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. 2021 Nov 23;12:747774. doi: 10.3389/fimmu.2021.747774

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Copyright © 2021 Zhou, Hansen, Pedersen, Grødeland and Cox

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Adjuvanted H5N1 vaccines elicit multifaceted antibody responses. (A, B) H5HA stalk specific antibodies were measured against the recombinant chimeric protein cH9/5HA, which consists of HA stalk domain from H5 A/Vietnam/1203/2004 strain and irrelevant H9HA head domain. Concentrations of total H5HA stalk specific antibody measured in ELISA (A) and fold-induction (FI) after vaccination (IgG FI,/D0, B) are shown. (C, D) Total neuraminidase (NA) specific antibodies were measured in ELISA against N1NA from A/Vietnam/1203/2004 virus. Antibody concentrations (C) and fold-induction after vaccination (IgG FI,/D0, D) are shown. (E, F) Pre- and post-vaccination sera were tested for inhibiting NA enzymatic activity in ELLA against reassortant NIBRG-73 virus, which expresses NA from H5N1 vaccine strain and irrelevant H7HA. The neuraminidase enzymatic activity inhibition titer (NI IC50, E) was calculated as the reciprocal dilutions of the sera that gave 50% inhibition. Fold-induction after vaccination (NI FI,/D0, F) is shown. (G, H) Sera were tested for inducing antibody-dependent cellular cytotoxicity (ADCC) in ADCC Reporter Bioassay against the vaccine strain (NIBRG-14). The ADCC titer (EC50, G) was calculated as the reciprocal dilutions of the sera that gave 50% of the maximal biological response. Fold-induction after vaccination (ADCC FI,/D0, H) is shown. The geometric mean titers are shown as bars, and each symbol represents one subject (A, C, E, G). The geometric means of fold-induction in each group ± geometric standard deviation as error bar are shown (B, D, F, H). *P < 0.05, **P < 0.01, ***P < 0.001 (Antibody titers and fold-inductions were Ln transformed in statistical analyses. Turkey’s multiple comparisons between pre-prime (D0) and post-prime (D7, D21, D28, D42), and between pre-boost (D21) and post-boost (D28 and D42) in each group were performed in two-way ANOVA in A, C, E, G. Turkey’s multiple comparisons between 30 μg + and 1.5 μg + after prime (D7 and D21), and between 30 μg + and 30 μg - after boost (D28 and D42) were performed in two-way ANOVA in B, D, F, H). The horizontal dotted lines indicate fold-induction of 1 (B, D, F, H). Duplicates were performed in all experiments.