Enhanced Immunogenicity of a Whole-Inactivated Influenza A Virus Vaccine Using Optimised Irradiation Conditions

Influenza A virus presents a constant pandemic threat due to the mutagenic nature of the virus and the inadequacy of current vaccines to protect against emerging strains. We have developed a whole-inactivated influenza vaccine using γ-irradiation (γ-Flu) that can protect against both vaccine-included strains as well as emerging pandemic strains. γ-irradiation is a widely used inactivation method and several γ-irradiated vaccines are currently in clinical or pre-clinical testing. To enhance vaccine efficacy, irradiation conditions should be carefully considered, particularly irradiation temperature. Specifically, while more damage to virus structure is expected when using higher irradiation temperatures, reduced radiation doses will be required to achieve sterility. In this study, we compared immunogenicity of γ-Flu irradiated at room temperature, chilled on ice or frozen on dry ice using different doses of γ-irradiation to meet internationally accepted sterility assurance levels. We found that, when irradiating at sterilising doses, the structural integrity and vaccine efficacy were well maintained in all preparations regardless of irradiation temperature. In fact, using a higher temperature and lower radiation dose appeared to induce higher neutralising antibody responses and more effective cytotoxic T cell responses. This outcome is expected to simplify irradiation protocols for manufacturing of highly effective irradiated vaccines.

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Influenza type A virus M protein expression on infected cells is responsible for cross-reactive recognition by cytotoxic thymus-derived lymphocytes

Infection and Immunity ◽

10.1128/iai.29.2.719-723.1980 ◽

1980 ◽

Vol 29 (2) ◽

pp. 719-723 ◽

Cited By ~ 1

Author(s):

C S Reiss ◽

J L Schulman

Keyword(s):

T Cell ◽

Influenza A Virus ◽

Influenza A ◽

Rabbit Antiserum ◽

Type A ◽

M Protein ◽

Cytotoxic T Cell ◽

Influenza A Viruses ◽

Infected Cells ◽

Cytotoxic T Cell Responses

M protein of influenza A virus was detected with rabbit antiserum by both indirect immunofluorescence and by antibody plus complement-mediated cytolysis on the cell surfaces of both productively and nonproductively infected cells. In contrast, antiserum to nucleoprotein failed to react with unfixed infected cells, but did bind to fixed infected cells, especially in the perinuclear area. Incorporation of antiserum to M protein in a T-cell-mediated cytotoxicity assay produced almost complete abrogation of lysis of H-2-compatible cells infected with an influenza A virus of a subtype which differed from that used to elicit the cytotoxic T cells. However, the antibody did not significantly block 51Cr release from cells infected with the homotypic type A influenza virus. These observations are in accord with the hypothesis that the cross-reactive cytotoxic T-cell responses seen with cells infected by heterotypic influenza A viruses are due to recognition of a common M protein.

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Induction of cytotoxic T-cell responses by gene gun DNA vaccination with minigenes encoding influenza A virus HA and NP CTL-epitopes

Vaccine ◽

10.1016/s0264-410x(99)00279-0 ◽

1999 ◽

Vol 18 (7-8) ◽

pp. 681-691 ◽

Cited By ~ 27

Author(s):

Anders Fomsgaard ◽

Henrik V Nielsen ◽

Nikolai Kirkby ◽

Karin Bryder ◽

Sylvie Corbet ◽

...

Keyword(s):

T Cell ◽

Influenza A Virus ◽

Influenza A ◽

T Cell Responses ◽

Dna Vaccination ◽

Cytotoxic T Cell ◽

Gene Gun ◽

Ctl Epitopes ◽

Cell Responses ◽

Cytotoxic T Cell Responses

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Cytotoxic T Cells Are the Predominant Players Providing Cross-Protective Immunity Induced by γ-Irradiated Influenza A Viruses

Journal of Virology ◽

10.1128/jvi.02508-09 ◽

2010 ◽

Vol 84 (9) ◽

pp. 4212-4221 ◽

Cited By ~ 58

Author(s):

Yoichi Furuya ◽

Jennifer Chan ◽

Matthias Regner ◽

Mario Lobigs ◽

Aulikki Koskinen ◽

...

