Exploiting a natural conformational switch to engineer an interleukin-2 'superkine'.
Exploiting a natural conformational switch to engineer an interleukin-2 'superkine'.
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DOI：
10.1038/nature10975
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发表时间：
2012-03-25
期刊：
影响因子：
64.8
通讯作者：
Garcia, K. Christopher
中科院分区：
文献类型：
--
作者：
Levin, Aron M.;Bates, Darren L.;Ring, Aaron M.;Krieg, Carsten;Lin, Jack T.;Su, Leon;Moraga, Ignacio;Raeber, Miro E.;Bowman, Gregory R.;Novick, Paul;Pande, Vijay S.;Fathman, C. Garrison;Boyman, Onur;Garcia, K. Christopher
免疫刺激细胞因子白细胞介素-2（IL-2）是一种广泛的白细胞生长因子，包括T细胞和自然杀伤（NK）细胞。已经投入相当大的努力使用IL-2作为从AIDS到癌症的各种免疫病症的治疗剂。然而，不良反应限制了其在临床上的应用。在活化的T细胞上，IL-2通过由IL-2、IL-2 R α（称为CD 25）、IL-2 R β和γc组成的四元“高亲和力”受体复合物发出信号。幼稚T细胞仅表达低密度的IL-2 R β和γc，因此对IL-2相对不敏感，但在CD 25表达后获得敏感性，其捕获细胞因子并将其呈递给IL-2 R β和γc。在这里，使用体外进化，我们通过工程改造IL-2“超级因子”（称为super-2），使其对IL-2 R β的结合亲和力增加，从而消除了IL-2对CD 25表达的功能需求。游离和受体结合形式的super-2的晶体结构表明，进化的突变主要位于细胞因子的核心，分子动力学模拟表明，进化的突变使IL-2（包括IL-2 R β结合位点中的柔性螺旋）稳定为优化的受体结合构象，类似于与CD 25结合时的构象。super-2中进化的突变通过引发STAT 5的有效磷酸化和与CD 25表达无关的T细胞的剧烈增殖来概括CD 25的功能作用。与IL-2相比，super-2诱导细胞毒性T细胞的上级扩增，导致体内抗肿瘤反应的改善，并引起调节性T细胞的比例较少的扩增和减少肺水肿。总的来说，我们表明，体外进化模拟了CD 25在增强IL-2效力和调节靶细胞特异性方面的功能作用，这对免疫治疗有影响。
The immunostimulatory cytokine interleukin-2 (IL-2) is a growth factor for a wide range of leukocytes, including T cells and natural killer (NK) cells. Considerable effort has been invested using IL-2 as a therapeutic agent for a variety of immune disorders ranging from AIDS to cancer. However, adverse effects have limited its use in the clinic. On activated T cells, IL-2 signals through a quaternary “high affinity” receptor complex consisting of IL-2, IL-2Rα (termed CD25), IL-2Rβ, and γc. Naïve T cells express only a low density of IL-2Rβ and γc, and are therefore relatively insensitive to IL-2, but acquire sensitivity after CD25 expression, which captures the cytokine and presents it to IL-2Rβ andγc. Here, using in vitro evolution, we eliminated IL-2’s functional requirement for CD25 expression by engineering an IL-2 “superkine” (termed super-2) with increased binding affinity for IL-2Rβ. Crystal structures of super-2 in free and receptor-bound forms showed that the evolved mutations are principally in the core of the cytokine, and molecular dynamics simulations indicated that the evolved mutations stabilized IL-2, including a flexible helix in the IL-2Rβ binding site, into an optimized receptor-binding conformation resembling that when bound to CD25. The evolved mutations in super-2 recapitulated the functional role of CD25 by eliciting potent phosphorylation of STAT5 and vigorous proliferation T cells irrespective of CD25 expression. Compared to IL-2, super-2 induced superior expansion of cytotoxic T cells, leading to improved anti-tumor responses in vivo, and elicited proportionally less expansion of T regulatory cells and reduced pulmonary edema. Collectively, we show that in vitro evolution has mimicked the functional role of CD25 in enhancing IL-2 potency and regulating target cell specificity, which has implications for immunotherapy.
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影响因子：
44.1
作者：
通讯作者：
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影响因子：
5.5
作者：
Hess, Berk;Kutzner, Carsten;Lindahl, Erik
通讯作者：
Lindahl, Erik
DOI：
10.1107/s0907444904019158
发表时间：
2004-12-01
影响因子：
2.2
作者：
Emsley, P;Cowtan, K
通讯作者：
Cowtan, K
DOI：
10.1073/pnas.0909384107
发表时间：
2010-02-02
影响因子：
11.1
作者：
Letourneau, Sven;van Leeuwen, Ester M. M.;Boyman, Onur
通讯作者：
Boyman, Onur
影响因子：
5.5
作者：
Hess, Berk
通讯作者：
Hess, Berk