Immunogenicity and structures of a rationally designed prefusion MERS-CoV spike antigen

Pallesen, Jesper; Wang, Nianshuang; Corbett, Kizzmekia S.; Wrapp, Daniel; Kirchdoerfer, Robert N.; Turner, Hannah L.; Cottrell, Christopher A.; Becker, Michelle M.; Wang, Lingshu; Shi, Wei; Kong, Wing-Pui; Andres, Erica L.; Kettenbach, Arminja N.; Denison, Mark R.; Chappell, James D.; Graham, Barney S.; Ward, Andrew B.; McLellan, Jason S.

doi:10.1073/pnas.1707304114

Cited by 994 publications

(1,228 citation statements)

References 88 publications

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“…Coronavirus entry into host cells is mediated by the trimeric transmembrane spike (S) glycoprotein. S is composed of two functional subunits responsible for binding to the host cell receptor (S 1 subunit) and fusion of the viral and cellular membranes (S 2 subunit) (Gui et al, 2017;Kirchdoerfer et al, 2016;Pallesen et al, 2017;Shang et al, 2017Shang et al, , 2018Walls et al, 2016aWalls et al, , 2016bWalls et al, , 2017Xiong et al, 2017;Yuan et al, 2017). We have previously determined structures of the mouse hepatitis virus (MHV) S ectodomain in the pre-fusion and post-fusion states, which provided snapshots of the start and end points of the membrane fusion reaction (Walls et al, 2016a.…”

Section: Introductionmentioning

confidence: 99%

Unexpected Receptor Functional Mimicry Elucidates Activation of Coronavirus Fusion

Walls

Xiong

Park

et al. 2019

Cell

616

765

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Graphical Abstract Highlights d MERS-CoV/SARS-CoV S composite glycan shields analyzed by cryo-EM and mass spectrometry d Structures of MERS-CoV/SARS-CoV S with neutralizing antibodies from survivors d LCA60 inhibits receptor binding by interacting with MERS-CoV S protein/glycans d S230 blocks receptor binding and triggers fusogenic rearrangements via functional mimicry In Brief Structural analysis of the SARS-CoV S and MERS-CoV S glycoproteins in complex with neutralizing antibodies from human survivors sheds light into the mechanisms of membrane fusion and neutralization Walls et al., SUMMARYRecent outbreaks of severe acute respiratory syndrome and Middle East respiratory syndrome, along with the threat of a future coronavirus-mediated pandemic, underscore the importance of finding ways to combat these viruses. The trimeric spike transmembrane glycoprotein S mediates entry into host cells and is the major target of neutralizing antibodies. To understand the humoral immune response elicited upon natural infections with coronaviruses, we structurally characterized the SARS-CoV and MERS-CoV S glycoproteins in complex with neutralizing antibodies isolated from human survivors. Although the two antibodies studied blocked attachment to the host cell receptor, only the anti-SARS-CoV S antibody triggered fusogenic conformational changes via receptor functional mimicry. These results provide a structural framework for understanding coronavirus neutralization by human antibodies and shed light on activation of coronavirus membrane fusion, which takes place through a receptor-driven ratcheting mechanism.

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Section: Introductionmentioning

confidence: 99%

Unexpected Receptor Functional Mimicry Elucidates Activation of Coronavirus Fusion

Walls

Xiong

Park

et al. 2019

Cell

616

765

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“…The mAbs developed by other groups, CDC-C2, m336, and REGN3051, also bind to RBD and potently neutralize MERS-CoV (de Wit et al, 2018;Pascal et al, 2015;Wang et al, 2018;Ying et al, 2014). Meanwhile, it has also been reported that non-RBD S1-specific mAbs, G2 and 5F9, and S2-specific mAb G4 neutralized MERS-CoV (Chen et al, 2017;Pallesen et al, 2017;Wang et al, 2015;Zhang et al, 2018).…”

Section: Discussionmentioning

confidence: 99%

“…The baculoviral expression system produced all recombinant S subunit proteins in this study. Hence, the baculovirus-expressed SΔTM can be considered as a promising MERS-CoV vaccine antigen given its ability to induce potent neutralizing antibodies, although it does not have trimerization tag for pre-fusion conformation (Pallesen et al, 2017) and may have different glycosylation patterns as compared to mammalian-expressed S proteins (Li et al, 2016).…”

