Recombinant Parainfluenza Virus 5 Vaccine Encoding the Influenza Virus Hemagglutinin Protects against H5N1 Highly Pathogenic Avian Influenza Virus Infection following Intranasal or Intramuscular Vaccination of BALB/c Mice

Department of Infectious Diseases, University of Georgia, College of Veterinary Medicine, Athens, GA 30622.
Journal of Virology (Impact Factor: 4.44). 10/2012; 87(1). DOI: 10.1128/JVI.02330-12
Source: PubMed


New approaches for vaccination to prevent influenza virus infection are needed. Emerging viruses, such as H5N1 highly pathogenic avian influenza (HPAI) virus, pose not only pandemic threats, but also challenges in vaccine development and production. Parainfluenza virus 5 (PIV5) is an appealing vector for vaccine development and we have previously shown that intranasal immunization with PIV5 expressing the hemagglutinin from influenza virus was protective against influenza virus challenge. While intranasal immunization is an appealing approach, PIV5 may have potential to be utilized in other formats, prompting us to test the efficacy of rPIV5-H5, which encodes the HA from H5N1 HPAI virus, in different vaccination schemes. In the BALB/c mouse model, a single intramuscular or intranasal immunization with a live rPIV5-H5 (ZL46) rapidly induced robust neutralizing serum antibody responses and protected against HPAI challenge, although mucosal IgA responses primed by intranasal immunization more effectively controlled virus replication in the lung. The rPIV5-H5 vaccine incorporated the H5 HA into the virion, and so we tested the efficacy of an inactivated format of the vaccine. Inactivated rPIV5-H5 primed neutralizing serum antibody responses and controlled H5N1 virus replication, although similar to other H5 antigen vaccines, it required a booster immunization to prime protective immune responses. Taken together, these results suggest that rPIV5-HA vaccines and H5-specific vaccines in particular, may be utilized in multiple formats and by multiple routes of administration. This could avoid potential contraindications based upon intranasal administration alone and provide opportunities for broader applications, with the use of a single vaccine vector.

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    • "Sulfuric acid was added to stop the TMB reaction and plates were read at 450 nm using a BioTek Powerwave plate reader (Bio-TEK, Winooski, Vermont, USA). For some samples, the virus titer was also estimated by measuring hemagglutinin (HA) of MDCK culture supernatants with 0.5% turkey red blood cells (Soboleski et al. 2011; Mooney et al. 2013). Briefly, 0.05 mL of supernatant from each well of the TCID 50 plate was added to 0.05 mL of 0.5% red blood cells (diluted in PBS) and assayed for agglutination within 1 h. "
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    ABSTRACT: Abstract Avian influenza has emerged as one of the most ubiquitous viruses within our biosphere. Wild aquatic birds are believed to be the primary reservoir of all influenza viruses; however, the spillover of H5N1 highly pathogenic avian influenza (HPAI) and the recent swine-origin pandemic H1N1 viruses have sparked increased interest in identifying and understanding which and how many species can be infected. Moreover, novel influenza virus sequences were recently isolated from New World bats. Crocodilians have a slow rate of molecular evolution and are the sister group to birds; thus they are a logical reptilian group to explore susceptibility to influenza virus infection and they provide a link between birds and mammals. A primary American alligator (Alligator mississippiensis) cell line, and embryos, were infected with four, low pathogenic avian influenza (LPAI) strains to assess susceptibility to infection. Embryonated alligator eggs supported virus replication, as evidenced by the influenza virus M gene and infectious virus detected in allantoic fluid and by virus antigen staining in embryo tissues. Primary alligator cells were also inoculated with the LPAI viruses and showed susceptibility based upon antigen staining; however, the requirement for trypsin to support replication in cell culture limited replication. To assess influenza virus replication in culture, primary alligator cells were inoculated with H1N1 human influenza or H5N1 HPAI viruses that replicate independent of trypsin. Both viruses replicated efficiently in culture, even at the 30 C temperature preferred by the alligator cells. This research demonstrates the ability of wild-type influenza viruses to infect and replicate within two crocodilian substrates and suggests the need for further research to assess crocodilians as a species potentially susceptible to influenza virus infection.
    Journal of wildlife diseases 01/2015; 51(1):187-198. DOI:10.7589/2013-12-321 · 1.36 Impact Factor
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    • "Additionally, a single-dose inoculation of PIV5 expressing hemagglutinin (HA) or the NP protein of influenza virus protects against lethal H5N1 challenge in mice [15] [16]. Importantly, intranasal administration of PIV5 is effective for eliciting robust mucosal immune responses [17], and is therefore ideal for vaccinating against respiratory pathogens. "
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    ABSTRACT: Human respiratory syncytial virus (RSV) is a leading cause of severe respiratory disease and hospitalizations in infants and young children. It also causes significant morbidity and mortality in elderly and immune compromised individuals. No licensed vaccine currently exists. Parainfluenza virus 5 (PIV5) is a paramyxovirus that causes no known human illness and has been used as a platform for vector-based vaccine development. To evaluate the efficacy of PIV5 as a RSV vaccine vector, we generated two recombinant PIV5 viruses - one expressing the fusion (F) protein and the other expressing the attachment glycoprotein (G) of RSV strain A2 (RSV A2). The vaccine strains were used separately for single-dose vaccinations in BALB/c mice. The results showed that both vaccines induced RSV antigen-specific antibody responses, with IgG2a/IgG1 ratios similar to those seen in wild-type RSV A2 infection. After challenging the vaccinated mice with RSV A2, histopathology of lung sections showed that the vaccines did not exacerbate lung lesions relative to RSV A2-immunized mice. Importantly, both F and G vaccines induced protective immunity. Therefore, PIV5 presents an attractive platform for vector-based vaccines against RSV infection.
    Vaccine 04/2014; 32(25). DOI:10.1016/j.vaccine.2014.03.049 · 3.62 Impact Factor
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    ABSTRACT: A safe and effective vaccine is the best way to prevent large-scale high pathogenic avian influenza virus (HPAI) H5N1 outbreaks in the human population. Currently FDA-approved H5N1 vaccine has serious limitations. A more efficacious H5N1 vaccine is urgently needed. Parainfluenza virus 5 (PIV5), a paramyxovirus, is not known to cause any illness in humans. PIV5 is an attractive vaccine vector. In our studies, a single dose of a live recombinant PIV5 expressing a HA gene of H5N1 (rPIV5-H5) from the H5N1 subtype provided sterilizing immunity against lethal dose of HPAI H5N1 infection in mice. Furthermore, we have examined the effect of insertion of H5N1 HA at different locations within the PIV5 genome on efficacy of PIV5-based vaccine. Interestingly, insertion of H5N1 HA between the leader sequence, the de facto promoter of PIV5, and the first viral gene, NP, did not lead to a viable virus. Insertion of H5N1 HA between NP and the next gene V/P led to a virus that was defective in growth. We have found that insertion of H5N1 HA at the junction between the SH gene and the HN gene gave the best immunity against HPAI H5N1 challenge: a dose as low as 1000 plaque forming unit (PFU) was sufficient to protect against lethal HPAI H5N1 challenge in mice. The work suggests that recombinant PIV5 expressing H5N1 HA has great potential as HPAI H5N1 vaccine.
    Journal of Virology 10/2012; 87(1). DOI:10.1128/JVI.02321-12 · 4.44 Impact Factor
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