Bacterial contamination rates in extracorporeal photopheresis.

BACKGROUND: Extracorporeal photopheresis (ECP) is an immunosuppressive treatment that involves leukocyte apheresis, psoralen and UV light treatment, and subsequent reinfusion. Patients treated with ECP are usually immunosuppressed. Bacterial contamination therefore poses a much unwanted risk, but incidence data are lacking. PATIENTS AND METHODS: We screened all 1922 consecutive ECP procedures scheduled within a roughly 3-year period for eligibility. Those with missing data on ECP method (inline or offline) or type of venous access (peripheral or central) were excluded. ECPs with complete aerobic and anaerobic microbial testing of baseline patient blood samples (n = 1637) and of ECP cell concentrates (n = 1814) were included in the analysis. RESULTS: A test for microbial contamination was positive for 1.82% of the cell concentrates, with central venous access was the most significant risk factor for the contamination (odds ratio = 19). Patient blood samples were positive in 3.85% of cases, but no patients became septic. Staphylococcus spp. were most abundant, and products with bacterial contamination did not cause side effects after reinfusion. There were no significant differences in contamination rates between inline and offline ECP. CONCLUSION: These findings stress the importance of sterile procedures and the benefits of using peripheral over central venous access for reducing the risk of bacterial contamination in ECP.

Granulocytapheresis with modified fluid gelatin versus high-molecular-weight hydroxyethyl starch: a matched-pair analysis.

BACKGROUND: Granulocyte apheresis requires a sedimentation agent. Usually, hydroxyethyl starch (HES) is administered to donors for this purpose and, as granulocyte concentrate (GC) ingredient, also to patients. Authorities recently recommended suspending market authorizations for starch-containing products due to side effects. Therefore, we tested the efficacy of modified fluid gelatin (MFG, Gelafundin 4%) versus hetastarch (Hespan) for GC apheresis. STUDY DESIGN AND METHODS: This retrospective matched-pair analysis compared MFG- and hetastarch-derived GCs. Each group consisted of 15 unrelated male donors mobilized with dexamethasone and granulocyte-colony-stimulating factor for apheresis on 1 or 2 days with the COBE Spectra's PMN program. RESULTS: In each group, 24 GCs were collected from 15 male donors and analyzed. None of the HES-derived products, but two of the MFG-derived products (8.3%), had aggregates and could not be used. The HES-derived products had significantly higher neutrophil counts on the first day (7.7 × 1010 /unit vs. 4.0 × 1010 /unit; p = 0.00005) as well as second day of apheresis (4.0 × 1010 /unit vs. 1.1 × 1010 /unit; p = 0.0002). Median white blood cell collection efficacies were lower with MFG than with HES on Day 1 (24% vs. 43%) and Day 2 (15% vs. 37%). Twenty-one percent of the MFG-derived products had less than 1 × 1010 granulocytes. CONCLUSION: These results indicate that granulocyte apheresis is feasible with MFG as well as with hetastarch and that the latter is superior for GC production, if used in the same dosage. In addition, aggregates in GC from the COBE Spectra were observed in the MFG group but not in the hetastarch group.

Multistep screening and selection of COVID-19 convalescent plasma donors at the early stage of the SARS-CoV-2 pandemic: A retrospective analysis.

Background and Aims: The COVID-19 pandemic reached Bavaria in February 2020. Almost simultaneously, Chinese physicians published reports on the first successful treatments with plasma from COVID-19 convalescent donors. With these silver linings on the horizon, we decided to establish the manufacturing of anti-severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) antibody-containing plasma from COVID-19 convalescent donors at our site. Here we describe our donor selection process, built from the ground up, which enabled us to cope with the immense resonance after our social media call for donors. Methods: As a first step, we created a specific questionnaire for telephone interviews applied by trained students to filter the wave of callers interested in plasma donation. Afterward, the medical staff evaluated the hotline questionnaires and chose eligible donors to be invited for on-site donor evaluation. Data documentation was performed with MS Excel, and statistical analyses were calculated with GraphPad Prism 8. A quantitative in-house ELISA was used to detect anti-SARS-CoV-2 antibodies and determine specific titers. Results: Out of 1465 calls from potential plasma donors, we could register 420 persons with a completed questionnaire. Evaluation of questionnaires identified 222 of 420 persons as eligible for donation, and 55 were directly asked for on-site donor qualification. Subsequently, as anti-SARS-CoV-2 antibody titers ≥1:800 were required, we invited 89 of 222 potential donors for an antibody screening. This procedure resulted in another 28 potential donors for an on-site evaluation. Finally, 12 donors qualified with a titer of 1:400 and 24 with ≥1:800. Conclusion: Identifying suitable COVID-19 convalescent plasma donors was expected to be highly time-consuming. Implementing a screening procedure with our hotline questionnaire helped us streamline the donor selection process and reduce the workload for the staff. We propose combining the described selection process with the later introduced on-site antibody screening as an effective strategy.

Manufacturing of convalescent plasma of COVID-19 patients: Aspects of quality.

The ongoing coronavirus disease 2019 (COVID-19) pandemic emerged in December 2019. Convalescent plasma represents a promising COVID-19 treatment. Here, we report on the manufacturing of a plasma-based product containing antibodies specific to SARS-CoV-2 obtained from recently recovered COVID-19 patients. Convalescent plasma donors were screened as follows: 1) previously confirmed SARS-CoV-2 infection (by real-time PCR (RT-PCR)); 2) a subsequent negative PCR test followed by a 2-week waiting period; 3) an additional negative PCR test prior to plasmapheresis; and 4) confirmation of the presence of SARS-CoV-2 specific antibodies. Convalescent plasma was stored fresh (2-6°C) for up to 5 days or frozen (-30°C) for long-term storage. Donor peripheral blood and final plasma product were assayed for binding antibodies targeting the SARS-CoV-2 S-protein receptor-binding domain (RBD) and their titers measured by an enzyme-linked immunosorbent assay (ELISA). We performed 72 plasmaphereses resulting in 248 final products. Convalescent plasma contained an RBD-specific antibody titer (IgG) ranging from 1:100 to 1:3200 (median 1:800). The titer was congruent to the titer of the blood (n = 34) before collection (1:100-1:6400, median 1:800). Levels of IL-8 and LBP of donors were slightly increased. Therapeutic products derived from a human origin must undergo rigorous testing to ensure uniform quality and patient safety. Whilst previous publications recommended RBD-specific binding antibody titers of ≥ 1:320, we selected a minimum titer of 1:800 in order to maximize antibody delivery. Production of highly standardized convalescent plasma was safe, feasible and was readily implemented in the treatment of severely ill COVID-19 patients.