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Related Experiment Videos

Inside-out vesicles prepared from complement lysed sheep red blood cells.

E W Rauterberg

    Complement (Basel, Switzerland)
    |January 1, 1985
    PubMed
    Summary

    Complement-mediated lysis of red blood cells produces vesicles. Most floating vesicles from complement lysis show an inside-out orientation, suggesting membrane lesions that don't impede flotation.

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    Area of Science:

    • Immunology
    • Cell Biology
    • Biochemistry

    Background:

    • Antibody-sensitized sheep red blood cells are lysed by human serum complement.
    • Complement lysis generates red blood cell 'ghosts' that can vesiculate.

    Purpose of the Study:

    • To investigate the properties of vesicles derived from complement-lysed red blood cells.
    • To determine the orientation and complement component content of these vesicles.

    Main Methods:

    • Sheep red blood cells sensitized with antibodies were lysed with diluted human serum.
    • Red blood cell ghosts were subjected to vesiculation using low ionic strength buffer and shearing.
    • Vesicles were separated using ultracentrifugation on a dextran gradient.
    • Antibody probes were used to determine vesicle membrane orientation (sideness).
    • Complement component 9 (C9) presence on vesicles was quantified.

    Main Results:

    • Approximately 68% lysis of red blood cells was achieved.
    • Vesiculation of red blood cell ghosts was induced.
    • About 45% of vesicles from complement-lysed cells floated on a dextran gradient.
    • 90% of floating vesicles exhibited an inside-out orientation.
    • Floating vesicles had significant C9 on their inverted outer membrane, suggesting intact C5b-9 complexes.

    Conclusions:

    • Complement-mediated lysis of red blood cells generates vesicles with a predominant inside-out orientation.
    • The flotation of these vesicles is not due to a lack of C5b-9.
    • Lysis may involve ineffective C5b-9 complexes or lesions with limited permeability that allow vesicle flotation.

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