One-shot vaccination with an insect cell-derived low-dose influenza A H7 virus-like particle preparation protects mice against H7N9 challenge

ABSTRACT The cytoplasmic tail of the influenza A virus M2 proton-selective ion channel has been shown to be important for virus replication. Previous analysis of M2 cytoplasmic tail truncation mutants demonstrated a defect in incorporation of viral RNA (vRNA) into virions, suggesting a role for M2 in the recruitment of M1-vRNA complexes. To further characterize the effect of the M2 cytoplasmic tail mutations on virus assembly and budding, we constructed a series of alanine substitution mutants of M2 with mutations in the cytoplasmic tail, from residues 71 to 97. Mutant proteins M2-Mut1 and M2-Mut2, with mutations of residues 71 to 73 and 74 to 76, respectively, appeared to have the greatest effect on virus-like particle and virus budding, showing a defect in M1 incorporation. Mutant viruses containing M2-Mut1 and M2-Mut2 failed to replicate in multistep growth analyses on wild-type (wt) MDCK cells and were able to form plaques only on MDCK cells stably expressing wt M2 protein. Compared to wt M2 protein, M2-Mut1 and M2-Mut2 were unable to efficiently coimmunoprecipitate with M1. Furthermore, statistical analysis of planar sheets of membrane from cells infected by virus containing M2-Mut1 revealed a reduction in M1-hemagglutinin (HA) and M2-HA clustering as well as a severe loss of clustering between M1 and M2. These results suggest an essential, direct interaction between the cytoplasmic tail of M2 and M1 that promotes the recruitment of the internal viral proteins and vRNA to the plasma membrane for efficient virus assembly to occur.

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Poly-γ-Glutamic Acid Nanoparticles and Aluminum Adjuvant Used as an Adjuvant with a Single Dose of Japanese Encephalitis Virus-Like Particles Provide Effective Protection from Japanese Encephalitis Virus

Clinical and Vaccine Immunology ◽

10.1128/cvi.05412-11 ◽

2011 ◽

Vol 19 (1) ◽

pp. 17-22 ◽

Cited By ~ 17

Author(s):

Shigefumi Okamoto ◽

Hironori Yoshii ◽

Masaaki Matsuura ◽

Asato Kojima ◽

Toyokazu Ishikawa ◽

...

Keyword(s):

Single Dose ◽

Glutamic Acid ◽

Japanese Encephalitis Virus ◽

Japanese Encephalitis ◽

Single Injection ◽

Neutralizing Antibody ◽

Encephalitis Virus ◽

Virus Like Particle ◽

Generation Vaccine ◽

Particle Preparation

ABSTRACTTo maintain immunity against Japanese encephalitis virus (JEV), a formalin-inactivated Japanese encephalitis (JE) vaccine should be administered several times. The repeated vaccination is not helpful in the case of a sudden outbreak of JEV or when urgent travel to a high-JEV-risk region is required; however, there are few single-injection JE vaccine options. In the present study, we investigated the efficacy of a single dose of a new effective JE virus-like particle preparation containing the JE envelope protein (JE-VLP). Although single administration with JE-VLP protected less than 50% of mice against lethal JEV infection, adding poly(γ-glutamic acid) nanoparticles (γ-PGA-NPs) or aluminum adjuvant (alum) to JE-VLP significantly protected more than 90% of the mice. A single injection of JE-VLP with either γ-PGA-NPs or alum induced a significantly greater anti-JEV neutralizing antibody titer than JE-VLP alone. The enhanced titers were maintained for more than 6 months, resulting in long-lasting protection of 90% of the immunized mice. Although the vaccine design needs further modification to reach 100% protection, a single dose of JE-VLP with γ-PGA-NPs may be a useful step in developing a next-generation vaccine to stop a JE outbreak or to immunize travelers or military personnel.

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Inhibition of the Ubiquitin-Proteasome System Affects Influenza A Virus Infection at a Postfusion Step

Journal of Virology ◽

10.1128/jvi.01048-10 ◽

2010 ◽

Vol 84 (18) ◽

pp. 9625-9631 ◽

Cited By ~ 55

Author(s):

Ivy Widjaja ◽

Erik de Vries ◽

Donna M. Tscherne ◽

Adolfo García-Sastre ◽

Peter J. M. Rottier ◽

...

