The Veesler Lab Profile picture
Jan 17 29 tweets 14 min read
Curious about #COVID19 #SARSCoV2 therapeutic antibodies, bivalent vaccines & immune imprinting?

Here is our latest work on these topics in collaboration with @DavideCorti6 @msdiamondlab

Led by @AminAddetia @LucaPiccoli9 Brett Case @YoungjunPark11

1/
biorxiv.org/content/10.110…
Currently circulating #SARSCoV2 variants are evolving from the Omicron BA.2 and BA.5 lineages, through independent accumulation of amino acid mutations at shared receptor-binding domain (RBD) hot spots, underscoring the importance of antibody-mediated selective pressure.

2/
We first studied how mutations in the #SARSCoV2 BQ.1.1, XBB.1(.5) and BA.2.75.2 variants affect function.
@AminAddetia @johnbowenbio in our lab previously showed that BA.5 affinity for ACE2 was >> Wu but fusogenicity was worse

3/
science.org/doi/10.1126/sc…
We found that the (dominant) BQ.1.1 RBD and the BA.2.75.2 RBD have similar ACE2 binding affinity to the BA.5 RBD whereas XBB.1 RBD has lower ACE2 affinity (similar to Wu RBD).
Modulation results from off-rate differences, as we have seen throughout the pandemic

4/
We also observed that spike-mediated membrane fusion (leading to viral entry) is more efficient with BQ.1.1, XBB.1(.5) and BA.2.75.2 than previous Omicron variants, almost reaching Wu levels

5/
BQ.1.1 and BA.2.75.2 viral fitness is therefore not limited by ACE2 binding whereas the lower ACE2-binding affinity of XBB.1 might have limited its spread.
Recent data from @yunlong_cao showing enhanced ACE2 binding of XBB.1.5 (S486P) support this!

6/
biorxiv.org/content/10.110…
BA.1, BA.2 and BA.5 had altered cell entry pathway vs previous #SARSCoV2 strains, with Omicron variants entering preferentially through the endosomal entry route (cathepsin-mediated) as opposed to plasma membrane fusion (TMPRSS2-mediated)
@GuptaR_lab

7/

nature.com/articles/s4158…
To assess the preferred cell entry route of currently circulating variants, we investigated the impact of protease inhibitors on spike-mediated entry into VeroE6-TMPRSS2 cells (enabling both plasma membrane and endosomal entry routes)

8/
Camostat and nafamostat (TMPRSS2 inhibitors) had limited impact on Omicron variants, including BQ.1.1, BA.2.75.2 and XBB.1, whereas E64d (cathepsin B/L inhibitor) reduced BA.1, BA.2 and BA.5 entry more than BA.2.75.2, BQ.1.1 or XBB.1, relative to Wu-G614

9/
Inefficient use of TMPRSS2 concurs with the identical BQ.1.1, BA.2.75.2 and XBB.1 S2 subunit sequences, including spike N969K which @PeacockFlu previously showed to account for preferred endosomal cell entry of prior Omicron variants (BA.2)

10/

biorxiv.org/content/10.110…
To reveal how amino acid substitutions in the BQ.1.1 and XBB.1 RBDs alter receptor recognition and key antigenic sites, we determined #cryoEM structures of the RBDs bound to ACE2 & S309 Fab (parent of the sotrovimab monoclonal antibody therapeutic)

11/
The R493Q reversion likely relieves repulsion with ACE2 residue K31 and restores a network of local interactions similar to that made with the Wu RBD

12/
These data provide a blueprint to contextualize deep-mutational scanning and mutagenesis data we recently reported in the context of the BA.1 and BA.2 RBD backgrounds with @tylernstarr @jbloom_lab

13/
journals.plos.org/plospathogens/…
Our #cryoEM structures reveal how the BQ.1.1 K444T and the BQ.1.1/XBB.1 R346T would abrogate key electrostatic interactions with the LY-CoV1404 (bebtelovimab parent) and COV2-2130 (cilgavimab parent, Evushield cocktail), explaining dampened binding and neutralization

14/
We show how S309 binds both the BQ.1.1 and XBB.1 RBDs and accommodates the XBB.1 H339 residue in the epitope! S309 binding pose is indistinguishable from that observed when bound to the Wu/BA.1 RBDs

