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Related Experiment Videos

A sensitive microassay for the murine alternative complement pathway.

K A Joiner, R Shahon, J A Gelfand

    Journal of Immunological Methods
    |January 1, 1979
    PubMed
    Summary

    A new microhemolytic assay accurately measures mouse alternative complement pathway activity. This sensitive method uses treated red blood cells and small serum volumes, revealing significant strain differences.

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    Area of Science:

    • Immunology
    • Biochemistry

    Background:

    • The complement system, particularly the alternative pathway, plays a crucial role in innate immunity.
    • Accurate measurement of alternative complement pathway activity is essential for immunological research, especially in murine models.
    • Existing methods for assessing murine alternative complement pathway activity can be limited in sensitivity or require large sample volumes.

    Purpose of the Study:

    • To develop and validate a novel, highly sensitive microhemolytic assay for quantifying murine alternative complement pathway activity.
    • To enable the measurement of alternative complement pathway activity using small serum volumes from individual mice.
    • To assess variations in alternative complement pathway activity among different inbred murine strains.

    Main Methods:

    • The assay utilizes the release of chromium-51 (51Cr) from neuraminidase-treated rabbit erythrocytes.

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  • Murine serum, chelated with Mg2+ and EGTA, is incubated with the treated erythrocytes.
  • Neuraminidase pretreatment enhances erythrocyte sensitivity, increasing assay sensitivity 8-10 fold.

    • Main Results:

    • The described microhemolytic assay provides a simple, sensitive, and reproducible method for measuring murine alternative complement pathway activity.
    • Significant differences in alternative complement pathway activity were observed between serum samples from various inbred murine lines.
    • The assay's increased sensitivity allows for the use of minimal serum volumes, facilitating studies on individual mice.

    Conclusions:

    • A robust and sensitive microhemolytic assay for murine alternative complement pathway activity has been established.
    • This assay facilitates the investigation of genetic and environmental factors influencing complement activity in mice.
    • The findings highlight the utility of this assay for comparative immunological studies across different murine strains.