Use of lentiviral pseudotypes as an alternative to reassortant or Triton X‐100‐treated wild‐type Influenza viruses in the neuraminidase inhibition enzyme‐linked lectin assay

BACKGROUND: Formulation of neuraminidase (NA) within influenza vaccines is gaining importance in light of recent human studies. The enzyme‐linked lectin assay (ELLA) is considered a reliable assay to evaluate human anti‐NA antibodies. OBJECTIVES: To overcome interference by hemagglutinin (HA)‐specific antibodies and detect neuraminidase inhibitory (NI) antibodies only, two different sources of antigen have been studied in ELLA: reassortant viruses with a mismatched avian origin‐HA or Triton X‐100 (Tx)‐treated wild‐type viruses. Pseudotypes or pseudovirus (PV), characterized by a lentivirus core bearing human influenza NA and avian influenza HA, were investigated as an alternative source of antigen and compared to HA‐mismatched and Tx‐treated viruses, since represent a safer product to be handled. METHODS: Two independent panels of sera were analyzed by ELLA to evaluate the anti‐NA response against N1 (A/California/07/2009 (H1N1pdm)) and N2 (A/Hong Kong/4801/2014 (H3N2)). The NA inhibition (NI) antibody titers measured as either the 50% end point or 50% inhibitory concentration (IC(50)) were compared for every source of antigen. RESULTS: The ELLA assay performed well with all three sources of antigen. NI titers measured using each antigen type correlated well when reported either as end point titers or as the IC(50). CONCLUSIONS: This study suggests that HA‐mismatched whole virus, Triton‐treated wild‐type virus or PV can be used to measure NI antibody titers of human sera, but further comparability/validation assays should be performed to assess statistical differences. The data support the use of PV as an attractive alternative source of antigen and justify further investigation to improve stability of this antigen source.