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Multiplatform mass spectrometry-based analysis of Leishmania donovani infected macrophages at different time points after infection

ABSTRACT: The project aims to measure targeted and non-targeted metabolite data of intracellular extracts of uninfected and Leishmania-donovani infected macrophages at 0, 12, 36 and 72 hours post infection using a multiplatform mass spectrometry approach combining CE-TOF/MS (polar metabolites), LC-QTOF/MS (non-polar metabolites) and LC-QqQ/MS (polar metabolites) to characterize the dynamics of metabolic alterations ocurring in the human macrophage upon L. donovani infection.

ORGANISM(S): Human Homo Sapiens Homo Sapiens + Leishmania Donovani

TISSUE(S): Cultured Cells

DISEASE(S): Parasitic Infection

SUBMITTER: Miguel Fernández García

PROVIDER: ST002285 | MetabolomicsWorkbench | Fri Jul 01 00:00:00 BST 2022

REPOSITORIES: MetabolomicsWorkbench

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Project description:The interplay between pathogens and hosts has been studied for decades using targeted approaches such as the analysis of mutants and host immunological responses. Although much has been learned from such studies, they focus on individual pathways and fail to reveal the global effects of infection on the host. To alleviate this issue, high-throughput methods such as transcriptomics and proteomics have been used to study host-pathogen interactions. Recently, metabolomics was established as a new method to study changes in the biochemical composition of host tissues. We report a metabolomics study of Salmonella enterica serovar Typhimurium infection. We used Fourier Transform Ion Cyclotron Resonance Mass Spectrometry with Direct Infusion to reveal that dozens of host metabolic pathways are affected by Salmonella in a murine infection model. In particular, multiple host hormone pathways are disrupted. Our results identify unappreciated effects of infection on host metabolism and shed light on mechanisms used by Salmonella to cause disease, and by the host to counter infection. Female C57BL/6 mice were infected with Salmonella enterica serovar Typhimurium SL1344 cells by oral gavage. Feces and livers were collected and metabolites extracted using acetonitrile. For experiments with feces, samples were collected from 4 mice before and after infection. For liver experiments, 11 uninfected and 11 infected mice were used. Samples were combined into 3 groups of 3-4 mice each, resulting in the analysis of 3 group samples of uninfected and 3 of infected mice. Extracts were infused into a 12-T Apex-Qe hybrid quadrupole-FT-ICR mass spectrometer equipped with an Apollo II electrospray ionization source, a quadrupole mass filter and a hexapole collision cell. Raw mass spectrometry data were processed as described elsewhere (Han et al. 2008. Metabolomics. 4:128-140 [PMID 19081807]). To identify differences in metabolite composition between uninfected and infected samples, we filtered the list of masses for metabolites which were present on one set of samples but not the other. Additionally, we calculated the ratios between averaged intensities of metabolites from uninfected and infected mice. To assign possible metabolite identities, monoisotopic neutral masses of interest were queried against MassTrix (http://masstrix.org). Masses were searched against the Mus musculus database within a mass error of 3 ppm. Data were analyzed by unpaired t tests with 95% confidence intervals.

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Multiplatform mass spectrometry-based analysis of Leishmania donovani infected macrophages at different time points after infection

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