Efficacy of Parainfluenza Virus 5 Mutants Expressing Hemagglutinin from H5N1 Influenza A Virus in Mice

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and may be good candidates for vaccine development. In this study, we examined immunity induced by rMuV⌬SH and rMuV⌬V in mice. Furthermore, we generated recombinant mumps viruses lacking expression of both the V protein and the SH protein (rMuV⌬SH⌬V). Analysis of rMuV⌬SH⌬V indicated that it was stable in tissue culture cell lines. Importantly, rMuV⌬SH⌬V was immunogenic in mice, indicating that it is a promising candidate for mumps vaccine development. Mumps is a human infectious disease characterized by lateral or bilateral nonsuppurative swelling of the parotid glands. In severe cases, mumps can lead to orchitis in postpuberty male patients and damage to the central nervous system. In the prevaccine era, 90% of the population turned seropositive for mumps virus (MuV) by 14 to 15 years of age, reflecting its highly contagious nature. Mumps virus is neurotropic and was one of the most common causes of aseptic meningitis before the implementation of mass mumps vaccination programs.At present, the Jeryl Lynn (JL) vaccine is the most commonly used mumps vaccine, administered as lyophilized live virus with measles and rubella vaccine components. The JL vaccine strain originated from an infectious isolate from a mumps patient in 1963 (1). The virus was attenuated through continuous passages in embryonic hen eggs and chicken embryos/chicken embryo cell cultures (1). The JL vaccine was licensed in the United States in 1967 and has been used for over 40 years. This vaccine has been efficacious and safe overall (2-6). However, several large mumps outbreaks have occurred recently in the United States and worldwide in populations that have been vaccinated with the JL vaccine (7-10). Major mumps outbreaks in the United States include the 2006 multistate mumps outbreak, reporting 6,584 suspected cases originating from the state of Iowa (11, 12) and the 2009 -2010 New York and New Jersey mumps outbreaks with a total of 2,078 suspected cases reported in 2010 (13). Both of the outbreaks occurred among highly vaccinated populations, raising questions about the efficacy of the current vaccination program in the United States. One possible causality is the antigenic differences between the genotype A vaccine strain and the genotype G circulating wild-type mumps viruses.In this study, we seek to develop a mumps vaccine candidate through genetic modification of a clinically isolated mumps virus. Mumps virus is a member of the family Paramyxoviridae, subfamily Paramyxovirinae, and genus Rubulavirus (6,14). It is an enveloped virus enclosing a negative-sense, single-stranded, nonsegmented RNA genome of 15,384 nucleotides in length which encodes 9 viral proteins (15-17). Studies of the function of the Paramyxovirus SH protein reveal that it blocks tumor necrosis factor alpha (TNF-␣) induction, signaling, caspase activation, and NF-B nuclear translocation in transfected and virus-infected cells (18)(19)(20)(21)(22)(23). The V protein is an accessory protein translated from the authentic transcript of the V/P gene (24,25). Mumps V protein...

Section: Discussionsupporting

confidence: 61%

Immunogenicity of Novel Mumps Vaccine Candidates Generated by Genetic Modification

Chen

Phan

et al. 2014

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“…Two recombinant PIV5 viruses containing the NA gene between the HN and L genes, ZL108 (rPIV5-H1N1-N1-HN/L) and ZL116 (rPIV5-H5N1-N1-HN/L), were generated as previously described (24)(25)(26)(27)(28). Plasmid pZL108 or pZL116 and three helper plasmids, pPIV5-NP, pPIV5-P, and pPIV5-L, encoding the NP, P, and L proteins, respectively, were cotransfected into BSR-T7 cells, and after 72 h of incubation at 37°C, the media were harvested and cell debris was pelleted by low-speed centrifugation (3,000 rpm for 10 min).…”

Section: Methodsmentioning

confidence: 99%

“…PIV5 titers were determined by a plaque assay on Vero cells as previously reported (24,26). Briefly, cells were incubated with serial dilutions of the virus for 2 h, the virus was removed, and cells were overlaid with 1:1 low-melting-point agarose and DMEM with 2% FBS plus antibiotic-antimycotic and incubated at 37°C for 5 to 6 days.…”

Section: Methodsmentioning

confidence: 99%

“…Recombinant PIV5 (rPIV5) is nonpathogenic, does not have a DNA phase in the life cycle (reducing the potential for integration), has a stable genome, and replicates to high titers without cytopathic effects in a variety of mammalian cell substrates (reviewed in reference 23). We previously demonstrated that rPIV5 vaccines expressing the HA gene from influenza A virus strain A/Udorn/72 (H3N2) (rPIV5-H3) or A/Vietnam/1203/04 (H5N1) (A/VN/1203-04) (rPIV5-H5) are safe and efficacious, providing protection against homologous influenza virus challenge in murine models (24)(25)(26)(27). Moreover, the expression of the internal nucleocapsid (NP) protein, a dominant T cell antigen in mice, by PIV5 (rPIV5-NP) elicited heterosubtypic immunity in mice and provided protection against H1N1 and H5N1 challenges (28).…”

mentioning

confidence: 99%

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Vaccination with Recombinant Parainfluenza Virus 5 Expressing Neuraminidase Protects against Homologous and Heterologous Influenza Virus Challenge

