Respiratory syncytial virus escape mutant derived in vitro resists palivizumab prophylaxis in cotton rats

Zhao, Xiaodong; Chen, Fu-Ping; Sullender, Wayne M.

doi:10.1016/j.virol.2003.10.018

Cited by 59 publications

(59 citation statements)

References 22 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…1D), Ile266 is positioned at the bottom of the antigenic site II helixloop-helix motif and is pointed toward the inner protein core, suggesting the residue disrupts the antigenic motif by allosteric effects. In the same study (11), selection with several murine mAbs produced MARM viruses with Lys272Asn, and similarly, selection with palivizumab in vitro or in vivo, generated similar MARM viruses with the following mutations: Lys272Met, Lys272Gln, and Asn268Ile (20,21). The Lys272Gln MARM virus completely resisted prophylactic palivizumab treatment (22).…”

Section: Resultsmentioning

confidence: 92%

Structural basis for nonneutralizing antibody competition at antigenic site II of the respiratory syncytial virus fusion protein

Mousa

Sauer

Sevy

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

Palivizumab was the first antiviral monoclonal antibody (mAb) approved for therapeutic use in humans, and remains a prophylactic treatment for infants at risk for severe disease because of respiratory syncytial virus (RSV). Palivizumab is an engineered humanized version of a murine mAb targeting antigenic site II of the RSV fusion (F) protein, a key target in vaccine development. There are limited reported naturally occurring human mAbs to site II; therefore, the structural basis for human antibody recognition of this major antigenic site is poorly understood. Here, we describe a nonneutralizing class of site II-specific mAbs that competed for binding with palivizumab to postfusion RSV F protein. We also describe two classes of site II-specific neutralizing mAbs, one of which escaped competition with nonneutralizing mAbs. An X-ray crystal structure of the neutralizing mAb 14N4 in complex with F protein showed that the binding angle at which human neutralizing mAbs interact with antigenic site II determines whether or not nonneutralizing antibodies compete with their binding. Fine-mapping studies determined that nonneutralizing mAbs that interfere with binding of neutralizing mAbs recognize site II with a pose that facilitates binding to an epitope containing F surface residues on a neighboring protomer. Neutralizing antibodies, like motavizumab and a new mAb designated 3J20 that escape interference by the inhibiting mAbs, avoid such contact by binding at an angle that is shifted away from the nonneutralizing site. Furthermore, binding to rationally and computationally designed site II helixloop-helix epitope-scaffold vaccines distinguished neutralizing from nonneutralizing site II antibodies.F protein | human respiratory syncytial virus | neutralizing antibodies R espiratory syncytial virus (RSV) is a highly contagious human pathogen, infecting the majority of infants before age 2 y, and is the leading cause of viral bronchiolitis and viral pneumonia in infants and children (1, 2). RSV remains a top priority for vaccine development, as thousands of deaths are recorded worldwide each year because of complications from infection (3). To date, there is no licensed RSV vaccine. A major focus of RSV vaccine development has been inclusion of the RSV fusion (F) protein, a class I fusion glycoprotein that is synthesized as a precursor and cleaved into two disulfide-linked fragments upon maturation into a trimer (4). Although the RSV virion contains two additional surface proteins, the highly-glycosylated attachment (G) protein and the small hydrophobic protein, the F protein is highly conserved among strains of RSV strains and is the major target of protective neutralizing antibodies.The F protein is known to adopt at least two major conformations: the metastable prefusion conformation and the postfusion conformation. Following attachment of the virion to a cell by the G protein, the F protein undergoes a dramatic structural rearrangement, resulting in fusion of the viral and cell membranes, and in cultured cells causes...

show abstract

Section: Resultsmentioning

confidence: 92%

Structural basis for nonneutralizing antibody competition at antigenic site II of the respiratory syncytial virus fusion protein

Mousa

Sauer

Sevy

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

show abstract

“…The antigenic structure of the RSV F trimer has been mapped by a variety of techniques (7,17,(26)(27)(28)(29)(30)(31)(32)(33) (Table S2 and Fig. S1).…”

Section: Dilutions Of Seramentioning

confidence: 99%

Structural basis for immunization with postfusion respiratory syncytial virus fusion F glycoprotein (RSV F) to elicit high neutralizing antibody titers

Swanson¹,

Settembre²,

Shaw³

et al. 2011

Proc. Natl. Acad. Sci. U.S.A.

