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Brocchi, M.

Publications and source records attributed to Brocchi, M..

2 recordsLinked to original sources

Impact of Bacterial Membrane Vesicles on Cellular Responses in Leishmania amazonensis-Infected Macrophages In Vitro

Leishmania amazonensis is an intracellular protozoan parasite and the main cause of Localized Cutaneous Leishmaniasis (LCL). A practical problem that can worsen the condition of infected individuals is secondary co-infection caused by opportunistic bacteria. This study evaluated the influence of Bacterial Membrane Vesicles (BMVs) from Pseudomonas aeruginosa and Staphylococcus aureus--both commonly associated with LCL--on macrophages previously infected with L. amazonensis. The diameter and concentration of BMVs were consistent with previous findings. We assessed infection dynamics, macrophage viability, and cytokine production. Results showed significant reductions in these parameters when L. amazonensis-infected macrophages were subsequently treated with BMVs, compared to the control group (infected but not treated with BMVs). Quantification of IL-2, IL-4, IL-6, IL-10, IL-17A, IFN-{gamma}, and TNF- revealed elevated levels of the last cytokine, suggesting an inflammatory response. Co-cultures treated with BMVs from P. aeruginosa (POMVs) exhibited a more pronounced inflammatory profile--marked by higher IL-1{beta} and TNF- concentrations--compared to those treated with BMVs from S. aureus (SEVs). IL-4 and IL-6 levels remained low relative to IL-1{beta} and TNF-. In conclusion, our data suggest that macrophage infection with L. amazonensis followed by exposure to bacterial MVs simulates a host-parasite-bacterial interaction, inducing strong immunogenic and inflammatory responses, which may represent potential targets for future vaccine strategies.

microbiology↗

Uptake and survival of Leishmania amazonensis in Acanthamoeba castellanii: An infection organism model?

Leishmaniasis is a neglected tropical disease. Parasite strategies and the evaluation of drugs and vaccines are inferred from studies carried out using mouse models and macrophages. The development of model organisms with no ethical restrictions will contribute to our knowledge of leishmaniasis. Acanthamoeba castellanii, a free-living protozoan, is known to interact with various microorganisms. In this study, the interaction between the amoeba A. castellanii and the trypanosomatid Leishmania amazonensis was investigated by combining quantitative kinetics analysis, optical, fluorescence, electronic, confocal, and live video microcopy. We sought to standardize protocols for the co-culture; the optimal experimental conditions were: RPMI medium + 10% SFB at 26{degrees}C. L. amazonensis invades A. castellanii through its acanthopods, and the promastigotes interact with the trophozoites via their flagellum, which also occurs when parasites infect mammalian macrophages. The forms of L. amazonensis inside the amoeba become rounded and lose their flagellum; they are similar to amastigotes. These round forms were isolated from trophozoites after 3 h of co-culture and differentiated into promastigotes, demonstrating their viability inside amoeba. The percentage of amoebas with L. amazonensis was reduced overtime. Thus, considering that A. castellanii can clear Leishmania, this interaction could serve as an effective model of cellular leishmanicidal mechanisms.

cell biology↗