Structural Analysis of the Genome of Respiratory Syncytial Virus

Collins, P L.

Abstract

We have been exploring methods of obtaining biological activity for cDNA- encoded synthetic genomic RNA (vRNA) of human respiratory syncytial virus (RSV). The ultimate goal is to produce infectious virus from a complete synthetic vRNA. This capability would provide an experimental system for analyzing the functions of RSV nucleic acid signals and proteins, and also would be invaluable for the production and characterization of engineered attenuated vaccine viruses. We have constructed cDNAs encoding vRNA analogs in which all of the viral genes have been deleted and replaced with the bacterial chloramphenicol acetyl transferase (CAT) marker gene. When transfected into tissue culture cells and complemented by infection with RSV, these truncated, negative-sense vRNA analogs were """"""""rescued"""""""" in that they were expressed, amplified and packaged into infectious particles. Substantial progress has been made in identifying and characterizing cis- acting nucleotide signals important in rescue.

Funding Agency

• Agency: National Institute of Health (NIH)
• Institute: National Institute of Allergy and Infectious Diseases (NIAID)
• Type: Intramural Research (Z01)
• Project #: 1Z01AI000368-09
• Application #: 3790741
• Support Year: 9
• Fiscal Year: 1992

Institution

• Name: National Institute of Allergy and Infectious Diseases
• Country: United States

Publications

Melendi, Guillermina A; Zavala, Fidel; Buchholz, Ursula J et al. (2007) Mapping and characterization of the primary and anamnestic H-2(d)-restricted cytotoxic T-lymphocyte response in mice against human metapneumovirus. J Virol 81:11461-7

Buchholz, Ursula J; Nagashima, Kunio; Murphy, Brian R et al. (2006) Live vaccines for human metapneumovirus designed by reverse genetics. Expert Rev Vaccines 5:695-706

Biacchesi, Stephane; Pham, Quynh N; Skiadopoulos, Mario H et al. (2006) Modification of the trypsin-dependent cleavage activation site of the human metapneumovirus fusion protein to be trypsin independent does not increase replication or spread in rodents or nonhuman primates. J Virol 80:5798-806

Biacchesi, Stephane; Skiadopoulos, Mario H; Yang, Lijuan et al. (2005) Rapid human metapneumovirus microneutralization assay based on green fluorescent protein expression. J Virol Methods 128:192-7

Buchholz, Ursula J; Biacchesi, Stephane; Pham, Quynh N et al. (2005) Deletion of M2 gene open reading frames 1 and 2 of human metapneumovirus: effects on RNA synthesis, attenuation, and immunogenicity. J Virol 79:6588-97

Pham, Quynh N; Biacchesi, Stephane; Skiadopoulos, Mario H et al. (2005) Chimeric recombinant human metapneumoviruses with the nucleoprotein or phosphoprotein open reading frame replaced by that of avian metapneumovirus exhibit improved growth in vitro and attenuation in vivo. J Virol 79:15114-22

Biacchesi, Stephane; Skiadopoulos, Mario H; Yang, Lijuan et al. (2004) Recombinant human Metapneumovirus lacking the small hydrophobic SH and/or attachment G glycoprotein: deletion of G yields a promising vaccine candidate. J Virol 78:12877-87

Biacchesi, Stephane; Skiadopoulos, Mario H; Tran, Kim C et al. (2004) Recovery of human metapneumovirus from cDNA: optimization of growth in vitro and expression of additional genes. Virology 321:247-59

Buchholz, Ursula J; Bukreyev, Alexander; Yang, Lijuan et al. (2004) Contributions of the structural proteins of severe acute respiratory syndrome coronavirus to protective immunity. Proc Natl Acad Sci U S A 101:9804-9

Biacchesi, Stephane; Skiadopoulos, Mario H; Boivin, Guy et al. (2003) Genetic diversity between human metapneumovirus subgroups. Virology 315:1-9