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目的分析人源可诱导共刺激分子(ICOS)可溶性融合蛋白(ICOSIg)与鼠源不成熟树突状细胞(DCs)是否发生特异性结合及其一般生物学特性。方法通过流式细胞术结合特异性抗体检测了解ICOSIg与不成熟DCs的结合作用;以AnnexinⅤ/PI双染色、活细胞染料CSFE和CCK-8研究ICOSIg对DCs的细胞毒性作用;以[3 H]掺入法研究ICOSIg对于混合淋巴细胞反应的阻断作用。结果 ICOSIg可结合小鼠DCs表面的ICOSL,ICOSIg不引起DCs早期和晚期凋亡,不影响DCs细胞的增殖,可以抑制不同基因小鼠脾细胞的混合淋巴细胞反应。结论本实验室构建的体外表达的ICOSIg具有较强的生物学活性,对鼠DCs无明显的生物学毒性作用,首次从实验角度证实人源ICOSIg可以特异性结合小鼠不成熟DCs表面配体ICOSL,可以用于进一步探讨ICOSL/ICOS共刺激通路的免疫作用。
Objective To analyze whether specific binding and general biological characteristics of ICOSIg and murine immature dendritic cells (DCs) are analyzed. Methods The binding of ICOSIg to immature DCs was detected by flow cytometry and specific antibodies. The cytotoxicity of ICOSIg on DCs was studied by AnnexinⅤ / PI double staining and viable cells dye CSFE and CCK-8. [3H] Binding studies ICOSIg on the mixed lymphocyte reaction blockade. Results ICOSIg could bind ICOSL on the surface of mouse DCs. ICOSIg did not induce early and late apoptosis of DCs, did not affect the proliferation of DCs, and inhibited the mixed lymphocyte reaction of spleen cells from different gene mice. CONCLUSION ICOSIg constructed in our laboratory has strong biological activity and no obvious biological toxicity to DCs. It is the first time from experimental point of view that human ICOSIg can specifically bind ICOSL of mouse immature DCs surface ligand , Can be used to further explore the ICOSL / ICOS costimulation pathway.