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Published on: April 21, 2015
Mechanisms of temperature sensitivity of attenuated Urabe mumps virus
Stephanie C Burke Schinkel1, Steven Rubin2, Kathryn E Wright1
1Department of Biochemistry, Microbiology and Immunology, University of Ottawa, 451 Smyth Road, Ottawa, K1H 8M5, Canada.
Abstract:
Temperature sensitivity is a phenotype often associated with attenuation of viruses. Previously, we purified several mumps variants from an incompletely attenuated Urabe strain live attenuated vaccine. Here we characterize one isolate that is sensitive to growth at high temperature. This virus was attenuated in a small animal model of mumps virulence, and we identified unique coding substitutions in the hemagglutinin-neuraminidase (HN), the viral polymerase (L) gene, and a non-coding substitution close to the anti-genome promoter sequences. At the non-permissive temperature, transcription of viral mRNAs and production of the replication intermediate were reduced compared to events at the permissive temperature and to a non-ts virulent Urabe virus. As well, synthesis of viral proteins was also reduced at the higher temperature. While the actual sequence substitutions in the ts virus were unique, the pattern of substitutions in HN, L and genome end sequences is similar to another attenuated Urabe virus previously described by us.
Insights
This study identifies a temperature-sensitive mumps virus variant with unique genetic changes. This variant shows reduced viral replication and protein synthesis at higher temperatures, indicating its potential as an attenuated vaccine candidate.
Area of Science:
- Virology
- Molecular Biology
- Vaccine Development
Background:
- Temperature sensitivity (ts) is a key phenotype linked to viral attenuation.
- Mumps virus variants were previously isolated from an attenuated Urabe strain vaccine.
- Understanding genetic determinants of attenuation is crucial for vaccine design.
Purpose of the Study:
- To characterize a specific temperature-sensitive mumps virus isolate.
- To investigate the genetic basis of its attenuated phenotype.
- To assess its potential for live attenuated vaccine development.
Main Methods:
- Isolation and purification of mumps virus variants.
- Phenotypic characterization of temperature sensitivity.
- Genetic sequencing to identify mutations in viral genes (HN, L) and non-coding regions.
- Assessment of viral replication, transcription, and protein synthesis at permissive and non-permissive temperatures.
- Evaluation of virulence in a small animal model.
Main Results:
- One mumps virus isolate exhibited significant temperature sensitivity.
- Unique coding substitutions were identified in the hemagglutinin-neuraminidase (HN) and viral polymerase (L) genes, along with a non-coding substitution near the anti-genome promoter.
- At non-permissive temperatures, the virus showed reduced viral mRNA transcription, replication intermediate production, and viral protein synthesis compared to permissive conditions and a virulent control.
- The characterized virus demonstrated attenuation in a small animal model of mumps virulence.
Conclusions:
- The identified temperature-sensitive mumps virus possesses unique genetic alterations contributing to its attenuated phenotype.
- The pattern of mutations in HN, L, and genome end sequences mirrors that of previously described attenuated Urabe viruses, suggesting conserved mechanisms of attenuation.
- This ts mumps virus isolate represents a promising candidate for further development as a live attenuated vaccine.
