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Related Experiment Videos

Human lymphoid cell lines as targets for DRw.

D M Strong, M A Pellegrino, S Ferrone

    In Vitro
    |December 1, 1978
    PubMed
    Summary

    Cultured human lymphoid cell lines (LCL) provide reliable HLA-DR antigen typing, comparable to patient B-cells. Careful complement selection minimizes assay discordancy for accurate immunologic studies.

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    Area of Science:

    • Immunology
    • Cell Biology
    • Genetics

    Background:

    • Cultured human lymphoid cell lines (LCL) are valuable tools in immunological assays.
    • Their application has expanded to serological characterization of human leukocyte antigen (HLA)-DR antigens.
    • Previous studies showed comparable HLA-DR typing results for LCL across laboratories, with discordancy below 10%.

    Purpose of the Study:

    • To assess the utility of cultured human lymphoid cell lines (LCL) for HLA-DR antigen typing.
    • To identify factors contributing to discordancy in cytotoxic assays, particularly complement source.
    • To evaluate methods for improving the reliability of LCL-based HLA-DR typing.

    Main Methods:

    • Serological characterization of HLA-DR antigens using cultured human lymphoid cell lines (LCL).
    • Comparison of HLA-DR typing results from multiple laboratories during the VII Histocompatibility Workshop.
    • Evaluation of different methods to mitigate the effects of natural antibodies in rabbit complement.
    • Correlation analysis between LCL and peripheral B-cell HLA-DR typing from the same donor.
    • Blocking assays using F(ab')2 fragments of anti-beta2-microglobulin antibodies.

    Main Results:

    • HLA-DR typing of LCL yielded comparable results across laboratories, with less than 10% discordancy.
    • Variability in complement source significantly impacts cytotoxic assay outcomes.
    • Methods like complement absorption, dilution, and screening effectively reduce natural antibody interference.
    • LCL typing showed good correlation with peripheral B-cell typing, though occasional extra reactions were observed in LCL.
    • Blocking assays enabled discrimination of anti-DRw activity from other HLA antibody reactions.

    Conclusions:

    • Cultured human lymphoid cell lines (LCL) are reliable for HLA-DR antigen typing.
    • Optimizing complement source and employing blocking strategies enhance typing accuracy.
    • LCL offer a consistent and reproducible method for HLA-DR serological characterization in immunological research.

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