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Related Experiment Videos

A simple system for studying antibiotic binding to leukocytes.

G D Gray, G M Ohlmann, C W Ford

    The Journal of Antibiotics
    |March 1, 1984
    PubMed
    Summary

    This study introduces a novel method to assess antibiotic effectiveness against Staphylococcus aureus within human immune cells (polymorphonuclear leukocytes). The new system simplifies testing by focusing on anti-staphylococcal activity, not requiring radiolabeled antibiotics.

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    Area of Science:

    • Pharmacology and Microbiology
    • Immunology and Cellular Biology

    Background:

    • Traditional methods for studying antibiotic-leukocyte interactions require radiolabeled antibiotics or assessment of antibiotic effects on leukocyte function.
    • A simplified, effective method is needed to evaluate antibiotic activity against intracellular pathogens within phagocytic cells.

    Purpose of the Study:

    • To develop and validate a novel system for assessing antibiotic efficacy against Staphylococcus aureus residing within polymorphonuclear leukocytes (PMLs).
    • To bypass the need for radiolabeled antibiotics or direct measurement of antibiotic effects on leukocyte function.

    Main Methods:

    • A system was developed involving incubation of PMLs with antibiotics, followed by removal of extracellular antibiotics via centrifugation.
    • Contact between Staphylococcus aureus and the phagocyte-antibiotic complex was facilitated by low-temperature centrifugation.

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  • Phagocytosis was induced at 37°C, and surviving bacteria were quantified using [3H]thymidine labeling and a cell harvester.

    • Main Results:

    • The developed system successfully identified several classes of antibiotics with anti-staphylococcal activity within PMLs.
    • Active antibiotics included naphthalenic ansamycins (rifamycins, streptovaricins), lincosaminides (clindamycins, pirlimycins), novobiocin, erythromycin, tetracycline, tyrocidine, and paulomycin.

    Conclusions:

    • This novel system provides an effective and simplified approach to evaluate antibiotic activity against intracellular Staphylococcus aureus.
    • The method is valuable for drug discovery and development, particularly for antibiotics targeting intracellular bacterial infections.