Codon stabilization analysis of the “248” temperature sensitive mutation for increased phenotypic stability of respiratory syncytial virus vaccine candidates.

Luongo, C., Yang, L., Winter, C.C., Spann, K. M., Murphy, B. R., Collins, P. L. and Buchholz, U. J. (2009) Codon stabilization analysis of the “248” temperature sensitive mutation for increased phenotypic stability of respiratory syncytial virus vaccine candidates.. Vaccine, 27 41: 5667-5676. doi:10.1016/j.vaccine.2009.07.022


Author Luongo, C.
Yang, L.
Winter, C.C.
Spann, K. M.
Murphy, B. R.
Collins, P. L.
Buchholz, U. J.
Title Codon stabilization analysis of the “248” temperature sensitive mutation for increased phenotypic stability of respiratory syncytial virus vaccine candidates.
Journal name Vaccine   Check publisher's open access policy
ISSN 0264-410X
Publication date 2009-09-18
Year available 2009
Sub-type Article (original research)
DOI 10.1016/j.vaccine.2009.07.022
Open Access Status Not yet assessed
Volume 27
Issue 41
Start page 5667
End page 5676
Total pages 10
Editor Spier, R. E.
Place of publication United Kingdom
Publisher Elsevier
Language eng
Subject 060502 Infectious Agents
060506 Virology
920115 Respiratory System and Diseases (incl. Asthma)
920109 Infectious Diseases
C1
Abstract Human respiratory syncytial virus (RSV) is the most important viral agent of serious pediatric respiratory tract illness worldwide. Presently, the most promising vaccine candidate is a live, attenuated, cDNA-derived virus, RSV rA2cp248/404/1030ΔSH, whose attenuation phenotype is based in large part on a series of point mutations including a glutamine to leucine (Q to L) substitution at amino acid residue 831 of the polymerase protein L, a mutation originally called “248”. This mutation specifies both a temperature sensitive (ts) and attenuation phenotype. Reversion of this mutation from leucine back to glutamine was detected in some samples in clinical phase 1 trials. To identify the most genetically stable “attenuating” codon at this position to be included in a more stable RSV vaccine, we sought to create and evaluate recombinant RSVs representing all 20 possible amino acid assignments at this position, as well as small insertions and deletions. The recoverable viruses constituted a panel representing 18 different amino acid assignments, and were evaluated for temperature sensitivity in vitro and attenuation in mice. The original leucine mutation was found to be the most attenuating, followed only by phenylalanine. The paucity of highly attenuating assignments limited the possibility of increasing genetic stability. Indeed, it was not possible to find a leucine or phenylalanine codon requiring more than a single nucleotide change to yield a “non-attenuating” codon, as is necessary for the stabilization strategy. Nonetheless, serial passage of the six possible leucine codons in vitro at increasing temperatures revealed differences, with slower reversion to non-attenuated phenotypes for a subset of codons. Thus, it should be possible to modestly increase the phenotypic stability of the rA2cp248/404/1030ΔSH vaccine virus by codon modification at the locus of the 248 mutation. In addition to characterizing the phenotypes associated with a particular locus in the RSV L protein, this manuscript provides insight into the problem of the instability of point mutations and the limitations of strategies to stabilize them. Copyright © 2009 Published by Elsevier Ltd
Keyword Pneumovirus
Temperature sensitive
Respiratory syncytial virus vaccine
Q-Index Code C1
Q-Index Status Confirmed Code
Institutional Status UQ

 
Versions
Version Filter Type
Citation counts: TR Web of Science Citation Count  Cited 23 times in Thomson Reuters Web of Science Article | Citations
Scopus Citation Count Cited 22 times in Scopus Article | Citations
Google Scholar Search Google Scholar
Created: Tue, 26 Jan 2010, 00:52:06 EST by Lesley Arnicar on behalf of Clinical Medical Virology Centre