Leishmania Parasite Quantification by Bioluminescence in Murine Models

Reverte, Marta

doi:10.21769/bioprotoc.3431

Cited by 5 publications

(7 citation statements)

References 7 publications

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“…with 200 mg/kg of Luminol sodium salt (Carbosynth) or 150 mg/kg of VivoGlo Luciferin (Promega) diluted in dPBS, respectively. Bioluminescence from mouse footpads was measured by In-vivo Xtreme II (BRUKER) and quantified using Molecular Imaging (MI) software (BRUKER) as described previously ( 40 ).…”

Section: Methodsmentioning

confidence: 99%

Sex-Biased Control of Inflammation and Metabolism by a Mitochondrial Nod-Like Receptor

et al. 2022

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Mitochondria regulate steroid hormone synthesis, and in turn sex hormones regulate mitochondrial function for maintaining cellular homeostasis and controlling inflammation. This crosstalk can explain sex differences observed in several pathologies such as in metabolic or inflammatory disorders. Nod-like receptor X1 (NLRX1) is a mitochondria-associated innate receptor that could modulate metabolic functions and attenuates inflammatory responses. Here, we showed that in an infectious model with the human protozoan parasite, Leishmania guyanensis, NLRX1 attenuated inflammation in females but not in male mice. Analysis of infected female and male bone marrow derived macrophages showed both sex- and genotype-specific differences in both inflammatory and metabolic profiles with increased type I interferon production, mitochondrial respiration, and glycolytic rate in Nlrx1-deficient female BMDMs in comparison to wild-type cells, while no differences were observed between males. Transcriptomics of female and male BMDMs revealed an altered steroid hormone signaling in Nlrx1-deficient cells, and a “masculinization” of Nlrx1-deficient female BMDMs. Thus, our findings suggest that NLRX1 prevents uncontrolled inflammation and metabolism in females and therefore may contribute to the sex differences observed in infectious and inflammatory diseases.

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Section: Methodsmentioning

confidence: 99%

Sex-Biased Control of Inflammation and Metabolism by a Mitochondrial Nod-Like Receptor

et al. 2022

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“…Immunocompromised mice, such as Ifng -/- and dKO mice, received constant pain medication (1g/L Dafalgan (Upsa) diluted in drinking water) during the second half of their infection. In vivo parasite burden was quantified by intra-peritoneally injecting VivoGlo Luciferin (Promega) at a concentration of 150 mg/kg and measuring the bioluminescence produced in the mouse footpads with In-Vivo Xtreme II (BRUKER) as previously described [ 94 ]. Alternatively, in vivo ROS levels were measured by intra-peritoneally injecting Luminol sodium salt (Carbosynth) at a concentration of 200 mg/kg and following the same procedure as parasite burden quantification.…”

Section: Methodsmentioning

confidence: 99%

“…The acquired images were analyzed using Molecular Imaging (MI) software (BRUKER). Regions of Interest (ROI) were set on the footpads and the bioluminescent signals were expressed in units of photons per second (P/s) [ 94 ].…”

Section: Methodsmentioning

confidence: 99%

The antioxidant response favors Leishmania parasites survival, limits inflammation and reprograms the host cell metabolism

et al. 2021

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The oxidative burst generated by the host immune system can restrict intracellular parasite entry and growth. While this burst leads to the induction of antioxidative enzymes, the molecular mechanisms and the consequences of this counter-response on the life of intracellular human parasites are largely unknown. The transcription factor NF-E2-related factor (NRF2) could be a key mediator of antioxidant signaling during infection due to the entry of parasites. Here, we showed that NRF2 was strongly upregulated in infection with the human Leishmania protozoan parasites, its activation was dependent on a NADPH oxidase 2 (NOX2) and SRC family of protein tyrosine kinases (SFKs) signaling pathway and it reprogrammed host cell metabolism. In inflammatory leishmaniasis caused by a viral endosymbiont inducing TNF-α in chronic leishmaniasis, NRF2 activation promoted parasite persistence but limited TNF-α production and tissue destruction. These data provided evidence of the dual role of NRF2 in protecting both the invading pathogen from reactive oxygen species and the host from an excess of the TNF-α destructive pro-inflammatory cytokine.

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“…Parasite burden was quantified in the infected mice by injecting VivoGlo D-Luciferin salt (Promega) at a concentration of 150 mg/kg, intraperitoneally (i.p. ), and parasite bioluminescence produced in the mouse footpads and other visible secondary lesions (like tail and snout) were measured with In-Vivo Xtreme II (BRUKER) as previously described (Reverte and Fasel, 2019). Additionally, in vivo read out of inflammation was measured similarly by injecting Luminol sodium salt (Carbosynth) at a concentration of 200 mg/kg i.p.…”

Section: Mice Infection and In Vivo Bioluminescence Imaging And Quant...mentioning

confidence: 99%

“…These acquired images were then analyzed using Molecular Imaging (MI) software (BRUKER), setting specific regions of interest (ROIs) on FP, tail, etc. and expressing the measured bioluminescence signals in units of photons per second (p/s) (Reverte and Fasel, 2019). Immunocompromised mice (Ifng −/− ) received constant painrelieving medication [1g/L DAFALGAN (Upsa) diluted in drinking water], from the appearance of big primary lesions (W3-4 p.i.)…”

Section: Mice Infection and In Vivo Bioluminescence Imaging And Quant...mentioning

confidence: 99%

In and out: Leishmania metastasis by hijacking lymphatic system and migrating immune cells

Jha

Reverte

Ronet

et al. 2022

Front. Cell. Infect. Microbiol.

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The lymphatic system plays a crucial role in mounting immune response against intracellular pathogens, and recent studies have documented its role in facilitating tumor dissemination linked largely with cancer cells. However, in mucocutaneous leishmaniasis (MCL) caused by Leishmania Viannia subgenus showing infectious metastasis and resulting in severe distant secondary lesions, the route of escape of these parasites to secondary sites has not yet been investigated in detail. Our results demonstrated that when infection was associated with inflammation and additionally exacerbated by the presence of dsRNA viral endosymbiont (LRV1), lymphatic vessels could serve as efficient routes for infected cells to egress from the primary site and colonize distant organs. We challenged this hypothesis by using the intracellular Leishmania protozoan parasites Leishmania guyanensis (Lgy) associated with or without a dsRNA viral endosymbiont, exacerbating the infection and responsible for a strong inflammatory response, and favoring metastasis of the infection. We analyzed possible cargo cells and the routes of dissemination through flow cytometry, histological analysis, and in vivo imaging in our metastatic model to show that parasites disseminated not only intracellularly but also as free extracellular parasites using migrating immune cells, lymph nodes (LNs), and lymph vessels, and followed intricate connections of draining and non-draining lymph node to finally end up in the blood and in distant skin, causing new lesions.

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Leishmania Parasite Quantification by Bioluminescence in Murine Models

Cited by 5 publications

References 7 publications

Sex-Biased Control of Inflammation and Metabolism by a Mitochondrial Nod-Like Receptor

Sex-Biased Control of Inflammation and Metabolism by a Mitochondrial Nod-Like Receptor

The antioxidant response favors Leishmania parasites survival, limits inflammation and reprograms the host cell metabolism

In and out: Leishmania metastasis by hijacking lymphatic system and migrating immune cells

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