Science - USA (2022-03-04)

SCIENCEscience.org 4 MARCH 2022¥VOL 375 ISSUE 6584 1043

WT

(0.16%)

WT S/S-2P

cross-reactive

(0.84%)

Vaccinated

A BC

pandemicPre- PBPB

0

5

10

15

mRNA-1273ChAdOx1:

% plasmablasts of total

CD19

+ B cells

ChAdOx1:ChAdOx1

0.98%0.95%

0.97%

3.3%

0.72%

S-2P | PE

WT S | APC^0

5

% total S-specific IgG

+ B cells

10

15

0.76%1.2%

1.1%

4.7%

0.06%

Pre-

pandemicPBPBChAdOx1:

mRNA-1273

ChAdOx1:

ChAdOx1

D E

F G

H I

22 19

0 1 2 3 4 5

# of variant RBDs

Wuhan-1 Beta Delta Kappa Lambda ChAdOx1 boost mRNA-1273 boost

ChAdOx1

boost

mRNA-1273

boost

0.0

0.5

1.0

1.5

2

3

4

5

0

20

40

60

80

100

ChAdOx1

prime

ChAdOx1

boost

mRNA-1273

boost

WT S

S-2P

NTD

RBD

HexaPro S2

HKU1 S

OC43 S

anti-S2 mAb

competition

DI-70A

643

ADI-70146ADI-69A DI-69A

116

DI-70A

846

DI-70A

098

DI-69A

084

DI-69A

738

DI-69A

746

DI-69A

834

DI-69A

518

DI-69A

657

DI-69A

665

DI-69A

787

DI-69A

696

DI-70A

485

DI-69A

134

DI-69A

725

DI-69A

823

DI-69A

745

DI-70A

757

DI-70A

035

DI-69A

095

DI-69A

760

DI-69A

702

DI-69A

682

DI-69A

530

DI-69A

554

DI-69A

534

DI-69A

824

DI-69A

821 067

DI-69A

950

DI-70

(^9580)
20
40
60
80
100
% Neutralization
Non-competitive
(pos. control)
Competitive
5-10
<5
10-50

50
0
KDApp (nM)
–10^3
103
104
105
Gated on CD19+ IgG+ B cells
Pre-pandemic
–10^30103104105 –10^30103104105
UM70UM85UM67UM84UM92UM88UM91UM75UM90UM58UM71UM72UM73UM86UM70UM85UM67UM84UM92UM88UM91UM75UM90UM58UM71UM72UM73UM86
0
20
40
60
80
100
Proportion of total S-specific
IgG

• B cells (%)
  Donor ID:
  ChAdOx1 prime ChAdOx1 boost
  UM110UM113UM102UM123UM121UM128UM103UM104UM115UM127UM139UM132UM106UM110UM113UM102UM123UM121UM128UM103UM104UM115UM127UM139UM132UM106
  0
  20
  40
  60
  80
  100
  ChAdOx1 prime mRNA1273 boost
  WT S-specific
  WT S/S-2P
  cross-reactive
  S-2P-specific
  WT S-specific
  WT S/S-2P
  cross-reactive
  S-2P-specific
  S-2P-specific
  (0.00%)
  S-2P-specific
  (0.00%)
  WT S/S-2P
  cross-reactive
  (0.00%)
  S-specific
  WT
  (0.00%)
  S-specific
  % RBD-specific IgG


• B cells
  Proportion of total S-specific
  IgG


• B cells (%)
  Fig. 2. SARS-CoV-2 S-specific B cell responses induced by homologous and
  heterologous prime-boost vaccination.(A) Frequencies of plasmablasts
  (defined as CD19+CD20–/loCD71+cells) among circulating CD19+B cells after
  prime and boost vaccination, as determined with flow cytometry. Prepandemic
  donor samples (n= 9 donors) were included for comparison. Median values are
  shown above data points. (B) Representative fluorescence-activated cell sorting
  (FACS) gating strategy used for identifying WT S-specific, S-2P–specific, and
  WT/S-2P cross-reactive IgG+B cells. (C) Frequencies of total (WT + S-2P)
  SARS-CoV-2 S-reactive B cells among circulating IgG+B cells, as determined with
  flow cytometry. Median frequencies are shown above data points. (D) Frequency
  of circulating IgG+B cells reactive with RBDs encoding mutations present in
  Beta, Delta, Kappa, and Lambda variants. The height of each bar indicates
  median frequency. (E) Proportion of donors with detectable B cell reactivity with
  the indicated number of variant RBDs. The total number of donors analyzed is
  indicated in the center of the pies. Statistical significance was determined by
  means of Fisher’s exact test and calculated according to the proportion of donors
  with B cells displaying reactivity to≥3 variant RBDs. (F) Proportions of WT
  S-specific, WT S/S-2P cross-reactive, and S-2P–specific B cells among total
  S-specific B cells after (left) homologous or (right) heterologous prime-boost
  immunization. Donors with S-specific B cell frequencies <1% of total IgG+B cells
  at either time point were excluded from this analysis. Donor identification
  numbers (IDs) are denoted above each bar. (G) Mean proportions of WT
  S-specific, WT S/S-2P cross-reactive, and S-2P–specific B cells across all donors
  within each cohort. Error bars indicate 95% confidence intervals. (H) Apparent
  binding affinities (KDApp) of WT S-specific monoclonal antibodies for WT S,
  S-2P, prefusion S subdomains (NTD, RBD, and prefusion-stabilized S2), HKU1 S,
  and OC43 S, as determined with biolayer interferometry (BLI). Competitive
  binding with an S2-directed antibody (ADI-69962), as determined with a
  BLI competitive sandwich assay, is indicated below the heatmap. (I) Neutralizing
  activity of WT S-specific antibodies against MLV-SARS-CoV-2 Wuhan-1 at a
  concentration of 1mg/ml. A previously described neutralizing antibody to RBD
  (ADG-2) was included as a positive control ( 29 ). Statistical comparisons between
  paired prime and boost samples were determined by means of [(A) and (C)]
  Wilcoxon pair-matched rank sum test. Statistical comparisons between
  vaccination cohorts were determined by means of (D) two-sided Mann-Whitney
  Utests and [(C) and (G)] two-sided Kruskal-Wallis test by ranks with subsequent
  Dunn’s multiple comparisons. P< 0.05, P< 0.01, P< 0.001, ****P<
  0.0001. P, Prime; B, Boost;KDApp, apparent equilibrium constant.
  RESEARCH | REPORTS