Culture, Isolation, and Labeling of Anaplasma phagocytophilum for Subsequent Infection of Human Neutrophils - Bacterial Pathogenesis: Methods and Protocols - page 150

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Culture, Isolation, and Labeling of Anaplasma
phagocytophilum for Subsequent Infection
of Human Neutrophils
Dori L. Borjesson
Summary
Anaplasma phagocytophilum is the etiologic agent of granulocytic anaplasmosis, a tick-
borne, zoonotic, emerging infectious disease. A. phagocytophilum is an obligate intracel-
lular pathogen that primarily resides within membrane-bound, cytoplasmic vacuoles of host
neutrophils. Closely related to Ehrlichial and Rickettsial organisms, A. phagocytophilum
is a small, fragile, Gram-negative bacterium that presents unique challenges for culture,
isolation, enumeration, and labeling. This chapter delineates pathogen-specific consider-
ations for culture and labeling of this organism for subsequent use in assays to examine
mechanisms of host cell-pathogen interactions.
Key Words: Anaplasma phagocytophilum ; granulocytic anaplasmosis; cell culture;
fluorescent labeling; neutrophils.
1. Introduction
Anaplasma phagocytophilum is the etiologic agent of granulocytic anaplas-
mosis, an emerging, zoonotic, tick-borne, infectious disease (1,2) . A. phago-
cytophilum is an obligate intracellular bacterium that principally inhabits
the cytoplasm of host neutrophils. This intracellular niche is particularly
noteworthy, as neutrophils are powerful cells that function as a first line of
defense against bacterial pathogens. Understanding the mechanisms by which
A. phagocytophilum circumvents neutrophil defense systems is facilitated by
optimum pathogen culture, isolation, and labeling techniques.