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Related Experiment Videos

Assessing B cell diversification by antigen receptor and precursor cell analysis.

N R Klinman, A R Pickard, N H Sigal

    Annales D'Immunologie
    |June 1, 1976
    PubMed
    Summary

    The murine B cell repertoire contains over 10 million clonotypes, developing in an antigen-independent manner from a limited neonatal repertoire. This study validates methods for analyzing B cell diversity and precursor frequencies.

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    Is tolerance the result of engaging surface Ig of B cells in cycle?

    Immunology today·2014

    Area of Science:

    • Immunology
    • B cell biology
    • Immune repertoire diversification

    Background:

    • Understanding B cell specificity diversification is crucial for immunology.
    • Two methods, antigen binding and antibody-forming cell clone production, assess B cell repertoires.
    • Our lab uses the latter (splenic focus assay) to determine B cell frequencies.

    Purpose of the Study:

    • To establish the frequency of B cells responsive to diverse antigenic determinants.
    • To correlate findings from antigen-binding cell assays and splenic focus assays.
    • To understand the development and diversity of the murine B cell repertoire.

    Main Methods:

    • Utilized the splenic focus assay to enumerate B cells responsive to antigenic stimulation.
    • Employed antigen-binding cell enumeration to assess B cell receptor interactions.

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  • Compared results from both methods across different antigens (e.g., DNP-BSA, PC-BSA).

    • Main Results:

    • The primary murine B cell repertoire likely exceeds 10^7 clonotypes.
    • Repertoire acquisition is antigen-independent; neonatal and adult repertoires differ significantly in size and diversity.
    • Splenic focus assay efficiency is 4-5%; antigen-binding cell analysis validates precursor frequency calculations, though some cells may not be stimulatible.

    Conclusions:

    • Murine B cell repertoire development is a highly ordered, antigen-independent process.
    • Antigen-binding cell assays confirm the validity of splenic focus assay precursor frequency estimations.
    • Discrepancies in some antigen-binding cell analyses suggest non-stimulatable B cell subclasses or low-affinity receptors.