A stable trimeric influenza hemagglutinin stem as a broadly protective immunogen

Impagliazzo, Antonietta; Milder, Fin J; Kuipers, Harmjan; Wagner, Michelle V.; Zhu, Xueyong; Hoffman, Ryan M. B.; Meersbergen, Ruud van; Huizingh, Jeroen; Wanningen, Patrick; Verspuij, Johan; Man, Martijn de; Ding, Zhaoqing; Apetri, Adrian; Kükrer, Başak; Sneekes-Vriese, Eveline; Tomkiewicz, Danuta; Laursen, N.S.; Lee, Peter S.; Zakrzewska, Anna; Dekking, Liesbeth; Tolboom, Jeroen; Tettero, Lisanne; Meerten, Sander van; Yu, Wenli; Koudstaal, Wouter; Goudsmit, Jaap; Ward, Andrew B.; Meijberg, Wim; Wilson, Ian A.; Radošević, Katarina

doi:10.1126/science.aac7263

Cited by 533 publications

(621 citation statements)

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“…Although it is known that human IAV-specific CD8 + T cells provide cross-protection against influenza (32), it is important to further understand how the stalk and catalytic domains of NA affect host immune response. We believe that our findings provide an understanding of glycosylation on influenza cell surface and an innovative direction for the design of a universal influenza vaccine (33,34).…”

Section: Discussionmentioning

confidence: 99%

Influenza A surface glycosylation and vaccine design

Lin

Tsai

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

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We have shown that glycosylation of influenza A virus (IAV) hemagglutinin (HA), especially at position N-27, is crucial for HA folding and virus survival. However, it is not known whether the glycosylation of HA and the other two major IAV surface glycoproteins, neuraminidase (NA) and M2 ion channel, is essential for the replication of IAV. Here, we show that glycosylation of HA at N-142 modulates virus infectivity and host immune response. Glycosylation of NA in the stalk region affects its structure, activity, and specificity, thereby modulating virus release and virulence, and glycosylation at the catalytic domain affects its thermostability; however, glycosylation of M2 had no effect on its function. In addition, using IAV without the stalk and catalytic domains of NA as a live attenuated vaccine was shown to confer a strong IAVspecific CD8+ T-cell response and a strong cross-strain as well as cross-subtype protection against various virus strains.influenza A virus | glycosylation | vaccine design I nfluenza A viruses (IAV) belong to the Orthomyxoviridae family and can circulate widely and cross interspecies barriers through the highly antigenic drift and shift of the 18 subtypes of HA and 11 subtypes of neuraminidase (NA) (1, 2). In addition, the posttranslational modification of the IAV surface proteins is important to circumvent host defense and support the virus life cycle (3).HA is a major surface glycoprotein of IAV and is involved in viral infection via binding to sialic acid (SA)-containing glycans on the surface of host cell (3). The other major glycoprotein of IAV, NA, is involved in the cleavage of SA on the host cell receptor to facilitate the release of viral particles to infect other cells (4). M2, the third surface protein of IAV, has ion channel activity to regulate virus penetration and uncoating (1). All surface proteins interact with M1 protein for virus assembly and release (5).The modification of HA and NA by N-glycosylation is important in the IAV life cycle (1, 6-8). Previously, we have shown that the glycosylation of HA affects its receptor binding, immune response, and structural stability, and glycosite 27 is essential for retaining the structural integrity of HA and its receptor binding (6, 9). In addition, using the monoglycosylated HA as immunogen, it showed a broader protection against various IAV subtypes compared with the fully glycosylated version (10). However, it is not clear whether the other specific glycosites and their glycan structures on HA regulate its functions. With regard to NA, the functional roles of its glycosylation are not well understood, though N-glycosylation was reported to stabilize the protein from protease digestion and may affect the enzyme activity (11,12). The aim of this study was to understand the functional effects of glycosylation on HA, NA, and M2, and how glycosylation of the surface proteins affects the life cycle of IAV. ResultsGlycosylation of HA at N-142. Because several glycosylation sites (glycosites 27, 40, 176, 303, and 497) on HA are ...

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Section: Discussionmentioning

confidence: 99%

Influenza A surface glycosylation and vaccine design

Lin

Tsai

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

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“…While both humoral and cellular immunity can mediate heterosubtypic responses, inducing antibodies against the more conserved stalk domain of hemagglutinin (HA) has been the recent focus of many vaccine programmes (10,11). However, multipronged approaches, targeting several antigens inducing both humoral and cellular responses may limit the generation of viral escape mutants compared to vaccines targeting a limited number of protective epitopes on the HA stalk .…”

Section: Introductionmentioning

confidence: 99%

Novel Bivalent Viral-Vectored Vaccines Induce Potent Humoral and Cellular Immune Responses Conferring Protection against Stringent Influenza A Virus Challenge