Keyword(s):

T Cells ◽

Influenza Virus ◽

Influenza A Virus ◽

Protective Immunity ◽

Influenza A ◽

Gene Knockout ◽

Influenza Viruses ◽

Cytotoxic T Cell ◽

Cell Responses ◽

Cytotoxic T Cell Responses

ABSTRACT We previously demonstrated that a single dose of nonadjuvanted intranasal γ-irradiated influenza A virus can provide robust protection in mice against both homologous and heterosubtypic challenges, including challenge with an H5N1 avian virus strain. We investigated the mechanism behind the observed cross-protection to define which arms of the adaptive immune response are involved in mediating this protection. Studies with gene knockout mice showed the cross-protective immunity to be mediated mainly by T cells and to be dependent on the cytolytic effector molecule perforin. Adoptive transfer of memory T cells from immunized mice, but not of memory B cells, protected naïve recipients against lethal heterosubtypic influenza virus challenge. Furthermore, γ-irradiated influenza viruses induced cross-reactive Tc-cell responses but not cross-neutralizing or cross-protective antibodies. In addition, histological analysis showed reduced lung inflammation in vaccinated mice compared to that in unvaccinated controls following heterosubtypic challenge. This reduced inflammation was associated with enhanced early recruitment of T cells, both CD4+ and CD8+, and with early influenza virus-specific cytotoxic T-cell responses. Therefore, cross-protective immunity induced by vaccination with γ-irradiated influenza A virus is mediated mainly by Tc-cell responses.

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In vitro human cytotoxic T cell responses against influenza A virus can be induced and selected by synthetic peptides

European Journal of Immunology ◽

10.1002/eji.1830201004 ◽

1990 ◽

Vol 20 (10) ◽

pp. 2171-2176 ◽

Cited By ~ 11

Author(s):

Frédéric Martinon ◽

Elisabeth Gomard ◽

Claude Hannoun ◽

Jean-Paul Lévy

Keyword(s):

T Cell ◽

Influenza A Virus ◽

Synthetic Peptides ◽

Influenza A ◽

T Cell Responses ◽

Cytotoxic T Cell ◽

Cell Responses ◽

Cytotoxic T Cell Responses

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Comparison of Adjuvanted-Whole Inactivated Virus and Live-Attenuated Virus Vaccines against Challenge with Contemporary, Antigenically Distinct H3N2 Influenza A Viruses

Journal of Virology ◽

10.1128/jvi.01323-18 ◽

2018 ◽

Vol 92 (22) ◽

Cited By ~ 5

Author(s):

Eugenio J. Abente ◽

Daniela S. Rajao ◽

Jefferson Santos ◽

Bryan S. Kaplan ◽

Tracy L. Nicholson ◽

...

Keyword(s):

Influenza A Virus ◽

Vaccine Efficacy ◽

Influenza A ◽

Rapid Evolution ◽

The United States ◽

Upper Respiratory Tract ◽

Influenza A Viruses ◽

Partial Protection ◽

Attenuated Virus ◽

The Impact

ABSTRACTInfluenza A viruses in swine (IAV-S) circulating in the United States of America are phylogenetically and antigenically distinct. A human H3 hemagglutinin (HA) was introduced into the IAV-S gene pool in the late 1990s, sustained continued circulation, and evolved into five monophyletic genetic clades, H3 clades IV-A to -E, after 2009. Across these phylogenetic clades, distinct antigenic clusters were identified, with three clusters (cyan, red, and green antigenic cluster) among the most frequently detected antigenic phenotypes (Abente EJ, Santos J, Lewis NS, Gauger PC, Stratton J, et al. J Virol 90:8266–8280, 2016,https://doi.org/10.1128/JVI.01002-16). Although it was demonstrated that antigenic diversity of H3N2 IAV-S was associated with changes at a few amino acid positions in the head of the HA, the implications of this diversity for vaccine efficacy were not tested. Using antigenically representative H3N2 viruses, we compared whole inactivated virus (WIV) and live-attenuated influenza virus (LAIV) vaccines for protection against challenge with antigenically distinct H3N2 viruses in pigs. WIV provided partial protection against antigenically distinct viruses but did not prevent virus replication in the upper respiratory tract. In contrast, LAIV provided complete protection from disease and virus was not detected after challenge with antigenically distinct viruses.IMPORTANCEDue to the rapid evolution of the influenza A virus, vaccines require continuous strain updates. Additionally, the platform used to deliver the vaccine can have an impact on the breadth of protection. Currently, there are various vaccine platforms available to prevent influenza A virus infection in swine, and we experimentally tested two: adjuvanted-whole inactivated virus and live-attenuated virus. When challenged with an antigenically distinct virus, adjuvanted-whole inactivated virus provided partial protection, while live-attenuated virus provided effective protection. Additional strategies are required to broaden the protective properties of inactivated virus vaccines, given the dynamic antigenic landscape of cocirculating strains in North America, whereas live-attenuated vaccines may require less frequent strain updates, based on demonstrated cross-protection. Enhancing vaccine efficacy to control influenza infections in swine will help reduce the impact they have on swine production and reduce the risk of swine-to-human transmission.

