Early response of monocyte-derived macrophages from vaccinated and non-vaccinated goats against in vitro infection with Mycobacterium avium subsp. paratuberculosis

Abstract Paratuberculosis is a disease of ruminants caused by Mycobacterium avium subsp. paratuberculosis (Map). Vaccination is the most cost-effective control method. However, despite the fact that macrophages are the main target cells for this pathogen, the precise mechanisms behind the response o...

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Main Authors: Noive Arteche-Villasol, Daniel Gutiérrez-Expósito, Raquel Vallejo, Jose Espinosa, Natalia Elguezabal, Iraia Ladero-Auñon, Marcos Royo, María del Carmen Ferreras, Julio Benavides, Valentín Pérez
Format: Article
Language:English
Published: BMC 2021-05-01
Series:Veterinary Research
Subjects:
Map
Online Access:https://doi.org/10.1186/s13567-021-00940-y
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spelling doaj-74f4296363254f98a884eb7fa76038da2021-05-16T11:30:20ZengBMCVeterinary Research1297-97162021-05-0152111210.1186/s13567-021-00940-yEarly response of monocyte-derived macrophages from vaccinated and non-vaccinated goats against in vitro infection with Mycobacterium avium subsp. paratuberculosisNoive Arteche-Villasol0Daniel Gutiérrez-Expósito1Raquel Vallejo2Jose Espinosa3Natalia Elguezabal4Iraia Ladero-Auñon5Marcos Royo6María del Carmen Ferreras7Julio Benavides8Valentín Pérez9Departamento de Sanidad Animal, Facultad de Veterinaria, Universidad de LeónDepartamento de Sanidad Animal, Facultad de Veterinaria, Universidad de LeónDepartamento de Sanidad Animal, Facultad de Veterinaria, Universidad de LeónDepartamento de Sanidad Animal, Facultad de Veterinaria, Universidad de LeónDepartamento de Sanidad Animal, NEIKER-Instituto Vasco de Investigación y Desarrollo AgrarioDepartamento de Sanidad Animal, NEIKER-Instituto Vasco de Investigación y Desarrollo AgrarioDepartamento de Sanidad Animal, Facultad de Veterinaria, Universidad de LeónDepartamento de Sanidad Animal, Facultad de Veterinaria, Universidad de LeónInstituto de Ganadería de Montaña (CSIC-ULE),Departamento de Sanidad Animal, Facultad de Veterinaria, Universidad de LeónAbstract Paratuberculosis is a disease of ruminants caused by Mycobacterium avium subsp. paratuberculosis (Map). Vaccination is the most cost-effective control method. However, despite the fact that macrophages are the main target cells for this pathogen, the precise mechanisms behind the response of the macrophage to Map infection and how it is modified by vaccination are yet poorly understood. The aim of this study was to investigate the effect of Silirum® vaccination in the early immune response of caprine monocyte-derived macrophages (CaMØs). Peripheral blood mononuclear cells (PBMCs) were obtained from vaccinated and non-vaccinated goats, cultured in vitro until differentiation to macrophages and infected with Map. After a 24 h incubation, Map viability and DNA were assessed in culture by viable colony count and real time quantitative polymerase chain reaction (qPCR). In addition, Map phagocytosis and expression of IL-10, IL-12, IFN-γ, TNF-α, IL-17A, IL-1β, iNOS, IL-6 and MIP-1β were also evaluated through immunofluorescence labelling and reverse transcriptase qPCR (RT-qPCR), respectively. A significant reduction of Map viability was observed in both supernatants (P < 0.05) and CaMØs (P < 0.001) from the vaccinated group. Similarly, the percentage of infected CaMØs and the number of internalized Map by CaMØs (P < 0.0001) was higher in the vaccinated group. Finally, iNOS (P < 0.01) and IL-10 were significantly up-regulated in CaMØs from vaccinated goats, whereas only MIP-1β was up-regulated in non-vaccinated animals (P < 0.05). These results show that vaccination modifies the immune response of CaMØs, suggesting that the phagocytosis and microbiocidal activity of macrophages against Map is enhanced after vaccination.https://doi.org/10.1186/s13567-021-00940-yMapVaccinationCaprine monocyte-derived macrophagesViabilityPhagocytosis
