Diphtheria toxin-based bivalent human IL-2 fusion toxin with improved efficacy for targeting human CD25(+) cells.
Diphtheria toxin-based bivalent human IL-2 fusion toxin with improved efficacy for targeting human CD25(+) cells.
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DOI：
10.1016/j.jim.2014.01.008
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发表时间：
2014-03
影响因子：
2.2
通讯作者：
Wang, Zhirui
中科院分区：
文献类型：
--
作者：
Peraino, Jaclyn Stromp;Zhang, Huiping;Rajasekera, Priyani V.;Wei, Min;Madsen, Joren C.;Sachs, David H.;Huang, Christene A.;Wang, Zhirui
调节性T细胞（Treg）构成抑制免疫应答的主要抑制性细胞群体。因此，Treg已被证明是诱导移植耐受、保护免受自身免疫性疾病和预防有效的抗肿瘤免疫反应发展的关键参与者。Treg表达高水平的高亲和力白细胞介素-2受体（IL-2 R），由IL-2 R α（CD 25）、IL-2 R β（CD 122）和共同的γ链（CD 132）组成。能够在体内耗尽Treg的有效试剂将促进更好的癌症治疗，并允许对Treg在移植耐受性和自身免疫性疾病的发展中的作用的机制研究。在这项研究中，我们开发了一种新的二价人IL-2融合毒素沿着Ontak®样单价人IL-2融合毒素，并在体外比较了这些试剂的功能能力。在这里，我们表明，基因串联连接两个人IL-2结构域，从而产生二价融合毒素，结果在体外靶向人CD 25+细胞的能力显着提高。通过流式细胞术的结合分析显示，二价人IL-2融合毒素对人CD 25+细胞具有显著增加的亲和力。体外功能分析表明，与单价人IL-2融合毒素相比，二价同种型在抑制人CD 25+细胞中的细胞增殖和蛋白质合成方面具有约2log的增加的效力。此外，我们进行了两次抑制测定，以验证融合毒素通过融合毒素的人IL-2结构域与细胞表面上的人IL-2受体的结合特异性靶向细胞。这些结果表明：1）通过流式细胞术，单价和二价人IL-2融合毒素都能够阻断生物素化的人IL-2与人CD 25的结合;和2）人IL-2在体外阻断融合毒素抑制蛋白质合成和细胞增殖，从而证实人IL-2融合毒素通过与人IL-2受体结合特异性靶向细胞。我们相信，二价人IL-2融合毒素将是比目前临床上使用的单价融合毒素（地尼白介素diftitox，Ontak®）更有效的，因此更适合于体内耗尽Treg的试剂。
Regulatory T cells (Treg) constitute a major inhibitory cell population which suppresses immune responses. Thus, Treg have proven to be key players in the induction of transplantation tolerance, protection from autoimmune disease and prevention of the development of effective anti-tumor immune reactions. Treg express high levels of the high affinity interleukin-2 receptor (IL-2R) consisting of IL-2Rα (CD25) together with IL-2Rβ (CD122) and the common γ-chain (CD132). An effective reagent capable of depleting Treg in vivo would facilitate better cancer treatment and allow mechanistic studies of the role of Treg in transplantation tolerance and the development of autoimmune disease. In this study, we have developed a novel bivalent human IL-2 fusion toxin along with an Ontak®-like monovalent human IL-2 fusion toxin and compared the functional ability of these reagents in vitro. Here we show that genetically linking two human IL-2 domains in tandem, thereby generating a bivalent fusion toxin, results in significantly improved capacity in targeting human CD25+ cells in vitro. Binding analysis by flow cytometry showed that the bivalent human IL-2 fusion toxin has notably increased affinity for human CD25+ cells. In vitro functional analysis demonstrated that the bivalent isoform has an increased potency of approximately 2 logs in inhibiting cellular proliferation and protein synthesis in human CD25+ cells compared to the monovalent human IL-2 fusion toxin. Additionally, we performed two inhibition assays in order to verify that the fusion toxins target the cells specifically through binding of the human IL-2 domain of the fusion toxin to the human IL-2 receptor on the cell surface. These results demonstrated that 1) both monovalent and bivalent human IL-2 fusion toxins are capable of blocking the binding of biotinylated human IL-2 to human CD25 by flow cytometry; and 2) human IL-2 blocked the fusion toxins from inhibiting protein synthesis and cellular proliferation in vitro, thus confirming that the human IL-2 fusion toxins target the cells specifically through binding to the human IL-2 receptor. We believe that the bivalent human IL-2 fusion toxin will be a more potent, and therefore, more optimal agent than the current clinically-used monovalent fusion toxin (denileukin diftitox, Ontak®) for in vivo depletion of Treg.
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影响因子：
4.2
作者：
Murphy JR
通讯作者：
Murphy JR
DOI：
10.1111/j.1600-0897.2005.00330.x
发表时间：
2005-12-01
影响因子：
3.6
作者：
Barnett, B;Kryczek, I;Curiel, TJ
通讯作者：
Curiel, TJ
调节性T细胞在类风湿关节炎和抗TNFALPHA治疗中的逆转功能受损。
影响因子：
15.3
作者：
Ehrenstein, MR;Evans, JG;Singh, A;Moore, S;Warnes, G;Isenberg, DA;Mauri, C
通讯作者：
Mauri, C
影响因子：
7.7
作者：
Lindley, S;Dayan, CM;Tree, TIM
通讯作者：
Tree, TIM
影响因子：
4.4
作者：
Yamada, Yohei;Aoyama, Akihiro;Kawai, Tatsuo
通讯作者：
Kawai, Tatsuo