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Development and characterization of monoclonal antibodies against Besnoitia besnoiti tachyzoites

Published online by Cambridge University Press:  16 August 2018

P. García-Lunar ,
G. Schares ,
A. Sanz-Fernández ,
A. Jiménez-Meléndez ,
I. García-Soto ,
J. Regidor-Cerrillo Open the ORCID record for J. Regidor-Cerrillo [Opens in a new window] ,
I. Pastor-Fernández ,
A. Hemphill Open the ORCID record for A. Hemphill [Opens in a new window] ,
M. Fernández-Álvarez  and
L. M. Ortega-Mora
...Show all authors
P. García-Lunar
Affiliation:
SALUVET, Animal Health Department, Faculty of Veterinary Sciences, Complutense University of Madrid, Ciudad Universitaria s/n, 28040 Madrid, Spain
G. Schares
Affiliation:
Friedrich-Loeffler-Institut, Federal Research Institute for Animal Health, Institute of Epidemiology, Greifswald-Insel Riems, Germany
A. Sanz-Fernández
Affiliation:
Inmunología y Genética Aplicada, SA (INGENASA), Calle de los Hermanos García Noblejas, 41, 28037, Madrid, Spain
A. Jiménez-Meléndez
Affiliation:
SALUVET, Animal Health Department, Faculty of Veterinary Sciences, Complutense University of Madrid, Ciudad Universitaria s/n, 28040 Madrid, Spain
I. García-Soto
Affiliation:
Inmunología y Genética Aplicada, SA (INGENASA), Calle de los Hermanos García Noblejas, 41, 28037, Madrid, Spain
J. Regidor-Cerrillo
Affiliation:
SALUVET, Animal Health Department, Faculty of Veterinary Sciences, Complutense University of Madrid, Ciudad Universitaria s/n, 28040 Madrid, Spain
I. Pastor-Fernández
Affiliation:
SALUVET, Animal Health Department, Faculty of Veterinary Sciences, Complutense University of Madrid, Ciudad Universitaria s/n, 28040 Madrid, Spain
A. Hemphill
Affiliation:
Institute of Parasitology, Vetsuisse Faculty, University of Bern, Länggass-Strasse 122, CH-3012 Bern, Switzerland
M. Fernández-Álvarez
Affiliation:
SALUVET, Animal Health Department, Faculty of Veterinary Sciences, Complutense University of Madrid, Ciudad Universitaria s/n, 28040 Madrid, Spain
L. M. Ortega-Mora
Affiliation:
SALUVET, Animal Health Department, Faculty of Veterinary Sciences, Complutense University of Madrid, Ciudad Universitaria s/n, 28040 Madrid, Spain
G. Álvarez-García*
Affiliation:
SALUVET, Animal Health Department, Faculty of Veterinary Sciences, Complutense University of Madrid, Ciudad Universitaria s/n, 28040 Madrid, Spain
*
Author for correspondence: G. Álvarez-García, E-mail: [email protected]
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Abstract

This is the first report on the development and characterization of eight monoclonal antibodies (MABs) generated against whole- and membrane-enriched tachyzoite extracts of the apicomplexan parasite Besnoitia besnoiti. Confocal laser scanning immunofluorescence microscopy was used to localize respective epitopes in B. besnoiti tachyzoites along the lytic cycle. A pattern compatible with dense granule staining was observed with MABs 2.A.12, 2.F.3 and 2.G.4, which could be confirmed by immunogold electron microscopy for MABs 2.A.12 and 2.F.3. In particular, MABs 2.F.3 and 2.G.4 were secreted during early invasion, proliferation and egress phases. MABs 3.10.8 and 5.5.11 labelled the tachyzoite surface, whilst MABs 1.17.8, 8.9.2 and 2.G.A recognized the apical tip, which is reminiscent for microneme localization. Besides, the epitopes recognized by the latter two (MABs 8.9.2 and 2.G.A) exhibited a redistribution from the anterior part across the parasite surface towards the posterior end during invasion. Most MABs developed were genus-specific. Indeed, the MABs cross-reacted neither with T. gondii nor with N. caninum tachyzoites. In summary, we have generated MABs that will be useful to study the key processes in the lytic cycle of the parasite and with additional promising diagnostic value. However, the molecular identity of the antigens recognized remains to be elucidated.

Keywords

B. besnoiticonfocal laser scanning microscopymonoclonal antibodytachyzoitetransmission electron microscopyWestern blot
Type
Research Article
Information
Parasitology , Volume 146 , Issue 2 , February 2019 , pp. 187 - 196
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Copyright
Copyright © Cambridge University Press 2018 

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