Julianna Han / Aaron J Schmitz / Sara T Richey / Ya-Nan Dai / Hannah L Turner / Bassem M Mohammed / Daved H Fremont / Ali H Ellebedy / Andrew B Ward /
PubMed Abstract
Novel influenza A virus (IAV) strains elicit recall immune responses to conserved epitopes, making them favorable antigenic choices for universal influenza virus vaccines. Evaluating these immunogens ...Novel influenza A virus (IAV) strains elicit recall immune responses to conserved epitopes, making them favorable antigenic choices for universal influenza virus vaccines. Evaluating these immunogens requires a thorough understanding of the antigenic sites targeted by the polyclonal antibody (pAb) response, which single-particle electron microscopy (EM) can sensitively detect. In this study, we employ EM polyclonal epitope mapping (EMPEM) to extensively characterize the pAb response to hemagglutinin (HA) after H5N1 immunization in humans. Cross-reactive pAbs originating from memory B cells immediately bound the stem of HA and persisted for more than a year after vaccination. In contrast, de novo pAb responses to multiple sites on the head of HA, targeting previously determined key neutralizing sites on H5 HA, expanded after the second immunization and waned quickly. Thus, EMPEM provides a robust tool for comprehensively tracking the specificity and durability of immune responses elicited by novel universal influenza vaccine candidates.
EMDB-22536: Polyclonal immune complex of Fab binding the stem of H5 HA from serum of subject 4 at day 21 Method: EM (single particle) / Resolution: 20.0 Å
EMDB-22537: Polyclonal immune complex of Fab binding the stem of H5 HA from serum of subject 4 at day 28 Method: EM (single particle) / Resolution: 20.0 Å
EMDB-22538: Polyclonal immune complex of Fab binding the side of the head of H5 HA from serum of subject 4 at day 28 Method: EM (single particle) / Resolution: 20.0 Å
EMDB-22539: Polyclonal immune complex of Fab binding the side of the head of H5 HA from serum of subject 4 at day 28 Method: EM (single particle) / Resolution: 20.0 Å
EMDB-22540: Polyclonal immune complex of Fab binding the receptor binding site region of H5 HA from serum of subject 4 at day 28 Method: EM (single particle) / Resolution: 20.0 Å
EMDB-22541: Polyclonal immune complex of Fab binding the top of the head of H5 HA from serum of subject 4 at day 28 Method: EM (single particle) / Resolution: 20.0 Å
EMDB-22542: Polyclonal immune complex of Fab binding the stem of H5 HA from serum of subject 4 at day 42 Method: EM (single particle) / Resolution: 20.0 Å
EMDB-22543: Polyclonal immune complex of Fab binding the stem of H5 HA from serum of subject 28 at day 7 Method: EM (single particle) / Resolution: 20.0 Å
EMDB-22544: Polyclonal immune complex of Fab binding the stem of H5 HA from serum of subject 28 at day 21 Method: EM (single particle) / Resolution: 20.0 Å
EMDB-22545: Polyclonal immune complex of Fab binding the stem of H5 HA from serum of subject 28 at day 28 Method: EM (single particle) / Resolution: 20.0 Å
EMDB-22546: Polyclonal immune complex of Fab binding the receptor binding site region of H5 HA from serum of subject 28 at day 28 Method: EM (single particle) / Resolution: 20.0 Å
EMDB-22547: Polyclonal immune complex of Fab binding the top of the head of H5 HA from serum of subject 28 at day 28 Method: EM (single particle) / Resolution: 20.0 Å
EMDB-22548: Polyclonal immune complex of Fab binding the stem of H5 HA from serum of subject 36 at day 0 Method: EM (single particle) / Resolution: 20.0 Å
EMDB-22549: Polyclonal immune complex of Fab binding the stem of H5 HA from serum of subject 36 at day 7 Method: EM (single particle) / Resolution: 20.0 Å
