Affiliation:
1. Department of Veterinary Biosciences, College of Veterinary Medicine, The Ohio State University, Columbus, Ohio 43210-1092
Abstract
ABSTRACT
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; α-adaptin; clathrin heavy chain; or β-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 C
6
-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.
Publisher
American Society for Microbiology
Subject
Infectious Diseases,Immunology,Microbiology,Parasitology
Cited by
105 articles.
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