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Infection and Immunity, August 2002, p. 4132-4141, Vol. 70, No. 8
0019-9567/02/$04.00+0     DOI: 10.1128/IAI.70.8.4132-4141.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Roles of p38 Mitogen-Activated Protein Kinase, NF-{kappa}B, and Protein Kinase C in Proinflammatory Cytokine mRNA Expression by Human Peripheral Blood Leukocytes, Monocytes, and Neutrophils in Response to Anaplasma phagocytophila

Hyung-Yong Kim and Yasuko Rikihisa*

Department of Veterinary Biosciences, The Ohio State University, Columbus, Ohio 43210-1093

Received 26 November 2001/ Returned for modification 15 February 2002/ Accepted 24 April 2002

Anaplasma phagocytophila, an obligately intracellular bacterium of granulocytes, causes human granulocytic ehrlichiosis. Within 2 h after addition of A. phagocytophila, interleukin-1ß (IL-1ß), tumor necrosis factor alpha (TNF-{alpha}), and IL-6 mRNAs are induced in human peripheral blood leukocytes (PBLs) or monocytes in vitro. However, neutrophils generate only IL-1ß mRNA. In the present study, signaling pathways for induction of these three cytokines were examined. TNF-{alpha} and IL-6 mRNA expression by PBLs was inhibited with SB 203580 (a p38 mitogen-activated protein kinase [MAPK] inhibitor), MG-132 (a proteasome inhibitor), and SN-50 (an NF-{kappa}B inhibitor). Activation of p38 MAPK and NF-{kappa}B mRNAs in monocytes was detectable within 15 to 30 min after addition of A. phagocytophila. Expression of these two cytokine mRNAs in PBLs and monocytes was also dependent on protein kinase C (PKC), protein kinase A (PKA), and protein tyrosine kinase (PTK). IL-1ß mRNA expression by neutrophils was not dependent on p38 MAPK, and p38 MAPK was not activated in neutrophils incubated with A. phagocytophila. IL-1ß mRNA induction by PBLs, monocytes, and neutrophils was dependent on PKC and PKA. Neutrophil expression of IL-1ß mRNA was dependent on transglutaminase, phospholipase C, and PTK, all of which are also required for internalization of A. phagocytophila. However, monocyte expression of IL-1ß mRNA was less dependent on these enzymes. These results suggest that A. phagocytophila transduces different signals between its host neutrophils and monocytes for proinflammatory cytokine generation.

* Corresponding author. Mailing address: Department of Veterinary Biosciences, College of Veterinary Medicine, The Ohio State University, 1925 Coffey Rd., Columbus, OH 43210-1093. Phone: (614) 292-5661. Fax: (614) 292-6473. E-mail: rikihisa.1{at}osu.edu.

Editor: R. N. Moore

Infection and Immunity, August 2002, p. 4132-4141, Vol. 70, No. 8
0019-9567/02/$04.00+0     DOI: 10.1128/IAI.70.8.4132-4141.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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