Raw values underlying the summary data displayed in the figures and tables.

(XLSX)</p

Autoantibodies targeting type I IFNs in the plasmas and tracheobronchial secretions of critically ill COVID-19 patients.

(A and B) Multiplexed bead-based assay to detect IgG, IgA, and IgM autoAbs against IFNα2 (A) or IFNω (B) in plasmas of patients in ICU with severe COVID-19 (Male: M = 179 samples corresponding to 80 patients, Female: F = 51 samples corresponding to 23 patients) or Healthy Donors (HD = 130 samples). MFI FC of signal derived from IFN-coated beads relative to the MFI of signal derived from albumin-coated beads is shown. Dashed lines indicate 10 SDs (A) or 5 SDs (B) from the mean calculated from HD values for each IFN and each isotype. Values above the dashed lines are considered positive. Percentage of positive patients (not samples) per analyzed group is indicated. (C and D) Multiplexed bead-based assay to detect IgG, IgA, and IgM autoAbs against IFNα2 (C) or IFNω (D) in TBSs of COVID-19 ICU patients described in (A). Pos (positivity) and Neg (negativity) for anti-IFNα2 IgG (C) or anti-IFNω IgG (D) in plasma samples from the same patient were used to stratify patients. MFI FC of signal derived from IFN-coated beads relative to the MFI of signal derived from albumin-coated beads is shown. In all panels, red dots indicate the patients/samples that were positive for anti-IFNα2 IgG autoAbs in plasma (A) and are denoted simply for reference. Data underlying this figure can be found in S1 Data. autoAbs, autoantibodies; COVID-19, Coronavirus Disease 2019; FC, fold change; HD, Healthy Donors; ICU, intensive care unit; IFN, interferon; MFI, median fluorescence intensity; SD, standard deviation; TBS, tracheobronchial secretion.</p

Analysis of autoantibodies targeting IFNβ in the plasmas and tracheobronchial secretions of critically ill COVID-19 patients.

Multiplexed bead-based assay to detect IgG, IgA, and IgM autoantibodies (autoAbs) against IFNβ in the plasmas (A) or TBSs of COVID-19 ICU patients described in Fig 1A. Pos (positivity) and Neg (negativity) for anti-IFNα2 IgG in plasma samples from the same patient (results from Fig 1A) were used to stratify patients. MFI FC of signal derived from IFN-coated beads relative to the MFI of signal derived from albumin-coated beads is shown. In all panels, red dots indicate the patients/samples that were positive for anti-IFNα2 IgG autoantibodies in plasma (Fig 1A) and are denoted simply for reference. Data underlying this figure can be found in S1 Data. COVID-19, Coronavirus Disease 2019; FC, fold change; ICU, intensive care unit; IFN, interferon; MFI, median fluorescence intensity; TBS, tracheobronchial secretion. (TIF)</p

Presence of autoantibodies targeting type I IFNs as a predictor of herpesvirus disease in critically ill COVID-19 patients.

Fifty-nine severe COVID-19 patients in ICU were tested for herpesvirus (HSV-1/2 and CMV) reactivations in their blood by PCR. Pos (positivity) and Neg (negativity) for CMV, HSV-1/2, and CMV and/or HSV-1/2 were used to stratify the results obtained when plasma samples from the same patient were assayed for IgG autoAbs targeting IFNα2 (see Fig 1). MFI FC of signal derived from IFNα2-coated beads relative to the MFI of signal derived from albumin-coated beads is shown for each individual patient. Values above the dashed line are considered positive (red). Data underlying this figure can be found in S1 Data. autoAbs, autoantibodies; CMV, cytomegalovirus; COVID-19, Coronavirus Disease 2019; FC, fold change; MFI, median fluorescence intensity; HSV-1/2, herpes simplex virus types 1 or 2; ICU, intensive care unit; IFN, interferon.</p

Longitudinal analysis of plasma autoantibodies targeting type I IFNs in individual critically ill COVID-19 patients and their neutralization capacities.

