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PMID: 11796624 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Rapid activation of protein tyrosine kinase and phospholipase C-gamma2 and increase in cytosolic free calcium are required by Ehrlichia chaffeensis for internalization and growth in THP-1 cells.

Infection and immunity ·Vol. 70 ·No. 2 ·2002-02-00 ·Pages 889-98

Lin M, Zhu MX, Rikihisa Y

Abstract

Ehrlichia chaffeensis, a bacterium that cannot survive outside the eukaryotic cell, proliferates exclusively in human monocytes and macrophages. In this study, signaling events required for ehrlichial infection of human monocytic cell line THP-1 were characterized. Entry and proliferation of E. chaffeensis in THP-1 cells were significantly blocked by various inhibitors that can regulate calcium signaling, including 8-(diethylamino)octyl-3,4,5-trimethoxybenzoate and 2-aminoethoxydiphenyl borate (intracellular calcium mobilization inhibitors), verapamil and 1-[beta-[3-(4-methoxyphenyl)propyl]-4-methoxyphenethyl]-1H-imidazole (SKF-96365) (calcium channel inhibitors), neomycin and 1-(6-[[17beta-3-methoxyestra-1,3,5(10)-trien-17-yl]amino]hexyl)-1H-pyrrole-2,5-dione (U-73122) (phospholipase C [PLC] inhibitors), monodansylcadaverine (a transglutaminase [TGase] inhibitor), and genistein (a protein tyrosine kinase [PTK] inhibitor). Addition of E. chaffeensis resulted in rapid increases in the level of inositol 1,4,5-trisphosphate (IP(3)) and the level of cytosolic free calcium ([Ca(2+)](i)) in THP-1 cells, which were prevented by pretreatment of THP-1 cells with inhibitors of TGase, PTK, and PLC. E. chaffeensis induced rapid tyrosine phosphorylation of PLC-gamma2, and the presence of a PLC-gamma2 antisense oligonucleotide in THP-1 cells significantly blocked ehrlichial infection. Furthermore, tyrosine-phosphorylated proteins and PLC-gamma2 were colocalized with ehrlichial inclusions, as determined by double-immunofluorescence labeling. The heat-sensitive component of viable E. chaffeensis cells was essential for these signaling events. E. chaffeensis, therefore, can recruit interacting signal-transducing molecules and induce the following signaling events required for the establishment of infection in host cells: protein cross-linking by TGase, tyrosine phosphorylation, PLC-gamma2 activation, IP(3) production, and an increase in [Ca(2+)](i).

MeSH Terms
Calcium/metabolism Calcium Signaling Cell Line Cytosol/metabolism Ehrlichia chaffeensis/drug effects,growth & development,physiology Endocytosis Enzyme Activation Humans Inositol 1,4,5-Trisphosphate/biosynthesis Isoenzymes/metabolism Phospholipase C gamma Phosphorylation Protein-Tyrosine Kinases/metabolism Time Factors Type C Phospholipases/metabolism Tyrosine/metabolism
Chemicals
Isoenzymes Tyrosine Inositol 1,4,5-Trisphosphate Protein-Tyrosine Kinases Type C Phospholipases Phospholipase C gamma Calcium
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Lin Mingqun
Department of Veterinary Biosciences. Neurobiotechnology Center, The Ohio State University, Columbus, Ohio 43210, USA.
Zhu Michael X
Rikihisa Yasuko
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Article Info
Journal
Infection and immunity
Abbr.
Infect Immun
ISSN
0019-9567
DOI
10.1128/IAI.70.2.889-898.2002
Published
2002-02-00
Pages
889-98
Language
English
Region
United States
NLM ID
0246127
PMCID
PMC127685
Subset
IM
Grants
NIAID NIH HHS · R01 AI030010 · United States
NIAID NIH HHS · R01 AI 30010 · United States
NIAID NIH HHS · R01 AI 40934 · United States
External Links
PubMed source DOI: 10.1128/IAI.70.2.889-898.2002 PMC Full Text View journal details
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