Monoclonal Antibody-Based Antigenic Mapping of Norovirus GII.4-2002

Lindesmith, Lisa C.; Debbink, Kari; Swanstrom, Jesica; Vinjé, Jan; Costantini, Verónica; Baric, Ralph S.; Donaldson, Eric F.

doi:10.1128/jvi.06200-11

Cited by 116 publications

(167 citation statements)

References 50 publications

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“…For example, in the case of HIV, with the aid of an extra long CDR3 loop, the neutralizing Nanobody D7 effectively competed for the CD4 binding site on gp120 protein [42]. Previously described Nanobodies and mAbs with therapeutic potential against human norovirus were also proposed to interfere with the HBGA binding site [20,[22][23][24][25][26][27]. MAb termed NV8812 bound to a conformational epitope on the GI.1 P domain and blocked the binding of norovirus VLPs to human and animal cell lines [24].…”

Section: Discussionmentioning

confidence: 99%

“…A recent study suggested that antibodies targeting the HBGA pocket could inhibit norovirus replication by steric interference with the GI.1 HBGA pocket [22]. A number of other studies have identified norovirus-specific monoclonal antibodies (mAbs) and single chain variable domains (VHH or Nanobodies) that could block norovirus VLP binding to HBGAs [20,[23][24][25][26][27]. However, most of these antibodies and Nanobodies are genotype specific, which limits their therapeutic potential [28].…”

Section: Introductionmentioning

confidence: 99%

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Nanobodies Targeting Norovirus Capsid Reveal Functional Epitopes and Potential Mechanisms of Neutralization

Koromyslova

Hansman

2018

Biophysical Journal

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Norovirus is the leading cause of gastroenteritis worldwide. Despite recent developments in norovirus propagation in cell culture, these viruses are still challenging to grow routinely. Moreover, little is known on how norovirus infects the host cells, except that histo-blood group antigens (HBGAs) are important binding factors for infection and cell entry. Antibodies that bind at the HBGA pocket and block attachment to HBGAs are believed to neutralize the virus. However, additional neutralization epitopes elsewhere on the capsid likely exist and impeding the intrinsic structural dynamics of the capsid could be equally important. In the current study, we investigated a panel of Nanobodies in order to probe functional epitopes that could trigger capsid rearrangement and/ or interfere with HBGA binding interactions. The precise binding sites of six Nanobodies (Nano-4, Nano-14, Nano-26, Nano-27, Nano-32, and Nano-42) were identified using X-ray crystallography. We showed that these Nanobodies bound on the top, side, and bottom of the norovirus protruding domain. The impact of Nanobody binding on norovirus capsid morphology was analyzed using electron microscopy and dynamic light scattering. We discovered that distinct Nanobody epitopes were associated with varied changes in particle structural integrity and assembly. Interestingly, certain Nanobody-induced capsid morphological changes lead to the capsid protein degradation and viral RNA exposure. Moreover, Nanobodies employed multiple inhibition mechanisms to prevent norovirus attachment to HBGAs, which included steric obstruction (Nano-14), allosteric interference (Nano-32), and violation of normal capsid morphology (Nano-26 and Nano-85). Finally, we showed that two Nanobodies (Nano-26 and Nano-85) not only compromised capsid integrity and inhibited VLPs attachment to HBGAs, but also recognized a broad panel of norovirus genotypes with high affinities. Consequently, Nano-26 and Nano-85 have a great potential to function as novel therapeutic agents against human noroviruses. PLOS Author summaryWe determined the binding sites of six novel human norovirus specific Nanobodies (Nano-4, Nano-14, Nano-26, Nano-27, Nano-32, and Nano-42) using X-ray crystallography. The unique Nanobody recognition epitopes were correlated with their potential neutralizing capacities. We showed that one Nanobody (Nano-26) bound numerous genogroup II genotypes and interacted with highly conserved capsid residues. Four Nanobodies (Nano-4, Nano-26, Nano-27, and Nano-42) bound to occluded regions on the intact particles and impaired normal capsid morphology and particle integrity. One Nanobody (Nano-14) bound contiguous to the HBGA pocket and interacted with several residues involved in binding HBGAs. We found that the Nanobodies delivered multiple inhibition mechanisms, which included steric obstruction, allosteric interference, and disruption of the capsid stability. Our data suggested that the HBGA pocket might not be an ideal target for drug development, since the surrounding regio...

