Nature - USA (2020-08-20)

466 | Nature | Vol 584 | 20 August 2020

Article

IL-6, which is linked to CRS, was elevated in patients with severe
disease^19. Circulating neutrophils did not show a significant increase
in our longitudinal analysis (Fig. 2f), although patients with severe
disease showed hallmarks of type 3 responses, including increased

plasma IL-17A and IL-22, as well as secretion of IL-17 by circulating CD4

T cells as assessed by intracellular cytokine staining (Fig. 2f, Extended

Data Fig. 5). These data identify broad elevations of type 1, type 2 and

type 3 signatures in severe cases of COVID-19, with differences in their

b

de f

0

1

2

3

1.0

1.5

2.0

2.5

0.0

0.5

1.0

1.5

2.0

1.0

1.5

2.0

2.5

3.0

1.0

1.5

2.0

2.5

3.0

3.5

0.5

1.0

1.5

2.0

−2

−1

0

1

3.0

3.5

4.0

4.5

2.0

2.5

3.0

0

1

2

3

4

−1

0

1

2

3

IL-33

1.0

1.5

2.0

2.5 IL-22

0.0

0.5

1.0

1.5

2.0 IL-17A

1

2

3 IL-13

−1

0

1

2 IL-12 (p70)

0

1

2

3 IL-6

0.0

0.5

1.0

1.5

2.0 IL-5

−2

−1

0

1 IL-4

3

4

IgE

0

1

2

IFN-a

1.0

1.5

2.0

2.5

3.0

Eotaxin-2

0

1

2

3

4

TSLP

log

pg ml 10

–1

log

pg ml 10

–1

log

pg ml 10

–1

Severe

Moderate

510152025

510152025

510152025

5101 5 20 25 510152025

510152025

510152025

510152025

510152025

51 0 15 2025

5101 5 20 25

510152025 51 0 15 2025

5101 5 20 25

510152025

1-56-10

11-1516-2021-25

1–56–1011–1516–2021–25 1–56–1011–1516–2021–25 1–56–1011–1516–2021–25

Monocytes Eosinophils Neutrophils

c

log

pg ml 10

–1

1.6

2.0

2.4

0.5

1.0

1.5

2.0

2.5

1.0

1.5

2.0

2.5

3.0

0.5

1.0

1.5

2.0

2.5 IL-18

0

1

2

IL-1β

1

2

3

IFNλ

1.0 Mean of HCW

1.5

2.0

2.5

3.0 IFNα SevereModerate

log

pg ml 10

–1

Days from symptom onsetDays from symptom onset

1–56–10

11–1516–2021–25

(^510152025) 1–5 5101 5 20 25 510152025 510152025
6–1011–1516–2021–25 1–56–1011–1516–2021–25 1–56–1011–1516–2021–25
1.0
2.0
3.0
0
Days from symptom onset
Days from symptom onsetDays from symptom onset Days from symptom onset
log
pg ml 10
–1
log
pg ml 10
–1
log
pg ml 10
–1
log
pg ml 10
–1
log
pg ml 10
–1
log
pg ml 10
–1
log
pg ml 10
–1
log
pg ml 10
–1
log
pg ml 10
–1
0
10
20
Per cent of live cells
10
20
(^0 0)
10
20
30
40
50
Per cent of live cells
10
20
30
40
50
0
2
4
6
8
0
2
4
6
8
Per cent of live cells
Late
Moderate Moderate
Sever
e
Sever
e
R R = 0.085, = 0.066, P P = 0.64= 0.38
R R = –0.25, = –0.15, P P = 0.15= 0.11
R R = 0.2, = 0.025, P = 0.26P = 0.79
R = 0.24, P = 0.16 R R = 0.22, = –0.044, P = 0.21P = 0.65
R = –0.18, P = 0.054
R R = 0.25, = 0.041, P P = 0.16= 0.67
R R = 0.21, = –0.031, P = 0.23P = 0.75
R R = 0.11, = 0.068, P P = 0.23= 0.71 R R = 0.077, = –0.028, P P = 0.65= 0.77
