Subject(s)
B-Lymphocyte Subsets/immunology , Betacoronavirus/pathogenicity , Coronavirus Infections/immunology , Dyspnea/immunology , Immunologic Memory , Pneumonia, Viral/immunology , Antibodies, Viral/blood , Antigens, CD/genetics , Antigens, CD/immunology , B-Lymphocyte Subsets/classification , B-Lymphocyte Subsets/virology , Betacoronavirus/immunology , COVID-19 , Case-Control Studies , Cell Proliferation , Convalescence , Coronavirus Infections/mortality , Coronavirus Infections/therapy , Coronavirus Infections/virology , Dyspnea/mortality , Dyspnea/therapy , Dyspnea/virology , Host-Pathogen Interactions/immunology , Humans , Immunization, Passive/methods , Immunophenotyping , Pandemics , Pneumonia, Viral/mortality , Pneumonia, Viral/therapy , Pneumonia, Viral/virology , Respiration, Artificial , SARS-CoV-2 , Severity of Illness Index , Survival Analysis , COVID-19 SerotherapyABSTRACT
INTRODUCTION: Allele-level donor-recipient match at HLA-A, HLA-B, HLA-C and HLA-DRB1 loci impacts the outcome after cord blood transplantation for hematologic malignancies and modifies the strategy of donor selection. High definition of both class I and II HLA loci at time of listing is a way to improve the attractiveness of cord blood bank inventories, reducing the time for donor search and procurement and simplifying donor choice, in particular, for patients of non-European heritage. METHODS: In 2014, Luminex® xMAP® technology was introduced in our laboratory practice and was applied to cord blood units typing. In this study, we evaluated the impact of this strategy in comparison with the platform in use until 2013, relying on LiPA reverse polymerase chain reaction-sequence-specific oligonucleotide (revPCR-SSO) plus polymerase chain reaction-sequence-specific primer (PCR-SSP). RESULTS: In 2014, the time for testing was shorter (141 vs 181 days on average), the number of test repetitions was lower (in particular for HLA-A locus, p = 0.026), and the cost reduced (240.7 vs 395.6 euros per unit on average) compared to 2013, demonstrating that Luminex xMAP technology is superior to the previous approach. CONCLUSION: Luminex xMAP platform has useful application in cord blood banking programs, to achieve high-definition HLA typing of cord blood units at the time of banking in a quick, accurate, and cost-effective manner.
Subject(s)
Fetal Blood/metabolism , Histocompatibility Testing/methods , Humans , Polymerase Chain ReactionABSTRACT
BACKGROUND: The current available techniques to wash out DMSO from thawed umbilical cord blood (UCB) units are based essentially on standard centrifugation in an open system with various degrees of cell loss. STUDY DESIGN AND METHODS: We evaluated the capacity of a new automated closed device (Cytomate, Baxter, IL) to wash out the DMSO from thawed UCB units, saving at the same time the progenitor and accessory cells in terms of CD34+ cells and MNCs. We modified the standard software of the device and calculated the cell recovery on 25 UCB units. Moreover, we set up a new gas chromatographic method to exactly detect the DMSO removal rate. RESULTS: To evaluate the efficiency of the Cytomate device, we considered the postthawing (prewashing) versus postwashing cell recovery. The average recovery (%) in terms of total nucleated cells was 63.30 (range, 40.12-89.00), CD34+ cells was 70.20 (range, 11.51-89.01), CD3+ cells was 61.01 (range, 28.80-87.08), CD4+ cells was 62.53 (range, 30.62-96.73), CD8+ cells was 57.4 (range, 26.87-94.72), CD19+ cells was 63.33 (range, 39.10-90.33), CD16+/56+ cells was 70.67 (range, 8.91-98.40), CFU-GM was 74.33 (range, 20.23-98.60), total CFUs was 82.34 (range, 14.83-247.12), and viability was 89.67(range, 70.74-98.30). The total working time required was, on average, 15 minutes (range, 7-20). CONCLUSIONS: The Cytomate device demonstrated a satisfying efficiency in cell recovery and in maintaining the clonogenic power of the UCB graft. The removal rate of DMSO was practically complete with evident advantages for the recipient. Finally, the entire manipulation performed in a closed system revealed to be safe, maintaining the sterility of the graft.