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Differential cellular immune response of Galleria mellonella to Actinobacillus pleuropneumoniae

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Abstract

In the present work, we have investigate the cellular immune response of Galleria mellonella larvae against three strains of the gram-negative bacterium Actinobacillus pleuropneumoniae: low-virulence (780), high-virulence (1022) and the serotype 8 reference strain (R8). Prohemocytes, plasmatocytes, granulocytes, oenocytoids and spherulocytes were distinguished according to their size and morphology, their molecular markers and dye-staining properties and their role in the immune response. Total hemocyte count, differential hemocyte count, lysosome activity, autophagic response, cell viability and caspase-3 activation were determined in circulating hemocytes of naive and infected larvae. The presence of the autophagosome protein LC3 A/B within the circulating hemocytes of G. mellonella was dependent on and related to the infecting A. pleuropneumoniae strain and duration of infection. Hemocytes treated with the high-virulence strain expressed higher levels of LC3 A/B, whereas treatment with the low-virulence strain induced lower expression levels of this protein in the cells. Moreover, our results showed that apoptosis in circulating hemocytes of G. mellonella larvae after exposure to virulent bacterial strains occurred simultaneously with excessive cell death response induced by stress and subsequent caspase-3 activation.

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Acknowledgments

This work was supported by Financiadora de Estudos e Projetos (Finep), Coordenação de Aperfeiçoamento de Pessoal de Nível Superior [CAPES/PVE 88881.030429/2013-01; PROEX], and Fundação de Amparo à Pesquisa do Estado de Minas Gerais (FAPEMIG). The authors are grateful to Núcleo de Microscopia e Microanálise (NMM) and the Laboratório de Sistemática Molecular (DBA/UFV) for their technical assistance and support in all analyses involving microscopy and cytometry. They also thank J.L. Arteaga Blanco for his technical support in obtaining hemocyte images, and Dr. J.E. Serrão for kindly providing the primary antibodies used.

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  1. Departamento de Biologia Geral, Universidade Federal de Viçosa, Viçosa, 36570-900, Brazil

    Luis Andrés Arteaga Blanco, Kenner Morais Fernandes, Leandro Licursi de Oliveira & Gustavo Ferreira Martins

  2. Departamento de Microbiologia, Laboratório de Genética Molecular de Bactérias, Instituto de Biotecnologia Aplicada à Agropecuária – BIOAGRO, Universidade Federal de Viçosa, Viçosa, 36570-900, Brazil

    Josicelli Souza Crispim, Monalessa Fábia Pereira & Denise Mara Soares Bazzolli

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  1. Luis Andrés Arteaga Blanco
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Arteaga Blanco, L.A., Crispim, J.S., Fernandes, K.M. et al. Differential cellular immune response of Galleria mellonella to Actinobacillus pleuropneumoniae . Cell Tissue Res 370, 153–168 (2017). https://doi.org/10.1007/s00441-017-2653-5

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