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A novel neutralization sensitive and subdominant RAP-1-related antigen (RRA) is expressed by Babesia bovis merozoites

Published online by Cambridge University Press:  18 April 2011

CARLOS E. SUAREZ ,
JACOB M. LAUGHERY ,
REGINALDO G. BASTOS ,
WENDELL C. JOHNSON ,
JUNZO NORIMINE ,
GUSTAVO ASENZO ,
WENDY C. BROWN ,
MONICA FLORIN-CHRISTENSEN  and
WILL L. GOFF
CARLOS E. SUAREZ*
Affiliation:
Program in Vector-Borne Diseases, Department of Veterinary Microbiology and Pathology, Washington State University, Pullman, WA, USA 99164-7040 Animal Disease Research Unit, Agricultural Research Service, United States Department of Agriculture, Pullman, WA, USA 99164-6630
JACOB M. LAUGHERY
Affiliation:
Program in Vector-Borne Diseases, Department of Veterinary Microbiology and Pathology, Washington State University, Pullman, WA, USA 99164-7040
REGINALDO G. BASTOS
Affiliation:
Program in Vector-Borne Diseases, Department of Veterinary Microbiology and Pathology, Washington State University, Pullman, WA, USA 99164-7040
WENDELL C. JOHNSON
Affiliation:
Animal Disease Research Unit, Agricultural Research Service, United States Department of Agriculture, Pullman, WA, USA 99164-6630
JUNZO NORIMINE
Affiliation:
Program in Vector-Borne Diseases, Department of Veterinary Microbiology and Pathology, Washington State University, Pullman, WA, USA 99164-7040
GUSTAVO ASENZO
Affiliation:
Instituto Nacional de Tecnología Agropecuaria, Centro de Investigación en Ciencias Veterinarias y Agronómicas, Castelar, Argentina
WENDY C. BROWN
Affiliation:
Program in Vector-Borne Diseases, Department of Veterinary Microbiology and Pathology, Washington State University, Pullman, WA, USA 99164-7040 School for Global Animal Health Washington State University, Pullman, WA, USA
MONICA FLORIN-CHRISTENSEN
Affiliation:
Instituto Nacional de Tecnología Agropecuaria, Centro de Investigación en Ciencias Veterinarias y Agronómicas, Castelar, Argentina
WILL L. GOFF
Affiliation:
Animal Disease Research Unit, Agricultural Research Service, United States Department of Agriculture, Pullman, WA, USA 99164-6630
*
*Corresponding author: Animal Disease Research Unit, Agricultural Research Service, United States Department of Agriculture, Pullman, WA, USA 99164-6630. Tel: +509 335 6341. Fax +509 335 8328. E-mail: [email protected]
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Summary

Objective. The Babesia bovis genome encodes a rap-1 related gene denominated RAP-1 related antigen (RRA). In this study, we analysed the pattern of expression, immunogenicity and functional relevance of RRA. Methods. Phylogenetic analysis was performed using the program Phylip. Expression of rra was analysed by Northern blots, RT-PCR, immunoprecipitation, Western blots and immunofluorescence. RRA antigenicity was tested by T-cell proliferation and Western blot analysis, and functional relevance was determined in an in vitro neutralization assay. Results. RRA is more closely related to RAP-1b of Babesia bigemina than to B. bovis RAP-1, and it is highly conserved among distinct strains. Transcriptional analysis suggests lower numbers of rra transcripts compared to rap-1. Immunoprecipitation of metabolically labelled B. bovis proteins with antibodies against synthetic peptides representing predicted antigenic regions of RRA confirmed the expression of a ∼43 kDa RRA in cultured merozoites. Antibodies present in B. bovis hyperimmune sera, but not in field-infected cattle sera, reacted weakly with recombinant RRA, and no significant stimulation was obtained using recombinant RRA as antigen in T-cell proliferation assays, indicating that RRA is a subdominant antigen. Antibodies against RRA synthetic peptides reacted with merozoites using immunofluorescence, and were able to significantly inhibit erythrocyte invasion in in vitro neutralization tests, suggesting functional relevance for parasite survival. Conclusion. B. bovis express a novel subdominant RAP-1-like molecule that may contribute to erythrocyte invasion and/or egression by the parasite.

Keywords

ApicomplexanBabesia bovisrhoptry-associated protein-1RAP-1
Type
Research Article
Information
Parasitology , Volume 138 , Issue 7 , June 2011 , pp. 809 - 818
Creative Commons
This is a work of the United States Government and is not subject to copyright protection in the United States
Copyright
Copyright © Cambridge University Press 2011. This is a work of the United States Government and is not subject to copyright protection in the United States.

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