A Live Attenuated Bovine Parainfluenza Virus Type 3 Vaccine Is Safe, Infectious, Immunogenic, and Phenotypically Stable in Infants and Children

The shipping fever (SF) and Kansas (Ka) strains of bovine parainfluenza virus type 3 (BPIV3) are restricted in their replication in rhesus monkeys 100-to 1,000-fold compared to human parainfluenza virus type 3 (HPIV3), and the Ka strain also was shown to be attenuated in humans. To initiate an investigation of the genetic basis of the attenuation of BPIV3 in primates, we produced viable chimeric HPIV3 recombinants containing the nucleoprotein (N) open reading frame (ORF) from either BPIV3 Ka or SF in place of the HPIV3 N ORF. These chimeric recombinants were designated cKa-N and cSF-N, respectively. Remarkably, cKa-N and cSF-N grew to titers comparable to those of their HPIV3 and BPIV3 parents in LLC-MK2 monkey kidney and Madin-Darby bovine kidney cells. Thus, the heterologous nature of the N protein did not impede replication in vitro. However, cKa-N and cSF-N were each restricted in replication in rhesus monkeys to a similar extent as Ka and SF, respectively. This identified the BPIV3 N protein as a determinant of the host range restriction of BPIV3 in primates. These chimeras thus combine the antigenic determinants of HPIV3 with the host range restriction and attenuation phenotype of BPIV3. Despite their restricted replication in rhesus monkeys, the chimeric viruses induced a level of resistance to HPIV3 challenge in these animals which was indistinguishable from that conferred by immunization with HPIV3. The infectivity, attenuation, and immunogenicity of these BPIV3/HPIV3 chimeras suggest that the modified Jennerian approach described in the present report represents a novel method to design vaccines to protect against HPIV3-induced disease in humans.Human parainfluenza virus type 3 (HPIV3) and bovine parainfluenza virus type 3 (BPIV3) are enveloped, nonsegmented negative-strand RNA viruses within the Respirovirus genus of the family Paramyxoviridae (8). HPIV3 is a common cause of serious lower respiratory tract infection in infants and children less than 1 year of age (8, 9, 23), but a vaccine has not yet been licensed for HPIV3-induced disease. Since BPIV3 and HPIV3 have been shown to be 25% related antigenically by reciprocal cross-neutralization studies (6) and share neutralizing epitopes on their HN and F surface glycoproteins (7, 21), BPIV3 is being evaluated for use as a live-virus vaccine to protect humans against HPIV3-induced disease. The use of a naturally occurring animal pathogen to immunize humans against the counterpart human pathogen was pioneered by Jenner 200 years ago to control smallpox. In the Jennerian approach to vaccine development, an antigenically related animal virus that is restricted in replication in humans is used as a vaccine to protect against the related human disease. BPIV3 strains Kansas (Ka) and shipping fever (SF) were restricted in their replication in the upper and lower respiratory tracts of rhesus monkeys relative to HPIV3, induced antibodies that cross-neutralized HPIV3, and conferred resistance to challenge with HPIV3 (6). BPIV3 Ka was found to be attenuated, infe...

Section: Discussionmentioning

confidence: 97%

“…BPIV3 wt strain Ka/15626/84 (clone 5-2-4, lot BPI3-1) was previously described (3,19); the virus preparation used in the present work is identical in passage level to that currently undergoing clinical evaluation. The prototype BPIV3 wt strain SF (ATCC 281-VR) was obtained from the American Type Culture Collection, Manassas, Va.…”

Section: Methodsmentioning

confidence: 99%

A Recombinant Human Parainfluenza Virus Type 3 (PIV3) in Which the Nucleocapsid N Protein Has Been Replaced by That of Bovine PIV3 Is Attenuated in Primates

Bailly

McAuliffe

Durbin

et al. 2000

“…It would be anticipated that a growth restriction in a nonnatural host would involve multiple genes and many sequence differences, and this is consistent with the present findings that the G and F glycoproteins contribute only part of this phenotype. Not surprisingly, experience with other viruses indicates that a host range restriction is a very stable phenotype (19,33). Also, host range restrictions typically are not associated with temperature sensitivity.…”

Section: Discussionmentioning

confidence: 99%

“…This strategy requires that the human virus of interest has an animal counterpart which is attenuated in the nonnatural human host, yet grows sufficiently well and is sufficiently related antigenically that it can induce effective protection against the human virus. As a current example, bovine parainfluenzavirus (BPIV) type 3 is under clinical evaluation as vaccine against human parainfluenzavirus (HPIV) type 3 and appears to have appropriate levels of attenuation and immunogenicity (19). The recently licensed quadrivalent rotavirus vaccine exemplifies both the Jennerian approach and a "modified Jennerian" approach (33).…”

mentioning

confidence: 99%

Chimeric Bovine Respiratory Syncytial Virus with Glycoprotein Gene Substitutions from Human Respiratory Syncytial Virus (HRSV): Effects on Host Range and Evaluation as a Live-Attenuated HRSV Vaccine

