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Related Experiment Videos

Immunoglobulin isotype expression. II. Frequency analysis in mitogen-reactive B cells

R Benner, A Coutinho, A M Rijnbeek

    European Journal of Immunology
    |October 1, 1981
    PubMed
    Summary

    Lipopolysaccharide (LPS) stimulation induces B cells to secrete various immunoglobulin (Ig) classes. Most LPS-reactive B cells initially produce IgM, with significant switching to IgG and IgA isotypes observed in their progeny.

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    Area of Science:

    • Immunology
    • Cell Biology
    • Molecular Biology

    Background:

    • B cells are crucial for adaptive immunity, producing antibodies of various immunoglobulin (Ig) classes.
    • Lipopolysaccharide (LPS) is a potent B cell activator used to study B cell differentiation and Ig class switching.
    • Understanding Ig class switching is vital for comprehending immune responses and developing targeted therapies.

    Purpose of the Study:

    • To quantify the frequency of B cells that differentiate into clones secreting different immunoglobulin (Ig) classes after lipopolysaccharide (LPS) stimulation.
    • To investigate the in vitro development of LPS-reactive B cells into diverse Ig-secreting clones across various lymphoid organs.
    • To determine the proportions of B cell clones switching to specific IgG subclasses and IgA following polyclonal activation.

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    Main Methods:

    • In vitro culture of B cells from BALB/c mice under conditions detecting all growth-inducible cells.
    • Assessment of Ig secretion using the protein A plaque assay with antisera specific for IgM, IgG1, IgG2a, IgG2b, IgG3, and IgA.
    • Analysis of B cells from spleen, bone marrow, mesenteric lymph nodes, and thoracic duct, including athymic nude mice.

    Main Results:

    • A significant proportion of B cells (5-20%) in all tested lymphoid organs were induced by LPS to form IgM-secreting clones.
    • On average, 7% of IgM-secreting clones switched to IgG1, 39% to IgG2, 41% to IgG3, and 1% to IgA.
    • Similar frequencies of B cells switching immunoglobulin heavy chain gene expression were observed in spleen cells of athymic nude mice.
    • No correlation was found between in vitro clonal frequencies and in vivo background Ig-secreting cells.

    Conclusions:

    • Polyclonal B cell activation by LPS in vitro leads to significant immunoglobulin class switching to various IgG subclasses and IgA.
    • The observed frequencies of immunoglobulin class switching are largely independent of T cell help, as evidenced by similar results in athymic nude mice.
    • The quality of stimulation and other regulatory influences likely play a critical role in modulating immunoglobulin heavy chain gene expression in B cells.