Keyword(s):

Influenza A Virus ◽

Influenza A ◽

Rna Synthesis ◽

Proteasome Inhibitors ◽

Ubiquitin Proteasome System ◽

Bafilomycin A1 ◽

Virus Like Particle ◽

Viral Particles ◽

Removal Experiments ◽

Ubiquitin Proteasome

ABSTRACT We have demonstrated that influenza A virus (IAV) RNA synthesis depends on the ubiquitin-proteasome system. IAV replication was reduced both by proteasome inhibitors and in E36ts20 cells, which contain the thermolabile ubiquitin-activating enzyme E1. While virus entry was not affected in E36ts20 cells, the proteasome inhibitor MG132 retained viral particles in the cytoplasm. Addition-removal experiments of MG132 in combination with bafilomycin A1, a well-established inhibitor of IAV entry and fusion, showed that MG132 affected IAV infection at a postfusion step. This was confirmed by the lack of inhibition of IAV entry by proteasome inhibitors in a virus-like particle fusion assay.

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Transdermal Vaccination with the Matrix-2 Protein Virus-like Particle (M2e VLP) Induces Immunity in Mice against Influenza A Virus

Vaccines ◽

10.3390/vaccines9111324 ◽

2021 ◽

Vol 9 (11) ◽

pp. 1324

Author(s):

Kimberly Braz Gomes ◽

Sucheta D’Sa ◽

Grace Lovia Allotey-Babington ◽

Sang-Moo Kang ◽

Martin J. D’Souza

Keyword(s):

T Cells ◽

Lymph Node ◽

Influenza A ◽

Lipid A ◽

Adaptive Immune ◽

Monophosphoryl Lipid A ◽

Virus Like Particle ◽

The Matrix ◽

A Cell ◽

Potential Vaccine

In this study, our goal was to utilize the extracellular domain matrix-2 protein virus-like particle (M2e VLP) that has been found to be highly conserved amongst all strains of influenza and could serve as a potential vaccine candidate against influenza. Previous studies have demonstrated that the VLP of the M2e showed increased activation of innate and adaptive immune responses. Therefore, to further explore its level of efficacy and protection, this vaccine was administered transdermally and tested in a pre-clinical mouse model. The M2e VLP was encapsulated into a polymeric matrix with the addition of Alhydrogel® and Monophosphoryl Lipid-A (MPL-A®), together referred to as AS04. The M2e VLP formulations induced IgG titers, with increased levels of IgG1 in the M2e VLP MP groups and further elevated levels of IgG2a were found specifically in the M2e VLP MP Adjuvant group. This trend in humoral immunity was also observed from a cell-mediated standpoint, where M2e VLP MP groups showed increased expression in CD4+ T cells in the spleen and the lymph node and high levels of CD8+ T cells in the lymph node. Taken together, the results illustrate the immunogenic potential of the matrix-2 protein virus-like particle (M2e VLP) vaccine.

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Low Dose Hyperoxia Primes Airways for Fibrosis in Mice after Influenza A Infection

10.1101/2020.06.17.157610 ◽

2020 ◽

Author(s):

Andrew M. Dylag ◽

Jeannie Haak ◽

Rachel Warren ◽

Min Yee ◽

Gloria S. Pryhuber ◽

...

Keyword(s):

Lung Function ◽

Viral Infection ◽

Preterm Infants ◽

Influenza A ◽

Low Dose ◽

Viral Infections ◽

Airway Hyperreactivity ◽

Normal Lung ◽

High Dose ◽

Neonatal Hyperoxia

AbstractIt is well known that supplemental oxygen used to treat preterm infants in respiratory distress is associated with permanently disrupting lung development and the host response to influenza A virus (IAV). However, many infants who go home with normally functioning lungs are also at risk for hyperreactivity after a respiratory viral infection suggesting neonatal oxygen may have induced hidden molecular changes that may prime to the lung for disease. We discovered that thrombospondin-1 (TSP-1) is elevated in adult mice exposed to high-dose neonatal hyperoxia that is known to cause alveolar simplification and fibrotic lung disease following IAV infection. TSP-1 was also elevated in a new, low-dose hyperoxia mouse model (40% for 8 days; 40×8) that we recently reported causes a transient change in lung function that resolves by 8 weeks of age. Elevated TSP-1 was also identified in human autopsy samples of BPD-affected former preterm infants. Consistent with TSP-1 being a master TGFβ regulator, an early transient activation of TGFβ signaling, increased airway hyperreactivity, and peribronchial inflammation and fibrosis were seen when 40×8 mice were infected with IAV, which was not seen in infected room air controls. These findings reveal low dose of neonatal hyperoxia that does not affect lung function or structure may still change expression of genes, such as TSP-1, that may prime the lung for disease following respiratory viral infections, and may help explain why former preterm infants who have normal lung function are susceptible to airway obstruction and increased morbidity after viral infection.

