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. 2001 Nov;75(22):11079-87.
doi: 10.1128/JVI.75.22.11079-11087.2001.

Replication-competent or attenuated, nonpropagating vesicular stomatitis viruses expressing respiratory syncytial virus (RSV) antigens protect mice against RSV challenge

J S Kahn  1 , A Roberts, C Weibel, L Buonocore, J K Rose
Affiliations

Replication-competent or attenuated, nonpropagating vesicular stomatitis viruses expressing respiratory syncytial virus (RSV) antigens protect mice against RSV challenge

J S Kahn et al. J Virol. 2001 Nov.

Abstract

Foreign glycoproteins expressed in recombinant vesicular stomatitis virus (VSV) can elicit specific and protective immunity in the mouse model. We have previously demonstrated the expression of respiratory syncytial virus (RSV) G (attachment) and F (fusion) glycoprotein genes in recombinant VSV. In this study, we demonstrate the expression of RSV F and G glycoproteins in attenuated, nonpropagating VSVs which lack the VSV G gene (VSVDeltaG) and the incorporation of these RSV proteins into recombinant virions. We also show that intranasal vaccination of mice with nondefective VSV recombinants expressing RSV G (VSV-RSV G) or RSV F (VSV-RSV F) elicited RSV-specific antibodies in serum (by enzyme-linked immunosorbent assay [ELISA]) as well as neutralizing antibodies to RSV and afford complete protection against RSV challenge. In contrast, VSVDeltaG-RSV F induced detectable serum antibodies to RSV by ELISA, but no detectable neutralizing antibodies, yet it still protected from RSV challenge. VSVDeltaG-RSV G failed to induce any detectable serum (by ELISA) or neutralizing antibodies and failed to protect from RSV challenge. The attenuated, nonpropagating VSVDeltaG-RSV F is a particularly attractive candidate for a live attenuated recombinant RSV vaccine.

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Figures

FIG. 1
FIG. 1
Schematic representation of RSV glycoprotein genes in recombinant VSVs lacking the VSV G gene (VSVΔG). Maps displaying gene order are in the antigenomic orientation. Maps of wild-type VSV and VSVΔG recombinants represent viruses used as controls. Maps of VSV-RSV G and VSV-RSV F were published previously (28). VSVΔG-RSV recombinants were derived from wild-type VSV-RSV cDNAs. Deletions of the VSV G gene in VSVΔG-RSV G and VSVΔG-RSV F recombinants are designated (Δ) beneath the respective maps. The newly created RSV G/VSV L and VSV M/RSV F gene junctions are displayed under the linear maps. cDNA sequences corresponding to the positive-sense RNA are shown in the 5′ to 3′ orientation. The transcription stop poly(A) signal and transcription start are labeled. The RSV G termination codon (TAG) and the RSV F initiation codon (ATG) are underlined and are represented in lowercase letters. The intergenic dinucleotide CT is italicized.
FIG. 2
FIG. 2
Expression of RSV glycoproteins in VSVΔG recombinant viruses. BHK cell monolayers were infected with either VSVΔG-RSV G or VSVΔG-RSV F at an MOI of 0.1. At 18 h postinfection, monolayers were fixed and screened by immunofluorescence. Primary antibody specificity is shown along the top of the figure and virus is indicated along the left of the figure. Objective magnification, 25×.
FIG. 3
FIG. 3
Western blot analysis of cell lysates and purified virions. (A) Proteins from uninfected and infected cell lysates were separated in a 12% polyacrylamide (SDS) gel and transferred to nitrocellulose. The blot was probed with a goat anti-RSV antiserum, and antibody binding was detected by ECL following binding with a horseradish conjugated-donkey anti-goat immunoglobulin G. The position of the heavily glycosylated RSV G is noted. Positions of molecular weight markers are noted on the left. (B) Proteins were separated in a 12% polyacrylamide (SDS) gel under nonreducing conditions. Samples were treated with SDS at room temperature prior to electrophoresis. Detection of the proteins bound by the goat anti-RSV antiserum is described above. The position of the RSV F 140-kDa dimer is noted, as are the positions of the molecular mass markers.
FIG. 4
FIG. 4
ELISA of serum from immunized mice. ELISA plates were coated with RSV-infected HEp-2 cell lysates. Serial dilutions of pooled serum from each group of immunized mice (recombinant wild-type VSV [A] and recombinant VSVΔG [B]) were incubated in coated wells. Bound antibody was detected by a colorimetric reaction following binding of a horseradish peroxidase-conjugated anti-mouse antibody. OD at 450 nm (OD450) are plotted against the inverse of the dilution factor. An RSV F-specific mouse monoclonal antibody (mAbαF) was used as a positive control.
FIG. 5
FIG. 5
RSV titers in BAL fluid and lung homogenates from immunized mice. BAL fluid and lung homogenates were prepared 4 days after challenge with RSV. BAL fluid titers (PFU/ml) and lung homogenate titers (PFU/g of tissue) are displayed in a log scale. RSV titers were determined on Vero cells by standard plaque assays. The lower level of detection was 50 PFU/ml of BAL fluid and 50 PFU/g of lung tissue. RSV titers in BAL fluid (∗) and lung homogenates (∗∗) were below the level of detection as indicated.
FIG. 6
FIG. 6
Lung histopathology following RSV challenge. Mice were immunized with the virus noted in each photomicrograph. Lung sections were obtained 4 days following RSV challenge. Bronchial airways (B) are noted. Objective magnification, 20×.
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References

    1. Abman S H, Ogle J W, Butler-Simon N, Rumack C M, Accurso F J. Role of respiratory syncytial virus in early hospitalizations for respiratory distress of young infants with cystic fibrosis. J Pediatr. 1988;113:826–830. - PubMed
    1. Anderson L J, Hierholzer J C, Bingham P G, Stone Y O. Microneutralization test for respiratory syncytial virus based on an enzyme immunoassay. J Clin Microbiol. 1985;22:1050–1052. - PMC - PubMed
    1. Anonymous. Acute respiratory infections: the forgotten pandemic. Bull W H O. 1998;76:101–103. , 105–107. - PMC - PubMed
    1. Anonymous. Palivizumab, a humanized respiratory syncytial virus monoclonal antibody, reduces hospitalization from respiratory syncytial virus infection in high-risk infants. The IMpact-RSV Study Group. Pediatrics. 1998;102:531–537. - PubMed
    1. Belshe R B, Anderson E L, Walsh E E. Immunogenicity of purified F glycoprotein of respiratory syncytial virus: clinical and immune responses to subsequent natural infection in children. J Infect Dis. 1993;168:1024–1029. - PubMed

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