Kaufmann Lukas, Syedbasha Mohammedyaseen, Vogt Dominik, Hollenstein Yvonne, Hartmann Julia, Linnik Janina E, Egli Adrian
Applied Microbiology Research, Department of Biomedicine, University of Basel.
Applied Microbiology Research, Department of Biomedicine, University of Basel; Department of Biosystems Science and Engineering, ETH Zurich; Swiss Institute of Bioinformatics.
J Vis Exp. 2017 Dec 1(130):55833. doi: 10.3791/55833.
Antibody titers are commonly used as surrogate markers for serological protection against influenza and other pathogens. Detailed knowledge of antibody production pre- and post-vaccination is required to understand vaccine-induced immunity. This article describes a reliable point-by-point protocol to determine influenza-specific antibody titers. The first protocol describes a method to specify the antigen amounts required for hemagglutination, which standardizes the concentrations for subsequent usage in the second protocol (hemagglutination assay, HA assay). The second protocol describes the quantification of influenza-specific antibody titers against different viral strains by using a serial dilution of human serum or cell culture supernatants (hemagglutination inhibition assay, HI assay). As an applied example, we show the antibody response of a healthy cohort, which received a trivalent inactivated influenza vaccine. Additionally, the cross-reactivity between the different influenza viruses is shown and methods to minimize cross-reactivity by using different types of animal red blood cells (RBCs) are explained. The discussion highlights advantages and disadvantages of the presented assays and how the determination of influenza-specific antibody titers can improve the understanding of vaccine-related immunity.
抗体滴度通常用作针对流感和其他病原体的血清学保护的替代标志物。为了理解疫苗诱导的免疫，需要详细了解接种疫苗前后的抗体产生情况。本文描述了一种可靠的逐点方案来确定流感特异性抗体滴度。第一个方案描述了一种确定血凝所需抗原量的方法，该方法标准化了后续在第二个方案（血凝试验，HA试验）中使用的浓度。第二个方案描述了通过对人血清或细胞培养上清液进行系列稀释（血凝抑制试验，HI试验）来定量针对不同病毒株的流感特异性抗体滴度。作为一个应用实例，我们展示了一组健康人群接种三价灭活流感疫苗后的抗体反应。此外，还展示了不同流感病毒之间的交叉反应性，并解释了通过使用不同类型的动物红细胞（RBC）来最小化交叉反应性的方法。讨论突出了所介绍试验的优缺点，以及流感特异性抗体滴度的测定如何能增进对疫苗相关免疫的理解。
