Mass Spectrometric Characterization of Narcolepsy-Associated Pandemic 2009 Influenza Vaccines

The onset of narcolepsy, an irreversible sleep disorder, has been associated with 2009 influenza pandemic (pH1N1) infections in China, and with ASO3-adjuvanted pH1N1 vaccinations using Pandemrix in Europe. Intriguingly, however, the increased incidence was only observed following vaccination with Pandemrix but not Arepanrix in Canada. In this study, the mutational burden of actual vaccine lots of Pandemrix (n = 6) and Arepanrix (n = 5) sourced from Canada, and Northern Europe were characterized by mass spectrometry. The four most abundant influenza proteins across both vaccines were nucleoprotein NP, hemagglutinin HA, matrix protein M1, with the exception that Pandemrix harbored a significantly increased proportion of neuraminidase NA (7.5%) as compared to Arepanrix (2.6%). Most significantly, 17 motifs in HA, NP, and M1 harbored mutations, which significantly differed in Pandemrix versus Arepanrix. Among these, a 6-fold higher deamidation of HA146 (p.Asn146Asp) in Arepanrix was found relative to Pandemrix, while NP257 (p.Thr257Ala) and NP424 (p.Thr424Ile) were increased in Pandemrix. DQ0602 binding and tetramer analysis with mutated epitopes were conducted in Pandemrix-vaccinated cases versus controls but were unremarkable. Pandemrix harbored lower mutational burden than Arepanrix, indicating higher similarity to wild-type 2009 pH1N1, which could explain differences in narcolepsy susceptibility amongst the vaccines.

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Mass Spectrometric Characterization of Narcolepsy-Associated Pandemic 2009 Influenza Vaccines

10.1101/2020.08.21.256180 ◽

2020 ◽

Author(s):

Aditya Ambati ◽

Guo Luo ◽

Elora Pradhan ◽

Jacob Louis ◽

Ling Lin ◽

...

Keyword(s):

Matrix Protein ◽

Influenza Pandemic ◽

Influenza Vaccines ◽

Northern Europe ◽

Wild Type ◽

Mutational Burden ◽

Matrix Protein M1 ◽

2009 Influenza Pandemic ◽

Actual Vaccine

AbstractThe onset of narcolepsy, an irreversible sleep disorder, has been associated with 2009 influenza pandemic (pH1N1) infections in China, and with ASO3-adjuvanted pH1N1 vaccinations using Pandemrix in Europe. Intriguingly, however, the increased incidence was only observed following vaccination with Pandemrix but not Arepanrix in Canada. In this study, the mutational burden of actual vaccine lots of Pandemrix (n=6) and Arepanrix (n=5) sourced from Canada, and Northern Europe were characterized by mass spectrometry. The four most abundant influenza proteins across both vaccines were nucleoprotein NP, hemagglutinin HA, matrix protein M1, with the exception that Pandemrix harbored a significantly increased proportion of neuraminidase NA (7.5%) as compared to Arepanrix (2.6%). Most significantly, 17 motifs in HA, NP, and M1 harbored mutations, which significantly differed in Pandemrix versus Arepanrix. Among these, a 6-fold higher deamidation of HA146 (N to D) in Arepanrix was found relative to Pandemrix, while NP257 (T to A) and NP424 (T to I) were increased in Pandemrix. DQ0602 binding and tetramer analysis with mutated epitopes were conducted in Pandemrix-vaccinated cases versus controls but were unremarkable. Pandemrix harbored lower mutational burden than Arepanrix, indicating higher similarity to wild-type 2009 pH1N1, which could explain differences in narcolepsy susceptibility amongst the vaccines.

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Endogenously expressed matrix protein M1 and nucleoprotein of influenza A are efficiently presented by class I and class II major histocompatibility complexes

Journal of General Virology ◽

10.1099/vir.0.029777-0 ◽

2011 ◽

Vol 92 (5) ◽

pp. 1162-1171 ◽

Cited By ~ 15

Author(s):

Jean-Daniel Doucet ◽

Marie-Andrée Forget ◽

Cécile Grange ◽

Ronan Nicolas Rouxel ◽

Nathalie Arbour ◽

...

Keyword(s):

Influenza A ◽

Matrix Protein ◽

Class Ii ◽

Influenza Vaccines ◽

Class I ◽

Mhc I ◽

Mhc Ii ◽

The Matrix ◽

Specific Effector ◽

Matrix Protein M1

Current influenza vaccines containing primarily hypervariable haemagglutinin and neuraminidase proteins must be prepared against frequent new antigenic variants. Therefore, there is an ongoing effort to develop influenza vaccines that also elicit strong and sustained cytotoxic responses against highly conserved determinants such as the matrix (M1) protein and nucleoprotein (NP). However, their antigenic presentation properties in humans are less defined. Accordingly, we analysed MHC class I and class II presentation of endogenously processed M1 and NP in human antigen presenting cells and observed expansion of both CD8+- and CD4+-specific effector T lymphocytes secreting gamma interferon and tumour necrosis factor. Further enhancement of basal MHC-II antigenic presentation did not improve CD4+ or CD8+ T-cell quality based on cytokine production upon challenge, suggesting that endogenous M1 and NP MHC-II presentation is sufficient. These new insights about T-lymphocyte expansion following endogenous M1 and NP MHC-I and -II presentation will be important to design complementary heterosubtypic vaccination strategies.

