免疫和功能性治疗（DeFT）版本的用于抗病毒治疗的长效重组

StephenW

Clin Immunol. 2017 Jan 10;176:31-41. doi: 10.1016/j.clim.2017.01.003. [Epub ahead of print]
De-immunized and Functional Therapeutic (DeFT) versions of a long lasting recombinant alpha interferon for antiviral therapy.Mufarrege EF1, Giorgetti S2, Etcheverrigaray M2, Terry F3, Martin W3, De Groot AS4.
Author information

• 1Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Laboratorio de Cultivos Celulares, FBCB, UNL, Santa Fe, Argentina. Electronic address: [email protected].
• 2Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Laboratorio de Cultivos Celulares, FBCB, UNL, Santa Fe, Argentina.
• 3EpiVax, Inc., Providence, RI, USA.
• 4EpiVax, Inc., Providence, RI, USA; Institute for Immunology and Informatics, University of Rhode Island, RI, USA.
  

AbstractInterferon α (IFN-α) exerts potent antiviral, immunomodulatory, and antiproliferative activity and have proven clinical utility in chronic hepatitis B and C virus infections. However, repeated IFN-α administration induces neutralizing antibodies (NAb) against the therapeutic in a significant number of patients. Associations between IFN-α immunogenicity and loss of efficacy have been described. So as to improve the in vivo biological efficacy of IFN-α, a long lasting hyperglycosylated protein (4N-IFN) derived from IFN-α2b wild type (WT-IFN) was developed. However, in silico analysis performed using established in silico methods revealed that 4N-IFN had more T cell epitopes than WT-IFN. In order to develop a safer and more efficient IFN therapy, we applied the DeFT (De-immunization of Functional Therapeutics) approach to producing functional, de-immunized versions of 4N-IFN. Using the OptiMatrix in silico tool in ISPRI, the 4N-IFN sequence was modified to reduce HLA binding potential of specific T cell epitopes. Following verification of predictions by HLA binding assays, eight modifications were selected and integrated in three variants: 4N-IFN(VAR1), (VAR2) and (VAR3). Two of the three variants (VAR1 and VAR3) retained anti-viral function and demonstrated reduced T-cell immunogenicity in terms of T-cell proliferation and Th1 and Th2 cytokine levels, when compared to controls (commercial NG-IFN (non-glycosylated), PEG-IFN, WT-IFN and 4N-IFN). It was previously demonstrated that N-glycosylation improved IFN-α pharmacokinetic properties. Here, we further reduce immunogenicity as measured in vitro using T cell assays and cytokine profiling by modifying the T cell epitope content of a protein (de-immunizing). Taking into consideration the present results and previously reported immunogenicity data for commercial IFN-α2b variants, 4N-IFN(VAR1) and 4N-IFN-4N(VAR3) appear to be promising candidates for improved IFN-α therapy of HCV and HBV.

Copyright © 2017 Elsevier Inc. All rights reserved.

KEYWORDS: De-immunization; Hepatitis therapy; IFN alpha; IFN-α; Immunogenicity; In silico prediction; T cell epitope; T-cell proliferation assay

PMID:28089609DOI:10.1016/j.clim.2017.01.003

StephenW

Clin Immunol。 2017 Jan 10; 176：31-41。 doi：10.1016 / j.clim.2017.01.003。 [打印前的电子版]
免疫和功能性治疗（DeFT）版本的用于抗病毒治疗的长效重组α干扰素。
Mufarrege EF1，Giorgetti S2，Etcheverrigaray M2，Terry F3，Martin W3，De Groot AS4。
作者信息

    国家农业研究中心（CONICET），Laboratorio de Cultivos Celulares，FBCB，UNL，Santa Fe，Argentina。电子地址：[email protected]。
    2Consejo Nacional de InvestigacionesCientíficasyTécnicas（CONICET），Laboratorio de Cultivos Celulares，FBCB，UNL，Santa Fe，Argentina。
    3EpiVax，Inc.，Providence，RI，USA。
    4EpiVax，Inc.，Providence，RI，USA;美国罗德岛大学免疫学和信息学研究所。

摘要

干扰素α（IFN-α）发挥有效的抗病毒，免疫调节和抗增殖活性，并已被证明在慢性乙型和丙型肝炎病毒感染中具有临床应用价值。然而，重复的IFN-α施用在显着数目的患者中诱导针对治疗剂的中和抗体（NAb）。已经描述了IFN-α免疫原性和功效丧失之间的关联。为了提高IFN-α的体内生物学功效，开发了源自​​IFN-α2b野生型（WT-IFN）的长效的高糖基化蛋白（4N-IFN）。然而，使用建立的计算机模拟方法进行的计算机分析揭示4N-IFN具有比WT-IFN更多的T细胞表位。为了开发更安全和更有效的IFN治疗，我们应用DeFT（功能性治疗的去免疫）方法来产生4N-IFN的功能性，去免疫化版本。使用在ISPRI中的OptiMatrix in silico工具，修饰4N-IFN序列以降低特异性T细胞表位的HLA结合潜力。在通过HLA结合测定验证预测后，选择8个修饰并整合入3个变体：4N-IFN（VAR1），（VAR2）和（VAR3）。当与对照（商业NG-IFN（非糖基化的））相比时，三种变体（VAR1和VAR3）中的两种保留了抗病毒功能并且表现出T-细胞增殖和Th1和Th2细胞因子水平方面的降低的T细胞免疫原性。 ，PEG-IFN，WT-IFN和4N-IFN）。先前证明N-糖基化改善IFN-α药代动力学性质。在这里，我们进一步降低免疫原性，如使用T细胞测定和通过修饰蛋白质的T细胞表位含量（去免疫）的细胞因子谱在体外测量的。考虑到本结果和先前报道的商业IFN-α2b变体的免疫原性数据，4N-IFN（VAR1）和4N-IFN-4N（VAR3）似乎是改善IFN和IFN-α治疗HCV和HBV的有希望的候选物。

版权所有©2017 Elsevier Inc.保留所有权利。
关键词：

免疫接种;肝炎治疗; IFNα; IFN-α;免疫原性;计算机预测; T细胞表位; T细胞增殖测定

PMID：
    28089609
DOI：
    10.1016 / j.clim.2017.01.003