Rational design of live-attenuated recombinant vaccine virus for human respiratory syncytial virus by reverse genetics.

P. L. Collins, S. S. Whitehead, Alexander Bukreyev, R. Fearns, M. N. Teng, K. Juhasz, R. M. Chanock, B. R. Murphy

Research output: Contribution to journalArticle

72 Citations (Scopus)

Abstract

RSV is a major cause of pediatric respiratory tract disease worldwide, but a vaccine is not yet available. It is now possible to prepare live infectious RSV completely from cDNA. This provides a method for introducing defined mutations into infectious virus, making possible the rational design of a live-attenuated vaccine virus for intranasal administration. This is particularly important for RSV, for which achieving the appropriate balance between attenuation and immunogenicity by conventional methods has proven elusive. We took advantage of the existence of a panel of biologically derived vaccine candidate viruses that were incompletely attenuated but well characterized biologically. The mutations in these viruses were identified by sequence analysis and characterized by insertion into recombinant virus, thereby providing a menu of known attenuating mutations. These included a series of amino acid point mutations, mostly in the L polymerase, and a nucleotide substitution in a transcription gene-start signal, a cis-acting RNA element. The second source of mutations was from experimental mutational analysis of recombinant virus and involves deletion of the NS1, NS2, or SH gene. We have reconstructed a previously tested, biologically derived attenuated virus, cpts248/404, in recombinant form and are now proceeding to introduce additional mutations from the menu to achieve stepwise increases in attenuation. The ability to modify the attenuation phenotype incrementally in a directed manner should result in an appropriate vaccine virus.

Original languageEnglish (US)
Pages (from-to)423-451
Number of pages29
JournalAdvances in Virus Research
Volume54
StatePublished - 1999
Externally publishedYes

Fingerprint

Human respiratory syncytial virus
Reverse Genetics
Attenuated Vaccines
Synthetic Vaccines
Viruses
Mutation
Vaccines
Respiratory Tract Diseases
Intranasal Administration
Point Mutation
Genes
Sequence Analysis
Nucleotides
Complementary DNA
RNA
Pediatrics
Phenotype
Amino Acids

ASJC Scopus subject areas

  • Virology
  • Biotechnology

Cite this

Collins, P. L., Whitehead, S. S., Bukreyev, A., Fearns, R., Teng, M. N., Juhasz, K., ... Murphy, B. R. (1999). Rational design of live-attenuated recombinant vaccine virus for human respiratory syncytial virus by reverse genetics. Advances in Virus Research, 54, 423-451.

Rational design of live-attenuated recombinant vaccine virus for human respiratory syncytial virus by reverse genetics. / Collins, P. L.; Whitehead, S. S.; Bukreyev, Alexander; Fearns, R.; Teng, M. N.; Juhasz, K.; Chanock, R. M.; Murphy, B. R.

In: Advances in Virus Research, Vol. 54, 1999, p. 423-451.

Research output: Contribution to journalArticle

Collins, PL, Whitehead, SS, Bukreyev, A, Fearns, R, Teng, MN, Juhasz, K, Chanock, RM & Murphy, BR 1999, 'Rational design of live-attenuated recombinant vaccine virus for human respiratory syncytial virus by reverse genetics.', Advances in Virus Research, vol. 54, pp. 423-451.
Collins, P. L. ; Whitehead, S. S. ; Bukreyev, Alexander ; Fearns, R. ; Teng, M. N. ; Juhasz, K. ; Chanock, R. M. ; Murphy, B. R. / Rational design of live-attenuated recombinant vaccine virus for human respiratory syncytial virus by reverse genetics. In: Advances in Virus Research. 1999 ; Vol. 54. pp. 423-451.
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