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Targeted surface expression of an exogenous antigen in stably transfected Babesia bovis. | LitMetric

Targeted surface expression of an exogenous antigen in stably transfected Babesia bovis.

PLoS One

Program in Vector-Borne Diseases, Department of Veterinary Microbiology and Pathology, Washington State University, Pullman, Washington, United States of America; Animal Disease Research Unit, Agricultural Research Service, United States Department of Agriculture, Pullman, Washington, United States of America.

Published: January 2015

AI Article Synopsis

  • Babesia bovis is a parasite causing serious illness in cattle, but vaccination with modified strains can help manage this disease.
  • New transfection technologies allow scientists to introduce multiple genes into B. bovis, potentially enhancing vaccine development by using the parasites as delivery systems for antigens.
  • This research demonstrated a novel approach to expressing multiple genes in B. bovis, including a protective tick antigen, which could improve future vaccine effectiveness.

Article Abstract

Babesia bovis is a tick-borne intraerythocytic protozoan responsible for acute disease in cattle which can be controlled by vaccination with attenuated B. bovis strains. Emerging B. bovis transfection technologies may increase the usefulness of these live vaccines. One use of transfected B. bovis parasites may be as a vaccine delivery platform. Previous transfection methods for B. bovis were limited by single expression sites and intracellular expression of transfected antigens. This study describes a novel transfection system in which two exogenous genes are expressed: one for selection and the other for a selected antigen designed to be delivered to the surface of the parasites. The strategy for duplicating the number of transfected genes was based on the use of the putative bidirectional promoter of the B. bovis 1.4 Kb ef-1α intergenic region. The ability of this region to regulate two independent expression sites was demonstrated using a luciferase assay on transiently transfected B. bovis parasites and then incorporated into a stable transfection plasmid to control independent expression of the selectable marker GFP-BSD and another gene of interest. A chimeric gene was synthetized using sequences from the protective B-cell epitopes of Rhipicephalus microplus tick antigen Bm86 along with sequences from the surface exposed B. bovis major surface antigen-1. This chimeric gene was then cloned into the additional expression site of the transfection plasmid. Transfection of the B. bovis Mo7 strain with this plasmid resulted in stable insertion into the ef-1α locus and simultaneous expression of both exogenous genes. Expression of the Bm86 epitopes on the surface of transfected merozoites was demonstrated using immunofluorescence analyses. The ability to independently express multiple genes by the inclusion of a bidirectional promoter and the achievement of surface expression of foreign epitopes advances the potential of transfected B. bovis as a future vaccine delivery platform.

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Source
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4026526PMC
http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0097890PLOS

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