重组人IgE Fc的制备及抗过敏应用

Preparation of the recombinant human IgE Fc and its application against allergic reaction

[目的]考察重组人Ig E Fc的抗过敏反应作用。[方法]建立阴离子交换和疏水层析为主的分离纯化工艺,从CHO细胞培养的上清中获得重组人Ig E Fc样品;利用流式细胞法测定重组人Ig E Fc与表达人FcεRIα的CHO3D10细胞的亲和力;利用表达人FcεRIαEG-RBL2H3的细胞评价重组人Ig E Fc对细胞活化的抑制;在猴体上考察重组人Ig E Fc阻止被动皮肤过敏反应的作用。[结果]制备的重组人Ig E Fc的纯度达到98%;重组人Ig E Fc与CHO3D10细胞的亲和常数为1.40×109(mmol/L)-1是人Ig E的5倍;重组人Ig E Fc的剂量为NP-Ig E的5或6倍时可完全抑制体外细胞活化和体内过敏反应。[结论]重组人Ig E Fc可以抑制Ig E介导的过敏反应,具有新型抗过敏药物开发的前景。

[Objective] To evaluate the potential of the recombinant human Ig E Fc against allergic reaction. [Methods] The recombinant human Ig E Fc expressed by the engineered CHO cells was obtained through anion exchange and hydrophobic chromatography. Its affinity to CHO3 D10 cells expressing human FcεRIα on their surface was analyzed on a flow cytometry. EG-RBL2 H3 cells,the rat basophil cells expressing human FcεRIα,was applied to in vitro evaluate the inhibition of the recombinant human Ig E Fc on the cell activation and the release of histamine mediated by the specific Ig E and allergen,and Ig E Fc was further in vivo evaluated in the passive cutaneous anaphylaxis on monkeys. [Results]The recombinant human Ig E Fc prepared was highly purified,reaching 98% in purity. The flow cytometry showed that the recombinant human IgE Fc had a 5-fold affinity coefficient to CHO3 D10 cells of the human Ig E,at 1. 40 × 10^9（ mmol/L）-1. The recombinant human Ig E Fc could dose dependently inhibit in vitro cell activation and histamine release,and in vivo passive cutaneous anaphylaxis on monkeys at the 5-or 6-fold concentration of NP-Ig E,respectively. [Conclusion] The recombinant human Ig E Fc could competitively inhibit the allergic reaction and has a promising part to play in the treatment of allergic diseases.