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Infection and Immunity, March 1999, p. 1368-1378, Vol. 67, No. 3
0019-9567/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
Human Granulocytic Ehrlichiosis Agent and Ehrlichia
chaffeensis Reside in Different Cytoplasmic Compartments in
HL-60 Cells
Jason
Mott,
Roy E.
Barnewall, and
Yasuko
Rikihisa*
Department of Veterinary Biosciences, College
of Veterinary Medicine, The Ohio State University, Columbus, Ohio
43210-1092
Received 19 August 1998/Returned for modification 20 October
1998/Accepted 18 November 1998
The human granulocytic ehrlichiosis (HGE) agent resides and
multiplies exclusively in cytoplasmic vacuoles of granulocytes. Double
immunofluorescence labeling was used to characterize the nature of the
HGE agent replicative inclusions and to compare them with inclusions
containing the human monocytic ehrlichia, Ehrlichia
chaffeensis, in HL-60 cells. Although both Ehrlichia spp. can coinfect HL-60 cells, they resided in separate inclusions. Inclusions of both Ehrlichia spp. were not labeled with
either anti-lysosome-associated membrane protein 1 or anti-CD63.
Accumulation of myeloperoxidase-positive granules were seen around HGE
agent inclusions but not around E. chaffeensis inclusions.
3-(2,4-Dinitroanilino)-3'-amino-N-methyldipropylamine and
acridine orange were not localized to either inclusion type. Vacuolar-type H+-ATPase was not colocalized with HGE agent
inclusions but was weakly colocalized with E. chaffeensis
inclusions. E. chaffeensis inclusions were labeled with the
transferrin receptor, early endosomal antigen 1, and rab5, but HGE
agent inclusions were not. Some HGE agent and E. chaffeensis inclusions colocalized with major histocompatibility complex class I and II antigens. These two inclusions were not labeled
for annexins I, II, IV, and VI;
-adaptin; clathrin heavy chain; or
-coatomer protein. Vesicle-associated membrane protein 2 colocalized
to both inclusions. The cation-independent mannose 6-phosphate receptor
was not colocalized with either inclusion type. Endogenously
synthesized sphingomyelin, from C6-NBD-ceramide, was not
incorporated into either inclusion type. Brefeldin A did not affect the
growth of either Ehrlichia sp. in HL-60 cells. These
results suggest that the HGE agent resides in inclusions which are
neither early nor late endosomes and does not fuse with lysosomes or
Golgi-derived vesicles, while E. chaffeensis resides in an
early endosomal compartment which accumulates the transferrin receptor.
*
Corresponding author. Mailing address: Department of
Veterinary Biosciences, College of Veterinary Medicine, The Ohio State University, 1925 Coffey Rd., Columbus, OH 43210-1092. Phone: (614) 292-9677. Fax: (614)-292-6473. E-mail address:
rikihisa.1{at}osu.edu.
Infection and Immunity, March 1999, p. 1368-1378, Vol. 67, No. 3
0019-9567/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
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