FIGURE LEGENDS
FIGURE 1. Comparisons of phenotypic features of monocytes and
neutrophils in patients with COVID-19 and controls a) HLA-DR expression
on monocytes from Covid-19 patients from all clinical courses is
significantly low compared to controls. *: p<0.01 b)MFI values of HLA-DR on monocytes from Covid-19 patients with the mild
clinical course is significantly different from the patients with
moderate and severe disease course (*: p=0.02; **: p=0.05 respectively).
All patients’ groups have decreased HLA-DR MFI compared to the control
group (#: p=0.02 for mild patients; ##: p<0.01 for moderate
and severe patients). c) A representative plot showing gate for
monocytes on FSC vs SSC and an overlaid histogram demonstrating CD16
expression on monocytes from a patient with COVID 19 and healthy
control. d) The expression of CD16 on monocytes gradually
decreases depending on the disease course (*:p=0.05 between the mild and
severe groups). But, all were significantly high in comparison to
controls (#: p<0.01). e) The expression of CD10 on
neutrophils is significantly low in COVID-19 patients with the severe
course (*: p<0.02). f) MFI values of CD10 on
neutrophils in patients with COVID-19 are low compared to controls. (*:
p<0.01) g) A representative plot showing gate for
neutrophils on FSC vs SSC and an histogram demonstrating CD16 expression
on neutrophils from a patient with COVID 19 and healthy control.h) CD16 MFI values of neutrophils from patients with COVID-19
with the different clinical course were decreased in comparison to
controls (*: p<0.01)
FIGURE 2. A representative figure for neutrophil and monocyte
oxidative burst in a patient with COVID-19. a) Neutrophils (N)
and monocytes (M) were gated on forward vs side scatter plot b)Neutrophil oxidative burst as DHR123 fluorescence (Us: unstimulated; PMA
stimulated) c) Monocyte oxidative burst as DHR123 fluorescence
(Us: unstimulated; PMA stimulated)
FIGURE 3. Increased CD3-CD8+CD56+ NK like innate lymphoid cells
in COVID-19: a) A representative CD3 vs CD8 plot from lymphocytes of a
patients with COVID-19. CD3-CD8+cells were gated. b) CD56 expression of
CD3-CD8+ cells. c) The
percentages of
CD3-CD8+CD56+cells were calculated on CD8 gated lymphocytes. This was significantly
high in all patient groups with COVID-19 in comparison to control
(#:p<0.01).
FIGURE 4. Representative flow cytometric plots for the main
lymphocyte subsets in a patient with COVID-19 and a
control. a) Lymphocytes (L) were gated on forward vs side
scatter plot b) Histograms for CD19+ B cells.c) Plots demonstrating NK
(CD3-CD16+CD56+)
and T
(CD3+CD16-CD56-)
cells. d) Representative plots for T helper
(CD3+CD4+) e) and T
cytotoxic/suppressor cells
(CD3+CD8+).
FIGURE 5. Spontaneous CD69 expression on CD4+ and CD8+ T cells
in patients with COVID-19 a, c) Representative CD69 vs CD4 and CD8
plots demonstrating spontaneous CD69 expression on lymphocytes from a
patient with COVID-19 b) The ratio of CD69 on CD4+ T cells was
significantly high in the severe patients (9.4% ±6.8) in comparison to
the mild (3.7% ±4.9) and moderate patients (3.6% ±4; *:
p<0.01). c) The ratio of CD8+CD69+ T cells was
significantly increased in the severe group (45% ±34.5) in comparison
to the mild group (13.3% ±12.4; *: p=0.01)
FIGURE 6. CD3+CD56+ NK like
T cells were high in COVID-19. a, b) Representative CD3 vs CD56 plots
from a patient with COVID-19 and a control. c) *:
p<0.01: The percentage of
CD3+CD56+ T cells was significantly
high in patients with COVID-19.
FIGURE 7. Suppressive T lymphocyte subsets increased in patients
with COVID-19. a) The ratio of double-negative T cells
(CD3+CD4-CD8-) was
significantly increased in patients with COVID-19 with moderate or
severe courses in comparison to the control group (*:
p<0.01). b) Representative CD4 vs CD8 plots gated on
CD3+ T cells in a patient with COVID-19 andc) healthy control. Lower left quadrant indicates
CD3+CD4-CD8- cellsd) #: p<0.01: HLA-DR expressing T cells
were increased in patients with COVID-19. e, f) CD3 vs HLA-DR
plots from a patient with COVID-19 and a control. g) #:
p<0.03: The percentage of
CD8+CD28- suppressive phenotype was
prominent in COVID-19 cases. h, i) CD8 vs CD28 plots in patient
with COVID-19 and a control. Upper left quadrant indicates CD28 negative
CD8+ cells.
FIGURE 8. Lymphocyte early, late apoptosis, and cell death . a)
Early apoptosis measured by the percentage of Phiphilux fluorescence was
high in patients with Covid-19 (*: p=0.02). Total cell death as a sum of
Phiphilux-PI-positive and PI only positive cells was elevated in
patients with Covid-19 in comparison to controls (**: p<0.05).
b) Fold increase in PhiPhilux fluorescence as an indicator of caspase-3
activation in lymphocytes was low in Covid-19 patients (#:
p<0.01). c) A representative histogram showing baseline
caspase 3 activations (PhiPhilux fluorescence intensity) in lymphocytes
from a healthy control (UsC: unstimulated control), a Covid-19 patient
(UsCv19: unstimulated Covid-19) and same Covid-19 patient following PMA
stimulation (PMA-Cv19).