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Computationally Optimized Broadly Reactive Hemagglutinin Elicits Hemagglutination Inhibition Antibodies against a Panel of H3N2 Influenza Virus Cocirculating Variants

Journal of Virology ◽

10.1128/jvi.01581-17 ◽

2017 ◽

Vol 91 (24) ◽

Cited By ~ 29

Author(s):

Terianne M. Wong ◽

James D. Allen ◽

Anne-Gaelle Bebin-Blackwell ◽

Donald M. Carter ◽

Timothy Alefantis ◽

...

Keyword(s):

Influenza Virus ◽

Influenza Vaccine ◽

Influenza Viruses ◽

Influenza Vaccines ◽

Influenza B ◽

Wild Type ◽

Protective Antibodies ◽

H3n2 Influenza ◽

H3n2 Influenza Virus

ABSTRACT Each influenza season, a set of wild-type viruses, representing one H1N1, one H3N2, and one to two influenza B isolates, are selected for inclusion in the annual seasonal influenza vaccine. In order to develop broadly reactive subtype-specific influenza vaccines, a methodology called computationally optimized broadly reactive antigens (COBRA) was used to design novel hemagglutinin (HA) vaccine immunogens. COBRA technology was effectively used to design HA immunogens that elicited antibodies that neutralized H5N1 and H1N1 isolates. In this report, the development and characterization of 17 prototype H3N2 COBRA HA proteins were screened in mice and ferrets for the elicitation of antibodies with HA inhibition (HAI) activity against human seasonal H3N2 viruses that were isolated over the last 48 years. The most effective COBRA HA vaccine regimens elicited antibodies with broader HAI activity against a panel of H3N2 viruses than wild-type H3 HA vaccines. The top leading COBRA HA candidates were tested against cocirculating variants. These variants were not efficiently detected by antibodies elicited by the wild-type HA from viruses selected as the vaccine candidates. The T-11 COBRA HA vaccine elicited antibodies with HAI and neutralization activity against all cocirculating variants from 2004 to 2007. This is the first report demonstrating broader breadth of vaccine-induced antibodies against cocirculating H3N2 strains compared to the wild-type HA antigens that were represented in commercial influenza vaccines. IMPORTANCE There is a need for an improved influenza vaccine that elicits immune responses that recognize a broader number of influenza virus strains to prevent infection and transmission. Using the COBRA approach, a set of vaccines against influenza viruses in the H3N2 subtype was tested for the ability to elicit antibodies that neutralize virus infection against not only historical vaccine strains of H3N2 but also a set of cocirculating variants that circulated between 2004 and 2007. Three of the H3N2 COBRA vaccines recognized all of the cocirculating strains during this era, but the chosen wild-type vaccine strains were not able to elicit antibodies with HAI activity against these cocirculating strains. Therefore, the COBRA vaccines have the ability to elicit protective antibodies against not only the dominant vaccine strains but also minor circulating strains that can evolve into the dominant vaccine strains in the future.

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The Hansenula polymorpha PER1 gene is essential for peroxisome biogenesis and encodes a peroxisomal matrix protein with both carboxy- and amino-terminal targeting signals.

The Journal of Cell Biology ◽

10.1083/jcb.127.3.737 ◽

1994 ◽

Vol 127 (3) ◽

pp. 737-749 ◽

Cited By ~ 145

Author(s):

H R Waterham ◽

V I Titorenko ◽

P Haima ◽

J M Cregg ◽

W Harder ◽

...

Keyword(s):

Matrix Protein ◽

Sequence Similarity ◽

Hansenula Polymorpha ◽

Matrix Proteins ◽

Regulatory Function ◽

Wild Type ◽

Amino Terminal ◽

Peroxisomal Targeting ◽

Targeting Signal

We describe the cloning of the Hansenula polymorpha PER1 gene and the characterization of the gene and its product, PER1p. The gene was cloned by functional complementation of a per1 mutant of H. polymorpha, which was impaired in the import of peroxisomal matrix proteins (Pim- phenotype). The DNA sequence of PER1 predicts that PER1p is a polypeptide of 650 amino acids with no significant sequence similarity to other known proteins. PER1 expression was low but significant in wild-type H. polymorpha growing on glucose and increased during growth on any one of a number of substrates which induce peroxisome proliferation. PER1p contains both a carboxy- (PTS1) and an amino-terminal (PTS2) peroxisomal targeting signal which both were demonstrated to be capable of directing bacterial beta-lactamase to the organelle. In wild-type H. polymorpha PER1p is a protein of low abundance which was demonstrated to be localized in the peroxisomal matrix. Our results suggest that the import of PER1p into peroxisomes is a prerequisite for the import of additional matrix proteins and we suggest a regulatory function of PER1p on peroxisomal protein support.