Chinnakannan

Mullarkey

Ulaszewska

et al. 2017

The Journal of Immunology

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Abstract:Seasonal influenza viruses (IAV) are a common cause of acute respiratory illness worldwide and generate a significant socio-economic burden. IAV rapidly mutate, necessitating annual vaccine reformulation as traditional vaccines do not typically induce broad-spectrum immunity. In addition to seasonal infections, emerging pandemic influenza viruses present a continued threat to global public health. Pandemic influenza viruses have consistently higher attack rates and are typically associated with greater mortality as compared to seasonal strains. Ongoing strategies to improve vaccine efficacy typically focus on providing broad-spectrum immunity, and while both B and T cells can mediate heterosubtypic responses, typical vaccine development will augment either humoral or cellular immunity. However, multipronged approaches, targeting several antigens, may limit the generation of viral escape mutants. There are few vaccine platforms that can deliver multiple antigens and generate robust cellular and humoral immunity. In this work, we describe a novel vaccination strategy, tested pre-clinically in mice, for the delivery of novel bivalent viral-vectored vaccines. Here, we show this strategy elicits potent T cell responses toward highly conserved internal antigens, whilst simultaneously inducing high levels of antibodies towards hemagglutinin (HA). Importantly, these humoral responses generate longlived plasma cells and generate antibodies capable of neutralising variant HA-expressing pseudotyped lentiviruses. Significantly, these novel viral-vectored vaccines induce strong immune responses capable of conferring protection in a stringent influenza A virus challenge.Thus, this vaccination regimen induces lasting efficacy toward influenza. Importantly, the simultaneous delivery of dual antigens may alleviate the selective pressure thought to potentiate antigenic diversity in avian influenza viruses.

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“…A number of studies have now firmly established that stalk-specific bnAbs, normally present in low quantities, can be boosted substantially in humans following exposure to HA subtypes with antigenically foreign head domains (2)(3)(4)(5)(6)(7)(8). Sequential vaccination of animals with chimeric HAs or with "headless" vaccine constructs have effectively recapitulated the boosting of bnAbs observed in humans after exposure to foreign HAs, and also are protective against heterologous and heterosubtypic IAV challenge (9)(10)(11)(12)(13)(14)(15). These strategies are now being tested as promising "universal" influenza virus vaccine candidates (16).…”

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confidence: 99%

Epitope specificity plays a critical role in regulating antibody-dependent cell-mediated cytotoxicity against influenza A virus

Tan

Mullarkey

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

165

163

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The generation of strain-specific neutralizing antibodies against influenza A virus is known to confer potent protection against homologous infections. The majority of these antibodies bind to the hemagglutinin (HA) head domain and function by blocking the receptor binding site, preventing infection of host cells. Recently, elicitation of broadly neutralizing antibodies which target the conserved HA stalk domain has become a promising "universal" influenza virus vaccine strategy. The ability of these antibodies to elicit Fc-dependent effector functions has emerged as an important mechanism through which protection is achieved in vivo. However, the way in which Fc-dependent effector functions are regulated by polyclonal influenza virus-binding antibody mixtures in vivo has never been defined. Here, we demonstrate that interactions among viral glycoprotein-binding antibodies of varying specificities regulate the magnitude of antibody-dependent cell-mediated cytotoxicity induction. We show that the mechanism responsible for this phenotype relies upon competition for binding to HA on the surface of infected cells and virus particles. Nonneutralizing antibodies were poor inducers and did not inhibit antibody-dependent cell-mediated cytotoxicity. Interestingly, anti-neuraminidase antibodies weakly induced antibody-dependent cell-mediated cytotoxicity and enhanced induction in the presence of HA stalk-binding antibodies in an additive manner. Our data demonstrate that antibody specificity plays an important role in the regulation of ADCC, and that cross-talk among antibodies of varying specificities determines the magnitude of Fc receptor-mediated effector functions.T he discovery and ongoing characterization of broadly neutralizing antibodies (bnAbs) that bind to the hemagglutinin (HA) stalk domain of influenza A viruses (IAVs) has galvanized promising new efforts to generate a universal influenza virus vaccine (1). A number of studies have now firmly established that stalk-specific bnAbs, normally present in low quantities, can be boosted substantially in humans following exposure to HA subtypes with antigenically foreign head domains (2-8). Sequential vaccination of animals with chimeric HAs or with "headless" vaccine constructs have effectively recapitulated the boosting of bnAbs observed in humans after exposure to foreign HAs, and also are protective against heterologous and heterosubtypic IAV challenge (9-15). These strategies are now being tested as promising "universal" influenza virus vaccine candidates (16).Whereas traditional strain-specific antibodies neutralize virus by inhibiting receptor binding, stalk-binding bnAbs neutralize virus by distinct postbinding mechanisms (17). A direct comparison of in vitro neutralization has revealed that strain-specific mAbs that inhibit receptor binding tend to be more potent at neutralizing virus compared with monoclonal stalk-binding bnAbs (18, 19); however, this difference is minimized in the context of a polyclonal response (18). In addition to virus neutralization,...

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A stable trimeric influenza hemagglutinin stem as a broadly protective immunogen

Cited by 533 publications

References 59 publications

Influenza A surface glycosylation and vaccine design

Influenza A surface glycosylation and vaccine design

Novel Bivalent Viral-Vectored Vaccines Induce Potent Humoral and Cellular Immune Responses Conferring Protection against Stringent Influenza A Virus Challenge

Epitope specificity plays a critical role in regulating antibody-dependent cell-mediated cytotoxicity against influenza A virus

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