Abstract
Corynebacterium pseudotuberculosis is a facultative intracellular pathogen that uses various mechanisms to survive within macrophages. In phagocytosis, this survival can be attributed to the ability to inhibit phagosome-lysosome fusion. In this fusion, some proteins, including Rabs GTPases, are involved in the maturation process and are responsible for regulating membrane vesicle trafficking. Thus, to better understand these mechanisms, the capacity of biofilm-producing and non biofilm-producing strains of Corynebacterium pseudotuberculosis for modulating the expression of endosomal proteins GTPases Rab 5 and Rab 7 was evaluated in an in vitro study of infection of goat macrophages. Blood was collected from ten Canindé goats, infected with biofilm-producing and non biofilm-producing strains of C. pseudotuberculosis. Blood cells were separated in colloidal silica-polyvinylpyrrolidone gradients (GE Healthcare®). These cells were maintained at 37 °C, with 5% of CO2. After differentiation, macrophages were infected with the mentioned strains. The bacterial pellets were marked with Rab 5 and Rab 7 antibodies, and their expression was observed by flow cytometry. Both strains of C. pseudotuberculosis (biofilm-producing and non biofilm-producing) were observed to be capable of altering the expression of Rab proteins in macrophages cultivated in vitro. Macrophages from the animals infected with the biofilm-producing strain had an increase in the expression of Rab 5 protein, mainly when these macrophages were treated with the non biofilm-producing strain. The same mechanism was shown to function with Rab 7 protein, however at a lower intensity of expression when compared with Rab 5.
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Acknowledgements
The authors are grateful to FAPEX (Fundação de Apoio á Pesquisa e a Extenção, UFBA – Universidade Federal da Bahia) for the financial support for the project and FAPESB (Fundação de Amparo á Pesquisa do Estado da Bahia) for granting the scholarship to the first author.
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This work was funded by the Fundação de Apoio à Pesquisa do Estado da Bahia (FAPESB) grant number Nº BOL0275/2014.
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MCSA, MMC, and RM designed the project and discussed the results of the experiments. MCSA, MEC, MCS, and JTRRF developed methodologies, and MMS and ASS performed flow cytometry. MCSA, MMC, and RM prepared the manuscript. All authors have read and approved the final manuscript.
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da Conceição Aquino de Sá, M., Filho, J.T.R.R., Alcantara, M.E. et al. Analysis of Gtpases Rab 5 and Rab 7 expression from macrophages infected with biofilm-producing and non biofilm-producing strains of Corynebacterium pseudotuberculosis. Braz J Microbiol 53, 447–453 (2022). https://doi.org/10.1007/s42770-021-00665-2
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DOI: https://doi.org/10.1007/s42770-021-00665-2