Section: Discussionmentioning

confidence: 99%

Characterization of novel monoclonal antibodies against MERS-coronavirus spike protein

et al. 2020

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Middle East Respiratory Syndrome coronavirus (MERS-CoV) causes severe pulmonary infection, with ∼35 % mortality. Spike glycoprotein (S) of MERS-CoV is a key target for vaccines and therapeutics because S mediates viral entry and membrane-fusion to host cells. Here, four different S subunit proteins, receptor-binding domain (RBD; 358-606 aa), S1 (1-751 aa), S2 (752-1296 aa), and SΔTM (1-1296 aa), were generated using the baculoviral system and immunized in mice to develop neutralizing antibodies. We developed 77 hybridomas and selected five neutralizing mAbs by immunization with SΔTM against MERS-CoV EMC/2012 strain S-pseudotyped lentivirus. However, all five monoclonal antibodies (mAb) did not neutralize the pseudotyped V534A mutation. Additionally, one mAb RBD-14F8 did not show neutralizing activity against pseudoviruses with amino acid substitution of L506 F or D509 G (England1 strain, EMC/2012 L506 F, and EMC/2012 D509 G), and RBD-43E4 mAb could not neutralize the pseudotyped I529 T mutation, while three other neutralizing mAbs showed broad neutralizing activity. This implies that the mutation in residue 506-509, 529, and 534 of S is critical to generate neutralization escape variants of MERS-CoV. Interestingly, all five neutralizing mAbs have binding affinity to RBD, although most mAbs generated by RBD did not have neutralizing activity. Additionally, chimeric antibodies of RBD-14F8 and RBD-43E4 with human Fc and light chain showed neutralizing effect against wild type MERS-CoV KOR/KNIH/002, similar to the original mouse mAbs. Thus, our mAbs can be utilized for the identification of specific mutations of MERS-CoV.

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“…In the case of sera tested for binding to prefusion hRSV F, the anti‐foldon antibodies were first depleted by incubation with Ni 2+ beads coated with a MERS‐CoV S1 protein containing a C‐terminal foldon domain and 8xHis‐tag (Pallesen et al , 2017). …”

Section: Methodsmentioning

confidence: 99%

Chimeric Pneumoviridae fusion proteins as immunogens to induce cross‐neutralizing antibody responses

et al. 2017

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Human respiratory syncytial virus (hRSV) and human metapneumovirus (hMPV), two members of the Pneumoviridae family, account for the majority of severe lower respiratory tract infections worldwide in very young children. They are also a frequent cause of morbidity and mortality in the elderly and immunocompromised adults. High levels of neutralizing antibodies, mostly directed against the viral fusion (F) glycoprotein, correlate with protection against either hRSV or hMPV. However, no cross‐neutralization is observed in polyclonal antibody responses raised after virus infection or immunization with purified F proteins. Based on crystal structures of hRSV F and hMPV F, we designed chimeric F proteins in which certain residues of well‐characterized antigenic sites were swapped between the two antigens. The antigenic changes were monitored by ELISA with virus‐specific monoclonal antibodies. Inoculation of mice with these chimeras induced polyclonal cross‐neutralizing antibody responses, and mice were protected against challenge with the virus used for grafting of the heterologous antigenic site. These results provide a proof of principle for chimeric fusion proteins as single immunogens that can induce cross‐neutralizing antibody and protective responses against more than one human pneumovirus.

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Immunogenicity and structures of a rationally designed prefusion MERS-CoV spike antigen

Cited by 994 publications

References 88 publications

Unexpected Receptor Functional Mimicry Elucidates Activation of Coronavirus Fusion

Unexpected Receptor Functional Mimicry Elucidates Activation of Coronavirus Fusion

Characterization of novel monoclonal antibodies against MERS-coronavirus spike protein

Chimeric Pneumoviridae fusion proteins as immunogens to induce cross‐neutralizing antibody responses

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