Potent neutralizing antibodies from COVID-19 patients define multiple targets of vulnerability

Brouwer, Philip J.M.; Caniels, Tom G.; Straten, Karlijn van der; Snitselaar, Jonne L.; Aldon, Yoann; Bangaru, Sandhya; Torres, Jonathan L.; Okba, Nisreen M.A.; Claireaux, Mathieu; Kerster, Gius; Bentlage, Arthur E. H.; Haaren, Marlies M. van; Guerra, Denise; Burger, Judith A.; Schermer, Edith E.; Verheul, Kirsten D.; Velde, Niels van der; Kooi, Alex van der; Schooten, Jelle van; Breemen, Mariëlle J. van; Bijl, Tom P. L.; Sliepen, Kwinten; Aartse, Aafke; Derking, Ronald; Bontjer, Ilja; Kootstra, Neeltje A.; Wiersinga, W. Joost; Vidarsson, Gestur; Haagmans, Bart L.; Ward, Andrew B.; Bree, Godelieve J. de; Sanders, Rogier W.; Gils, Marit J. van

doi:10.1101/2020.05.12.088716

Cited by 345 publications

(617 citation statements)

References 53 publications

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“…From the perspective of immunogen development, the constructs developed here present an opportunity to examine the ability of differentially stabilized S-protein particles to induce two different, yet important antibody responses. First, the disulfide-linked 'down' state locked double mutant (rS2d) would presumably eliminate receptor binding site targeting antibodies which make up the majority of observed responses 30,31 . Indeed, a study of MERS responses suggests non-RBD responses (particularly NTD and S2 epitopes) will play an important role in vaccine induced protection 32 .…”

Section: Discussionmentioning

confidence: 99%

Controlling the SARS-CoV-2 Spike Glycoprotein Conformation

Henderson

Edwards

Mansouri³

et al. 2020

Preprint

119

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The coronavirus (CoV) viral host cell fusion spike (S) protein is the primary immunogenic target for virus neutralization and the current focus of many vaccine design efforts. The highly flexible S-protein, with its mobile domains, presents a moving target to the immune system. Here, to better understand S-protein mobility, we implemented a structure-based vector analysis of available β-CoV S-protein structures. We found that despite overall similarity in domain organization, different β-CoV strains display distinct S-protein configurations. Based on this analysis, we developed two soluble ectodomain constructs in which the highly immunogenic and mobile receptor binding domain (RBD) is locked in either the all-RBDs 'down' position or is induced to display a previously unobserved in SARS-CoV-2 2-RBDs 'up' configuration. These results demonstrate that the conformation of the S-protein can be controlled via rational design and provide a framework for the development of engineered coronavirus spike proteins for vaccine applications. IntroductionThe ongoing global pandemic of the novel SARS-CoV-2 (SARS-2) coronavirus presents an urgent need for the development of effective preventative and treatment therapies. The viral S-protein is a prime target for such therapies owing to its critical role in the virus lifecycle. The S-protein is divided into two regions: an N-terminal S1 domain that caps the C-terminal S2 fusion domain. Binding to host receptor via the Receptor Binding Domain (RBD) in S1 is followed by proteolytic cleavage of the spike by host proteases 1 . Large conformational changes in the S-protein result in S1 shedding and exposure of the fusion machinery in S2.Class I fusion proteins, such as the CoV-2 S-protein, undergo large conformational changes during the fusion process and must, by necessity, be highly flexible and dynamic. Indeed, cryo-electron microscopy (cryo-EM) structures of SARS-2 spike reveal considerable flexibility and dynamics in the S1 domain 1,2 , especially around the RBD that exhibits two discrete conformational states -a 'down' state that is shielded from receptor binding, and an 'up' state that is receptor-accessible.The wealth of structural information for β-CoV spike proteins, including the recently determined cryo-EM structures of the SARS-2 spike 1-11 , has provided a rich source of detailed geometric information from which to begin precise examination of the macromolecular transitions underlying triggering of this fusion machine.

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Section: Discussionmentioning

confidence: 99%

Controlling the SARS-CoV-2 Spike Glycoprotein Conformation

Henderson

Edwards

Mansouri³

et al. 2020

Preprint

119

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“…were over-represented . Other studies have also reported anti-SARS-CoV-2 mAbs derived from these genes (Brouwer et al, 2020;Cao et al, 2020;Chi et al, 2020;Ju et al, 2020;Rogers et al, 2020;Seydoux et al, 2020;Wu et al, 2020c;Zost et al, 2020).…”

Section: A Cryo-em Structure Of a Monoclonal Fab-s Protein Complex Rementioning

confidence: 95%

Structures of human antibodies bound to SARS-CoV-2 spike reveal common epitopes and recurrent features of antibodies

