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Flow sorting of antigen-binding B cell subsets.

J L Greenstein, J Leary, P Horan

    Journal of Immunology (Baltimore, Md. : 1950)
    |March 1, 1980
    PubMed
    Summary
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    Functional B cell heterogeneity was explored using antigen binding. Spleen cells binding fluorescent antigen were enriched for antibody precursors, revealing distinct B cell responses based on antigen carrier molecules like LPS, SRBC, and Ficoll.

    Area of Science:

    • Immunology
    • Cell Biology
    • B cell immunology

    Background:

    • B cell heterogeneity is crucial for adaptive immunity.
    • Understanding B cell subsets responding to different antigens is key to immune response modulation.

    Purpose of the Study:

    • To define functional B cell heterogeneity using antigen binding as a probe.
    • To investigate the antibody precursor frequencies in B cells with varying antigen binding capacities.

    Main Methods:

    • Spleen cells were stained with fluorescent trinitrophenylated bovine serum albumin (FL-TNP-BSA).
    • Cells were sorted into fluorescence-negative and fluorescence-positive populations using a multiparameter cell sorter.
    • Limiting dilution analysis was employed to assay precursor frequency for antibody responses.

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    Main Results:

    • Fluorescence-positive cells (bound FL-TNP-BSA) were enriched for antibody-forming precursors to TNP-lipopolysaccharide (LPS), TNP-sheep red blood cells (SRBC), and TNP- or DNP-Ficoll.
    • B cells were further sorted into moderate and high FL-TNP-BSA binding populations.
    • B cells responding to TNP-LPS and TNP-SRBC were found in both moderate and high binding groups, while those responding to TNP- or DNP-Ficoll were only in the moderate binding group.

    Conclusions:

    • A distinct population of B cells in adult mouse spleen binds significant amounts of antigen.
    • These B cells can respond to antigens presented on LPS or SRBC carriers.
    • Responses to antigens carried on Ficoll are limited to B cells with moderate antigen binding capacity, suggesting differential antigen recognition pathways.