Establishment of a novel double-monoclonal antibody sandwich enzyme-linked immunosorbent assay (ELISA): tool for human B7-H4 detection in autoimmune diseases
Establishment of a novel double-monoclonal antibody sandwich enzyme-linked immunosorbent assay (ELISA): tool for human B7-H4 detection in autoimmune diseases
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新型双单克隆抗体夹心酶联免疫吸附测定（ELISA）的建立：自身免疫性疾病中人 B7-H4 检测的工具
DOI：
10.1111/cei.13610
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发表时间：
2021-06-06
影响因子：
4.6
通讯作者：
Liu, Cuiping
中科院分区：
文献类型：
--
作者：
Ding, Sisi;Zhou, Hengxin;Liu, Cuiping
B7-H4是一种免疫调节蛋白，通过抑制T细胞增殖和细胞因子产生而发挥抑制作用。然而，可溶性B7-H4（sB 7-H4）在自身免疫性疾病中的意义尚不清楚。在我们的研究中，我们开发了两种新的小鼠抗人B7-H4单克隆抗体（mAb）（克隆8D 4和7 E1），用于流式细胞术，免疫印迹和免疫荧光。我们将7 E1鉴定为具有拮抗活性的功能性抗体，其可以促进T细胞增殖并调节细胞因子的产生。此外，基于不同抗原表位的特异性，我们建立了一种新的酶联免疫吸附试验（ELISA），可以灵敏、特异地检测sB 7-H4。使用该ELISA试剂盒，在高比例的自身免疫性疾病患者中观察到sB 7-H4。我们发现系统性红斑狼疮（SLE）、I型糖尿病（T1 D）和格雷夫斯病（GD）患者的sB 7-H4水平显着升高。总之，人血清中的sB 7-H4不仅被认为是T细胞活化的调节剂，而且也可能是自身免疫性疾病的诊断标志物。
B7-H4, one of the immunoregulatory proteins, plays an inhibitory role by inhibiting T cell proliferation and cytokine production. Nevertheless, the significance of soluble B7-H4 (sB7-H4) in autoimmune diseases is unclear. In our study, we developed two novel mouse anti-human B7-H4 monoclonal antibodies (mAbs) (clones 8D4 and 7E1) with utilities for flow cytometry, immunoblotting and immunofluorescence. We characterized 7E1 as a functional antibody with antagonistic activity, which could promote T cell proliferation and regulate cytokine production. Furthermore, based on the different epitope specificities, we established a novel enzyme-linked immunosorbent assay (ELISA) which could detect sB7-H4 sensitively and specifically. Using this ELISA kit, sB7-H4 was observed in a high proportion of autoimmune diseases patients. We found that the levels of sB7-H4 were significantly higher in patients with systemic lupus erythematosus (SLE), type I diabetes (T1D) and Graves' disease (GD). Together, sB7-H4 in human serum is regarded not only as a regulator of T cell activation but may also be a diagnostic marker of autoimmune diseases.