Barnes

West

Huey-Tubman

et al. 2020

Preprint

220

338

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Neutralizing antibody responses to coronaviruses focus on the trimeric spike, with most against the receptor-binding domain (RBD). Here we characterized polyclonal IgGs and Fabs from COVID-19 convalescent individuals for recognition of coronavirus spikes. Plasma IgGs differed in their degree of focus on RBD epitopes, recognition of SARS-CoV, MERS-CoV, and mild coronaviruses, and how avidity effects contributed to increased binding/neutralization of IgGs over Fabs. Electron microscopy reconstructions of polyclonal plasma Fab-spike complexes showed recognition of both S1 A and RBD epitopes. A 3.4Å cryo-EM structure of a neutralizing monoclonal Fab-S complex revealed an epitope that blocks ACE2 receptor-binding on "up" RBDs. Modeling suggested that IgGs targeting these sites have different potentials for interspike crosslinking on viruses and would not be greatly affected by identified SARS-CoV-2 spike mutations. These studies structurally define a recurrent anti-SARS-CoV-2 antibody class derived from VH3-53/VH3-66 and similarity to a SARS-CoV VH3-30 antibody, providing criteria for evaluating vaccine-elicited antibodies.

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“…SARS-Cov-2-specific antibodies were purified using antigen-coated plates (NUCN, Roskilde, Denmark). Plates were coated (over-night, 4°C) with recombinant trimerized spike protein produced as described recently ( 28 ) or N protein (accession number MN908947, produced in HEK cells with HAVT20 leader peptide, 10xhis tag and a Brit tag as in ( 23 )) in PBS(5 µg/mL and 1 µg/mL, respectively). Plates were washed 3× with PBS supplemented with 0.05 % TWEEN 20 ® (PBS-T) and plasma (20 µ L) diluted in PBS-T (180 µ L) was incubated on the plates(1 hour, 37 °C, shaking).…”

Section: Methodsmentioning

confidence: 99%

Afucosylated immunoglobulin G responses are a hallmark of enveloped virus infections and show an exacerbated phenotype in COVID-19

Larsen

Graaf

Sonneveld

et al. 2020

Preprint

Self Cite

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27IgG antibodies are crucial for protection against invading pathogens. A highly conserved N-28 linked glycan within the IgG-Fc-tail, essential for IgG function, shows variable composition in 29 humans. Afucosylated IgG variants are already used in anti-cancer therapeutic antibodies for 30 their elevated binding and killing activity through Fc receptors (FcγRIIIa). Here, we report that 31 afucosylated IgG which are of minor abundance in humans (~6% of total IgG) are specifically 32 formed against surface epitopes of enveloped viruses after natural infections or 33 immunization with attenuated viruses, while protein subunit immunization does not elicit 34 this low fucose response. This can give beneficial strong responses, but can also go awry, 35 resulting in a cytokine-storm and immune-mediated pathologies. In the case of COVID-19, 36 the critically ill show aggravated afucosylated-IgG responses against the viral spike protein. In 37 contrast, those clearing the infection unaided show higher fucosylation levels of the anti-38 spike protein IgG. Our findings indicate antibody glycosylation as a potential factor in 39 inflammation and protection in enveloped virus infections including COVID-19.40Main Text: 41 Antibodies have long been considered functionally static, mostly determined by their isotype 42 and subclass. The presence of a conserved N-linked glycan at position 297, in the so called 43 constant Fc-domain of IgG, is essential for effector functions (1-3). Moreover, it is now 44 generally accepted that the composition of this glycan is highly variable and has functional 45 consequences (2-4). This is especially true for the core fucose attached to the Fc glycan. The 46 discovery that IgG variants without core fucosylation cause elevated antibody dependent 47 cellular cytotoxicity (ADCC), via increased IgG-Fc-receptor IIIa (FcγRIIIa) affinity (5, 6), 48 resulted in next-generation glyco-engineered monoclonal antibodies (mAb) without core 49 fucosylation for targeting tumors (7). 50Generally, changes in the Fc glycans are associated with age, sex and autoimmune diseases 51 (8). Serum IgG are highly fucosylated at birth and slightly decrease to ~94% fucosylation at 52 adulthood (9). Until now, no strong clues on how IgG core fucosylation is controlled have 53 come forward. 54 We have previously observed that alloantibodies against red blood cells (RBC) and platelets 55 show remarkably low IgG-Fc-fucosylation in most patients, even down to 10% in several 56 cases (10-12), whereas the overall serum IgG Fc-fucosylation show consistently normal high 57 levels. Moreover, we have reported the lowered IgG-Fc fucosylation to be one of the factors 58 determining disease severity in pregnancy associated alloimmunizations, resulting in 59 excessive thrombocytopenia's and blood cell destruction when targeted by afucosylated 60 antibodies (11)(12)(13). In addition to the specific afucosylated-IgG response against platelets 61 and RBC antigens, this response has also been identified against HIV and Dengue virus (...

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Potent neutralizing antibodies from COVID-19 patients define multiple targets of vulnerability

Cited by 345 publications

References 53 publications

Controlling the SARS-CoV-2 Spike Glycoprotein Conformation

Controlling the SARS-CoV-2 Spike Glycoprotein Conformation

Structures of human antibodies bound to SARS-CoV-2 spike reveal common epitopes and recurrent features of antibodies

Afucosylated immunoglobulin G responses are a hallmark of enveloped virus infections and show an exacerbated phenotype in COVID-19

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