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Updated: Jul 7, 2026
Generation and Multi-phenotypic High-content Screening of Coxiella burnetii Transposon Mutants
Published on: May 13, 2015
Infection of human monocyte-derived macrophages with Coxiella burnetii
Jeffrey G Shannon1, Robert A Heinzen
1Coxiella Pathogenesis Section, Laboratory of Interacellular Parasites, Rocky Mountain Laboratoties, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Hamilton, MT, USA.
Abstract:
Coxiella burnetii, the agent of Q fever, is an obligate intracellular bacterium that has a tropism for cells of the mononuclear phagocyte system. Following internalization, C. burnetii remains in a phagosome that ultimately matures into a vacuole with lysosomal characteristics that supports pathogen replication. Most in vitro investigations of Coxiella - macrophage interactions have employed continuous cell lines. Although these studies have been informative, genetic alterations of immortalized cells may result in attenuated biological responses to infection relative to primary cells. Consequently, primary macrophages are preferred as in vitro model systems. Here, we describe procedures for propagation and isolation of C. burnetii from cell culture and the use of these preparations to infect primary macrophages derived from human peripheral blood monocytes. Both virulent phase I and avirulent phase II C. burnetii productively infect human monocyte-derived macrophages (MDMs) and replicate with approximately the same kinetics, thereby providing a more physiologically relevant in vitro model system to study the infectious process of this pathogen.
Insights
This study details methods for culturing Coxiella burnetii and infecting primary human macrophages. Both virulent and avirulent C. burnetii strains replicate similarly in these cells, offering a relevant model for Q fever research.
Area of Science:
- Microbiology
- Immunology
- Infectious Diseases
Background:
- Coxiella burnetii causes Q fever, an infectious disease.
- It is an obligate intracellular bacterium targeting mononuclear phagocyte system cells.
- Previous in vitro studies used immortalized cell lines, which may not accurately reflect in vivo responses.
Purpose of the Study:
- To describe procedures for C. burnetii propagation and isolation.
- To establish a more physiologically relevant in vitro model using primary human macrophages.
- To investigate C. burnetii infection dynamics in primary human monocyte-derived macrophages (MDMs).
Main Methods:
- Isolation and propagation of C. burnetii from cell culture.
- Generation of primary macrophages from human peripheral blood monocytes.
- Infection of primary MDMs with virulent (phase I) and avirulent (phase II) C. burnetii.
Main Results:
- Successful propagation and isolation of C. burnetii.
- Productive infection of human MDMs by both phase I and phase II C. burnetii.
- Similar replication kinetics observed for both virulent and avirulent strains in primary MDMs.
Conclusions:
- Primary human MDMs provide a relevant in vitro model for studying C. burnetii infection.
- This model allows for the investigation of Q fever pathogenesis with greater physiological relevance.
- The described procedures facilitate research into Coxiella-macrophage interactions.
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