collection DOAJ
language English
format Article
sources DOAJ
author Noive Arteche-Villasol
Daniel Gutiérrez-Expósito
Raquel Vallejo
Jose Espinosa
Natalia Elguezabal
Iraia Ladero-Auñon
Marcos Royo
María del Carmen Ferreras
Julio Benavides
Valentín Pérez
spellingShingle Noive Arteche-Villasol
Daniel Gutiérrez-Expósito
Raquel Vallejo
Jose Espinosa
Natalia Elguezabal
Iraia Ladero-Auñon
Marcos Royo
María del Carmen Ferreras
Julio Benavides
Valentín Pérez
Early response of monocyte-derived macrophages from vaccinated and non-vaccinated goats against in vitro infection with Mycobacterium avium subsp. paratuberculosis
Veterinary Research
Map
Vaccination
Caprine monocyte-derived macrophages
Viability
Phagocytosis
author_facet Noive Arteche-Villasol
Daniel Gutiérrez-Expósito
Raquel Vallejo
Jose Espinosa
Natalia Elguezabal
Iraia Ladero-Auñon
Marcos Royo
María del Carmen Ferreras
Julio Benavides
Valentín Pérez
author_sort Noive Arteche-Villasol
title Early response of monocyte-derived macrophages from vaccinated and non-vaccinated goats against in vitro infection with Mycobacterium avium subsp. paratuberculosis
title_short Early response of monocyte-derived macrophages from vaccinated and non-vaccinated goats against in vitro infection with Mycobacterium avium subsp. paratuberculosis
title_full Early response of monocyte-derived macrophages from vaccinated and non-vaccinated goats against in vitro infection with Mycobacterium avium subsp. paratuberculosis
title_fullStr Early response of monocyte-derived macrophages from vaccinated and non-vaccinated goats against in vitro infection with Mycobacterium avium subsp. paratuberculosis
title_full_unstemmed Early response of monocyte-derived macrophages from vaccinated and non-vaccinated goats against in vitro infection with Mycobacterium avium subsp. paratuberculosis
title_sort early response of monocyte-derived macrophages from vaccinated and non-vaccinated goats against in vitro infection with mycobacterium avium subsp. paratuberculosis
publisher BMC
series Veterinary Research
issn 1297-9716
publishDate 2021-05-01
description Abstract Paratuberculosis is a disease of ruminants caused by Mycobacterium avium subsp. paratuberculosis (Map). Vaccination is the most cost-effective control method. However, despite the fact that macrophages are the main target cells for this pathogen, the precise mechanisms behind the response of the macrophage to Map infection and how it is modified by vaccination are yet poorly understood. The aim of this study was to investigate the effect of Silirum® vaccination in the early immune response of caprine monocyte-derived macrophages (CaMØs). Peripheral blood mononuclear cells (PBMCs) were obtained from vaccinated and non-vaccinated goats, cultured in vitro until differentiation to macrophages and infected with Map. After a 24 h incubation, Map viability and DNA were assessed in culture by viable colony count and real time quantitative polymerase chain reaction (qPCR). In addition, Map phagocytosis and expression of IL-10, IL-12, IFN-γ, TNF-α, IL-17A, IL-1β, iNOS, IL-6 and MIP-1β were also evaluated through immunofluorescence labelling and reverse transcriptase qPCR (RT-qPCR), respectively. A significant reduction of Map viability was observed in both supernatants (P < 0.05) and CaMØs (P < 0.001) from the vaccinated group. Similarly, the percentage of infected CaMØs and the number of internalized Map by CaMØs (P < 0.0001) was higher in the vaccinated group. Finally, iNOS (P < 0.01) and IL-10 were significantly up-regulated in CaMØs from vaccinated goats, whereas only MIP-1β was up-regulated in non-vaccinated animals (P < 0.05). These results show that vaccination modifies the immune response of CaMØs, suggesting that the phagocytosis and microbiocidal activity of macrophages against Map is enhanced after vaccination.
topic Map
Vaccination
Caprine monocyte-derived macrophages
Viability
Phagocytosis
url https://doi.org/10.1186/s13567-021-00940-y
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