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1.
PLoS One ; 7(11): e50852, 2012.
Article in English | MEDLINE | ID: mdl-23226404

ABSTRACT

Respiratory Syncytial Virus (RSV) is an important viral agent causing severe respiratory tract disease in infants and children as well as in the elderly and immunocompromised individuals. The lack of a safe and effective RSV vaccine represents a major unmet medical need. RSV fusion (F) surface glycoprotein was modified and cloned into a baculovirus vector for efficient expression in Sf9 insect cells. Recombinant RSV F was glycosylated and cleaved into covalently linked F2 and F1 polypeptides that formed homotrimers. RSV F extracted and purified from insect cell membranes assembled into 40 nm protein nanoparticles composed of multiple RSV F oligomers arranged in the form of rosettes. The immunogenicity and protective efficacy of purified RSV F nanoparticles was compared to live and formalin inactivated RSV in cotton rats. Immunized animals induced neutralizing serum antibodies, inhibited virus replication in the lungs, and had no signs of disease enhancement in the respiratory track of challenged animals. RSV F nanoparticles also induced IgG competitive for binding of palivizumab neutralizing monoclonal antibody to RSV F antigenic site II. Antibodies to this epitope are known to protect against RSV when passively administered in high risk infants. Together these data provide a rational for continued development a recombinant RSV F nanoparticle vaccine candidate.


Subject(s)
Glycoproteins/immunology , Immunity/immunology , Nanoparticles/chemistry , Respiratory Syncytial Virus Infections/prevention & control , Respiratory Syncytial Viruses/immunology , Sigmodontinae/immunology , Viral Fusion Proteins/immunology , Amino Acid Sequence , Animals , Antibodies, Monoclonal, Humanized/immunology , Antibodies, Viral/immunology , Chromatography, High Pressure Liquid , Disease Models, Animal , Humans , Light , Lung/pathology , Lung/virology , Male , Mutant Proteins/chemistry , Mutant Proteins/immunology , Nanoparticles/ultrastructure , Palivizumab , Protein Structure, Tertiary , Respiratory Syncytial Virus Infections/immunology , Scattering, Radiation , Sf9 Cells , Sigmodontinae/virology , Surface Plasmon Resonance , Viral Fusion Proteins/chemistry , Viral Fusion Proteins/isolation & purification
2.
Vaccine ; 29(38): 6606-13, 2011 Sep 02.
Article in English | MEDLINE | ID: mdl-21762752

ABSTRACT

SARS-CoV was the cause of the global pandemic in 2003 that infected over 8000 people in 8 months. Vaccines against SARS are still not available. We developed a novel method to produce high levels of a recombinant SARS virus-like particles (VLPs) vaccine containing the SARS spike (S) protein and the influenza M1 protein using the baculovirus insect cell expression system. These chimeric SARS VLPs have a similar size and morphology to the wild type SARS-CoV. We tested the immunogenicity and protective efficacy of purified chimeric SARS VLPs and full length SARS S protein vaccines in a mouse lethal challenge model. The SARS VLP vaccine, containing 0.8 µg of SARS S protein, completely protected mice from death when administered intramuscular (IM) or intranasal (IN) routes in the absence of an adjuvant. Likewise, the SARS VLP vaccine, containing 4 µg of S protein without adjuvant, reduced lung virus titer to below detectable level, protected mice from weight loss, and elicited a high level of neutralizing antibodies against SARS-CoV. Sf9 cell-produced full length purified SARS S protein was also an effective vaccine against SARS-CoV but only when co-administered IM with aluminum hydroxide. SARS-CoV VLPs are highly immunogenic and induce neutralizing antibodies and provide protection against lethal challenge. Sf9 cell-based VLP vaccines are a potential tool to provide protection against novel pandemic agents.


Subject(s)
Membrane Glycoproteins/genetics , Membrane Glycoproteins/immunology , Severe Acute Respiratory Syndrome/prevention & control , Severe acute respiratory syndrome-related coronavirus/immunology , Viral Envelope Proteins/genetics , Viral Envelope Proteins/immunology , Viral Matrix Proteins/genetics , Viral Vaccines/immunology , Adjuvants, Immunologic/administration & dosage , Aluminum Hydroxide/administration & dosage , Animals , Antibodies, Neutralizing/blood , Antibodies, Viral/blood , Baculoviridae/genetics , Body Weight , Disease Models, Animal , Female , Genetic Vectors , Insecta , Lung/virology , Membrane Glycoproteins/metabolism , Mice , Mice, Inbred BALB C , Protein Multimerization , Recombinant Proteins/genetics , Recombinant Proteins/immunology , Recombinant Proteins/metabolism , Rodent Diseases/prevention & control , Severe acute respiratory syndrome-related coronavirus/genetics , Spike Glycoprotein, Coronavirus , Survival Analysis , Vaccines, Virosome/genetics , Vaccines, Virosome/immunology , Viral Envelope Proteins/metabolism , Viral Load , Viral Matrix Proteins/metabolism , Viral Vaccines/genetics
3.
Vaccine ; 28(30): 4771-6, 2010 Jul 05.
Article in English | MEDLINE | ID: mdl-20470801

