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Infection and Immunity, February 2002, p. 642-648, Vol. 70, No. 2
0019-9567/01/$04.00+0     DOI: 10.1128/IAI.70.2.642-648.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Expression of Major Surface Protein 2 Variants with Conserved T-Cell Epitopes in Anaplasma centrale Vaccinates

Varda Shkap,1 Thea Molad,1 Kelly A. Brayton,2 Wendy C. Brown,2 and Guy H. Palmer2,2*

Department of Parasitology, Kimron Veterinary Institute, Bet Dagan, Israel,1 Program in Vector-Borne Diseases, Washington State University, Pullman, Washington2

Received 22 June 2001/ Returned for modification 6 August 2001/ Accepted 5 November 2001

Major surface protein 2 (MSP-2), identified as a protection-inducing immunogen against Anaplasma marginale challenge, is an immunodominant outer membrane protein with orthologues in all examined Anaplasma species. Although immunization with live Anaplasma centrale has long been used to induce protection against acute disease upon challenge with virulent A. marginale, its MSP-2 structure and whether MSP-2 variants are generated during persistence of the vaccine strain was unknown. In this study, we showed that the A. centrale vaccine strain persisted for a minimum of 4 years postvaccination and generated sequential MSP-2 variants. Comparison of amino acid sequences encoded by A. centrale msp-2 transcripts from the initial postimmunization period and from sequential time points during persistence of the vaccine strain revealed a central hypervariable domain flanked by conserved amino and carboxy-terminal regions. This structure corresponded to that shown in A. marginale MSP-2, where the central hypervariable region encodes variant B-cell epitopes in the extracellular domain and the flanking transmembrane domains are rich in CD4+-T-cell epitopes. Importantly, at least four CD4+-T-cell epitopes are conserved between the two species, a finding consistent with A. marginale challenge triggering a recall response of CD4+ T cells induced by A. centrale vaccination. The genomic arrangement is conserved between A. centrale and A. marginale with multiple msp-2 pseudogenes and a single operon-linked expression site for the full-length msp-2. This conservation of both genomic structure for generating MSP-2 variants and the CD4+-T-cell epitopes between these two genetically distinct Anaplasma species indicates that they present a similar repertoire of MSP-2 epitopes to the immune system and that this similarity may be responsible for all or part of the A. centrale vaccine efficacy.


* Corresponding author. Mailing address: Department of Veterinary Microbiology and Pathology, Washington State University, Pullman, WA 99164-7040. Phone: (509) 335-6033. Fax: (509) 335-8529. E-mail: gpalmer{at}vetmed.wsu.edu.

Editor: E. I. Tuomanen


Infection and Immunity, February 2002, p. 642-648, Vol. 70, No. 2
0019-9567/01/$04.00+0     DOI: 10.1128/IAI.70.2.642-648.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.




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