IL-15:IL-15 receptor alpha superagonist complex: high-level co-expression in recombinant mammalian cells, purification and characterization.
IL-15:IL-15 receptor alpha superagonist complex: high-level co-expression in recombinant mammalian cells, purification and characterization.
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DOI：
10.1016/j.cyto.2011.09.028
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发表时间：
2011-12
期刊：
影响因子：
3.8
通讯作者：
Wong, Hing C.
中科院分区：
文献类型：
--
作者：
Han, Kai-Ping;Zhu, Xiaoyun;Liu, Bai;Jeng, Emily;Kong, Lin;Yovandich, Jason L.;Vyas, Vinay V.;Marcus, Warren D.;Chavaillaz, Pierre-Andre;Romero, Christian A.;Rhode, Peter R.;Wong, Hing C.
关键词：
IL-15是一种很有前途的细胞因子，可通过IL-15受体(IL-15R)α链反式呈现在T细胞和自然杀伤(NK)细胞表面的IL-15Rβγc复合体上。我们以前报道过，在IL-15的第72位氨基酸(N72D)上将天冬酰胺替换为天冬氨酸，与天然分子相比，生物活性提高了4-5倍。在本报告中，我们描述了由IL-15N72D超激动剂和二聚体IL-15Rα寿司结构域-IgG1Fc融合蛋白组成的可溶性复合体(IL-15N72D：IL-15RαSU/Fc)在中国仓鼠卵巢(CHO)细胞中的表达。开发了一种简单但容易扩展的亲和和离子交换层析方法，以高度纯化两个IL-15结合位点完全占据的络合物。通过细胞增殖实验证实了该复合体的免疫刺激作用。单次静脉注射IL-15N72D：IL-15RαSU/Fc后，小鼠外周血中CD8+T细胞和NK细胞显著增加，而IL-15治疗后未见明显升高。药代动力学分析表明，该复合体在小鼠体内的半衰期为25小时，比IL-15的40分钟半衰期要长得多。因此，IL-15N72D：IL-15RαSU/Fc复合体的活性增强可能是由于IL-15N72D与IL-15Rβγc结合活性增加，IL-15RαSU结构域优化了细胞因子的反式递送，细胞因子结构域的二聚性及其体内半衰期比IL-15延长所致。这些发现表明，这种IL-15超激动剂复合体可以作为一种更好的免疫刺激治疗药物。
IL-15, a promising cytokine for treating cancer and viral diseases, is presented in trans by the IL-15 receptor (IL-15R) alpha-chain to the IL-15Rβγc complex displayed on the surface of T cells and natural killer (NK) cells. We previously reported that an asparagine to aspartic acid substitution at amino acid 72 (N72D) of IL-15 provides a 4–5 fold increase in biological activity compared to the native molecule. In this report, we describe Chinese hamster ovary (CHO) cell expression of a soluble complex (IL-15N72D:IL-15RαSu/Fc) consisting of the IL-15 N72D superagonist and a dimeric IL-15Rα sushi domain-IgG1 Fc fusion protein. A simple but readily scalable affinity and ion exchange chromatography method was developed to highly purify the complex having both IL-15 binding sites fully occupied. The immunostimulatory effects of this complex were confirmed using cell proliferation assays. Treatment of mice with a single intravenous dose of IL-15N72D:IL-15RαSu/Fc resulted in a significant increase in CD8+ T cells and NK cells that was not observed following IL-15 treatment. Pharmacokinetic analysis indicated that the complex has a 25-hour half-life in mice which is considerably longer than <40-minute half-life of IL-15. Thus, the enhanced activity of the IL-15N72D:IL-15RαSu/Fc complex is likely the result of the increased binding activity of IL-15N72D to IL-15Rβγc, optimized cytokine trans-presentation by the IL-15RαSu domain, the dimeric nature of the cytokine domain and its increased in vivo half-life compared to IL-15. These findings indicate that this IL-15 superagonist complex could serve as a superior immunostimulatory therapeutic agent.
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影响因子：
5.7
作者：
Bessard, Anne;Sole, Veronique;Jacques, Yannick
通讯作者：
Jacques, Yannick
影响因子：
4.2
作者：
Hughes, MS;Yu, YYL;Morgan, RA
通讯作者：
Morgan, RA
影响因子：
1.6
作者：
Ward, Alison;Anderson, Malcolm;Mallinder, Philip R.
通讯作者：
Mallinder, Philip R.
影响因子：
5.6
作者：
Bouchaud, Gregory;Garrigue-Antar, Laure;Plet, Ariane
通讯作者：
Plet, Ariane
DOI：
10.1073/pnas.0600240103
发表时间：
2006-06-13
影响因子：
11.1
作者：
Rubinstein, Mark P.;Kovar, Marek;Sprent, Jonathan
通讯作者：
Sprent, Jonathan