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ISCOMATRIX™ adjuvant promotes epitope spreading and antibody affinity maturation of influenza A H7N9 virus like particle vaccine that correlate with virus neutralization in humans

Vaccine ◽

10.1016/j.vaccine.2015.06.047 ◽

2015 ◽

Vol 33 (32) ◽

pp. 3953-3962 ◽

Cited By ~ 32

Author(s):

Ka Yan Chung ◽

Elizabeth M. Coyle ◽

Dewal Jani ◽

Lisa R. King ◽

Rukmini Bhardwaj ◽

...

Keyword(s):

Influenza A ◽

Affinity Maturation ◽

Virus Neutralization ◽

H7n9 Virus ◽

Antibody Affinity ◽

Epitope Spreading ◽

Virus Like Particle

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Kinetics and phenotype of the draining lymph node and pulmonary B cell response to an influenza A virus-like particle vaccine

10.17077/etd.vfwi8r7i ◽

2013 ◽

Author(s):

Lea Nichole Goldman

Keyword(s):

Lymph Node ◽

B Cell ◽

Influenza A Virus ◽

Cell Response ◽

Influenza A ◽

Virus Like Particle ◽

Draining Lymph Node ◽

B Cell Response

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An Intranasal Virus-Like Particle Vaccine Broadly Protects Mice from Multiple Subtypes of Influenza A Virus

mBio ◽

10.1128/mbio.01044-15 ◽

2015 ◽

Vol 6 (4) ◽

Cited By ~ 49

Author(s):

Louis M. Schwartzman ◽

Andrea L. Cathcart ◽

Lindsey M. Pujanauski ◽

Li Qi ◽

John C. Kash ◽

...

Keyword(s):

Public Health ◽

Influenza A Virus ◽

Influenza A ◽

Influenza Viruses ◽

Significant Protection ◽

Virus Infections ◽

Influenza A Viruses ◽

Virus Like Particle ◽

Practical Strategy ◽

Universal Influenza Vaccine

ABSTRACTInfluenza virus infections are a global public health problem, with a significant impact of morbidity and mortality from both annual epidemics and pandemics. The current strategy for preventing annual influenza is to develop a new vaccine each year against specific circulating virus strains. Because these vaccines are unlikely to protect against an antigenically divergent strain or a new pandemic virus with a novel hemagglutinin (HA) subtype, there is a critical need for vaccines that protect against all influenza A viruses, a so-called “universal” vaccine. Here we show that mice were broadly protected against challenge with a wide variety of lethal influenza A virus infections (94% aggregate survival following vaccination) with a virus-like particle (VLP) vaccine cocktail. The vaccine consisted of a mixture of VLPs individually displaying H1, H3, H5, or H7 HAs, and vaccinated mice showed significant protection following challenge with influenza viruses expressing 1918 H1, 1957 H2, and avian H5, H6, H7, H10, and H11 hemagglutinin subtypes. These experiments suggest a promising and practical strategy for developing a broadly protective “universal” influenza vaccine.IMPORTANCEThe rapid and unpredictable nature of influenza A virus evolution requires new vaccines to be produced annually to match circulating strains. Human infections with influenza viruses derived from animals can cause outbreaks that may be associated with high mortality, and such strains may also adapt to humans to cause a future pandemic. Thus, there is a large public health need to create broadly protective, or “universal,” influenza vaccines that could prevent disease from a wide variety of human and animal influenza A viruses. In this study, a noninfectious virus-like particle (VLP) vaccine was shown to offer significant protection against a variety of influenza A viruses in mice, suggesting a practical strategy to develop a universal influenza vaccine.

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