(A and B) Longitudinal analysis of plasma anti-IFNα2 (A) and anti-IFNω (B) IgG, IgA, and IgM autoAbs in selected critically ill COVID-19 patients positive for plasma anti-IFNα2 IgG. Samples were collected on day of admission to ICU (d1) and as indicated thereafter. MFI FC of signal derived from IFN-coated beads relative to the MFI of signal derived from albumin-coated beads is shown. Dashed lines indicate 10 SDs (IFNα2) or 5 SDs (IFNω) from the mean calculated from HD values for each IFN and each isotype in Fig 1A and are used as threshold values for positivity (filled circles). Internal patient identifier numbers (P) are shown, together with the individual’s gender (male, M; female, F) and age (years, y). (C) Schematic representation of the luciferase reporter-based neutralization assay. HEK293T cells are cotransfected with a pGL3-Mx1P-FFLuc reporter (FF-Luc) plasmid and a constitutively active pRL-TK-Renilla (Ren-Luc) plasmid. After 24 h, cells are incubated with IFNα2 or IFNω that have been preincubated with patient plasmas. After a further 24 h, cells are lysed, and IFN-stimulated luminescence intensity (FF-Luc) is measured and made relative to the constitutively active Ren-Luc. Schematic created with BioRender.com. (D) Results for the neutralization of 10, 1, or 0.2 ng/mL of IFNα2 or IFNω in the presence of 1/50 diluted patient plasmas from ICU COVID-19 patients positive for anti-IFNα2 IgG (n = 12), ICU COVID-19 patients negative for anti-IFNα2 IgG (n = 6), or HD (n = 6). FF-Luc values were made relative to Ren-Luc values and then normalized to the median luminescence intensity of control samples without IFN. Some individual patient (P) and sampling day (d) identifiers (corresponding to Fig 2A and 2B) are shown for comparison with their IFNα2 or IFNω binding data. Data underlying this figure can be found in S1 Data. autoAbs, autoantibodies; COVID-19, Coronavirus Disease 2019; FC, fold change; HD, Healthy Donors; IFN, interferon; MFI, median fluorescence intensity; SD, standard deviation.</p

A multiplexed bead-based assay to detect IFN-binding antibodies.

(A) Schematic representation of the assay principle. Magnetic beads are covalently coated with the indicated IFNs or albumin as a negative control. Samples are then incubated with the coated beads for 1 h at room temperature to allow binding of any anti-IFN antibodies present. Following wash steps, PE-labeled secondary antibodies specific for antibody isotypes of interest (IgG, IgA, or IgM) are incubated with the beads. After washing, MFI values of bound PE secondary antibodies are measured for each “bead region” on a FlexMap 3D instrument. Schematic created with BioRender.com. (B, C, and D) Assay assessment using mouse monoclonal antibodies. IFNα2, IFNβ, IFNω, and albumin-coated beads mixed 1:1:1:1 were incubated with serial dilutions of mouse monoclonal antibodies raised against IFNα2 (B), IFNβ (C), or IFNω (D). Following the assay procedure described in (A), MFI values from IFN-coated beads were obtained and calculated relative to MFI values from albumin-coated beads. Data are representative of at least 2 independent experiments. (E) Assay assessment using human plasma samples. IFNα2, IFNβ, IFNω, and albumin-coated beads mixed 1:1:1:1 were incubated with serial dilutions of a pool of healthy donor plasmas (left panel) or a human plasma known to have anti-IFNα2 antibodies (right panel). Following the assay procedure described in (A), MFI values from IFN-coated beads were obtained and calculated relative to MFI values from albumin-coated beads. Data are representative of at least 2 independent experiments. Data underlying this figure can be found in S1 Data. FC, fold change; IFN, interferon; MFI, median fluorescence intensity; PE, phycoerythrin. (TIF)</p

Description of patient outcomes.

Description of patient outcomes.</p

Description of herpesvirus detection.

Description of herpesvirus detection.</p

Patient baseline characteristics.

Patient baseline characteristics.</p

Data_Sheet_1_Differing Visual Behavior Between Inexperienced and Experienced Critical Care Nurses While Using a Closed-Loop Ventilation System—A Prospective Observational Study.docx

Introduction: Closed-loop ventilation modes are increasingly being used in intensive care units to ensure more automaticity. Little is known about the visual behavior of health professionals using these ventilation modes. The aim of this study was to analyze gaze patterns of intensive care nurses while ventilating a patient in the closed-loop mode with Intellivent adaptive support ventilation® (I-ASV) and to compare inexperienced with experienced nurses.Materials and Methods: Intensive care nurses underwent eye-tracking during daily care of a patient ventilated in the closed-loop ventilation mode. Five specific areas of interest were predefined (ventilator settings, ventilation curves, numeric values, oxygenation Intellivent, ventilation Intellivent). The main independent variable and primary outcome was dwell time. Secondary outcomes were revisits, average fixation time, first fixation and fixation count on areas of interest in a targeted tracking-time of 60 min. Gaze patterns were compared between I-ASV inexperienced (n = 12) and experienced (n = 16) nurses.Results: In total, 28 participants were included. Overall, dwell time was longer for ventilator settings and numeric values compared to the other areas of interest. Similar results could be obtained for the secondary outcomes. Visual fixation of oxygenation Intellivent and ventilation Intellivent was low. However, dwell time, average fixation time and first fixation on oxygenation Intellivent were longer in experienced compared to inexperienced intensive care nurses.Discussion: Gaze patterns of intensive care nurses were mainly focused on numeric values and settings. Areas of interest related to traditional mechanical ventilation retain high significance for intensive care nurses, despite use of closed-loop mode. More visual attention to oxygenation Intellivent and ventilation Intellivent in experienced nurses implies more routine and familiarity with closed-loop modes in this group. The findings imply the need for constant training and education with new tools in critical care, especially for inexperienced professionals.</p