求路虎揽胜车架号SALGA2FV3HA353658详细解析。谢谢！_百度知道
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):10819-28.Effects of human metapneumovirus and respiratory syncytial virus antigen insertion in two 3' proximal genome positions of bovine/human parainfluenza virus type 3 on virus replication and immunogenicity.1, , , , , , , , , .1MedImmune Vaccines, Inc., Mountain View, California 94043, USA.AbstractA live attenuated bovine parainfluenza virus type 3 (PIV3), harboring the fusion (F) and hemagglutinin-neuraminidase (HN) genes of human PIV3, was used as a virus vector to express surface glycoproteins derived from two human pathogens, human metapneumovirus (hMPV) and respiratory syncytial virus (RSV). RSV and hMPV are both paramyxoviruses that cause respiratory disease in young children, the elderly, and immunocompromised individuals. RSV has been known for decades to cause acute lower respiratory tract infections in young children, which often result in hospitalization, while hMPV has only been recently identified as a novel human respiratory pathogen. In this study, the ability of bovine/human PIV3 to express three different foreign transmembrane surface glycoproteins and to induce a protective immune response was evaluated. The RNA-dependent RNA polymerase of paramyxoviruses binds to a single site at the 3' end of the viral RNA genome to initiate transcription of viral genes. The genome position of the viral gene determines its level of gene expression. The promoter-proximal gene is transcribed with the highest frequency, and each downstream gene is transcribed less often due to attenuation of transcription at each gene junction. This feature of paramyxoviruses was exploited using the PIV3 vector by inserting the foreign viral genes at the 3' terminus, at position 1 or 2, of the viral RNA genome. These locations were expected to yield high levels of foreign viral protein expression stimulating a protective immune response. The immunogenicity and protection results obtained with a hamster model showed that bovine/human PIV3 can be employed to generate bivalent PIV3/RSV or PIV3/hMPV vaccine candidates that will be further evaluated for safety and efficacy in primates.PMID:
[Indexed for MEDLINE] Schematic representation of the b/h PIV3 RNA genome, which serves as a vector backbone for insertion of RSV or hMPV surface glycoprotein genes. The RSV F, RSV G, and hMPV F gene insertions at position 1 or 2 of the PIV3 genome are indicated by gray boxes. The foreign genes were inserted at position 1 by using an AvrII restriction enzyme site introduced at nt 104 in the PIV3 genome or at position 2 at an AvrII site at nt 1774. The inserted transcriptional units contain the bPIV3 N-P intergenic region with the gene stop and start sequences utilized for transcription of the gene downstream of the inserted cassette.J Virol. ):.(A) Multicycle growth curves of b/h PIV3/RSV F1, F2, G1, G2, and b/h PIV3 carried out in Vero cells at an MOI of 0.1. (B) Multicycle growth curves of b/h PIV3/hMPV F1, F2, and b/h PIV3 in Vero cells at an MOI of 0.1. Cells and supernatants were collected at 0, 24, 48, 72, 96, and 120 hpi. The titers of the samples were determined by plaque assays on Vero cells.J Virol. ):.RSV or hMPV protein expression by the chimeric b/h PIV3 in Vero cells. (A) Immunoblot of Vero cell lysates infected with b/h PIV3/RSV F1 or F2 or wild-type RSV A2, b/h PIV3 (at an MOI of 0.1), or mock-infected cells at 48 hpi probed with RSV F-specific monoclonal antisera. An ~50-kDa band corresponding to the processed RSV F1 protein was observed for b/h PIV3/RSV F1 and F2 lysates as well as for RSV A2. A larger band of ~90 kDa, the uncleaved F0 precursor, is also present in relatively smaller quantities in lanes 1 and 2, indicating that RSV F protein processing occurs efficiently in the chimeric viruses. (B) Western blot of cell lysates infected with b/h PIV3/RSV G1 or G2, RSV A2, b/h PIV3 (at an MOI of 0.1) and mock-infected cell lysates at 48 hpi. The Western blot was probed with RSV G polyclonal antisera and displays mature glycosylated and immature forms of the RSV G protein expressed by b/h PIV3/RSV G1 or G2 or RSV A2. As expected, a signal was not observed in lysates derived from Vero cells infected with b/h PIV3 or in mock-infected cells. (C) b/h PIV3/hMPV F1- or F2-, b/h PIV3-, or mock-infected 35S-labeled cell lysates were harvested 48 hpi and immunoprecipitated with hMPV-specific polyclonal guinea pig antisera. A band of ~80 kDa was observed in cell lysates derived from b/h PIV3/hMPV F1 and F2 that was not present in b/h PIV3- or mock-infected lysates. This size is consistent with an uncleaved F0 hMPV protein.J Virol. ):.Publication typeMeSH termsSubstancesGrant supportFull Text SourcesOther Literature Sources
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