248

246

View full text Add to dashboard Cite

Respiratory syncytial virus (RSV), the main cause of infant bronchiolitis, remains a major unmet vaccine need despite more than 40 years of vaccine research. Vaccine candidates based on a chief RSV neutralization antigen, the fusion (F) glycoprotein, have foundered due to problems with stability, purity, reproducibility, and potency. Crystal structures of related parainfluenza F glycoproteins have revealed a large conformational change between the prefusion and postfusion states, suggesting that postfusion F antigens might not efficiently elicit neutralizing antibodies. We have generated a homogeneous, stable, and reproducible postfusion RSV F immunogen that elicits high titers of neutralizing antibodies in immunized animals. The 3.2-Å X-ray crystal structure of this substantially complete RSV F reveals important differences from homology-based structural models. Specifically, the RSV F crystal structure demonstrates the exposure of key neutralizing antibody binding sites on the surface of the postfusion RSV F trimer. This unanticipated structural feature explains the engineered RSV F antigen's efficiency as an immunogen. This work illustrates how structural-based antigen design can guide the rational optimization of candidate vaccine antigens.subunit | epitope

show abstract

“…After 6 weeks, the recovery of virus from nasal washes was sporadic. Viruses recovered in cells provided RNA for RT-PCR amplification and nucleotide sequence determination of the F gene nucleotides 660 to 1210, encompassing previously identified PZ resistance mutations and most of the point mutations described in antigenic site II of the F protein (8,35,36). At 6 weeks post-initial infection, one animal (A-4) revealed a mixed population at position 827 of A and C, with A representing the parent A2 virus and C identical to a mutant (MS412) completely resistant to PZ (35) (Fig.…”

Section: Resultsmentioning

confidence: 99%

“…Viral RNA was extracted from infected HEp-2 cells and ground lung tissue with Trizol reagent (Invitrogen), as described previously (36). The region from nucleotides 622 to 1225 in the RSV F gene was amplified by RT-PCR with primers F622For (5Ј-GTTACCTATTG TGAACAAGC-3Ј), corresponding to A2 F gene nucleotides 622 to 641, and F1225Rev (5Ј-GCTGCTTACATCTGTTTTTG-3Ј), which complements A2 F gene nucleotides 1206 to 1225, to yield a 603-bp fragment.…”

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

In Vivo Selection of Respiratory Syncytial Viruses Resistant to Palivizumab

Zhao¹,

Sullender

2005

J Virol

Self Cite

View full text Add to dashboard Cite

Palivizumab (PZ) is the only monoclonal antibody currently available for use in humans against an infectious disease. PZ is administered prophylactically for respiratory syncytial virus (RSV) infections. RSV selected in cell culture for growth in the presence of PZ develops F gene mutations and can be resistant to PZ prophylaxis in cotton rats. Here, we evaluated the potential for PZ-resistant RSV mutants to arise in vivo. Cotton rats were immunosuppressed with cyclophosphamide, administered PZ, and inoculated intranasally with RSV. Lungs were harvested 12 weeks after RSV infection, reverse transcription-PCR-amplified F gene fragments were cloned into plasmids, and the nucleotide sequences of the cloned cDNAs were determined. Three of the five animals had mixed populations of lung virus, and over 50% of the clones from the three animals revealed F gene mutations associated with resistance to PZ. A virus completely resistant to PZ neutralization was recovered from the lung homogenate of a rat that had received PZ. Thus, prolonged pulmonary replication of RSV in the presence of PZ was followed by the appearance of viruses resistant to PZ. The potential for the development of resistance is a consideration as the antibody is used prophylactically against RSV and as passively administered antibodies are under development for other infections, including emerging viruses and agents of biodefense.Respiratory syncytial virus (RSV) is the principal cause of viral respiratory infections among infants and young children and causes disease in adults, with the elderly at particular risk for severe disease (8,14,26). In addition, immunocompromised patients may suffer serious morbidity and even mortality due to RSV infections (13, 16). Palivizumab (PZ), is the first commercially available antibody for use against an infectious disease. PZ, a humanized neutralizing monoclonal antibody reactive with an epitope on the F glycoprotein of RSV, is used prophylactically for high-risk children with preterm birth or underlying cardiorespiratory disorders (1).RSV escape mutants from monoclonal and polyclonal antibodies against the F and G proteins have been derived in cell culture (5,9,15,23,31,33). We selected RSV resistant to PZ by replication of virus in the presence of PZ in cell culture (36). Point mutations occurred at two sites in the F gene. At positions 828 (A-T, virus MP4) and 827 (A-C, virus MS412) changes resulted in two different amino acid changes at position 272 in the F1 subunit (Lys to Met or Gln, respectively). Both changes were associated with resistance to PZ neutralization in vitro. In addition, viruses with these point mutations were completely resistant to the prophylactic effects of PZ in cotton rats. A point mutation at another site, 816 (A-T), led to an amino acid substitution from Asn to Ile at position 268 in the F1 subunit. This virus, F212, was partially resistant to PZ neutralization but remained fully susceptible to PZ prophylaxis (15 mg/kg of body weight) in cotton rats. Interestingly, F212 grew to lower t...

show abstract

Respiratory syncytial virus escape mutant derived in vitro resists palivizumab prophylaxis in cotton rats

Cited by 59 publications

References 22 publications

Structural basis for nonneutralizing antibody competition at antigenic site II of the respiratory syncytial virus fusion protein

Structural basis for nonneutralizing antibody competition at antigenic site II of the respiratory syncytial virus fusion protein

Structural basis for immunization with postfusion respiratory syncytial virus fusion F glycoprotein (RSV F) to elicit high neutralizing antibody titers

In Vivo Selection of Respiratory Syncytial Viruses Resistant to Palivizumab

Contact Info

Product

Resources

About