Phenotypic characterization of swine peripheral blood monocyte-derived macrophages and ex vivo infection with Salmonella enterica serovar Typhimurium

  • Carlos Eduardo Real Pereira
  • Márcio Sobreira Silva Araújo
  • Juliana Pinto da Silva Mol
  • José Paulo Hiroji Sato
  • Amanda Gabrielle de Souza Daniel
  • Olindo Assis Martins-Filho
  • Renato Lima Santos
  • Roberto Mauricio Carvalho GuedesEmail author
Veterinary Microbiology - Research Paper


Macrophages are critical mediators of the inflammatory process, playing a relevant role in the pathogenesis of Salmonella Typhimurium. The protocols for isolation, culture, and differentiation of monocytes into macrophages and their interaction with Salmonella are well established in humans and murine models, but little information is available in swine. The aims of this study were to establish an efficient protocol for macrophage culture and to evaluate the interaction of the invA mutant strain and the wild type (WT) Salmonella Typhimurium with porcine macrophages. Peripheral blood monocyte-derived macrophages from pigs were obtained, separated by density-gradient centrifugation, and cultured in Teflon vials for 10 days. After the differentiation period, cultures consisted of 92.4% CD14+ cells. In addition, these cells showed phagocytic ability, demonstrated by the presence of the same amount of WT and invA mutant Salmonella Typhimurium 1 h after interaction with macrophages. The early cytotoxic effect was Salmonella pathogenicity island (SPI)-[1]dependent, in which log-phase WT strains were more efficient (p < 0.01) than the invA mutant strain at inducing the death of macrophages.


Salmonella Typhimurium Pigs Macrophage Cytotoxicity 


Funding sources

This work was supported by the FAPEMIG (grant numbers: APQ-02189-13) and PRPq-UFMG. RMCG, RLS, and OAMF are recipients of fellowships from the Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq, Brazil). The authors thank the Program for Technological Development in Tools for Health-RPT-FIOCRUZ for the use of its facilities.

Compliance with ethical standards

This experimental protocol has been reviewed and approved by the Ethics in Animal Use Committee at the Universidade Federal de Minas Gerais (CEUA/UFMG protocol number 249/2015).


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Copyright information

© Sociedade Brasileira de Microbiologia 2019

Authors and Affiliations

  • Carlos Eduardo Real Pereira
    • 1
  • Márcio Sobreira Silva Araújo
    • 2
  • Juliana Pinto da Silva Mol
    • 1
  • José Paulo Hiroji Sato
    • 1
  • Amanda Gabrielle de Souza Daniel
    • 1
  • Olindo Assis Martins-Filho
    • 2
  • Renato Lima Santos
    • 1
  • Roberto Mauricio Carvalho Guedes
    • 1
    Email author
  1. 1.Departmento de Clínica e Cirúrgia Veterinária, Escola de VeterináriaUniversidade Federal de Minas GeraisBelo HorizonteBrazil
  2. 2.Laboratório de Biomarcadores de Diagnóstico e MonitoraçãoCentro de Pesquisas René Rachou-FiocruzBelo HorizonteBrazil

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