Recombinant human parainfluenza virus type 2 vaccine candidates containing a 3′ genomic promoter mutation and L polymerase mutations are attenuated and protective in non-human primates

Nolan, Sheila M.; Skiadopoulos, Mario H.; Bradley, Konrad C.; Kim, Olivia S.; Bier, Stacia; Amaro-Carambot, Emérito; Surman, Sonja R.; Davis, Stephanie D.; Elkins, Randy; Collins, Peter L.; Murphy, Brian R.; Schaap-Nutt, Anne

doi:10.1016/j.vaccine.2007.06.028

Cited by 18 publications

(34 citation statements)

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“…The virus has a 3′ genomic promoter mutation and L polymerase mutations that result in virus attenuation. 41 In African green monkeys, immunization with the virus conferred a high level of protection in the upper and lower respiratory tracts of the challenged animals with wild-type hPIV2. 41 Here, we advocate hPIV2ΔF as another potential vaccine candidate against the wild-type hPIV2 because of its property limiting to single round infectivity with no cell-to-cell spread.…”

Section: Discussionmentioning

confidence: 99%

Vero/BC-F: an efficient packaging cell line stably expressing F protein to generate single round-infectious human parainfluenza virus type 2 vector

et al. 2014

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A stable packaging cell line (Vero/BC-F) constitutively expressing fusion (F) protein of the human parainfluenza virus type 2 (hPIV2) was established for production of the F-defective and single round-infectious hPIV2 vector in a strategy for recombinant vaccine development. The F gene expression has not evoked cytostatic or cytotoxic effects on the Vero/BC-F cells and the F protein was physiologically active to induce syncytial formation with giant polykaryocytes when transfected with a plasmid expressing hPIV2 hemagglutinin-neuraminidase (HN). Transduction of the F-defective replicon RNA into the Vero/BC-F cells led to the release of the infectious particles that packaged the replicon RNA (named as hPIV2ΔF) without detectable mutations, limiting the infectivity to a single round. The maximal titer of the hPIV2ΔF was 6.0 × 10(8) median tissue culture infections dose per ml. The influenza A virus M2 gene was inserted into hPIV2ΔF, and the M2 protein was found to be highly expressed in a human lung cancer cell line after transduction. Furthermore, in vivo airway infection experiments revealed that the hPIV2ΔF was capable of delivering transgenes to hamster tracheal cells. Thus, non-transmissible or single round-infectious hPIV2 vector will be potentially applicable to human gene therapy or recombinant vaccine development.

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Section: Discussionmentioning

confidence: 99%

Vero/BC-F: an efficient packaging cell line stably expressing F protein to generate single round-infectious human parainfluenza virus type 2 vector

et al. 2014

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“…The HPIVs collectively are the second leading cause of pediatric hospitalizations for viral respiratory disease, behind RSV and ahead of influenza (31,33,47). A licensed vaccine is currently not available for the prevention of HPIV disease, but experimental live attenuated candidate vaccines are under development for HPIV1, -2, and -3, with those for HPIV3 in clinical trials (3,5,22,32,34,51).…”

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confidence: 99%

Human Parainfluenza Virus Type 1 C Proteins Are Nonessential Proteins That Inhibit the Host Interferon and Apoptotic Responses and Are Required for Efficient Replication in Nonhuman Primates

Bartlett

Cruz

Esker

et al. 2008

J Virol

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Recombinant human parainfluenza virus type 1 (rHPIV1) was modified to create rHPIV1-P(C؊), a virus in which expression of the C proteins (C, C, Y1, and Y2) was silenced without affecting the amino acid sequence of the P protein. Infectious rHPIV1-P(C؊) was readily recovered from cDNA, indicating that the four C proteins were not essential for virus replication. Early during infection in vitro, rHPIV1-P(C؊) replicated as efficiently as wild-type (wt) HPIV1, but its titer subsequently decreased coincident with the onset of an extensive cytopathic effect not observed with wt rHPIV1. rHPIV1-P(C؊) infection, but not wt rHPIV1 infection, induced caspase 3 activation and nuclear fragmentation in LLC-MK2 cells, identifying the HPIV1 C proteins as inhibitors of apoptosis. In contrast to wt rHPIV1, rHPIV1-P(C؊) and rHPIV1-C F170S , a mutant encoding an F170S substitution in C, induced interferon (IFN) and did not inhibit IFN signaling in vitro. However, only rHPIV1-P(C؊) induced apoptosis. Thus, the anti-IFN and antiapoptosis activities of HPIV1 were separable: both activities are disabled in rHPIV1-P(C؊), whereas only the anti-IFN activity is disabled in rHPIV1-C F170S . In African green monkeys (AGMs), rHPIV1-P(C؊) was considerably more attenuated than rHPIV1-C F170S , suggesting that disabling the anti-IFN and antiapoptotic activities of HPIV1 had additive effects on attenuation in vivo. Although rHPIV1-P(C؊) protected against challenge with wt HPIV1, its highly restricted replication in AGMs and in primary human airway epithelial cell cultures suggests that it might be overattenuated for use as a vaccine. Thus, the C proteins of HPIV1 are nonessential but have anti-IFN and antiapoptosis activities required for virulence in primates.

