Antiviral Biologic Produced in DNA Vaccine/Goose Platform Protects Hamsters Against Hantavirus Pulmonary Syndrome When Administered Post-exposure.
Antiviral Biologic Produced in DNA Vaccine/Goose Platform Protects Hamsters Against Hantavirus Pulmonary Syndrome When Administered Post-exposure.
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DOI：
10.1371/journal.pntd.0003803
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发表时间：
2015
影响因子：
3.8
通讯作者：
Bradley DS
中科院分区：
文献类型：
--
作者：
Haese N;Brocato RL;Henderson T;Nilles ML;Kwilas SA;Josleyn MD;Hammerbeck CD;Schiltz J;Royals M;Ballantyne J;Hooper JW;Bradley DS
安第斯病毒 (ANDV) 和 ANDV 样病毒是南美洲大多数汉坦病毒肺综合征 (HPS) 病例的罪魁祸首。智利最近的研究表明，被动转移恢复期人血浆有望成为 HPS 的一种可能治疗方法。不幸的是，这种致命疾病幸存者的恢复期血浆非常有限。我们有兴趣探索使用 DNA 疫苗技术生产抗病毒生物制剂的概念，包括用于人类的多克隆中和抗体。鹅产生 IgY 和一种选择性剪接形式 IgYΔFc，可以从蛋黄中纯化出高浓度的 IgYΔFc。 IgY 缺乏哺乳动物 Fc 的特性，而哺乳动物 Fc 使马、羊和兔子产生的抗体对人类产生反应性。鹅接种了编码病毒包膜糖蛋白的 ANDV DNA 疫苗。所有鹅在第二次疫苗接种后均产生了高滴度的中和抗体，并通过伪病毒粒子中和测定（PsVNA）测量，保持高水平的中和抗体超过一年。加强疫苗接种产生了极高水平的中和抗体（即 PsVNA80 滴度 >100,000）。通过表位作图对 IgY 和 IgYΔFc 进行的分析表明，这些抗体对 ANDV 包膜糖蛋白的特定氨基酸序列具有高度反应性。我们检查了鹅源抗体在致命性 HPS 仓鼠模型中的保护功效。在 IM 攻击 ANDV (25 LD50) 后 5 天开始，将 α-ANDV 免疫血清或从鸡蛋中纯化的 IgY/IgYΔFc 被动转移至仓鼠皮下。免疫血清和鸡蛋来源的纯化 IgY/IgYΔFc 分别保护了 8 只仓鼠中的 8 只和 8 只仓鼠中的 7 只。相比之下，所有接受从正常鹅中纯化的 IgY/IgYΔFc (n=8) 或不接受治疗 (n=8) 的仓鼠都出现了致命的 HPS。这些发现表明，DNA 疫苗/鹅平台可用于生产候选抗病毒生物产品，在暴露后施用时能够预防致命疾病。我们的研究表明，将 DNA 疫苗接种与鹅平台相结合，可用于开发多克隆禽类抗体，用作候选医疗对策。我们证明这些抗体在细胞培养物中具有有效的抗病毒中和活性，并且当作为暴露后预防剂施用时，可有效预防叙利亚仓鼠的汉坦病毒肺综合征。如果在病程后期施用，多克隆抗安第斯病毒抗体是无效的，这表明有效使用基于禽类多克隆抗体的方法来预防汉坦病毒病将需要对出现汉坦病毒病症状的人进行快速诊断和治疗。
Andes virus (ANDV) and ANDV-like viruses are responsible for most hantavirus pulmonary syndrome (HPS) cases in South America. Recent studies in Chile indicate that passive transfer of convalescent human plasma shows promise as a possible treatment for HPS. Unfortunately, availability of convalescent plasma from survivors of this lethal disease is very limited. We are interested in exploring the concept of using DNA vaccine technology to produce antiviral biologics, including polyclonal neutralizing antibodies for use in humans. Geese produce IgY and an alternatively spliced form, IgYΔFc, that can be purified at high concentrations from egg yolks. IgY lacks the properties of mammalian Fc that make antibodies produced in horses, sheep, and rabbits reactogenic in humans. Geese were vaccinated with an ANDV DNA vaccine encoding the virus envelope glycoproteins. All geese developed high-titer neutralizing antibodies after the second vaccination, and maintained high-levels of neutralizing antibodies as measured by a pseudovirion neutralization assay (PsVNA) for over 1 year. A booster vaccination resulted in extraordinarily high levels of neutralizing antibodies (i.e., PsVNA80 titers >100,000). Analysis of IgY and IgYΔFc by epitope mapping show these antibodies to be highly reactive to specific amino acid sequences of ANDV envelope glycoproteins. We examined the protective efficacy of the goose-derived antibody in the hamster model of lethal HPS. α-ANDV immune sera, or IgY/IgYΔFc purified from eggs, were passively transferred to hamsters subcutaneously starting 5 days after an IM challenge with ANDV (25 LD50). Both immune sera, and egg-derived purified IgY/IgYΔFc, protected 8 of 8 and 7 of 8 hamsters, respectively. In contrast, all hamsters receiving IgY/IgYΔFc purified from normal geese (n=8), or no-treatment (n=8), developed lethal HPS. These findings demonstrate that the DNA vaccine/goose platform can be used to produce a candidate antiviral biological product capable of preventing a lethal disease when administered post-exposure. Our studies show the utility of combining DNA vaccination with the goose platform for the development of polyclonal avian antibodies for use as candidate medical countermeasures. We demonstrate that these antibodies have potent anti-viral neutralizing activity in cell culture and are efficacious in preventing hantavirus pulmonary syndrome in Syrian hamsters when administered as a post-exposure prophylactic. The polyclonal anti-Andes virus antibodies were not effective if administered late in the disease course indicating that the effective use of an avian polyclonal antibody-based approach to preventing hantavirus disease will require rapid diagnosis and treatment of persons presenting signs of hantavirus disease.
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