A live single-cycle RSV vaccine expressing prefusion F protein

Lamichhane, Pramila; Schmidt, Megan E.; Terhüja, Megolhubino; Varga, Steven M; Snider, Timothy A.; Rostad, Christina A.; Oomens, Antonius G. P.

doi:10.1016/j.virol.2022.10.003

Cited by 4 publications

(38 citation statements)

References 60 publications

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“…MAb L9 [34] was provided by Ed Walsh (University of Rochester School of Medicine, Rochester, NY, USA). Rabbit polyclonal anti-M and anti-F peptide sera were reported previously [18,35]. Anti-N antibody was acquired from AbD Serotec.…”

Section: Cells and Antibodiesmentioning

confidence: 99%

“…The resulting F variant, F/K191R, was then cloned using conventional techniques into a wt RSV cDNA lacking the F ORF and containing, instead, a cloning cassette with BsmBI sites. The construction of an HRP-expressing RSV (RSV-HRP) was described previously [18].…”

Section: Construction Of Recombinant Rsv Cdnasmentioning

confidence: 99%

“…Infectious preF-expressing viruses were recovered from cDNA as described previously [18,35,36] with minor modifications. Briefly, BHK-21 cells expressing T7 polymerase were transfected with engineered cDNAs and internal ribosome entry site-containing support plasmids expressing nucleoprotein N, phosphoprotein P, transcription elongation factor M2-1, and polymerase L. In addition, a plasmid-expressing baculovirus GP64 containing an F CT (GP 64/F ) was included to enable a progeny virus generated in the transfected cells, to infect Vbac cells [21,22].…”

Section: Recovery Of Viruses From Cdna and Production Of Virus Stocksmentioning

confidence: 99%

“…We previously generated a live vaccine, RSV-preF ∆CT , in which SH and the native F open reading frame (ORF) were removed and replaced with that of a DS-Cav1-based prefusion-stabilized membrane-anchored F protein (preF), and GFP [3,18,19]. To enhance the surface expression, and to ensure that F would not regain functionality when incorporated in a live virus, the F cytoplasmic tail (CT) was removed, and the resulting protein was termed preF ∆CT [18,19]. The preF ∆CT gene was placed in the sixth genome position to further enhance expression, and a GFP ORF was placed in the natural F (eighth) position.…”

Section: Introductionmentioning

confidence: 99%

“…A surprising finding emerged: despite containing the mG ORF, RSV-preF ∆CT induced only very low levels of anti-G Abs [18]. A large body of work has shown the importance of anti-G Abs in the protection against pathology and the reduced severity of RSV disease in animal models and human infants [23][24][25][26][27][28][29][30][31][32][33].…”

Section: Introductionmentioning

confidence: 99%

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Enhancing Anti-G Antibody Induction by a Live Single-Cycle Prefusion F—Expressing RSV Vaccine Improves In Vitro and In Vivo Efficacy

Lamichhane¹,

Terhüja²,

Snider³

et al. 2022

Viruses

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The human respiratory syncytial virus (RSV) is a major cause of severe respiratory tract disease, and a vaccine is not available. We previously reported a novel live vaccine expressing prefusion-stabilized fusion protein (preF) in place of the native F protein (RSV-preFΔCT). As preF is non-functional, RSV-preFΔCT was amplified in a production line expressing a functional substitute, and exhibited a single-cycle replication phenotype, which holds several unique potential advantages. RSV-preFΔCT prevented shedding and lung pathology after viral challenge in mice, but induced low levels of anti-attachment protein (G) antibodies (Abs). Given the significant contributions of anti-G Abs toward disease prevention, we generated modifications to RSV-preFΔCT in an effort to induce higher anti-G Ab levels. The Ab levels were monitored after the prime-boost vaccination of mice with modified vaccines. The most successful modification for enhancing induced anti-G Abs was seen with the placement of G in the first genome position. This vaccine also reduced the pathology after challenge with a high dose of wt RSV, and outperformed the sera from wt RSV-vaccinated mice in in vitro neutralization. Thus, raising the anti-G Ab levels induced by RSV-preFΔCT enhanced efficacy in vitro and in vivo, and constitutes an important next step in developing a live, single-cycle, efficacious vaccine for the human population.

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Section: Cells and Antibodiesmentioning

confidence: 99%

Section: Construction Of Recombinant Rsv Cdnasmentioning

confidence: 99%