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Bacteriophage T4 Vaccine Platform for Next-Generation Influenza Vaccine Development

Frontiers in Immunology ◽

10.3389/fimmu.2021.745625 ◽

2021 ◽

Vol 12 ◽

Author(s):

Mengling Li ◽

Pengju Guo ◽

Cen Chen ◽

Helong Feng ◽

Wanpo Zhang ◽

...

Keyword(s):

Matrix Protein ◽

Vaccine Development ◽

Influenza Pandemic ◽

Bacteriophage T4 ◽

Influenza Vaccines ◽

Next Generation ◽

Complete Protection ◽

Vaccine Platform ◽

Virus Challenge ◽

Cellular Immune

Developing influenza vaccines that protect against a broad range of viruses is a global health priority. Several conserved viral proteins or domains have been identified as promising targets for such vaccine development. However, none of the targets is sufficiently immunogenic to elicit complete protection, and vaccine platforms that can enhance immunogenicity and deliver multiple antigens are desperately needed. Here, we report proof-of-concept studies for the development of next-generation influenza vaccines using the bacteriophage T4 virus-like particle (VLP) platform. Using the extracellular domain of influenza matrix protein 2 (M2e) as a readout, we demonstrate that up to ~1,281 M2e molecules can be assembled on a 120 x 86 nanometer phage capsid to generate M2e-T4 VLPs. These M2e-decorated nanoparticles, without any adjuvant, are highly immunogenic, stimulate robust humoral as well as cellular immune responses, and conferred complete protection against lethal influenza virus challenge. Potentially, additional conserved antigens could be incorporated into the M2e-T4 VLPs and mass-produced in E. coli in a short amount of time to deal with an emerging influenza pandemic.

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Bacteriophage T4 Vaccine Platform for Next-generation Influenza Vaccine Development

10.1101/2021.06.14.448336 ◽

2021 ◽

Author(s):

Mengling Li ◽

Pengju Guo ◽

Cen Chen ◽

Helong Feng ◽

Wanpo Zhang ◽

...

Keyword(s):

Influenza Vaccine ◽

Matrix Protein ◽

Vaccine Development ◽

Influenza Pandemic ◽

Bacteriophage T4 ◽

Influenza Vaccines ◽

Next Generation ◽

Vaccine Platform ◽

Virus Challenge ◽

Virus Like Particle

Developing influenza vaccines that protect against a broad range of viruses is a public health priority, and several conserved viral proteins or domains have been identified as promising targets for such vaccine development. However, none of the targets is immunogenic, and vaccine platforms that can incorporate multiple antigens with enhanced immunogenicity are desperately needed. In this study, we provided proof-of-concept for the development of next-generation influenza vaccine using T4 phage virus-like particle (VLP) platform. With extracellular domain of influenza matrix protein 2 (M2e) as a readout, we showed that more than 1,280 M2e molecules can be assembled on a 120×90 nanometer phage capsid to form T4-M2e VLPs, which are highly immunogenic and induced complete protection against influenza virus challenge without any addition adjuvant. Potentially, additional conserved antigens or molecular adjuvants could be incorporated into the T4-M2e VLPs to customize influenza vaccines to address different issues. All the components of T4 VLP vaccines can be mass-produced in E. coli in a short time, therefore, providing a rapid approach to deal with the potential influenza pandemic.

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Sequence Characterization of matrix protein (M1) in influenza A viruses (H1, H3 and H5)

Microbiology Research ◽

10.4081/mr.2011.e16 ◽

2011 ◽

Vol 2 (1) ◽

pp. 16

Author(s):

Jun Zhang ◽

Xiao Ling Yu ◽

Lei Xu ◽

Fang Zhi Li ◽

Yong Gang Li

Keyword(s):

Amino Acid ◽

Nuclear Localization Signal ◽

Influenza A ◽

Matrix Protein ◽

Amino Acid Sequences ◽

Influenza A Viruses ◽

Sequence Characterization ◽

Matrix Protein M1 ◽

Localization Signal

<p><strong> </strong>This study brings the analysis of amino acid sequences of matrix protein (M1) from the influenza virus A (H1N1, H3N2 and H5N1) during 2007-2208. 741 sequences of M1 were compared, of them, H1N1 388; H3N2 251 and H5N1 102. Even though, the M1 is relatively conserved among the influenza A viruses, we found some variations in the M1 among the viruses, H1N1, H3N2 and H5N1. The nuclear localization signal at amino acid 101 to 105 is RKLKR for H1N1 and H3N2, but for H5N1 is KKLKR. All differences of amino acid in M1 of H1, H3 and H5 were listed. 80 sequences of M1 of H1N1 H3N2 and H5N1 were used for phylogenetic analysis. There is no reasontantment found in the M1 among these subtypes. Further study is needed to study the differences of the function of M1 among H1N1, H3N2 and H5N1. The M1 of H5N1 may contribute to the high pathogenesis to this virus.</p>

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