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Infection and Immunity, March 2002, p. 1230-1234, Vol. 70, No. 3
0019-9567/02/$04.00+0     DOI: 10.1128/IAI.70.3.1230-1234.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Genetic Variability and Stability of Anaplasma phagocytophila msp2 (p44)

Karen Caspersen,1,2 Jin-Ho Park,1 Surekha Patil,1 and J. Stephen Dumler1*

Department of Pathology Division of Medical Microbiology,,1 Division of Comparative Medicine, The Johns Hopkins University School of Medicine, Baltimore, Maryland2

Received 20 September 2001/ Returned for modification 1 November 2001/ Accepted 20 November 2001

Anaplasma (Ehrlichia) phagocytophila's major immunodominant surface protein antigen, Msp2 (P44, 44-kDa antigen), is encoded by a family of paralogous genes characterized by conserved sequences flanking a hypervariable region. The antigenic profiles of most strains of A. phagocytophila are different, and the differences are principally related to Msp2 expression. To date, multiple unique msp2 gene paralogs have been found in A. phagocytophila isolates, but the overall number in the genome of a single strain is not yet known. Changes in msp2 expression may be related to antigenic variability; thus, we examined the minimal complement of msp2 genes or pseudogenes in two strains of A. phagocytophila and the number of transcriptionally active msp2 gene paralogs during low-passage, steady-state, in vitro propagation. Of 15 BDS strain clones, 1 had a hypervariable region identical to the region in a clone obtained from a BDS strain genomic library previously prepared from organisms after only two horse passages. When 124 Webster strain clones were examined, 18 unique hypervariable regions were identified. Of 64 Webster strain cDNA clones, 56 (87.5%) were derived from a single gene, and transcripts from six additional msp2 genes were also identified. The sequences of several hypervariable regions that were >=97% similar to regions present in other strains were identified by performing a BLAST analysis of sequences deposited in the GenBank database. These findings suggest that antigenic variability results from transcription of one or a few of the multiple paralogs and not from genetic instability that results in random accumulated mutations, although the possibility that gene recombination plays a role cannot be eliminated. The predominant Msp2 pattern in vitro is determined by transcription from a single gene.

* Corresponding author. Mailing address: Division of Medical Microbiology, Department of Pathology, The Johns Hopkins Medical Institutions, Meyer B1-193, 600 North Wolfe Street, Baltimore, MD 21287. Phone: (410) 955-5077. Fax: (410) 614-8087. E-mail: sdumler{at}jhmi.edu.

Infection and Immunity, March 2002, p. 1230-1234, Vol. 70, No. 3
0019-9567/02/$04.00+0     DOI: 10.1128/IAI.70.3.1230-1234.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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