Properly folded bacterially expressed H1N1 hemagglutinin globular head and ectodomain vaccines protect ferrets against H1N1 pandemic influenza virus.
Properly folded bacterially expressed H1N1 hemagglutinin globular head and ectodomain vaccines protect ferrets against H1N1 pandemic influenza virus.
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DOI：
10.1371/journal.pone.0011548
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发表时间：
2010-07-12
期刊：
影响因子：
3.7
通讯作者：
Golding H
中科院分区：
文献类型：
--
作者：
Khurana S;Verma S;Verma N;Crevar CJ;Carter DM;Manischewitz J;King LR;Ross TM;Golding H
面对即将来临的流感大流行，快速生产疫苗和大规模接种疫苗是预防与1918年“西班牙流感”相关的大规模死亡和发病的最有效方法。传统的在鸡蛋中生产流感疫苗的过程是耗时的，并且可能不满足减少流感大流行所需的快速全球疫苗接种的需求。重组技术可用于在包括哺乳动物、昆虫和细菌细胞在内的多种系统中表达新出现的流感病毒株的血凝素（HA）。在本研究中，从大肠杆菌中表达并纯化了来自当前流行的新型H1N1 A/加州/07/2009病毒的两种形式的HA蛋白，HA 1（1-330）和HA（1-480）。在受控的氧化还原重折叠条件下，有利于正确的蛋白质折叠。然而，只有重组HA 1（1-330）蛋白形成寡聚体，包括结合受体并引起人红细胞凝集的功能性三聚体。在用A/加州/07/2009病毒攻击之前，这些蛋白质用于接种雪貂。这两种蛋白质诱导中和抗体，并减少鼻洗液中的病毒载量。然而，与HA（1-480）相比，具有较高多聚体形式含量的HA 1（1-330）蛋白在较低疫苗剂量下提供了更好的发热和体重减轻保护。HA 1（1-330）的蛋白质产量范围约为40 mg/L，而HA（1-480）产量为0.4-0.8 mg/L。这是第一个研究，描述了正确折叠的功能性球状HA 1结构域三聚体的细菌系统中的生产，缺乏HA 2跨膜蛋白，引发有效的中和抗体反应后接种疫苗，并保护雪貂从体内挑战。细菌表达系统与已建立的质量控制方法相结合，可以为面对流感大流行威胁的流感疫苗的快速大规模生产提供机制。
In the face of impending influenza pandemic, a rapid vaccine production and mass vaccination is the most effective approach to prevent the large scale mortality and morbidity that was associated with the 1918 “Spanish Flu”. The traditional process of influenza vaccine production in eggs is time consuming and may not meet the demands of rapid global vaccination required to curtail influenza pandemic. Recombinant technology can be used to express the hemagglutinin (HA) of the emerging new influenza strain in a variety of systems including mammalian, insect, and bacterial cells. In this study, two forms of HA proteins derived from the currently circulating novel H1N1 A/California/07/2009 virus, HA1 (1–330) and HA (1–480), were expressed and purified from E. coli under controlled redox refolding conditions that favoured proper protein folding. However, only the recombinant HA1 (1–330) protein formed oligomers, including functional trimers that bound receptor and caused agglutination of human red blood cells. These proteins were used to vaccinate ferrets prior to challenge with the A/California/07/2009 virus. Both proteins induced neutralizing antibodies, and reduced viral loads in nasal washes. However, the HA1 (1–330) protein that had higher content of multimeric forms provided better protection from fever and weight loss at a lower vaccine dose compared with HA (1–480). Protein yield for the HA1 (1–330) ranged around 40 mg/Liter, while the HA (1–480) yield was 0.4–0.8 mg/Liter. This is the first study that describes production in bacterial system of properly folded functional globular HA1 domain trimers, lacking the HA2 transmembrane protein, that elicit potent neutralizing antibody responses following vaccination and protect ferrets from in vivo challenge. The combination of bacterial expression system with established quality control methods could provide a mechanism for rapid large scale production of influenza vaccines in the face of influenza pandemic threat.
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影响因子：
3.7
作者：
Rowe T;León AJ;Crevar CJ;Carter DM;Xu L;Ran L;Fang Y;Cameron CM;Cameron MJ;Banner D;Ng DC;Ran R;Weirback HK;Wiley CA;Kelvin DJ;Ross TM
通讯作者：
Ross TM
影响因子：
3.7
作者：
Song L;Nakaar V;Kavita U;Price A;Huleatt J;Tang J;Jacobs A;Liu G;Huang Y;Desai P;Maksymiuk G;Takahashi V;Umlauf S;Reiserova L;Bell R;Li H;Zhang Y;McDonald WF;Powell TJ;Tussey L
通讯作者：
Tussey L
影响因子：
15.8
作者：
Khurana S;Suguitan AL Jr;Rivera Y;Simmons CP;Lanzavecchia A;Sallusto F;Manischewitz J;King LR;Subbarao K;Golding H
通讯作者：
Golding H
影响因子：
64.8
作者：
WILEY, DC;WILSON, IA;SKEHEL, JJ
通讯作者：
SKEHEL, JJ
影响因子：
1.6
作者：
Curtis-Fisk, Jaime;Spencer, Ryan M.;Weliky, David P.
通讯作者：
Weliky, David P.