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Journal of Virology, June 2004, p. 5773-5783, Vol. 78, No. 11
0022-538X/04/$08.00+0     DOI: 10.1128/JVI.78.11.5773-5783.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Recombinant Respiratory Syncytial Viruses Lacking the C-Terminal Third of the Attachment (G) Protein Are Immunogenic and Attenuated In Vivo and In Vitro

Matthew B. Elliott,¹ Karin S. Pryharski,¹ Qingzhong Yu,^2, Christopher L. Parks,² Todd S. Laughlin,^1, C. Kanta Gupta,² Robert A. Lerch,² Valerie B. Randolph,² Natisha A. LaPierre,¹ Kristen M. Heers Dack,^1, and Gerald E. Hancock^1*

Departments of Immunology,¹ Viral Vaccine Research, Wyeth Vaccines Research, Pearl River, New York 10965²

Received 14 November 2003/ Accepted 29 January 2004

The design of attenuated vaccines for respiratory syncytial virus (RSV) historically focused on viruses made sensitive to physiologic temperature through point mutations in the genome. These prototype vaccines were not suitable for human infants primarily because of insufficient attenuation, genetic instability, and reversion to a less-attenuated phenotype. We therefore sought to construct novel attenuated viruses with less potential for reversion through genetic alteration of the attachment G protein. Complete deletion of G protein was previously shown to result in RSV strains overly attenuated for replication in mice. Using reverse genetics, recombinant RSV (rRSV) strains were engineered with truncations at amino acid 118, 174, 193, or 213 and respectively designated rA2cpG118, rA2cpG174, rA2cpG193, and rA2cpG213. All rA2cpG strains were attenuated for growth in vitro and in the respiratory tracts of BALB/c mice but not restricted for growth at 37°C. The mutations did not significantly affect nascent genome synthesis in human lung epithelial (A549) cells, but infectious rA2cpG virus shed into the culture medium was dramatically diminished. Hence, the data suggested that a site within the C-terminal 85 amino acids of G protein is important for efficient genome packaging or budding of RSV from the infected cell. Vaccination with the rA2cpG strains also generated efficacious immune responses in mice that were similar to those elicited by the temperature-sensitive cpts248/404 strain previously tested in human infants. Collectively, the data indicate that the rA2cpG strains are immunogenic, not likely to revert to the less-attenuated phenotype, and thus candidates for further development as vaccines against RSV.

Present address: USDA, Southeast Poultry Research Laboratory, Athens, GA 30605.

Present address: Ortho-Clinical Diagnostics, Rochester, NY 14626.

Present address: Division of Pediatric Hematology/Oncology, Golisano's Children's Hospital at Strong, Rochester, NY 14642.

This article has been cited by other articles:

• Elliott, M. B., Pryharski, K. S., Yu, Q., Boutilier, L. A., Campeol, N., Melville, K., Laughlin, T. S., Gupta, C. K., Lerch, R. A., Randolph, V. B., LaPierre, N. A., Dack, K. M. H., Hancock, G. E. (2004). Characterization of Recombinant Respiratory Syncytial Viruses with the Region Responsible for Type 2 T-Cell Responses and Pulmonary Eosinophilia Deleted from the Attachment (G) Protein. J. Virol. 78: 8446-8454 [Abstract] [Full Text]