Abstract
Leishmania (Viannia) braziliensis is the main species responsible for American tegumentary leishmaniasis in Brazil. Nevertheless, the use of this parasite species to study Leishmania infection in the murine model has been less conducted when compared with other Leishmania species. The control of murine infection with Leishmania has been associated with nitric oxide (NO) produced by inducible NO synthase (iNOS) from M1 macrophages, while arginase expressed by M2 macrophages is related to Leishmania proliferation. Here we use three different strains of L. (V.) braziliensis and one strain of L. (L.) major to study a 9-day infection of macrophages in vitro. Wild-type bone marrow–derived macrophages (BMDM) supported the proliferation of L. (L) major amastigotes from the 3rd day after infection, while all strains of L. (V.) braziliensis did not proliferate even inside IL-4-treated or iNOS knockout (KO) macrophages. The arginase activity was higher in iNOS KO than IL-4-treated macrophage showing that the absence of proliferation is independent of arginase. Importantly, L. (V.) braziliensis was able to cause uncontrolled disease in iNOS KO mice in vivo demonstrating that murine macrophages present at the site of infection have additional changes beyond inhibition of NO production or stimulation of arginase activity to support parasite proliferation.
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Acknowledgements
We are grateful to Dr. Daniel Silva Goulard, Ms Marco Vitor Silva de Melo, and Iraci Maria Cardoso for animal care.
Funding
This work was supported by Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES), Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq), Fundação de Amparo à Pesquisa do Estado de Goiás (FAPEG), and Instituto Nacional de Ciência e Tecnologia da Interação Patógeno Hospedeiro (IPH). MAPO and CMAS are Fellows from Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq).
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Milton Adriano Pelli de Oliveira: conceived, discussed, analyzed, interpreted the data, and finalized the manuscript
Mirian Vieira Teixeira: conceived, discussed, analyzed, interpreted the data, performed the experiments, and finalized the manuscript
Santiago Aguiar Spellet Soares: conceived, discussed, analyzed, interpreted the data, performed the experiments, and finalized the manuscript
Lilian Cristiane Baeza conceived, discussed, analyzed, interpreted the data, and finalized the manuscript
Vagniton Amelio Souza: performed the experiments, analyzed, interpreted the data
André Murilo de Suoza Marques: performed the experiments, analyzed, interpreted the data
Celia Maria de Almeida Soares: discussed, analyzed, interpreted the data
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The authors declare that all the experimental procedures were conducted according to the guidelines of the Animal Research Ethical Committee (CEUA) of the Federal University of Goiás, approved under protocol 047/2017.
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Supplementary Fig. 1
Survival of L. (V.) braziliensis and L. (L.). major inside of Thioglycolateelicited C57BL/6 and BALB/c mice macrophages. Thioglycolate elicited macrophages from C57BL/6 or BALB/cmice were infected with stationary phase promastigotes of L. (V.) braziliensis (JCJ8c) or L.( L.) major (Friedlin)at the ratio 10:1 (promastigotes: macrophage). Cells were cultured withpromastigotes for 3 h, washed and cultured for until 9 days. The data representthe mean ± SD of the percentage of infected cells (A) or the number ofamastigote/infected cells (B) of from four independent experiments. * indicatedifference among 3 hour and later time points by two-way ANOVA followed by Tukeytest (p <0.05) 75.1 KB
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Teixeira, M.V., Soares, S.A.E., Souza, V.A. et al. Murine macrophages do not support the proliferation of Leishmania (Viannia) braziliensis amastigotes even in absence of nitric oxide and presence of high arginase activity. Parasitol Res 121, 2891–2899 (2022). https://doi.org/10.1007/s00436-022-07614-4
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DOI: https://doi.org/10.1007/s00436-022-07614-4