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“…The strategy of deleting one or several codons to make attenuating mutations has been successful in some instances with parainfluenza virus types 1 and 2 (33,34). The expectation is that deletion of 3 nucleotides (per codon) would be more refractory to reversion than a single missense nucleotide substitution.…”

Section: Discussionmentioning

confidence: 99%

Respiratory Syncytial Virus Modified by Deletions of the NS2 Gene and Amino Acid S1313 of the L Polymerase Protein Is a Temperature-Sensitive, Live-Attenuated Vaccine Candidate That Is Phenotypically Stable at Physiological Temperature

Luongo

Winter

Collins

et al. 2013

J Virol

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Human respiratory syncytial virus (RSV) is the leading viral cause of lower respiratory tract disease in infants and children worldwide. In previous work to develop point mutations in RSV with improved genetic stability, we observed that an attenuating mutation at amino acid position 1321 in the L polymerase protein was subject to deattenuation by a spontaneous second-site compensatory mutation at position 1313 (C. Luongo, C. C. Winter, P. L. Collins, and U. J. Buchholz, J. Virol. 86:10792-10804, 2012). In the present study, we found that deletion of position 1313 (⌬1313), irrespective of the presence of an attenuating mutation at position 1321, provided a new attenuating mutation. RSV bearing ⌬1313 replicated in cell culture as efficiently as wildtype virus at 32°C, was restricted for replication at 37°C, and was restricted 50-fold and 150-fold in the upper and lower respiratory tracts, respectively, of mice. We combined the ⌬1313 deletion with the previously described, attenuating NS2 gene deletion (⌬NS2) to produce the recombinant live-attenuated RSV vaccine candidate ⌬NS2/⌬1313. During in vitro stress tests involving serial passage at incrementally increasing temperatures, a second-site compensatory mutation was detected in close proximity of ⌬1313, namely, I1314T. This site was genetically and phenotypically stabilized by an I1314L substitution. Combination of I1314L with ⌬NS2/⌬1313 yielded a virus, ⌬NS2/⌬1313/1314L, with genetic stability at physiological temperature. This stabilized vaccine candidate was moderately temperature sensitive and had a level of restriction in chimpanzees comparable to that of MEDI-559, a promising RSV vaccine candidate that presently is in clinical trials but lacks stabilized attenuating mutations. The level of attenuation and genetic stability identify ⌬NS2/⌬1313/1314L as a promising candidate for evaluation in pediatric phase I studies.H uman respiratory syncytial virus (RSV) is the leading viral cause of lower respiratory tract infection in infants and young children worldwide. RSV is an enveloped, nonsegmented negative-strand RNA virus that is a member of the subfamily Pneumovirinae, genus Pneumovirus, of family Paramyxoviridae. Its genome is approximately 15.2 kb in length and encodes 11 major proteins, including nucleoprotein N, phosphoprotein P, matrix protein M, polymerase protein L, polymerase factors M2-1 and M2-2, fusion glycoprotein F, attachment glycoprotein G, small hydrophobic protein SH, and nonstructural proteins NS1 and NS2. The viral gene order is 3=-NS1-NS2-N-P-M-SH-G-F-M2-1/ M2-2-L-5=.RSV causes an estimated 34 million episodes of acute lower respiratory tract infections in children under 5 years of age and 199,000 pediatric deaths annually worldwide (1). The only prophylaxis available to prevent RSV disease is a commercial neutralizing antibody to RSV F that is administered to high-risk infants as passive immunization monthly during the RSV season (2, 3). There are no effective antivirals, and a licensed vaccine is not available, even though the need for a...

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Recombinant human parainfluenza virus type 2 vaccine candidates containing a 3′ genomic promoter mutation and L polymerase mutations are attenuated and protective in non-human primates

Cited by 18 publications

References 50 publications

Vero/BC-F: an efficient packaging cell line stably expressing F protein to generate single round-infectious human parainfluenza virus type 2 vector

Vero/BC-F: an efficient packaging cell line stably expressing F protein to generate single round-infectious human parainfluenza virus type 2 vector

Human Parainfluenza Virus Type 1 C Proteins Are Nonessential Proteins That Inhibit the Host Interferon and Apoptotic Responses and Are Required for Efficient Replication in Nonhuman Primates

Respiratory Syncytial Virus Modified by Deletions of the NS2 Gene and Amino Acid S1313 of the L Polymerase Protein Is a Temperature-Sensitive, Live-Attenuated Vaccine Candidate That Is Phenotypically Stable at Physiological Temperature

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