R R = 0.46, = 0.31, P P = 0.0056= 0.00092
R R = –0.0021, = –0.061, P P = 0.52= 0.99
R R = –0.048, = –0.01, P P = 0.92= 0.79
R R = 0.011, = –0.37, P = 0.95
P = 5.1 × 10–5
R R = 0.23, = –0.1, P P = 0.18= 0.27
R R = 0.36, = 0.0093, P = 0.04P = 0.93
R R = –0.17, = 0.19, P P = 0.27= 0.07 R R = 0.22, = 0.044, P P = 0.22= 0.64 R = –0.48, P = 0.79
R = 0.055, P = 0.56
R R = 0.045, = –0.031, P P = 0.8= 0.00097
a Early
VEGF
A
EGFPDGFAA
PDGFsCD40LAB/B
B
CXCL1IL-7CCL4IL-8
TGFEotaxin2CCL17α
CXCL5IL-17FTPOIL-33
SDF1a/1BIL-20LIFIFNL2
IL-18CCL5IgE
CCL27CCL22CXCL13
TRAILCCL15IL-1RA
CXCL9CCL2IL-6
CCL1CX3CL1TNFα
IL-15MCSFCCL8
IL-10CXCL10CCL21
GCSFIL-12p40IL-1a
IL-17AFGF2IL-1b
IL-22IL-4
IL-17E/IL-25IL-12p70IL-2GMCSFIL-13
TNFIFNCCL7βγ
CCL13EotaxinEotaxin3
IFNα
2
IL-3FLT3LIL-5
IL-27TSLPIL23
IL-16IL-21IL-9
CCL3SCF
VEGF
A
PDGFEGFAA
PDsCD40LGFA B/BBCXCL1IL-7
CCL4IL-8TGFα
Eotaxin2CCL17CXCL5IL-17F
TPOIL-33
SDF1a/1BIL-20LIFIFNL2
IL-18CCL5IgE
CCL27CCL22CXCL13
TRAILCCL15IL-1RA
CXCL9CCL2IL-6CCL1
CX3CL1TNFIL-15MCSFα
CCL8IL-10
CXCL10CCL21GCSF
IL-12p40IL-1aIL-17AFGF2
IL-1bIL-22IL-4
IL-17E/IL-25IL-2IL-12p70GMCSFIL-13
TNFIFNβCCL7γ
CCL13EotaxinEotaxin3
IFNα
2
IL-3FLT3LIL-5
IL-27TSLPIL23
IL-16IL-21IL-9
CCL3SCF
−1.0
−0.8
−0.6
−0.4
−0.2
0.0
0.2
0.4
0.6
0.8
1.0
−1
0
1
2
1
2
Fig. 2 | Longitudinal immune prof iling of moderate and severe COVID-19
patients. a, Correlation matrices of 71 cytokines from patient blood
comparing cytokine concentrations in patients with moderate or severe
disease during the early phase (<10 DfSO) or late phase (>10 DfSO) of disease.
Only significant correlations (<0.05) are represented as dots, and Pearson’s
correlation coefficient from comparisons of cytokine measurements within
each patient is visualized by colour intensity. b, c, Anti-viral interferons (b) and
inf lammasome-related cytokines (c) plotted as log 10 concentrations over time
and grouped by disease severity. d–f, Cellular and cytokine measurements
representative of type 1 (d), type 2 (e) and type 3 (f) immune responses
reported over time in intervals of days (left) and continuously as linear
regressions (right). Left, each dot represents a distinct patient and time point
arranged in intervals of 5 days until 25 DfSO; dark blue, moderate disease
(n = 112), pink, severe disease (n = 40). Dark blue or pink lines pass through the
mean at each time interval; error bars denote the s.e.m. Dashed green line,
mean from healthy HCWs. Right, regression lines are indicated by the dark blue
(moderate) or red (severe) solid lines. Associated Pearson’s correlation
coefficients and linear regression significance are coloured accordingly;
shading represents 95% CI.