Vaccination with recombinant fusion proteins incorporating Toll-like receptor ligands induces rapid cellular and humoral immunity
Vaccination with recombinant fusion proteins incorporating Toll-like receptor ligands induces rapid cellular and humoral immunity
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DOI：
10.1016/j.vaccine.2006.08.013
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发表时间：
2007-01-08
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.;Jacobs, Andrea R.;Powell, T. J.
对特定病原体相关分子模式 (PAMP) 的识别主要由 Toll 样受体 (TLR) 家族的成员介导。通过这些受体的刺激会导致抗原呈递和细胞激活发生量和质的变化，从而将先天免疫和适应性免疫联系起来。因此，将 TLR 配体掺入疫苗中可能会产生更有效的疫苗。为了检验这一假设，我们采用了重组融合蛋白策略，将 TLR5 配体鞭毛蛋白与特定抗原融合以促进保护性免疫。这些纯化的重组融合蛋白在体外表现出有效的 TLR5 特异性 NF-κB 依赖性活性。用重组鞭毛蛋白-OVA融合蛋白STF2.OVA对小鼠进行免疫，产生了有效的抗原特异性T细胞和B细胞反应，该反应等于或优于弗氏完全佐剂中乳化的OVA诱导的反应。其中包括在免疫后 7 天内可检测到的快速且一致的抗原特异性 IgG(1) 和 IgG(2a) 抗体反应，以及保护性 CD8 T 细胞反应的发展。此外，OVA 增强的免疫原性取决于与鞭毛蛋白的直接融合，因为 OVA 与未连接的鞭毛蛋白的共同递送未能增强体内抗原特异性反应。使用由鞭毛蛋白和新的多肽序列组成的重组融合蛋白获得了类似的结果，该多肽序列含有源自单核细胞增生李斯特氏菌抗原p60和李斯特菌溶血素O的两个免疫保护表位。用该重组蛋白免疫的动物在受到有毒力的单核细胞增生李斯特氏菌攻击时表现出显着的抗原特异性CD8 T细胞反应和保护。我们得出结论，在缺乏补充佐剂的情况下，用PAMP：抗原融合蛋白进行免疫诱导快速且有效的抗原特异性反应。总的来说，我们的数据表明 PAMP：抗原融合蛋白为开发重组蛋白疫苗提供了重要前景。 (c) 2006 Elsevier Ltd. 保留所有权利。
Recognition of specific pathogen associated molecular patterns (PAMPs) is mediated primarily by members of the Toll-like receptor (TLR) family. Stimulation through these receptors results in quantitative and qualitative changes in antigen presentation and cellular activation, thereby linking innate and adaptive immunity. Consequently, the incorporation of TLR-ligands into vaccines could result in more potent and efficacious vaccines. To test this hypothesis, we employed a recombinant fusion protein strategy using the TLR5 ligand flagellin fused to specific antigens to promote protective immunity. These purified recombinant fusion proteins demonstrated potent TLR5-specific NF-KB dependent activity in vitro. Immunization of mice with the recombinant-flagellin-OVA fusion protein STF2.OVA resulted in potent antigen-specific T and B cell responses that were equal to or better than responses induced by OVA emulsified in Complete Freund's adjuvant. These included rapid and consistent antigen-specific IgG(1) and IgG(2a) antibody responses that were detectable within 7 days of immunization, and the development of protective CD8 T cell responses. Moreover, the enhanced immonogenicity to OVA is dependant on the direct fusion to flagellin, as co-delivery of OVA with flagellin unlinked failed to augment antigen-specific responses in vivo. Similar results were obtained using a recombinant fusion protein comprised of flagellin and a novel polypetide sequence containing two immuno-protective epitopes derived b from the Listeria monocytogenes antigens p60 and listeriolysin O. Animals immunized with this recombinant protein demonstrated significant antigen-specific CD8 T cell responses and protection upon challenge with virulent L. monocytogenes. We conclude that immunization with PAMP:antigen fusion proteins induce rapid and potent antigen-specific responses in the absence Of Supplemental adjuvants. Collectively our data demonstrate that PAMP:antigen fusion proteins offer significant promise for developing recombinant protein vaccines. (c) 2006 Elsevier Ltd. All rights reserved.