Assessment of antiCIFNC antibodies was done by ELISA (Thermo Fisher Scientific, Waltham, MA) following manufacturer’s specifications. 2.8.4. super-donors and donors according to antibody levels. Subjects with IgG antibody titers 1:3200 and IgA antibody titers 1:800 to SARS-CoV-2 were considered super-donors and were chosen for plasmapheresis and further therapeutic transfusion. Subjects who did not reach those titers were CAY10505 considered as donors, and were discard for plasmapheresis, but its serum composition was analyzed in this study. Approximately, 800?mL of plasma were collected from super-donors. Prior freezing, pathogens inactivation with Riboflavin followed by UV light exposure was performed [18]. 2.3. Inclusion criteria for COVID-19 patients Inclusion criteria were the following: (1) signed informed consent; (2) aged at least 18 years; (3) COVID-19 diagnosis based on RT-PCR testing; (4) hospitalized patients; (5) Sequential Organ Failure Assessment score (SOFA)??1000 copies/ml), two consecutive viral load measurements within a 3-month interval; (7) subjects with other confirmed infection that explains clinical manifestations; (8) end-stage kidney disease (i.e., glomerular filtration rate <15?ml/min/1.73 m2); (9) Child Pugh C stage liver cirrhosis; (10) high Rabbit Polyclonal to OR2T2 cardiac output diseases; (11) autoimmune diseases or immunoglobulin A nephropathy; (12) and subjects not willing to participate. 2.5. Convalescent plasma transfusion Each transfusion dose of CP was 250?mL, patients received two doses CAY10505 for a total of 500?mL within 48?h after study inclusion. Each CP unit was kept separate from other super-donors units. The transfused CP ABO type was compatible with the recipient’s ABO type in 8 out of 10 transfused patients. Each recipient received CP units from the same super-donor. CP transfusion was administered at 3?mL/min with CAY10505 close monitoring for the first 30?min, and regular monitoring over the following 6?h. 2.6. Standard therapy Standard treatment consisted of symptomatic control and supportive care for COVID-19. This treatment was based upon recommendations from the Colombian Association of Infectology and institutional protocols, which included management with antibiotics, corticosteroids, oxygen, and anticoagulants [19]. Both plasma recipient and standard therapy groups received this treatment. 2.7. Patient monitoring and evaluation Sociodemographic and pathological factors were evaluated on day 0. The biological baseline included cytokines, lymphocyte populations, IgA and IgG antibodies for SARS-CoV-2, viral load, blood gases, laboratory surrogate of possible thrombotic process (i.e., D-Dimer), hematological, inflammatory, hepatic and renal parameters. These measurements were repeated on days 4, 7, 14 and 28. In addition, the SOFA scale and the 4C mortality score (i.e., score for prediction of mortality 28 days after hospitalization) were evaluated on admission [20]. Clinical and paraclinical parameters were obtained using a standardized form. The former comprised all the variables that were included in the global COVID-19 clinical platform from the World Health Organization (WHO). 2.8. Biological parameters 2.8.1. Viral load The viral load was measured using the Ampliphi ? RT-qPCR SARS-CoV-2 Viral Load Kit (www.ampliphi.co). 2.8.2. Antibody detection against SARS-CoV-2 The Euroimmun anti-SARS-CoV-2 ELISA (Euroimmun, Luebeck, Germany) was used for serological detection of human IgG and IgA antibodies against the SARS-CoV-2 S1 structural protein, in accordance with the manufacturer’s instructions. The ratio interpretation was <0.8?=?negative, 0.8 to <1.1?=?borderline, 1.1?=?positive [17,21,22]. Antibody titration was performed using serial dilutions of serum samples from 1:100 to 1 1:1,638,400. 2.8.3. Autoantibodies Detection of IgM rheumatoid factor (RF), IgG anti-cyclic citrullinated peptide third-generation (CCP3),.