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Infection and Immunity, September 2000, p. 5277-5283, Vol. 68, No. 9
0019-9567/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

ankA: an Ehrlichia phagocytophila Group Gene Encoding a Cytoplasmic Protein Antigen with Ankyrin Repeats

Patrizio Caturegli,1 Kristin M. Asanovich,1 Jennifer J. Walls,1,2 Johan S. Bakken,3 John E. Madigan,4 Vsevolod L. Popov,5 and J. Stephen Dumler1,2,*

Department of Pathology, The Johns Hopkins University School of Medicine,1 and Department of Pathology, The University of Maryland School of Medicine,2 Baltimore, Maryland; Section of Infectious Diseases, Saint Mary's Duluth Clinic Health System, Duluth, Minnesota3; Department of Medicine and Epidemiology, Veterinary Medical School, University of California, Davis, California4; and Department of Pathology, The University of Texas Medical Branch, Galveston, Texas5

Received 22 February 2000/Returned for modification 2 April 2000/Accepted 10 June 2000

Human granulocytic ehrlichiosis (HGE) is a potentially fatal, tick-borne disease caused by a bacterium related or identical to Ehrlichia phagocytophila. To identify and characterize E. phagocytophila group-specific protein antigen genes, we prepared and screened HGE agent and Ehrlichia equi genomic DNA expression libraries using polyclonal equine E. equi antibodies. Two clones, one each from HGE agent and E. equi, that were recognized specifically by antibodies to the E. phagocytophila group ehrlichiae had complete open reading frames of 3,693 and 3,615 nucleotides, respectively. The two clones were 96.6% identical and predicted a protein with at least 11 tandemly repeated ankyrin motifs. Thus, the gene was named ank (for ankyrin). When the encoded protein, named AnkA, was expressed in Escherichia coli, it was recognized by antibodies from rabbits and mice immunized with the HGE agent, sera from humans convalescent from HGE, and sera from horses convalescent from HGE and E. equi infection. Monospecific AnkA antibodies reacted with proteins in HGE agent immunoblots, and AnkA monoclonal antibodies detected cytoplasmic antigen in E. phagocytophila group bacteria and also detected antigen associated with chromatin in infected but not uninfected HL-60 cell cultures. These results suggest that this Ehrlichia protein may influence host cell gene expression.

* Corresponding author. Mailing address: Division of Medical Microbiology, Department of Pathology, The Johns Hopkins Medical Institutions, Meyer B1-193, 600 North Wolfe St., Baltimore, MD 21201. Phone: (410) 955-5077. Fax: (410) 614-8087. E-mail: sdumler{at}jhmi.edu.

Infection and Immunity, September 2000, p. 5277-5283, Vol. 68, No. 9
0019-9567/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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