Extracellular vesicles in infectious diseases caused by protozoan parasites in buffaloes

 

Leticia Gomes de Pontes1, Wanessa Fernanda Altei2, Asier Galan3, Petra Bilić3, Nicolas Guillemin3, Josipa Kuleš3, Anita Horvatić3, Lígia Nunes de Morais Ribeiro4 , Eneida de Paula4, Virgínia Bodelão Richini Pereira5 , Simone Baldini Lucheis6 , Vladimir Mrljak3, Peter David Eckersall3,7, Rui Seabra Ferreira Jr1,8,9, Lucilene Delazari dos Santos1,8,9

 

1 Graduate Program in Tropical Diseases, Botucatu Medical School (FMB), São Paulo State University (UNESP), Botucatu, SP, Brazil.

2 Laboratory of Biochemistry and Molecular Biology, Department of Physiological Sciences, Federal University of São Carlos (UFSCar), São Carlos, SP, Brazil.

3 ERA Chair Team (VetMedZg), Clinic for Internal Diseases, Faculty of Veterinary Medicine, University of Zagreb, Zagreb, Croatia.

4 Department of Biochemistry and Tissue Biology, Institute of Biology, University of Campinas (UNICAMP), Campinas, SP, Brazil.

5 Adolfo Lutz Institute, Center of Regional Laboratories II, Bauru, SP, Brazil.

6 Paulista Agency of Agribusiness Technology (APTA), Bauru, SP, Brazil.

7 Institute of Biodiversity, Animal Health and Comparative Medicine, University of Glasgow, Glasgow, United Kingdom, UK.

8 Graduate Program in Clinical Research, Botucatu Medical School (FMB), São Paulo State University (UNESP), Botucatu, SP, Brazil.

9 Center for the Study of Venoms and Venomous Animals (CEVAP), São Paulo State University (UNESP), Botucatu, SP, Brazil.

 

Abstract

Background: Extracellular vesicles (EVs) are small membrane-bound vesicles of growing interest in vetetinary parasitology. The aim of the present report was to provide the first isolation, quantification and protein characterization of EVs from buffalo (Bubalus bubalis) sera infected with Theileria spp.

 

Methods: Infected animals were identified through optical microscopy and PCR. EVs were isolated from buffalo sera by size-exclusion chromatography and characterized using western blotting analysis, nanoparticle tracking analysis and transmission electron microscopy. Subsequently, the proteins from isolated vesicles were characterized by mass spectrometry.

 

Results: EVs from buffalo sera have shown sizes in the 124-140 nm range and 306 proteins were characterized. The protein-protein interaction analysis has evidenced biological processes and molecular function associated with signal transduction, binding, regulation of metabolic processes, transport, catalytic activity and response to acute stress. Five proteins have been shown to be differentially expressed between the control group and that infected with Theileria spp., all acting in the oxidative stress pathway.

 

Conclusions: EVs from buffaloes infected with Theileria spp. were successfully isolated and characterized. This is an advance in the knowledge of host-parasite relationship that contributes to the understanding of host immune response and theileriosis evasion mechanisms. These findings may pave the way for searching new EVs candidate-markers for a better production of safe biological products derived from buffaloes.

 

Keywords: Extracellular vesicles Theileria spp. Protozoan parasites Nanoparticle tracking analysis Proteomic analysis

 

Correspondence: lucilenebio@gmail.com

 

Received: 11 September 2019; Accepted: 01 May 2020; Published online: 29 May 2020.