Development of a novel monoclonal antibody to B7-H4: characterization and biological activity.
Development of a novel monoclonal antibody to B7-H4: characterization and biological activity.
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DOI：
10.1186/2047-783x-16-7-295
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发表时间：
2011-07-25
影响因子：
4.2
通讯作者：
Yao HP
中科院分区：
文献类型：
--
作者：
Qian Y;Shen L;Xu C;Wu Z;Brockmeyer NH;Altmeyer P;Wu N;Yao HP
B7-H4是免疫调节受体B7家族的成员，可能参与细胞介导的免疫的负调节。B7-H4在某些肿瘤中存在异常表达，在肿瘤的发生、发展中起一定作用。本研究的目的是阐明B7-H4的功能和结构特性。我们用表达外源性B7-H4的3 T3-mB 7-H4细胞免疫Balb/c小鼠，制备了抗B7-H4胞外区的单克隆抗体（mAb）。建立了稳定的杂交瘤细胞株。通过酶联免疫吸附试验（ELISA）、免疫沉淀试验（IP）、免疫印迹试验（Western blotting）、免疫组化试验（IHC）等方法对mAb的特性进行分析，并对mAb的生物学活性进行检测。ELISA、IP和western blotting分析表明，mAb特异性识别B7-H4。此外，流式细胞术表明，该单克隆抗体表现出良好的反应性时，适用于白血病细胞。IHC染色显示，当应用于某些肿瘤组织时，mAb以弥漫性质膜或细胞质模式染色。初步生物学活性测定结果表明，该mAb能有效抑制B7-H4对T细胞的抑制作用，并能促进T细胞的增殖和IL-2、IL-4、IL-10和IFN-γ的分泌。该单克隆抗体为进一步研究B7-H4的功能奠定了基础。
B7-H4, a member of the B7 family of immunoregulatory receptors, may participate in the negative regulation of cell-mediated immunity. Aberrant B7-H4 expression is detected in some tumors and it plays a role in the occurrence and development of tumors. The aim of this study was to elucidate the functional and structural properties of B7-H4. We developed a monoclonal antibody (mAb) against the extracellular domain of B7-H4 through immunization of Balb/c mice with 3T3-mB7-H4 cells which expressed extrinsic B7-H4. A stable hybridoma cell line was established. Then, we analysised the characterization of the mAb through Enzyme linked immunosorbent assay (ELISA), Immunoprecipitation (IP), western blotting, Immunohistochemical (IHC), and tested the biological activity of the mAb. ELISA, IP, and western blotting analyses indicated that the mAb specifically recognized B7-H4. In addition, flow cytometry demonstrated that the mAb exhibits excellent reactivity when applied to leukemic cells. IHC staining revealed that the mAb stained in a predominantly diffuse plasmalemmal or cytoplasmic pattern when applied to certain tumor tissues. The preliminary results of the mAb's biological activity showed that the mAb could effectively inhibit the function of B7-H4 in the inhibition of T cell, while promotingg the growth of T cells and the secretion of Interleukin-2 (lL-2), Interleukin-4 (IL-4), Interleukin10 (IL-10) and Interferon-γ (IFN-γ). This mAb will be a valuable tool for the further investigation of B7-H4 function.
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DOI：
10.1111/igc.0b013e3181ad0fa2
发表时间：
2009-12-01
影响因子：
4.8
作者：
Cheng, Lei;Jiang, Jie;Kong, Beihua
通讯作者：
Kong, Beihua
影响因子：
4.7
作者：
Tringler, B;Liu, WH;Shroyer, KR
通讯作者：
Shroyer, KR
影响因子：
3.6
作者：
Yao, Hang-Ping;Luo, Yu-Lan;Wang, Ming-Hai
通讯作者：
Wang, Ming-Hai
DOI：
10.1073/pnas.0709802104
发表时间：
2007-12-04
影响因子：
11.1
作者：
Zang, Xingxing;Thompson, R. Houston;Allison, James P.
通讯作者：
Allison, James P.
DOI：
10.1084/jem.20050930
发表时间：
2006-04-17
期刊：
The Journal of experimental medicine
影响因子：
--
作者：
Kryczek I;Zou L;Rodriguez P;Zhu G;Wei S;Mottram P;Brumlik M;Cheng P;Curiel T;Myers L;Lackner A;Alvarez X;Ochoa A;Chen L;Zou W
通讯作者：
Zou W