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Cross-protection in mice infected with influenza A virus by the respiratory route is correlated with local IgA antibody rather than serum antibody or cytotoxic T cell reactivity

European Journal of Immunology ◽

10.1002/eji.1830140414 ◽

1984 ◽

Vol 14 (4) ◽

pp. 350-356 ◽

Cited By ~ 212

Author(s):

Foo Y. Liew ◽

Stuart M. Russell ◽

Geoffrey Appleyard ◽

Colin M. Brand ◽

John Beale

Keyword(s):

T Cell ◽

Influenza A Virus ◽

Influenza A ◽

Serum Antibody ◽

Cross Protection ◽

Cytotoxic T Cell ◽

Cell Reactivity ◽

Iga Antibody ◽

T Cell Reactivity

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Rectification of Age-Associated Deficiency in Cytotoxic T Cell Response to Influenza A Virus by Immunization with Immune Complexes

The Journal of Immunology ◽

10.4049/jimmunol.179.9.6153 ◽

2007 ◽

Vol 179 (9) ◽

pp. 6153-6159 ◽

Cited By ~ 31

Author(s):

Biao Zheng ◽

Yongxin Zhang ◽

Hongxia He ◽

Ekaterina Marinova ◽

Kirsten Switzer ◽

...

Keyword(s):

T Cell ◽

Influenza A Virus ◽

Immune Complexes ◽

Cell Response ◽

Influenza A ◽

T Cell Response ◽

Cytotoxic T Cell

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HIV Type 1 Vaccine-Induced Cytotoxic T Cell Responses: Potential Role in Vaccine Efficacy

AIDS Research and Human Retroviruses ◽

10.1089/aid.1997.13.205 ◽

1997 ◽

Vol 13 (3) ◽

pp. 205-210 ◽

Cited By ~ 23

Author(s):

GORDON L. ADA ◽

M. JULIANA McELRATH

Keyword(s):

T Cell ◽

Vaccine Efficacy ◽

Potential Role ◽

T Cell Responses ◽

Cytotoxic T Cell ◽

Cell Responses ◽

Hiv Type 1 ◽

Cytotoxic T Cell Responses

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Crystal structure of an orthomyxovirus matrix protein reveals mechanisms for self-polymerization and membrane association

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.1701747114 ◽

2017 ◽

Vol 114 (32) ◽

pp. 8550-8555 ◽

Cited By ~ 14

Author(s):

Wenting Zhang ◽

Wenjie Zheng ◽

Yukimatsu Toh ◽

Miguel A. Betancourt-Solis ◽

Jiagang Tu ◽

...

Keyword(s):

Crystal Structure ◽

Crystal Lattice ◽

Influenza A Virus ◽

Nuclear Export ◽

Influenza A ◽

Electrostatic Interactions ◽

Matrix Protein ◽

Structural Integrity ◽

Viral Envelope ◽

Protein Layer

Many enveloped viruses encode a matrix protein. In the influenza A virus, the matrix protein M1 polymerizes into a rigid protein layer underneath the viral envelope to help enforce the shape and structural integrity of intact viruses. The influenza virus M1 is also known to mediate virus budding as well as the nuclear export of the viral nucleocapsids and their subsequent packaging into nascent viral particles. Despite extensive studies on the influenza A virus M1 (FLUA-M1), only crystal structures of its N-terminal domain are available. Here we report the crystal structure of the full-length M1 from another orthomyxovirus that infects fish, the infectious salmon anemia virus (ISAV). The structure of ISAV-M1 assumes the shape of an elbow, with its N domain closely resembling that of the FLUA-M1. The C domain, which is connected to the N domain through a flexible linker, is made of four α-helices packed as a tight bundle. In the crystal, ISAV-M1 monomers form infinite 2D arrays with a network of interactions involving both the N and C domains. Results from liposome flotation assays indicated that ISAV-M1 binds membrane via electrostatic interactions that are primarily mediated by a positively charged surface loop from the N domain. Cryoelectron tomography reconstruction of intact ISA virions identified a matrix protein layer adjacent to the inner leaflet of the viral membrane. The physical dimensions of the virion-associated matrix layer are consistent with the 2D ISAV-M1 crystal lattice, suggesting that the crystal lattice is a valid model for studying M1–M1, M1–membrane, and M1–RNP interactions in the virion.

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