Characterization of a live, attenuated human parainfluenza type 3 virus candidate vaccine strain

Ray, Ranjit; Meyer, Keith; Newman, Frances K.; Belshe, Robert B.

doi:10.1128/jvi.69.3.1959-1963.1995

Cited by 11 publications

(4 citation statements)

References 35 publications

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“…Alternatively, mutations present in a current candidate vaccine strain of HPIV-3 could be inserted into pHPIV-3. One such HPIV-3 strain was passaged serially at low temperatures and found to contain mutation(s) within the L gene that attenuate growth at 37°C (30,31). The precise mutation or mutations resulting in the attenuated phenotype are not known, but by inserting specific alterations into pHPIV-3, it should be possible to distinguish between silent mutations and the attenuating mutation(s).…”

Section: Discussionmentioning

confidence: 99%

An infectious clone of human parainfluenza virus type 3

Hoffman

Banerjee

1997

J Virol

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A full-length clone of the human parainfluenza virus type 3 (HPIV-3) genome (called pHPIV-3) was constructed, and recombinant, infectious HPIV-3 was generated by transfecting pHPIV-3 and support plasmids encoding the HPIV-3 NP, P, and L proteins into HeLa cells infected with a vaccinia virus recombinant which expresses T7 RNA polymerase. T7 RNA polymerase promoters on the transfected plasmids direct the synthesis of transcripts encoding the NP, P, and L proteins and a full-length, positive-sense copy of the HPIV-3 genome. Generation of virus was dependent on transfection of pHPIV-3 and the HPIV-3 P-and L-encoding plasmids. However, a plasmid encoding the NP protein was not required since NP was expressed from pHPIV-3. Recovered virus was neutralized by anti-HPIV-3 antisera and shown to contain specific base substitutions characteristic of pHPIV-3. Recombination was shown to occur during recovery, as viruses with two distinct genotypes and phenotypes were isolated. The ability to produce infectious HPIV-3 engineered to contain specific alterations within the HPIV-3 genes and cis-acting elements expedites the study of all aspects of the virus replication cycle. Additionally, analysis of mutations may lead to the identification of attenuating genotypes, a key step in the development of a live virus vaccine.

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Section: Discussionmentioning

confidence: 99%

An infectious clone of human parainfluenza virus type 3

Hoffman

Banerjee

1997

J Virol

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“…The ts phenotype is generally associated with attenuation of virus-associated disease in host animals (18). Work on a ca, ts, attenuated candidate vaccine strain (cp45) against human parainfluenza type 3, another important cause of viral respiratory disease in the pediatric population, has shown that the ts phenotype of the virus is associated with poor transcriptional activity at the nonpermissive temperature (16,17). 2B33F and 2B20L are mutants that are both ts and ca in vitro, as well as attenuated for growth in animal models (18).…”

Section: Discussionmentioning

confidence: 99%

Biochemical characterizations of two temperature-sensitive and attenuated strains of respiratory syncytial virus subgroup B

Broughan

Randolph

Tatem

1997

J Virol

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Cold-adapted, temperature-sensitive (ts), attenuated strains of respiratory syncytial virus have been developed from a B subgroup clinical isolate for potential use as vaccine candidates. The replication of two B subgroup ts mutant viruses (2B33F and 2B20L) at the permissive and nonpermissive temperatures have been compared with that of the parental 2B virus to establish differences that may account for their ts and/or attenuated phenotypes. We have shown that the ts restriction at 39°C in the replication of the two mutant viruses in tissue culture occurs at a step after virus adsorption but before or including initiation of virusspecific mRNA transcription. At the permissive temperature of 32°C a 12-to 24-h delay in the accumulation of mRNA for both mutant viruses in comparison to that of the parental 2B virus was exhibited. This effect was mirrored by equivalent delays in viral protein synthesis and production of infectious virus. By 36 h postinfection both mutants had produced levels of viral mRNA, protein, and infectious virus that were similar to those of the parent virus at 32°C. ts؉ revertant viruses derived from both mutants have also reverted in their viral mRNA, protein, and infectious virus production kinetics at 32°C to rates more like those exhibited by the parental 2B virus. This suggests a positive correlation between the ts step in the replication of the mutant viruses and the initial delay in mRNA production that occurs at the permissive temperature.

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“…Successive passage at lower temperatures has been shown to attenuate HPIV. The most promising vaccine candidate using this approach, cp45, is designated by the number of times the virus was passaged at low temperature in African green monkey kidney cells (Belshe and Hissom, 1982;Ray et al, 1995). The vaccine was safe and immunogenic in adults, seropositive and seronegative children, and infants 1 month of age or older (Belshe et al, 2004b).…”

Section: Hpivmentioning

confidence: 99%