15/
The S371F mutation (BA.2, BA.5, BQ.1.1, XBB.1 and BA.2.75.2) changes conformation of the RBD helix 364-372 which becomes sterically incompatible with the glycan N343 conformation observed in S309-bound spike structures

16/

science.org/doi/10.1126/sc…
The BQ.1.1 helix has a conformation similar to that in the S309-bound BA.1 structure, but distinct from apo BA.2 /BA.5 structures, and is disordered for XBB.1, indicative of conformational frustration upon S309 binding which could explain reduced neutralizing activity

17/
We found that S309 retains detectable neutralizing activity of currently circulating variants, including the rising XBB.1.5 and the dominant BQ.1.1, with potencies correlating with 1:1 Fab binding affinity to variant RBDs

18/
Sotrovimab (S309 derivative) IgG binds avidly to cell surface-expressed currently dominant Omicron #SARSCoV2 spike variants & promoted antibody-dependent cell cytotoxicity (ADCC) using primary natural killer effector cells (independently of 1:1 Fab affinity)!

19/
Prophylactic administration of S309 protected K18-hACE2 transgenic mice from weight loss and reduced both viral RNA and infectious virus titers in the lungs.
Despite marked reduction of in vitro neutralizing activity, S309 can protect mice from BQ.1.1 challenge!

20/
To assess the impact of the BQ1.1, XBB.1 and BA.2.75.2 S mutations on vaccine-elicited antibody responses, we quantified plasma neutralizing activity in various human cohorts infected and/or vaccinated with monovalent and bivalent mRNA vaccines

21/
Wu/BA.5 or Wu/BA.1 bivalent mRNA vaccinated subjects had detectable neutralizing activity vs vaccine-mismatched XBB.1, BA.2.75.2 and BQ.1.1 (w/wo prior infection) whereas little to no neutralization of these variants was detected for the Wu-only vaccinated subjects

22/
These data suggest that bivalent (Wu/BA.1 or Wu/BA.5) mRNA vaccination elicits more potent and broader antibody responses against vaccine-matched and mismatched Omicron variants than monovalent Wu mRNA vaccination in healthy subjects

23/
We further observed than vaccine-elicited polyclonal plasma antibodies retain comparable binding titers across all Omicron variants and that in some subjects they are triggering Fc effector functions against circulating Omicron variants.

24/
Analysis of memory B cell responses in these human cohorts showed that bivalent Wu/BA.5 mRNA vaccination enriches for MBCs that are cross-reactive with the vaccine-matched and mismatched RBD variants, relative to monovalent Wu mRNA vaccination, in line with neutralization.

25/
Surprisingly, few de novo elicited Omicron-specific memory B cells were detected even after two exposures to an Omicron spike via breakthrough infection followed by Wu/BA.5 bivalent mRNA vaccination.

26/
We did not detect any de novo elicited Omicron-specific memory B cells after Wu/BA.1 bivalent mRNA vaccination (w/wo Omicron infection) and most RBD-directed IgGs secreted by stimulated memory B cells cross-react with BA.1 and (to a lesser extent) with BQ.1.1 and XBB.1 RBDs

27/
Collectively, these data suggest that two Omicron spike exposures are not sufficient to overcome the immunological imprinting induced by repeated Wu spike exposures but does continue to enrich for memory B cells cross-reacting with multiple RBD variants

28/
Thank you so much to @sprouse_kaitlin Cameron Stewart, Courtney Yoshiyama & all our wonderful collaborators including @LanzavecchiaB @DrLisaPurcell @HelenChuMD @HaVanDang2712 @SkiClimbSciNick

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More from @veeslerlab

Dec 7, 2022
Can coronaviruses keep surprising us?
Yes - we joined forces with Xiangxi Wang (Chinese Acad of Sci, Beijing) & Huan Yan (Wuhan Univ) to show that close relatives of MERS-CoV found in bats use ACE2 as host cell receptor!
@HHMINEWS
@UWBiochemistry

1/18
nature.com/articles/s4158…
MERS-CoV (Middle-East respiratory syndrome coronavirus), is a pathogen that emerged in 2012 and caused several outbreaks in the Middle-East and South Korea with a 35% fatality rate

2/18

nejm.org/doi/full/10.10…
MERS-CoV uses dipeptidyl-peptidase 4 (DDP4) as host cell receptor, as is the case for a few related viruses (e.g. HKU4) from the same merbecovirus subgenus

3/18

nature.com/articles/natur…
Read 18 tweets
Nov 10, 2022
The peer-reviewed version of our manuscript describing the influence of the #SARSCoV2 spike glycoprotein conformation on vaccine- and infection-elicited antibody responses in humans is out!

