The application of a new quantitative assay for the monitoring of integrin associated protein CD47 on red blood cells during storage and comparison with expression of CD47 and phosphatidylserine using flow cytometry

A. Stewart, M. L. Turner, Stanislaw Urbaniak, H. Bessos

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    56 Citations (Scopus)

    Abstract

    BACKGROUND: After the introduction of universal leukoreduction, the role of factors other than white blood cells in red cell (RBC) storage lesion is attracting increasing attention. These include changes in the levels of CD47 and phosphatidylserine (PS) markers on RBCs during storage. The aim of this study was to monitor these changes with both flow cytometry (FACS) and a newly developed quantitative enzyme-linked immunosorbent assay (ELISA).

    STUDY DESIGN AND METHODS: A new quantitative ELISA (monoclonal antibody immobilization of RBC antigens [MAIRA]) was developed. The assay yielded consistent linear curves that enabled the measurement of CD47 expression on RBCs. In addition, FACS was used to measure both CD47 expression and PS on RBCs (n = 3 units) during storage (Days 4, 10, 24, and 31).

    RESULTS: A significant reduction in CD47 expression was observed both by MAIRA assay and by FACS by Days 24 and 31 (p < 0.01), and the correlation between the two assays was significant (p < 0.01). In addition, a significant increase in PS was observed by the same storage days with FACS (p < 0.01).

    CONCLUSIONS: The MAIRA assay appears to be suitable for the quantitative measurement of RBC markers during storage. Significant changes in CD47 and PS levels were observed during storage, which may have detrimental immunomodulatory and hemostatic effects on the transfused RBCs.

    Original languageEnglish
    Pages (from-to)1496-1503
    Number of pages7
    JournalTransfusion
    Volume45
    DOIs
    Publication statusPublished - 2005

    Keywords

    • ANNEXIN V-FERROFLUID
    • ERYTHROCYTE PHOSPHATIDYLSERINE
    • CLINICAL-TRIAL
    • TRANSFUSION
    • EXPOSURE
    • HEMOGLOBIN
    • SURVIVAL
    • ACTIVATION
    • ADHESION
    • ABSENCE

    Cite this

    @article{a90a6b3271754891b9502e9521f489be,
    title = "The application of a new quantitative assay for the monitoring of integrin associated protein CD47 on red blood cells during storage and comparison with expression of CD47 and phosphatidylserine using flow cytometry",
    abstract = "BACKGROUND: After the introduction of universal leukoreduction, the role of factors other than white blood cells in red cell (RBC) storage lesion is attracting increasing attention. These include changes in the levels of CD47 and phosphatidylserine (PS) markers on RBCs during storage. The aim of this study was to monitor these changes with both flow cytometry (FACS) and a newly developed quantitative enzyme-linked immunosorbent assay (ELISA).STUDY DESIGN AND METHODS: A new quantitative ELISA (monoclonal antibody immobilization of RBC antigens [MAIRA]) was developed. The assay yielded consistent linear curves that enabled the measurement of CD47 expression on RBCs. In addition, FACS was used to measure both CD47 expression and PS on RBCs (n = 3 units) during storage (Days 4, 10, 24, and 31).RESULTS: A significant reduction in CD47 expression was observed both by MAIRA assay and by FACS by Days 24 and 31 (p < 0.01), and the correlation between the two assays was significant (p < 0.01). In addition, a significant increase in PS was observed by the same storage days with FACS (p < 0.01).CONCLUSIONS: The MAIRA assay appears to be suitable for the quantitative measurement of RBC markers during storage. Significant changes in CD47 and PS levels were observed during storage, which may have detrimental immunomodulatory and hemostatic effects on the transfused RBCs.",
    keywords = "ANNEXIN V-FERROFLUID, ERYTHROCYTE PHOSPHATIDYLSERINE, CLINICAL-TRIAL, TRANSFUSION, EXPOSURE, HEMOGLOBIN, SURVIVAL, ACTIVATION, ADHESION, ABSENCE",
    author = "A. Stewart and Turner, {M. L.} and Stanislaw Urbaniak and H. Bessos",
    year = "2005",
    doi = "10.1111/j.1537-2995.2005.00564.x",
    language = "English",
    volume = "45",
    pages = "1496--1503",
    journal = "Transfusion",
    issn = "0041-1132",
    publisher = "Wiley-Blackwell",

    }

    TY - JOUR

    T1 - The application of a new quantitative assay for the monitoring of integrin associated protein CD47 on red blood cells during storage and comparison with expression of CD47 and phosphatidylserine using flow cytometry

    AU - Stewart, A.

