期刊
INFLUENZA AND OTHER RESPIRATORY VIRUSES
卷 17, 期 4, 页码 -出版社
WILEY
DOI: 10.1111/irv.13141
关键词
hemagglutination; influenza virus; neutralization; serology
A new luciferase microneutralization (LMN) assay was developed, combining the advantages of conventional microneutralization assay with the ease of hemagglutinin inhibition assay. It provides a more feasible, standardized and applicable method for measuring serological responses to influenza vaccination or infection.
BackgroundSerological responses from influenza vaccination or infection are typically measured by hemagglutinin inhibition (HAI) or microneutralization (MN). Both methods are limited in feasibility, standardization, and generalizability to recent strains. We developed a luciferase MN (LMN) assay that combines the advantages of the conventional MN assay with the ease of the HAI assay. MethodsSera were obtained from the HIVE study, a Michigan household cohort. Reverse genetics was used to generate recombinant influenza viruses expressing the hemagglutinin and neuraminidase of test strains, all other viral proteins from an A/WSN/1933 backbone, and a NanoLuc reporter. Serum neutralization of luciferase-expressing targets was quantified as a reduction in light emission from infected cells. Neutralization titers were measured for cell- and egg-adapted versions of A/Hong Kong/4801/2014 and A/Singapore/INFIMH-16-0019/2016 and compared to HAI titers against egg-grown antigens. ResultsThree hundred thirty-three sera were collected from 259 participants between May 2016 and July 2018. Sampled participants were 7-68 years of age, and >80% were vaccinated against influenza. HAI and LMN titers were correlated for A/Hong Kong/4801/2014 (rho = 0.52, p <= 0.01) and A/Singapore/INFIMH-16-0019/2016 (rho = 0.79, p <= 0.01). LMN titers were lower for cell strains compared to egg strains (A/Hong Kong/4801/2014 mean log(2) fold change = -2.66, p <= 0.01 and A/Singapore/INFIMH-16-0019/2016 mean log(2) fold change = -3.15, p <= 0.01). ConclusionsThe LMN assay was feasible using limited sample volumes and able to differentiate small antigenic differences between egg-adapted and cell-derived strains. The correspondence of these results with the commonly used HAI confirms the utility of this assay for high-throughput studies of correlates of protection and vaccine response.
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