Carnell George William, Ferrara Francesca, Grehan Keith, Thompson Craig Peter, Temperton Nigel James
Viral Pseudotype Unit, Medway School of Pharmacy, The Universities of Greenwich and Kent at Medway , Chatham Maritime, Kent , UK.
Viral Pseudotype Unit, Medway School of Pharmacy, The Universities of Greenwich and Kent at Medway , Chatham Maritime, Kent , UK ; Department of Zoology, University of Oxford , Oxford , UK ; The Jenner Institute Laboratories, University of Oxford , Oxford , UK.
Front Immunol. 2015 Apr 29;6:161. doi: 10.3389/fimmu.2015.00161. eCollection 2015.
The use of vaccination against the influenza virus remains the most effective method of mitigating the significant morbidity and mortality caused by this virus. Antibodies elicited by currently licensed influenza vaccines are predominantly hemagglutination-inhibition (HI)-competent antibodies that target the globular head of hemagglutinin (HA) thus inhibiting influenza virus entry into target cells. These antibodies predominantly confer homosubtypic/strain specific protection and only rarely confer heterosubtypic protection. However, recent academia or pharma-led R&D toward the production of a "universal vaccine" has centered on the elicitation of antibodies directed against the stalk of the influenza HA that has been shown to confer broad protection across a range of different subtypes (H1-H16). The accurate and sensitive measurement of antibody responses elicited by these "next-generation" influenza vaccines is, however, hampered by the lack of sensitivity of the traditional influenza serological assays HI, single radial hemolysis, and microneutralization. Assays utilizing pseudotypes, chimeric viruses bearing influenza glycoproteins, have been shown to be highly efficient for the measurement of homosubtypic and heterosubtypic broadly neutralizing antibodies, making them ideal serological tools for the study of cross-protective responses against multiple influenza subtypes with pandemic potential. In this review, we will analyze and compare literature involving the production of influenza pseudotypes with particular emphasis on their use in serum antibody neutralization assays. This will enable us to establish the parameters required for optimization and propose a consensus protocol to be employed for the further deployment of these assays in influenza vaccine immunogenicity studies.
接种流感病毒疫苗仍然是减轻该病毒所致严重发病和死亡的最有效方法。目前已获许可的流感疫苗所引发的抗体主要是具有血凝抑制（HI）活性的抗体，这些抗体靶向血凝素（HA）的球状头部，从而抑制流感病毒进入靶细胞。这些抗体主要提供同亚型/同毒株特异性保护，很少提供异亚型保护。然而，近期学术界或制药企业主导的研发“通用疫苗”的工作，主要集中在诱导产生针对流感HA茎部的抗体，这种抗体已被证明能对一系列不同亚型（H1 - H16）提供广泛保护。然而，这些“下一代”流感疫苗所引发的抗体反应的准确和灵敏测量，受到传统流感血清学检测方法（HI、单向辐射溶血和微量中和）灵敏度不足的阻碍。利用假病毒颗粒（携带流感糖蛋白的嵌合病毒）的检测方法，已被证明在测量同亚型和异亚型广泛中和抗体方面非常高效，使其成为研究针对多种具有大流行潜力的流感亚型的交叉保护反应的理想血清学工具。在本综述中，我们将分析和比较涉及流感假病毒颗粒生产的文献，特别强调它们在血清抗体中和检测中的应用。这将使我们能够确定优化所需的参数，并提出一个共识方案，以便在流感疫苗免疫原性研究中进一步应用这些检测方法。