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Nanobodies Targeting Norovirus Capsid Reveal Functional Epitopes and Potential Mechanisms of Neutralization

Koromyslova

Hansman

2018

Biophysical Journal

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“…11 Residues of the P2 subdomain of GII.4 are described to be under selective pressure and consequently leading to antigenic drift resulting in escape of the immune responses. 28,29,38,40 In summary, antigenic drift, strain recombination, 35 antigenic shift and the polymerase fidelity of NoVs 43 clearly contribute to the continuous propagation of GII.4. The evolution of GII.4, described as being epochal (long periods of status quo followed by outbursts of variation) over time generates escape mutants that are periodically selected for by herd immunity.…”

Section: Viral Diversitymentioning

confidence: 99%

“…28,29 Expression of the VP1 capsid as a recombinant protein independently of other viral parts leads to self-assembly into a virus-like particle (VLP). The latter has structural and antigenic characteristics that cannot be distinguished from the virus.…”

Section: Viral Diversitymentioning

confidence: 99%

“…The surface exposed, highly variable sites of the P2 subdomain (epitopes A to E) undergo changes that were associated with the emergence of new strains causing NoV outbreaks. 28,36,37 These sites are situated around the HBGA pocket. The binding of NoV to specific HBGA as well as antibody binding is affected by these putative epitopes as well as flanking binding sites.…”

Section: Viral Diversitymentioning

confidence: 99%

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Norovirus vaccines: Correlates of protection, challenges and limitations

Melhem

2016

Human Vaccines & Immunotherapeutics

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Norovirus (NoV) is responsible for at least 50% of all gastroenteritis outbreaks worldwide. NoVs are classified into 6 different genogroups (GGI-GGVI) based on the viral capsid protein with NoV genogroup II genotype 4 (GII.4) being the predominant strain causing human diseases. Supportive therapy involving reversal of dehydration and electrolyte deficiency is the main treatment of NoV gastroenteritis. However, the worldwide increased recognition of NoV as an important agent of diarrheal gastroenteritis prompted researchers to focus on establishing preventive strategies conferring long-lasting immunity. This review describes the current status of animal and human vaccine models/studies targeting NoV and addresses the factors hampering the development of a broadly effective vaccine.

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Molecular epidemiology of norovirus in Singapore, 2004-2011

et al. 2013

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Norovirus (NoV) is the most common cause of sporadic and epidemic gastroenteritis, globally. This study aimed to investigate the molecular epidemiology of NoV-associated acute gastroenteritis in Singapore by classifying circulating NoV genotypes and genogroup II, genotype 4 (GII.4) variants between September 2004 and February 2011. The temporal dominance and antigenic variation within the circulating epidemic NoV GII.4 variants was also examined, in order to compare the trends in Singapore to those observed globally during the same period. A total of 312 of 1,060 fecal specimens were positive for NoV RNA, using a quantitative RT-PCR. In a subset (125 of 312) of NoV positive samples, the 5' end of ORF2 (region C) of the GI or GII NoV genome was amplified and sequenced. Subsequent phylogenetic analysis identified GII.4 was the most commonly identified genotype representing 80.8% (101/125) of NoV sequenced in this study. The predominant GII.4 variants in circulation during the 2004-2011 epidemic periods were Hunter 2004 (2004-2005), Den Haag 2006b (2006-2009), and New Orleans 2009 (2009-2011). Amino acid variation within the P2 domain of the major capsid protein, VP1, was followed longitudinally within the GII.4 lineage. A constant turnover of variant-specific amino acid change was observed, particularly within the antigenic epitopes A, C and E. In conclusion, this study has characterized the NoV strains in circulation in Singapore between 2004 and 2011. The molecular epidemiology and persistence of GII.4 pandemic NoV lineages in Singapore was similar to trends seen globally, with a noted absence of the Asia 2003 variant.

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Monoclonal Antibody-Based Antigenic Mapping of Norovirus GII.4-2002

Cited by 116 publications

References 50 publications

Nanobodies Targeting Norovirus Capsid Reveal Functional Epitopes and Potential Mechanisms of Neutralization

Nanobodies Targeting Norovirus Capsid Reveal Functional Epitopes and Potential Mechanisms of Neutralization

Norovirus vaccines: Correlates of protection, challenges and limitations

Molecular epidemiology of norovirus in Singapore, 2004-2011

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