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

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 pa...

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Autores principales:	Jacob M Laughery, Donald P Knowles, David A Schneider, Reginaldo G Bastos, Terry F McElwain, Carlos E Suarez
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2014
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/4c233785c2184ea7a7a57a7bc40ecb9d
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spelling	oai:doaj.org-article:4c233785c2184ea7a7a57a7bc40ecb9d2021-11-18T08:18:50ZTargeted surface expression of an exogenous antigen in stably transfected Babesia bovis.1932-620310.1371/journal.pone.0097890https://doaj.org/article/4c233785c2184ea7a7a57a7bc40ecb9d2014-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/24840336/pdf/?tool=EBIhttps://doaj.org/toc/1932-6203Babesia 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.Jacob M LaugheryDonald P KnowlesDavid A SchneiderReginaldo G BastosTerry F McElwainCarlos E SuarezPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 9, Iss 5, p e97890 (2014)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Jacob M Laughery Donald P Knowles David A Schneider Reginaldo G Bastos Terry F McElwain Carlos E Suarez Targeted surface expression of an exogenous antigen in stably transfected Babesia bovis.
description	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.
format	article
author	Jacob M Laughery Donald P Knowles David A Schneider Reginaldo G Bastos Terry F McElwain Carlos E Suarez
author_facet	Jacob M Laughery Donald P Knowles David A Schneider Reginaldo G Bastos Terry F McElwain Carlos E Suarez
author_sort	Jacob M Laughery
title	Targeted surface expression of an exogenous antigen in stably transfected Babesia bovis.
title_short	Targeted surface expression of an exogenous antigen in stably transfected Babesia bovis.
title_full	Targeted surface expression of an exogenous antigen in stably transfected Babesia bovis.
title_fullStr	Targeted surface expression of an exogenous antigen in stably transfected Babesia bovis.
title_full_unstemmed	Targeted surface expression of an exogenous antigen in stably transfected Babesia bovis.
title_sort	targeted surface expression of an exogenous antigen in stably transfected babesia bovis.
publisher	Public Library of Science (PLoS)
publishDate	2014
url	https://doaj.org/article/4c233785c2184ea7a7a57a7bc40ecb9d
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Targeted surface expression of an exogenous antigen in stably transfected Babesia bovis.

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