Measuring the neutralization potency of influenza A virus hemagglutinin stalk/stem-binding antibodies in polyclonal preparations by microneutralization assay
Measuring the neutralization potency of influenza A virus hemagglutinin stalk/stem-binding antibodies in polyclonal preparations by microneutralization assay
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DOI：
10.1016/j.ymeth.2015.04.037
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发表时间：
2015-11-15
期刊：
影响因子：
4.8
通讯作者：
Miller, Matthew S.
中科院分区：
文献类型：
--
作者：
He, Wenqian;Mullarkey, Caitlin E.;Miller, Matthew S.
与血凝素茎/茎结构域结合的广泛中和抗体的发现为开发“通用”流感病毒疫苗和治疗剂开辟了令人兴奋的新途径。与结合血凝素头部结构域并抑制受体结合的菌株特异性抗体不同，结合茎部结构域的抗体起抑制感染后期的作用。血凝抑制试验长期以来一直是评价血清中中和血凝素特异性抗体滴度的标准。该测定具有相对快速、易于执行并且需要非常少的专门设备的有益特性。历史上，针对给定流感病毒株的40或以上的血凝抑制滴度被认为与群体水平上的保护相关。不幸的是，该测定法不能用于测量血凝素茎特异性抗体的滴度，因为它们缺乏血凝抑制活性。这使得有必要开发能够灵敏和特异性检测多克隆混合物中的广泛中和HA茎结合抗体的新型试剂和测定法。在这里，我们描述了一种新的基于微量中和的测定，利用重组甲型流感病毒表达嵌合血凝素分子和“外来”神经氨酸酶来测量多克隆制剂中广泛中和抗体的滴度。(C)2015 Elsevier Inc. All rights reserved.
The discovery of broadly-neutralizing antibodies that bind to the hemagglutinin stalk/stem domain has opened exciting new avenues for the development of "universal" influenza virus vaccines and therapeutics. Unlike strain-specific antibodies which bind to the hemagglutinin head domain and inhibit receptor binding, antibodies that bind to the stalk domain function to inhibit later stages of infection. The hemagglutination inhibition assay has long been the standard for evaluating titers of neutralizing hemagglutinin-specific antibodies in serum. The assay has the beneficial properties of being relatively rapid, easy-to-perform, and requires very little specialized equipment. Historically, hemagglutination inhibition titers of 40 or above against a given strain of influenza has been considered a correlate of protection on a population level. Unfortunately, this assay cannot be used to measure titers of hemagglutinin stalk-specific antibodies due to their lack of hemagglutination inhibiting activity. This has necessitated the development of novel reagents and assays capable of sensitive and specific detection of broadly-neutralizing HA stalk-binding antibodies in polyclonal mixtures. Here, we describe a novel microneutralization-based assay that utilizes recombinant influenza A viruses expressing chimeric hemagglutinin molecules and 'exotic' neuraminidase to measure titers of broadly-neutralizing antibodies in polyclonal preparations. (C) 2015 Elsevier Inc. All rights reserved.