Hu Xiaohan, Liu Cuiping, An Jingnan, Shen Yu, Hu Yumin, Jiang Juean, Wu Jian, Zhang Xueguang
1 Jiangsu Province Key Laboratory of Stem Cell Research, Soochow University , Suzhou, China .
2 Jiangsu Institute of Clinical Immunology, the First Affiliated Hospital of Soochow University , Suzhou, China .
Monoclon Antib Immunodiagn Immunother. 2016 Feb;35(1):18-24. doi: 10.1089/mab.2015.0059. Epub 2016 Jan 20.
CD275 (B7-H2, ICOSL), a co-stimulatory molecule of the B7 superfamily, plays a critical role in immune response. In this report, a novel mouse anti-human CD275 monoclonal antibody (MAb) was prepared using hybridoma technology, and immunological characteristics of the MAb were determined. The results showed that the MAb (clone 13D11) was IgG2(κ) and bound specifically to human CD275. By mutual competition, we found that the antibody recognized different epitopes of CD275 antigen compared with commercial antibodies and could block ICOS-CD275 interaction. Crosslinking of CD275 with MAb 13D11 markedly blocked ICOS positive signal and inhibited T cell proliferation and cytokine production. In addition, the 13D11 MAb was suitable for indirect ELISA detection. Thus, the MAb against human CD275 with high specificity and different activity would be useful for further study of this molecule.
CD275（B7-H2，ICOSL）是B7超家族的一种共刺激分子，在免疫反应中起关键作用。在本报告中，利用杂交瘤技术制备了一种新型小鼠抗人CD275单克隆抗体（MAb），并测定了该MAb的免疫学特性。结果表明，该MAb（克隆号13D11）为IgG2(κ)，能特异性结合人CD275。通过相互竞争，我们发现该抗体与市售抗体相比识别CD275抗原的不同表位，并且能够阻断ICOS-CD275相互作用。用MAb 13D11交联CD275可明显阻断ICOS阳性信号，并抑制T细胞增殖和细胞因子产生。此外，13D11 MAb适用于间接ELISA检测。因此，这种具有高特异性和不同活性的抗人CD275 MAb将有助于对该分子的进一步研究。