    AU - Turner, M. L.

    AU - Urbaniak, Stanislaw

    AU - Bessos, H.

    PY - 2005

    Y1 - 2005

    N2 - BACKGROUND: After the introduction of universal leukoreduction, the role of factors other than white blood cells in red cell (RBC) storage lesion is attracting increasing attention. These include changes in the levels of CD47 and phosphatidylserine (PS) markers on RBCs during storage. The aim of this study was to monitor these changes with both flow cytometry (FACS) and a newly developed quantitative enzyme-linked immunosorbent assay (ELISA).STUDY DESIGN AND METHODS: A new quantitative ELISA (monoclonal antibody immobilization of RBC antigens [MAIRA]) was developed. The assay yielded consistent linear curves that enabled the measurement of CD47 expression on RBCs. In addition, FACS was used to measure both CD47 expression and PS on RBCs (n = 3 units) during storage (Days 4, 10, 24, and 31).RESULTS: A significant reduction in CD47 expression was observed both by MAIRA assay and by FACS by Days 24 and 31 (p < 0.01), and the correlation between the two assays was significant (p < 0.01). In addition, a significant increase in PS was observed by the same storage days with FACS (p < 0.01).CONCLUSIONS: The MAIRA assay appears to be suitable for the quantitative measurement of RBC markers during storage. Significant changes in CD47 and PS levels were observed during storage, which may have detrimental immunomodulatory and hemostatic effects on the transfused RBCs.

    AB - BACKGROUND: After the introduction of universal leukoreduction, the role of factors other than white blood cells in red cell (RBC) storage lesion is attracting increasing attention. These include changes in the levels of CD47 and phosphatidylserine (PS) markers on RBCs during storage. The aim of this study was to monitor these changes with both flow cytometry (FACS) and a newly developed quantitative enzyme-linked immunosorbent assay (ELISA).STUDY DESIGN AND METHODS: A new quantitative ELISA (monoclonal antibody immobilization of RBC antigens [MAIRA]) was developed. The assay yielded consistent linear curves that enabled the measurement of CD47 expression on RBCs. In addition, FACS was used to measure both CD47 expression and PS on RBCs (n = 3 units) during storage (Days 4, 10, 24, and 31).RESULTS: A significant reduction in CD47 expression was observed both by MAIRA assay and by FACS by Days 24 and 31 (p < 0.01), and the correlation between the two assays was significant (p < 0.01). In addition, a significant increase in PS was observed by the same storage days with FACS (p < 0.01).CONCLUSIONS: The MAIRA assay appears to be suitable for the quantitative measurement of RBC markers during storage. Significant changes in CD47 and PS levels were observed during storage, which may have detrimental immunomodulatory and hemostatic effects on the transfused RBCs.

    KW - ANNEXIN V-FERROFLUID

    KW - ERYTHROCYTE PHOSPHATIDYLSERINE

    KW - CLINICAL-TRIAL

    KW - TRANSFUSION

    KW - EXPOSURE

    KW - HEMOGLOBIN

    KW - SURVIVAL

    KW - ACTIVATION

    KW - ADHESION

    KW - ABSENCE

    U2 - 10.1111/j.1537-2995.2005.00564.x

    DO - 10.1111/j.1537-2995.2005.00564.x

    M3 - Article

    VL - 45

    SP - 1496

    EP - 1503

    JO - Transfusion

    JF - Transfusion

    SN - 0041-1132

    ER -