Led by @johnbowenbio
@HHMINEWS
@UWBiochemistry

science.org/doi/10.1126/sc…

1/
The original thread describing the @biorxivpreprint can be found here:



2/
During the review process, we expanded our panel of vaccinee plasma samples to also include subjects that received 2 doses of Jansen Ad26.COV2.S. or of Novavax NVX-CoV2373, thereby covering all 4 vaccines authorized or approved in the US (and several others used worldwide)

3/
Read 12 tweets
Jul 19, 2022
The peer-reviewed version of our manuscript assessing human neutralizing antibodies against key #Omicron variant sublineages elicited by a comprehensive panel of vaccines is now out

Led by @johnbowenbio & @AminAddetia

@HHMINEWS
@UWBiochemistry

1/19

science.org/doi/10.1126/sc…
The thread describing our @biorxivpreprint submission can be found here and we added a large amount of data during the reviewing process (cf. thread below).

2/19

To understand how the constellations of spike mutations present in each #SARSCoV2 #Omicron variant impact entry into cells, we compared the functional properties of the Wuhan-Hu-1/G614, Delta, BA.1, BA.2, BA.2.12.1, and BA.4/5 variants.

3/19
Read 19 tweets
May 27, 2022
The peer-reviewed version of our manuscript characterizing the 8th human-infecting #coronavirus designated CCoV-HuPn-2018 is now published!

Led by the unstoppable @aletortorici

@HHMINEWS
@UWBiochemistry

1/8

cell.com/cell/fulltext/…
The original thread describing our results can be found here (with a link to our original @biorxivpreprint)



2/8
During the reviewing process, we determined a crystal structure of the CCoV-HuPn-2018 RBD/B domain bound to canine APN revealing that receptor recognition involves similar interactions to PRCV RBD bound to porcine APN.
With stunning & key contacts with the APN N747 glycan!

3/8 Image
Read 8 tweets
May 10, 2022
We analyzed human plasma, mucosal and memory antibody responses against #SARSCoV2 and found a pan-variant neutralizing antibody!

Led by @YoungjunPark11 Dora Pinto @coronalexington @JZhuoming

Collab. with @DavideCorti6

@HHMINEWS
@Vir_Biotech

1/24

biorxiv.org/content/10.110…
We assessed plasma neutralizing activity from humans previously infected in 2020 (WA-1-like) and then vaxed 2x or 3x, or vaxed before Delta or #Omicron BA.1 breakthrough infection or vaxed-'only' 3x

2/24
#Omicron breakthrough cases had highest neutralizing activity against #SARSCoV2 variants, possibly due to exposure to BA.1 spike whereas neutralization of SARS-CoV-1 was low for all cohorts, due to genetic and antigenic divergence of the latter spike

3/24
Read 24 tweets
Mar 17, 2022
Excited to share our latest serological data evaluating plasma neutralizing antibody responses and immune evasion associated with the constellation of spike mutations present in the #Omicron BA.1 and BA.2 variants.

Led by @johnbowenbio

1/13

biorxiv.org/content/10.110…
We compared side-by-side the plasma neutralizing activity elicited in humans by seven #COVID19 vaccines (Moderna, Pfizer, Novavax, Janssen, AstraZeneca, Sputnik V and Sinopharm) or SARS-CoV-2 Washington-1 infection. We cover mRNA, Ad-vectored and protein subunit vaccines!

2/13
We found that the large number of BA.1 and BA.2 spike mutations severely dampen plasma neutralizing activity elicited by infection or primary vaccine series, with a more marked effect for BA.1 compared to BA.2 across all groups.

3/13
Read 13 tweets

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