EMDB-22550: Polyclonal immune complex of Fab binding the stem of H5 HA from serum of subject 36 at day 21 Method: EM (single particle) / Resolution: 20.0 Å
EMDB-22551: Polyclonal immune complex of Fab binding the receptor binding site region of H5 HA from serum of subject 36 at day 28 Method: EM (single particle) / Resolution: 20.0 Å
EMDB-22552: Polyclonal immune complex of Fab binding the stem of H5 HA from serum of subject 36 at day 500 Method: EM (single particle) / Resolution: 20.0 Å
EMDB-22553: Polyclonal immune complex of Fab binding the stem of H5 HA from serum of subject 43 at day 7 Method: EM (single particle) / Resolution: 20.0 Å
EMDB-22554: Polyclonal immune complex of Fab binding the stem of H5 HA from serum of subject 43 at day 7 Method: EM (single particle) / Resolution: 20.0 Å
EMDB-22555: Polyclonal immune complex of Fab binding the stem of H5 HA from serum of subject 43 at day 21 Method: EM (single particle) / Resolution: 20.0 Å
EMDB-22556: Polyclonal immune complex of Fab binding the stem of H5 HA from serum of subject 43 at day 28 Method: EM (single particle) / Resolution: 20.0 Å
EMDB-22557: Polyclonal immune complex of Fab binding the stem of H5 HA from serum of subject 43 at day 28 Method: EM (single particle) / Resolution: 20.0 Å
EMDB-22558: Polyclonal immune complex of Fab binding the receptor binding site region of H5 HA from serum of subject 43 at day 28 Method: EM (single particle) / Resolution: 20.0 Å
EMDB-22559: Polyclonal immune complex of Fab binding the stem of H5 HA from serum of subject 43 at day 100 Method: EM (single particle) / Resolution: 20.0 Å
EMDB-22560: Polyclonal immune complex of Fab binding the receptor binding site region of H5 HA from serum of subject 43 at day 100 Method: EM (single particle) / Resolution: 20.0 Å
EMDB-22561: Monoclonal immune complex of Fab 1A06 from participant 43 at day 7 binding the stem of H5 HA Method: EM (single particle) / Resolution: 20.0 Å
EMDB-22562: Monoclonal immune complex of Fab 1B02 from participant 43 at day 7 binding the stem of H5 HA Method: EM (single particle) / Resolution: 20.0 Å
EMDB-22563: Monoclonal immune complex of Fab 1B04 from participant 43 at day 7 binding the stem of H5 HA Method: EM (single particle) / Resolution: 20.0 Å
EMDB-22564: Monoclonal immune complex of Fab 1A02 from participant 43 at day 28 binding the stem of H5 HA Method: EM (single particle) / Resolution: 20.0 Å
EMDB-22565: Monoclonal immune complex of Fab 1A08 from participant 43 at day 28 binding the stem of H5 HA Method: EM (single particle) / Resolution: 20.0 Å
EMDB-22566: Monoclonal immune complex of Fab 1H01 from participant 43 at day 28 binding the stem of H5 HA Method: EM (single particle) / Resolution: 20.0 Å
EMDB-22567: Monoclonal immune complex of Fab 1E01 from participant 43 at day 28 binding the stem of H5 HA Method: EM (single particle) / Resolution: 20.0 Å
EMDB-22568: Polyclonal immune complex of Fab binding the stem of H1 HA from serum of subject 43 at day 21 Method: EM (single particle) / Resolution: 20.0 Å
EMDB-22569: CryoEM polyclonal immune complex of Fab binding the stem of H5 HA from serum of subject 4 at day 28 Method: EM (single particle) / Resolution: 4.7 Å
EMDB-22570: CryoEM polyclonal immune complex of Fab binding the receptor binding site region of H5 HA from serum of subject 4 at day 28 Method: EM (single particle) / Resolution: 5.9 Å
EMDB-22571: CryoEM polyclonal immune complex of Fab binding the lateral patch of H5 HA from serum of subject 4 at day 28 Method: EM (single particle) / Resolution: 6.0 Å
EMDB-22572: CryoEM polyclonal immune complex of Fab binding the vestigial esterase domain of H5 HA from serum of subject 4 at day 28 Method: EM (single particle) / Resolution: 8.24 Å
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