Sequence analysis of the Washington/1964 strain of human parainfluenza virus type 1 (HPIV1) and recovery and characterization of wild-type recombinant HPIV1 produced by reverse genetics.
A complete consensus sequence was determined for the genomic RNA of human parainfluenza virus type 1 (HPIV1) strain Washington/20993/1964 (HPIV1 WASH/64), a clinical isolate that previously was shown to be virulent in adults. The sequence exhibited a high degree of relatedness to both Sendai virus, a PIV1 virus recovered from mice, and human PIV3 (HPIV3) with regard to cis-acting regulatory regions and protein-coding sequences. This consensus sequence was used to generate a full-length antigenomic cDNA and to recover a recombinant wild-type HPIV1 (rHPIV1). Interestingly, the rHPIV1 could be rescued from full-length antigenomic rHPIV1 cDNA using HPIV3 support plasmids, HPIV1 support plasmids, or a mixture thereof. The replication of rHPIV1 in vitro and in the respiratory tract of hamsters was similar to that of its biologically derived parent virus. The similar biological properties of rHPIV1 and HPIV1 WASH/64 in vitro and in vivo, together with the previous demonstration of the virulence of this specific isolate in humans, authenticates the rHPIV1 sequence as that of a wild-type virus. This rHPIV1 can now be used to study the biological properties of HPIV1 and as a substrate to introduce attenuating mutations for the generation of live-attenuated HPIV1 vaccine candidates.
Newman
Jason T
JT
National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD 20892-0720, USA. jnewman@niaid.nih.gov
Surman
Sonja R
SR
Riggs
Jeffrey M
JM
Hansen
Chris T
CT
Collins
Peter L
PL
Murphy
Brian R
BR
Skiadopoulos
Mario H
MH
D001483:Base Sequence; D002460:Cell Line; D016679:Genome, Viral; D006801:Humans; D008969:Molecular Sequence Data; D010222:Parainfluenza Virus 1, Human / Q000145:classification / Q000235:genetics*; D010252:Paramyxoviridae / Q000235:genetics*; D010957:Plasmids; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016415:Sequence Alignment; D017421:Sequence Analysis