Potent immunogenicity and efficacy of a universal influenza vaccine candidate comprising a recombinant fusion protein linking influenza M2e to the TLR5 ligand flagellin
Potent immunogenicity and efficacy of a universal influenza vaccine candidate comprising a recombinant fusion protein linking influenza M2e to the TLR5 ligand flagellin
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DOI：
10.1016/j.vaccine.2007.10.062
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发表时间：
2008-01-10
期刊：
影响因子：
5.5
通讯作者：
Powell, T. J.
中科院分区：
文献类型：
--
作者：
Huleatt, James W.;Nakaar, Valerian;Powell, T. J.
Toll 样受体 (TLR) 家族成员对特定病原体相关分子模式 (PAMP) 的识别对于适应性免疫反应的激活至关重要。因此，在没有佐剂或复杂制剂的情况下，将 PAMP 掺入疫苗中应该会产生更有效的保护性抗原特异性反应。在这里，我们描述了一种甲型流感疫苗，它对血凝素和神经氨酸酶的遗传不稳定性具有抵抗力，并且包含先天免疫反应的触发因素以增强免疫原性和功效。包含与保守流感基质蛋白 M2 (M2e) 的胞外域的四个串联拷贝融合的 TLR5 配体鞭毛蛋白的重组蛋白在大肠杆菌中表达并纯化至同质。这种蛋白质 STF2.4xM2e 保留了 TLR5 活性，并显示出由单克隆抗体 14C2 定义的 M2e 保护性表位。使用水性缓冲液中的 STF2.4xM2e（不含佐剂或其他制剂添加剂）免疫小鼠，产生了有效的 M2e 特异性抗体反应，其在数量和质量上均优于使用明矾中递送的 M2e 肽观察到的反应。抗体反应取决于抗原与鞭毛蛋白的物理连接，并识别单克隆抗体 14C2 定义的表位，该表位已被证明可以保护小鼠免受甲型流感病毒的攻击。此外，用每只小鼠0.3μg剂量的STF2.4xM2e进行免疫可以保护小鼠免受甲型流感病毒的致命攻击，并显着减少体重减轻和临床症状。这些数据表明，特定 TLR 配体与流感 M2e 的连接产生了一种候选疫苗，该候选疫苗为针对多种甲型流感病毒株的广泛保护提供了重要希望。 (c) 2007 Elsevier Ltd. 保留所有权利。
The recognition of specific pathogen associated molecular patterns (PAMPs) by members of the Toll-like receptor (TLR) family is critical for the activation of the adaptive immune response. Thus, incorporation of PAMPs into vaccines should result in more potent, protective antigen-specific responses in the absence of adjuvants or complex formulations. Here we describe an influenza A vaccine that is refractory to the genetic instability of hemagglutinin and neuraminidase and includes a trigger of the innate immune response to enhance immunogenicity and efficacy. A recombinant protein comprising the TLR5 ligand flagellin fused to four tandem copies of the ectodomain of the conserved influenza matrix protein M2 (M2e) was expressed in Escherichia coli and purified to homogeneity. This protein, STF2.4xM2e, retained TLR5 activity and displayed the protective epitope of M2e defined by a monoclonal antibody, 14C2. Mice immunized with STF2.4xM2e in aqueous buffer, without adjuvants or other formulation additives, developed potent M2e-specific antibody responses that were quantitatively and qualitatively superior to those observed with M2e peptide delivered in alum. The antibody response was dependent on the physical linkage of the antigen to flagellin and recognized the epitope defined by monoclonal antibody 14C2, which has been shown to protect mice from challenge with influenza A virus. Moreover, immunization with STF2.4xM2e at a dose of 0.3 mu g per mouse protected mice from a lethal challenge with influenza A virus, and significantly reduced weight loss and clinical symptoms. These data demonstrate that the linkage of specific TLR ligand with influenza M2e yields a vaccine candidate that offers significant promise for widespread protection against multiple influenza A virus strains. (c) 2007 Elsevier Ltd. All rights reserved.