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Related Experiment Videos

Does complement kill E. coli by producing transmural pores?

J Born, S Bhakdi

    Immunology
    |September 1, 1986
    PubMed
    Summary
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    The complement system

    Area of Science:

    • Microbiology
    • Immunology
    • Cell Biology

    Background:

    • The terminal complement pathway, involving the C5b-9 complex, is crucial for innate immunity against bacterial infections.
    • Serum-sensitive bacteria like E. coli K 12 are susceptible to complement-mediated lysis.
    • The precise mechanism by which C5b-9 induces bacterial cell death remains under investigation.

    Purpose of the Study:

    • To elucidate the mechanism of cell killing by the C5b-9 complex in serum-sensitive E. coli K 12.
    • To investigate the role of pore formation across both bacterial membranes in C5b-9-mediated lethality.
    • To determine the threshold of C5b-9 complex deposition required for bacterial cell death.

    Main Methods:

    • Assessing bacterial membrane permeability to o-nitrophenyl-beta-D-galactoside (ONPG) after C5b-9 complex formation.

    Related Experiment Videos

  • Quantifying bacterial viability and C5b-9 complex levels through CFU counts and serial dilutions.
  • Measuring intracellular potassium (K+) efflux from bacteria upon C5b-9 treatment.

    • Main Results:

    • C5b-9-treated E. coli K 12 cells become permeable to ONPG, indicating pore formation.
    • A small number of C5b-9 complexes (as low as 8-30 per CFU) are sufficient to cause significant bacterial killing.
    • Immediate release of intracellular K+ is observed following C5b-9 treatment, correlating with cell death.

    Conclusions:

    • C5b-9 induces bacterial cell death by forming functional pores spanning both the outer and inner bacterial membranes.
    • These transmural pores facilitate rapid K+ efflux, leading to membrane potential collapse and cell death.
    • The study provides direct evidence for the lytic mechanism of the terminal complement pathway in E. coli.