Buchholz¹,

Granzow

Schuldt³

et al. 2000

We recently developed a system for the generation of infectious bovine respiratory syncytial virus (BRSV) from cDNA. Here, we report the recovery of fully viable chimeric recombinant BRSVs (rBRSVs) that carry human respiratory syncytial virus (HRSV) glycoproteins in place of their BRSV counterparts, thus combining the replication machinery of BRSV with the major antigenic determinants of HRSV. A cDNA encoding the BRSV antigenome was modified so that the complete G and F genes, including the gene start and gene end signals, were replaced by their HRSV A2 counterparts. Alternatively, the BRSV F gene alone was replaced by that of HRSV Long. Each antigenomic cDNA directed the successful recovery of recombinant virus, yielding rBRSV/A2 and rBRSV/LongF, respectively. The HRSV G and F proteins or the HRSV F in combination with BRSV G were expressed efficiently in cells infected with the appropriate chimeric virus and were efficiently incorporated into recombinant virions. Whereas BRSV and HRSV grew more efficiently in bovine and human cells, respectively, the chimeric rBRSV/A2 exhibited intermediate growth characteristics in a human cell line and grew better than either parent in a bovine line. The cytopathology induced by the chimera more closely resembled that of BRSV. BRSV was confirmed to be highly restricted for replication in the respiratory tract of chimpanzees, a host that is highly permissive for HRSV. Interestingly, the rBRSV/A2 chimeric virus was somewhat more competent than BRSV for replication in chimpanzees but remained highly restricted compared to HRSV. This showed that the substitution of the G and F glycoproteins alone was not sufficient to induce efficient replication in chimpanzees. Thus, the F and G proteins contribute to the host range restriction of BRSV but are not the major determinants of this phenotype. Although rBRSV/A2 expresses the major neutralization and protective antigens of HRSV, chimpanzees infected with this chimeric virus were not significantly protected against subsequent challenge with wild-type HRSV. This suggests that the growth restriction of rBRSV/A2 was too great to provide adequate antigen expression and that the capacity of this chimeric vaccine candidate for replication in primates will need to be increased by the importation of additional HRSV genes. Bovine respiratory syncytial virus (BRSV) andHuman respiratory syncytial virus (HRSV) are closely related members of the Pneumovirus genus within the subfamily Pneumovirinae, family Paramyxoviridae, order Mononegavirales (6, 36). BRSV is a major cause of respiratory tract disease in cattle (32, 44). HRSV is the most important causative agent of viral pediatric respiratory disease worldwide (6). A licensed vaccine against HRSV is not available, though several promising candidates for attenuated live vaccines recently have been developed (references 17, 46, 47, and 48, and references therein).The genomes of HRSV and of BRSV are single-stranded, negative-sense RNAs of 15,222 nucleotides (nt) (HRSV A2) and 15,140 nt (BRSV ATue51...

“…The CDC reports that, in humans, most children have antibodies against human PIV-3 (HPIV-3) by 5 years of age (http://www.cdc.gov/parainfluenza /hcp/clinical.html). There is currently no vaccine available for control of HPIV-3 infection; however, a few studies have examined the use of an attenuated bovine PIV-3 (BPIV-3) to protect against HPIV-3 because of the homology between bovine and human strains (1)(2)(3). Given the lack of an efficacious vaccine for HPIV-3, there is a critical need to understand the mechanisms of HPIV-3-induced disease and the molecular pathways associated with viral modulation of the host antiviral defenses.…”

Section: Importancementioning

confidence: 99%

Parainfluenza Virus 3 Blocks Antiviral Mediators Downstream of the Interferon Lambda Receptor by Modulating Stat1 Phosphorylation

Eberle

McGill

Reinhardt

et al. 2016

Parainfluenza viruses are known to inhibit type I interferon (IFN) production; however, there is a lack of information regarding the type III IFN response during infection. Type III IFNs signal through a unique heterodimeric receptor, IFN-R1/interleukin-10R2 (IL-10R2), which is primarily expressed by epithelial cells. Parainfluenza virus 3 (PIV-3) infection is highly restricted to the airway epithelium. We therefore sought to examine type III IFN signaling pathways during PIV-3 infection of epithelial cells. We used three strains of PIV-3: human PIV-3 (HPIV-3), bovine PIV-3 (BPIV-3), and dolphin PIV-1 (Tursiops truncatus PIV-1, or TtPIV-1). Here, we show that message levels of IL-29 are significantly increased during PIV-3 infection, yet downstream antiviral signaling molecules are not upregulated to levels similar to those of the positive control. IMPORTANCEParainfluenza virus (PIV) in humans is associated with bronchiolitis and pneumonia and can be especially problematic in infants and the elderly. Also seen in cattle, bovine PIV-3 causes respiratory infections in young calves. In addition, PIV-3 is one of a number of pathogens that contribute to the bovine respiratory disease complex (BRDC). As their name suggests, interferons (IFNs) are produced by cells to interfere with viral replication. Paramyxoviruses have previously been shown to block production and downstream signaling of type I IFNs. For the first time, it is shown here that PIV-3 can induce protective type III IFNs in epithelial cells, the primary site of PIV-3 infection. However, we found that PIV-3 modulates signaling pathways downstream of the type III IFN receptor to block production of several specific molecules that aid in a productive antiviral response. Importantly, this work expands our understanding of how PIV-3 effectively evades host innate immunity. P arainfluenza virus 3 (PIV-3) causes a prominent respiratory infection in both cattle and humans. The CDC reports that, in humans, most children have antibodies against human PIV-3 (HPIV-3) by 5 years of age (http://www.cdc.gov/parainfluenza /hcp/clinical.html). There is currently no vaccine available for control of HPIV-3 infection; however, a few studies have examined the use of an attenuated bovine PIV-3 (BPIV-3) to protect against HPIV-3 because of the homology between bovine and human strains (1-3). Given the lack of an efficacious vaccine for HPIV-3, there is a critical need to understand the mechanisms of HPIV-3-induced disease and the molecular pathways associated with viral modulation of the host antiviral defenses.Paramyoxviruses are negative-sense single-stranded RNA viruses which are part of the Paramyxoviridae family and Paramyxovirinae subfamily (4). PIV-3 is found within the genus Respirovirus. PIV-3 is a respiratory virus that primarily infects the epithelial cells of the lung. Symptoms of HPIV-3 infection include bronchiolitis and pneumonia, and HPIV-3 is especially problematic in infants (4). Airway epithelial cells recognize viral infection through pattern recog...