Mooney

Gabbard

et al. 2017

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Seasonal human influenza virus continues to cause morbidity and mortality annually, and highly pathogenic avian influenza (HPAI) viruses along with other emerging influenza viruses continue to pose pandemic threats. Vaccination is considered the most effective measure for controlling influenza; however, current strategies rely on a precise vaccine match with currently circulating virus strains for efficacy, requiring constant surveillance and regular development of matched vaccines. Current vaccines focus on eliciting specific antibody responses against the hemagglutinin (HA) surface glycoprotein; however, the diversity of HAs across species and antigenic drift of circulating strains enable the evasion of virus-inhibiting antibody responses, resulting in vaccine failure. The neuraminidase (NA) surface glycoprotein, while diverse, has a conserved enzymatic site and presents an appealing target for priming broadly effective antibody responses. Here we show that vaccination with parainfluenza virus 5 (PIV5), a promising live viral vector expressing NA from avian (H5N1) or pandemic (H1N1) influenza virus, elicited NA-specific antibody and T cell responses, which conferred protection against homologous and heterologous influenza virus challenges. Vaccination with PIV5-N1 NA provided cross-protection against challenge with a heterosubtypic (H3N2) virus. Experiments using antibody transfer indicate that antibodies to NA have an important role in protection. These findings indicate that PIV5 expressing NA may be effective as a broadly protective vaccine against seasonal influenza and emerging pandemic threats.IMPORTANCE Seasonal influenza viruses cause considerable morbidity and mortality annually, while emerging viruses pose potential pandemic threats. Currently licensed influenza virus vaccines rely on the antigenic match of hemagglutinin (HA) for vaccine strain selection, and most vaccines rely on HA inhibition titers to determine efficacy, despite the growing awareness of the contribution of neuraminidase (NA) to influenza virus vaccine efficacy. Although NA is immunologically subdominant to HA, and clinical studies have shown variable NA responses to vaccination, in this study, we show that vaccination with a parainfluenza virus 5 recombinant vaccine candidate expressing NA (PIV5-NA) from a pandemic influenza (pdmH1N1) virus or highly pathogenic avian influenza (H5N1) virus elicits robust, cross-reactive protection from influenza virus infection in two animal models. New vaccination strategies incorporating NA, including PIV5-NA, could improve seasonal influenza virus vaccine efficacy and provide protection against emerging influenza viruses.KEYWORDS influenza, vaccine, neuraminidase, parainfluenza virus 5, highly pathogenic avian influenza virus

“…PIV5 can infect humans and animals without causing any known symptoms or diseases (3,4). It has been used as a vector for vaccine development (5)(6)(7)(8)(9)(10)(11).…”

mentioning

confidence: 99%

Regulation of Viral RNA Synthesis by the V Protein of Parainfluenza Virus 5

Yang

Zengel

Sun

et al. 2015

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Paramyxoviruses include many important animal and human pathogens. The genome of parainfluenza virus 5 (PIV5), a prototypical paramyxovirus, encodes a V protein that inhibits viral RNA synthesis. In this work, the mechanism of inhibition was investigated. Using mutational analysis and a minigenome system, we identified regions in the N and C termini of the V protein that inhibit viral RNA synthesis: one at the very N terminus of V and the second at the C terminus of V. Furthermore, we determined that residues L16 and I17 are critical for the inhibitory function of the N-terminal region of the V protein. IMPORTANCEWe identified two regions of the V protein that interact with NP and determined that one of these regions enhances viral RNA transcription via its interaction with NP. Our data suggest that a common host factor may be involved in the regulation of paramyxovirus replication and could be a target for broad antiviral drug development. Understanding the regulation of paramyxovirus replication will enable the rational design of vaccines and potential antiviral drugs. The family Paramyxoviridae belongs to the Mononegavirales and has two subfamilies, the Paramyxovirinae and the Pneumovirinae. The subfamily Paramyxovirinae contains five genera: Rubulavirus, Respirovirus, Morbillivirus, Avulavirus, and Henipavirus. The subfamily Pneumovirinae contains two genera: Pneumovirus and Metapneumovirus. This virus family includes many wellknown human and animal pathogens, such as mumps virus (MuV), which belongs to the genus Rubulavirus; Sendai virus (SeV) and human parainfluenza virus 3 (PIV3), which belong to the genus Respirovirus; measles virus (MV), which belongs to the genus Morbillivirus; and emerging viruses like Nipah virus (NiV), which belongs to the genus Henipavirus (1). Parainfluenza virus 5 (PIV5), formerly known as simian virus 5 (SV5) (2), is a prototypical paramyxovirus and belongs to the Rubulavirus genus in the Paramyxoviridae family. PIV5 can infect humans and animals without causing any known symptoms or diseases (3, 4). It has been used as a vector for vaccine development (5-11).The PIV5 genome consists of a nonsegmented negativestranded RNA which is encapsidated by the nucleocapsid (NP) protein (1). The viral RNA polymerase (vRNAP) complex that contains the large (L) protein and the phosphoprotein (P) transcribes the NP-encapsidated genome RNA into 5=-capped and 3=-polyadenylated mRNAs. vRNAP starts viral RNA synthesis at the 3= end of the genome, and it transcribes the genes into mRNAs in a sequential (and polar) manner by terminating and reinitiating transcription at each of the gene junctions (1). vRNAP also replicates the RNA genome by making an exact copy of the genome from negative-sense RNA to positive-sense RNA and from positive-sense RNA back to negative-sense RNA. Viral RNA transcription occurs first after infection to allow the production of viral proteins, and viral RNA replication follows (1). PIV5 is an enveloped virus and enters the cell by fusing with the plasma mem-