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Prozone effects in indirect immunofluorescence.

E Linder, A Miettinen

    Scandinavian Journal of Immunology
    |January 1, 1976
    PubMed
    Summary
    This summary is machine-generated.

    A prozone effect in indirect immunofluorescence (IFL) occurs when high antibody concentrations prevent detection. This phenomenon, observed with anti-brush border (BB) antibodies, is linked to antigen determinant density on target tissues.

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    Area of Science:

    • Immunology
    • Cell Biology
    • Histology

    Background:

    • Indirect immunofluorescence (IFL) is a common technique for detecting antigens in tissues.
    • The prozone effect, where antibodies are undetectable at high concentrations, can interfere with IFL results.
    • Understanding factors influencing the prozone effect is crucial for accurate immunofluorescence assays.

    Purpose of the Study:

    • To investigate the cause of a prozone effect observed with rabbit antisera against rat renal proximal tubular epithelial brush border (BB) antigens using indirect IFL.
    • To determine the relationship between antigen determinant density and the occurrence of the prozone effect.

    Main Methods:

    • Indirect immunofluorescence (IFL) assays were performed using rabbit antisera against rat renal proximal tubular epithelial brush border (BB) antigens.

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  • Direct IFL was used for comparison.
  • Detection of cross-reacting antigens in small-intestinal epithelial brush border was also assessed.

    • Main Results:

    • A significant prozone effect was observed with indirect IFL using anti-BB antibodies; antibodies were undetectable at high concentrations but detectable upon dilution.
    • The prozone effect persisted despite demonstrated binding of anti-BB antibodies to target antigens.
    • No prozone effect was seen with direct IFL, and cross-reacting antigens in the small intestine were detectable by indirect IFL without a prozone effect.

    Conclusions:

    • The prozone effect in this study is attributed to the inability of anti-immunoglobulin conjugates to access tightly clustered immunoglobulin molecules on the brush border antigens.
    • The density of antigen determinants in target tissues appears to be a primary factor influencing the occurrence of prozone effects in indirect immunofluorescence.