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Published on: May 11, 2012
Laboratory Maintenance of Anaplasma phagocytophilum
1University of Kentucky College of Medicine, Lexington, Kentucky, USA.
Abstract:
Anaplasma phagocytophilum is the etiologic agent of human granulocytic anaplasmosis (formerly human granulocytic ehrlichiosis), an emerging and potentially deadly disease in the United States, Europe, and Asia. A. phagocytophilum is an obligate intracellular bacterium that displays a unique tropism for neutrophils. Studying this fascinating organism not only provides insight into microbial invasion and intracellular survival strategies, but also offers a unique approach to understanding neutrophil biology and host defense mechanisms. This unit describes the inoculation and maintenance of A. phagocytophilum from an infected blood sample into eukaryotic cell culture or laboratory mice. Cytological staining and immunofluorescent methods for assessing A. phagocytophilum infection are also presented. In addition, this unit describes isolation of viable, host cell-free bacterial preparations from infected cells, as well as the cryopreservation of infected cultures. Lastly, fluorescent labeling of live A. phagocytophilum for the purpose of tracking infection is provided.
Insights
Anaplasma phagocytophilum causes a serious emerging disease. This study details methods for culturing, maintaining, and tracking this intracellular bacterium in lab settings.
Area of Science:
- Microbiology
- Infectious Diseases
- Immunology
Background:
- Anaplasma phagocytophilum is the cause of human granulocytic anaplasmosis, an emerging disease in multiple continents.
- This obligate intracellular bacterium specifically targets neutrophils, offering insights into host-pathogen interactions and neutrophil function.
Purpose of the Study:
- To provide standardized protocols for the laboratory study of Anaplasma phagocytophilum.
- To facilitate research into the invasion mechanisms, intracellular survival strategies, and host immune responses related to A. phagocytophilum infections.
Main Methods:
- Inoculation and maintenance of A. phagocytophilum in eukaryotic cell cultures and laboratory mice.
- Cytological staining and immunofluorescence assays for infection detection.
- Isolation of cell-free bacteria, cryopreservation of infected cultures, and fluorescent labeling for infection tracking.
Main Results:
- Established methods for propagating and preserving Anaplasma phagocytophilum in vitro and in vivo.
- Demonstrated techniques for reliable detection and monitoring of bacterial infection.
- Developed protocols for generating pure bacterial preparations and tracking live A. phagocytophilum.
Conclusions:
- The described methods enable robust laboratory investigation of Anaplasma phagocytophilum.
- These protocols support research into neutrophil biology, host defense, and the pathogenesis of anaplasmosis.
- Facilitates further study of this significant emerging infectious agent.
