Xn 2000 hematology system

Hemoglobin, platelet, white blood cell (WBC), and hematocrit counts were obtained from the collected blood samples using a Sysmex XN-2000 hematology system (Sysmex Corp., Norderstedt, Germany). The estimated glomerular filtration rate (eGFR), calcium, creatinine, potassium, sodium, urea, and blood urea nitrogen (BUN) levels were measured from the same blood samples using a Cobas 8000 autoanalyzer (Roche Diagnostics, Basel, Basel-Stadt, Switzerland). The kidney tissue interleukin-1β (IL-1β), IL-6, tumour necrosis factor-α (TNF-α), and interferon-γ (IF-γ) (E-Bioscience, Vienna, Austria) levels were measured using commercial ELISA rat kits with an MVGt Lamda scan 200 (Bio-Tek Instruments, Winooski, VT, USA).

During the third wave of COVID-19 pandemic occurred between December 2020 and April 2021, we enrolled all SARS-CoV-2+ patients hospitalized at University Hospital “Maggiore della Carità” (Novara, Italy) with blood samples available and who signed the informed consent to participate in the study. SARS-CoV-2 infection was confirmed by reverse-transcriptase polymerase chain reaction (RT-PCR). Blood was withdrawn into sodium citrate collection tubes at two different time points: at the time of admission at the emergency room (T0) and after one week of hospitalization (T7). Blood parameters were measured by using the Sysmex XN-2000™ Hematology System (Sysmex, Kobe, Japan) at Hospital “Maggiore della Carità” at the time of enrollment, while EV analysis was performed within few hours after blood withdrawal at the indicated time points above. Moreover, information on gender, age, comorbidities, and prehospitalization treatment with steroids and heparin was collected. The 4C score was also calculated to evaluate the mortality risk at T0 of the enrolled patients.
A sample of healthy controls (HC), identified among hospital workers, was also included in the study.
The study was approved by local ethic committee (CE67/20); written informed consent was obtained from the patients or their legal representative.

From April 2020 to November 2020, 131 patients with suspected Sars-Cov-2 infection were enrolled at Maggiore della Carità University Hospital in Novara (Italy). Patients were hospitalized for acute respiratory symptoms or influenza-like illness symptoms during the two waves of Sar-Cov-2 infection that occurred in Italy and stratified based on the results of molecular analysis on nasal swabs (1st wave: April/May 2020, 33 Sars-Cov-2− and 23 Sars-Cov-2+; 2nd wave: October/November 2020, 29 Sars-Cov-2− and 46 Sars-Cov-2+ patients). SARS-CoV-2 infection was confirmed by reverse-transcriptase polymerase chain reaction (RT-PCR). Healthy individuals (n = 10) were enrolled as controls. The blood parameters were measured by using the Sysmex XN-2000™ Hematology System (Sysmex, Kobe, Japan) at Hospital Maggiore della Carita’ at the time of enrollment as well as EVs analysis was performed within few hours after blood withdrawal. The study was approved by local ethic committee (CE67/20); written informed consent was obtained from the patients or their legal representative.