A Computationally Optimized Broadly Reactive Antigen (COBRA) Elicits Broadly Neutralizing Antibodies Against a Conserved Influenza Virus Hemagglutinin B-Cell Epitope

Abstract

Abstract The development of broadly protective influenza vaccines will represent the main countermeasure to overcome influenza virus spread and improve the coverage offered by the current standard of care. A computationally optimized broadly reactive influenza virus hemagglutinin (HA) antigen (COBRA), named P1, elicits a broadly reactive antibody (Ab) response against multiple H1N1 strains. In order to understand the mechanism of P1-conferred breadth of response, we first characterized the breadth of the Ab response at the B-cell level. Specifically, the reactivity of secreted Abs from mice immunized with P1 or seasonal/pandemic vaccine strains was assessed against a panel of H1N1 recombinant HA. Interestingly, while Abs from seasonal/pandemic immunized mice showed a predominant homologous response, those from P1- immunized animals exhibited a broader recognition. In order to dissect the antibody response, a panel of P1-specific B-cell hybridomas was generated. Subsequently, the corresponding purified monoclonal Abs (mAbs) were assessed for the breadth of HA binding and hemagglutination inhibition (HAI) activity against different H1N1 viruses. Collectively, P1-specific mAbs exhibited a broad spectrum of binding and functional activities, spanning from strain-specific to broadly reactive and neutralizing mAbs, while seasonal/pandemic-specific mAbs displayed a narrower spectrum of activity. Epitope mapping studies revealed that P1-elicited broadly reactive and neutralizing Abs recognize unique conserved epitopes within the HA receptor binding site. Collectively, these studies shed light on the mechanism of breadth conferred by P1 and leverage its optimization for the development of a more effective influenza vaccine.