ABSTRACT

The pandemic virus of 2009 (2009 H1N1) continues to cause illness worldwide, especially in younger age groups. The widespread H1N1 virus infection further emphasizes the need for vaccine strategies that are effective against emerging pandemic viruses and are not dependent on the limitations of traditional egg-based technology. This report describes a recombinant influenza virus-like particle (VLP) vaccine consisting of hemagglutinin (HA), neuraminidase (NA), and matrix (M1) proteins of influenza A/California/04/2009 (H1N1) virus. Influenza H1N1 VLPs with a diameter of approximately 120nm were released into the culture medium from Sf9 insect cells infected with recombinant baculovirus coexpressing HA, NA, and M1 proteins. Purified recombinant H1N1 VLPs morphologically resembled influenza virions and exhibited biological characteristics of influenza virus, including HA and NA activities. In the ferret challenge model, 2009 influenza H1N1 VLPs elicited high-titer serum hemagglutination inhibition (HI) antibodies specific for the 2009 H1N1 virus and inhibited replication of the influenza virus in the upper and lower respiratory tract tissues following A/Mexico/4482/09 (H1N1) virus challenge. Moreover, a single 15mug dose of H1N1 VLPs resulted in complete virus clearance in the ferret lung. These results provide support for the use of recombinant influenza VLP vaccine as an effective strategy against pandemic H1N1 virus.


Subject(s)
Ferrets/immunology , Influenza A Virus, H1N1 Subtype/immunology , Influenza Vaccines/immunology , Orthomyxoviridae Infections/immunology , Orthomyxoviridae Infections/prevention & control , Animals , Disease Outbreaks , Hemagglutination Tests , Influenza A Virus, H1N1 Subtype/ultrastructure , Lung/pathology , Microscopy, Electron, Transmission , Neuraminidase/analysis , Orthomyxoviridae Infections/pathology , Plasmids , Vaccination , Vaccines, Synthetic , Virion/immunology , Virus Replication
4.
Vaccine ; 25(21): 4283-90, 2007 May 22.
Article in English | MEDLINE | ID: mdl-17403562

ABSTRACT

The development of safe and effective vaccines for avian influenza viruses is a priority for pandemic preparedness. Adjuvants improve the efficacy of vaccines and may allow antigen sparing during a pandemic. We have previously shown that influenza virus-like particles (VLPs) comprised of HA, NA, and M1 proteins represent a candidate vaccine for avian influenza H9N2 virus [Pushko P, Tumpey TM, Fang Bu, Knell J, Robinson R, Smith G. Influenza virus-like particles comprised of the HA, NA, and M1 proteins of H9N2 influenza virus induce protective immune responses in BALB/c mice. Vaccine 2005;23(50):5751-9]. In this study, an H9N2 VLP vaccine and recombinant HA (rH9) vaccine were evaluated in three animal models. The H9N2 VLP vaccine protected mice and ferrets from challenge with A/Hong Kong/1073/99 (H9N2) virus. Novasome adjuvant improved immunogenicity and protection. Positive effect of the adjuvant was also detected using the rH9 vaccine. The results have implications for the development of safe and effective vaccines for avian influenza viruses with pandemic potential.


Subject(s)
Adjuvants, Immunologic/pharmacology , Influenza A Virus, H9N2 Subtype/immunology , Influenza Vaccines/immunology , Influenza, Human/prevention & control , Virosomes/immunology , Animals , Antibodies, Viral/blood , Body Weight , Disease Models, Animal , Female , Ferrets , Hemagglutination Inhibition Tests , Humans , Influenza, Human/immunology , Male , Mice , Mice, Inbred BALB C , Microscopy, Electron, Transmission , Rats , Spodoptera/cytology , Vaccines, Synthetic/immunology , Virosomes/ultrastructure
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