Goelet al.,Science 374 , eabm0829 (2021) 3 December 2021 4of17
Spike+ IgG+ Spike+ IgM+ Spike+ IgA+(^0) 0.50.75 (^1360) 0.50.75 (^1360) 0.50.75 136
0.001
0.010
0.100
1.000
0.001
0.010
0.100
1.000
Months Post−Vaccine
% of Total B
Isotype−Specific Memory B
RBD+ IgG+ RBD+ IgM+ RBD+ IgA+
(^0) 0.50.75 (^1360) 0.50.75 (^1360) 0.50.75 136
0.001
0.010
0.100
0.001
0.010
0.100
Months Post−Vaccine
% of Total B
Isotype−Specific Memory B
IgG+ IgM+ IgA+IgG+IgM+IgA+
0
25
50
75
100
0
25
50
75
100
% Isotype+
6 Months
(^0) 0.50.75 136
0
20
40
60
80
0
20
40
60
80
Months Post−Vaccine
% of Spike+
% CD71+
0.054
ns
- Naive
Recovered
Naive
Recovered
Naive
Recovered
Naive
Recovered
Spike+ RBD+
0.01
0.10
1.00
(^050100150200)
Days Post−Vaccine
% of Total B
Spike+ Memory B
0.03
0.10
0.30
0 2 4 6
Months
Spike+ Memory B
0.001
0.010
0.100
(^050100150200)
Days Post−Vaccine
% of Total B
Spike+ RBD+ Memory B
0.003
0.010
0.030
0.100
0 2 4 6
Months
Spike+ RBD+ Memory B
0.01
0.10
1.00
(^050100150200)
Days Post−Vaccine
% of Total B
HA+ Memory B
0.01
0.10
1.00
0 2 4 6
Months
HA+ Memory B
Spike-BV421
HA-PE
IgG
IgM
IgG
IgM
RBD-APC
HA-PE
Spike-BV421
HA-PE
RBD-APC
HA-PE
IgA
IgD
IgA
IgD
Pre-Immune Post-Vaccine
Spike+ Spike+
Spike+ RBD+ Spike+ RBD+
IgG+
IgM+
IgG+
IgM+
IgA+
IgA+
Spike/RBD-specific Isotypes
Flu HA+ Flu HA+
AC
D
Memory B Cells (non-naive CD20+ CD38lo/int)
E
H
B
Streptavidin
SARS-CoV-2
Spike/RBD
Fluorophore
Biotin
Multimerize
B Cell Tetramers
Label
Antigen-Specific B Cells
Quantify
Frequency/Phenotype
Flow Cytometry
1e+02
1e+03
1e+04
1e+05
1e+06
(^0 2 4 6 810)
Days Post−Stimulation
U/mL
anti−Spike IgG
Naïve Recovered
Contro
l
R8^48 +
IL^2 Control
R8^48 +
IL^2
1e+03
1e+05
1e+07
Stimulation
U/mL
anti−Spike IgG
=0.56,p= 0.00029
3e+04
1e+05
3e+05
1e+06
3e+06
0.03 0.10 0.30
% Spike+ of Total B
U/mL
anti−Spike IgG
=0.52,p= 0.001
0
25
50
75
100
0.003 0.010 0.030 0.100
% RBD+ of Total B
% Binding Inhibition
IJ WT RBD Inhibition
R848 + IL-2
Control
KL
F
43/44
15/16
37/37
8/10
44/4412/16 37/379/10
0.0760.054 ns*
ns
**
nsns nsns
nsns
SARS-CoV-2 Naive
N = 45
SARS-CoV-2 Recovered
N = 16
- mRNA
Vaccine
Pfizer
Moderna
** ns
Naïve Recovered
1
10
100
FRNT50
PSV Neutralization
=0.57,p= 0.00047
1
10
100
0.003 0.010 0.030 0.100
% RBD+ of Total B
FRNT50
B.1.351
=0.57,p= 0.00033
1
10
100
0.003 0.010 0.030 0.100
% RBD+ of Total B
FRNT50
B.1.617.2
6 Months Post-Vaccine
Memory B Cell
Differentiate into
Antibody Secreting Cell
In Vitro
R848 + IL-2
10 days
Quantify Memory B Cell-
Derived Antibodies
- anti-Spike IgG
- hACE2-RBD Binding Inhibition3) PSV Neutralization
MNO
ns ns
0.062
G
ns ns
ns ns
**
ns
***ns
ns
ns
****ns
ns
ns
ns
nsns
ns
ns
ns
nsns
**
ns
ns
ns
ns
*ns
0.068ns ns
ns
ns
ns
- Variant:
R848 + IL-2: -+-+-+-+
B.1.351 B.1.617.2 B.1.351 B.1.617.2
Fig. 2. SARS-CoV-2 mRNA vaccines generate durable and functional
memory B cell responses.(AandB) Experimental design (A) and gating
strategy (B) for quantifying the frequency and phenotype of SARS-CoV-2–
specific memory B cells by flow cytometry. Antigen specificity was determined
on the basis of binding to fluorophore-labeled spike, RBD, and influenza
HA tetramers. (C) Frequencies of SARS-CoV-2 spike+, spike+RBD+, and
influenza HA+memory B cells over time in PBMC samples from vaccinated
individuals. Data are represented as a percentage of total B cells, black
triangles indicate time of vaccine doses, fractions below plots indicate the
number of individuals above their individual baseline at memory time points,
and summary plots show mean values with the 95% confidence interval.
(DandE) Frequency of isotype-specific spike+(D) and spike+RBD+(E)
memory B cells over time. IgA was assessed on a subset of subjects.
(F) Percent IgG+, IgM+, or IgA+of SARS-CoV-2–specific memory B cells at
6 months postvaccination. (G) Percent CD71+of total spike+memory B cells
over time. (H) Experimental design for in vitro differentiation of memory
B cells into antibody-secreting cells. (I) Anti-spike IgG levels in culture
supernatants over time from PBMCs stimulated with PBS control or R848 +
IL-2 (n= 4). (J) Anti-spike IgG levels in culture supernatants after 10 days
of stimulation (K) Correlation of spike+memory B cell frequencies by flow
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