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. 2020 Dec 17;9:e63430. doi: 10.7554/eLife.63430

Figure 3. Kinetics of the adaptive and innate immune response in a COVID-19 patient during ICU treatment.

(a–d) Representative flow cytometry plots showing the expression of activation and proliferation markers on CD4+ and CD8+ T cells (top and bottom panels, respectively in a, b), TCR-γδ T cells (c) and NK cells (d: plots shown for NK CD56dim cells) in a healthy control and the COVID-19 patient (at days 23 and 58). (e-f) Immune activation and proliferation levels are summarized for healthy controls (HC, n = 7) and five longitudinal time points of the COVID-19 patient and are assessed as co-expression of HLA-DR and CD38 (e) and expression of Ki67 (f) in CD4+ and CD8+ T cells, co-expression of CD38 and Ki67 by TCR-γδ T cells (g), and co-expression of HLA-DR and CD38 in NK CD56dim cells (h). Gating strategies for each population are included in Figure 3—figure supplement 1. All data is obtained by flow cytometry and samples were acquired on a Becton Dickinson LSR Fortessa X-20.

Figure 3.

Figure 3—figure supplement 1. The gating strategies and flow cytometry plots are shown for each immune population.

Figure 3—figure supplement 1.

(a) T and NK cell populations; (b–c) T cell activation and proliferation markers; (d) TCR-γδ T cells; (e) TCR-γδ activation and proliferation markers; (f) NK CD56dim and CD56bright cell activation markers.

Figure 3—figure supplement 2. Kinetics of the innate immune response in the COVID-19 patient.

Figure 3—figure supplement 2.

(a) Percentages of CD14+ CD16- classical monocytes (cMo), CD14+ CD16+ intermediate monocytes (iMo) and CD14- CD16+ non classical monocyes (ncMo). Percentages are calculated in live, CD3-, CD66b-, HLA-DR+ cells in the COVID-19 patient and in healthy controls (n = 6). (b) Percentages of blood monocyte-derived M1 (CD14+ CD80+ CD68+) and M2 (CD14+ CD163+ CD68+) macrophages are shown for the COVID-19 patient and healthy controls. (c–e) Neutrophil degranulation measured as MFI of CD11b (secretory vesicles), CD66b (secondary granules) and CD63 (primary granules) is shown for the patient and healthy controls (n = 4). Neutrophils were immunostained in whole blood, followed by erythrocyte lysis with ACK buffer and fixation with 4% PFA prior for analysis by flow cytometry. Blood monocytes and monocyte-derived macrophages were stained from PBMCs isolated by Ficoll gradient as described in the methods section.

Figure 3—figure supplement 3. Antibody response towards Pseudomonas aeruginosa and SARS-CoV-2.

Figure 3—figure supplement 3.

(a) Serum IgG response to Pseudomonas aeruginosa (PA) protein outer membrane porin F (OprF) was assessed in healthy controls (n = 2), in the COVID-19 patient (n = 1) and in patients with diagnosis of Cystic Fibrosis and PA infection in the lung (PA patient, n = 2). Data are presented as mean ± SD. (b) Plasma IgG responses to SARS-CoV-2 Spike and N were assessed in the COVID-19 patient at five timepoints. For reference we also show levels from serum of healthy control donors (HC; n = 5), healthy pre-pandemic control donors (PP; n = 6) and a pooled serum (PS; n = 3) control per plate (collected from 3 RT-PCR-confirmed convalescent donors). The optical density (OD) at 492 nm was corrected for background (620 nm) and for the blank values. Data are presented as mean ± SD.