细胞黏附分子1(ELAM-1)抗体,经流式细胞术检测ELAM-1的表达量;用IL-12和双功能抗体处理经IL-2活化的人外周血单个核细胞(PBMC),并用ELISA检测上清液中γ干扰素(IFN-γ)的量;体外实验采用双功能抗体治疗IL-12和TNF-α诱导的银屑病小鼠,HE染色检测病变皮肤厚度。结果3种双功能抗体对其相应的抗原都有较强的亲和力,能明显抑制ELAM-1蛋白的表达和IFN-γ的分泌,并且明显抑制小鼠的银屑病形成,其效果与现有的抗体药阿达木单抗和优特克单抗相当或更优。结论新构建的3种双功能抗体Bi AU003、Bi AU022和Bi AU023,是TNF-α和IL-12/IL-23 p40的阻断剂,能有效阻断小鼠银屑病形成。
[Abstract]:Objective to construct a bifunctional antibody with simultaneous blocking of interleukin 12 (IL-12) / IL-23 p40 and tumor necrosis factor 伪 (TNF- 伪), and to identify its biological function and to observe its blocking effect on psoriasis in mice. Methods A novel bifunctional antibody, Bi AU003,Bi AU022 and Bi AU023., was designed, constructed and prepared according to the existing Adamumab (adalimumab) and UTC McAb (ustekinumab) protein sequences. Antigen-antibody binding ability was detected by ELISA. Human umbilical vein endothelial cells (HUVEC),) were treated with TNF- 伪 and bifunctional antibodies to FITC labeled endothelial leukocyte adhesion molecule-1 (ELAM-1) antibody. The expression of ELAM-1 was detected by flow cytometry. Human peripheral blood mononuclear cells (PBMC),) activated by IL-2 were treated with IL-12 and bifunctional antibody, and the amount of interferon 纬 (IFN- 纬) in supernatant was detected by ELISA. In vitro, bifunctional antibodies were used to treat psoriasis induced by IL-12 and TNF- 伪. The thickness of diseased skin was detected by HE staining. Results all of the three bifunctional antibodies had strong affinity to the corresponding antigens, and could significantly inhibit the expression of ELAM-1 protein and the secretion of IFN- 纬, as well as the formation of psoriasis in mice. The effect is comparable to or better than that of the existing antibodies adamumab and eutectoma. Conclusion three newly constructed bifunctional antibodies, Bi AU003,Bi AU022 and Bi AU023, are antagonists of TNF- 伪 and IL-12/IL-23 p40, which can effectively block the formation of psoriasis in mice.
【作者单位】：广东医科大学中美肿瘤研究所;深圳市第二人民医院检验科;
【基金】：广东省特支计划项目(4TZ16001)
【分类号】：R392-33

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