Novel human interleukin-15 agonists.
Novel human interleukin-15 agonists.
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DOI：
10.4049/jimmunol.0901244
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发表时间：
2009-09-15
期刊：
影响因子：
--
通讯作者：
Wong HC
中科院分区：
文献类型：
--
作者：
Zhu X;Marcus WD;Xu W;Lee HI;Han K;Egan JO;Yovandich JL;Rhode PR;Wong HC
IL-15是一种免疫刺激性细胞因子，通过IL-15受体α链反式呈递至T细胞和NK细胞表面上展示的共有IL-2/IL-15R β和共有γ链。为了进一步确定该细胞因子的功能重要区域，对通过定点诱变产生的人IL-15突变蛋白进行活性和结合研究。发现位于螺旋C末端的位置72处的天冬酰胺残基的氨基酸取代提供部分激动剂和超激动剂活性，其中各种非保守取代提供增强的活性。特别地，基于用携带人IL-15R β和共同γ链的细胞进行的增殖测定，与天然分子相比，N72D取代提供IL-15突变蛋白的生物活性增加4 - 5倍。IL-15N72D突变蛋白通过改善与人IL-15R β链的结合能力而表现出超激动剂活性。然而，在表达小鼠IL-15R α-IL-15R β-γ c复合物的细胞中未观察到IL-15N72D的增强效力，表明该效应对人IL-15受体具有特异性。与野生型IL-15相比，IL-15N72D增强的生物活性与Jak1和Stat5更强的磷酸化以及更好的抗凋亡活性相关。IL-15 N72D超激动剂活性在与单链T细胞受体结构域连接以产生肿瘤特异性融合蛋白时也被保留。因此，人IL-15超激动剂突变蛋白和融合物可以创造机会来构建具有临床实用性的更有效的免疫抑制剂。
IL-15 is an immunostimulatory cytokine trans-presented with the IL-15 receptor α chain to the shared IL-2/IL-15Rβ and common γ chains displayed on the surface of T cells and NK cells. To further define the functionally important regions of this cytokine, activity and binding studies were conducted on human IL-15 muteins generated by site-directed mutagenesis. Amino acid substitutions of the asparagine residue at position 72, which is located at the end of helix C, were found to provide both partial agonist and superagonist activity, with various non-conservative substitutions providing enhanced activity. Particularly, the N72D substitution provided a 4–5 fold increased in biological activity of the IL-15 mutein compared to the native molecule based on proliferations assays with cells bearing human IL-15Rβ and common γ chains. The IL-15N72D mutein exhibited superagonist activity through improved binding ability to the human IL-15Rβ chain. However, the enhanced potency of IL-15N72D was not observed with cells expressing the mouse IL-15Rα-IL-15Rβ-γc complex suggesting that this effect is specific to human IL-15 receptor. The enhanced biological activity of IL-15N72D was associated with more intense phosphorylation of Jak1 and Stat5 and better anti-apoptotic activity compared to the wild-type IL-15. IL-15N72D superagonist activity was also preserved when linked to a single-chain T cell receptor domain to generate a tumor-specific fusion protein. Thus, the human IL-15 superagonist muteins and fusions may create opportunities to construct more efficacious immunotherapeutic agents with clinical utility.
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DOI：
10.1073/pnas.91.11.4940
发表时间：
1994-05-24
影响因子：
11.1
作者：
BAMFORD, RN;GRANT, AJ;WALDMANN, TA
通讯作者：
WALDMANN, TA
影响因子：
30.5
作者：
Chirifu, Mami;Hayashi, Chiharu;Ikemizu, Shinji
通讯作者：
Ikemizu, Shinji
影响因子：
4.2
作者：
Hughes, MS;Yu, YYL;Morgan, RA
通讯作者：
Morgan, RA
影响因子：
3.8
作者：
Eisenman, J;Ahdieh, M;Park, LS
通讯作者：
Park, LS
影响因子：
5.6
作者：
Bouchaud, Gregory;Garrigue-Antar, Laure;Plet, Ariane
通讯作者：
Plet, Ariane