Murine cutaneous leishmaniasis investigated by MALDI mass spectrometry imaging

Negrão, Fernanda; Rocha, Daniel Guimarães Pedro; Jaeeger, Caroline F.; Rocha, Francisca Janaína Soares; Eberlin, Marcos N.; Giorgio, Selma

doi:10.1039/c7mb00411g

Cited by 10 publications

(12 citation statements)

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“…In vitro and in vivo assays, protein extraction, and digestion were performed as previously described. , The following sections describe our findings on the modulation of proteins in the host (macrophages and mice) following Leishmania infection or LPS stimulation. Some of our previous work has demonstrated that infection caused by L. amazonensis induces the formation of strongly vacuolized macrophages, each of them containing multiple amastigotes, and that infection by L. amazonensis progresses more aggressively than infection caused by L. major . ,, The correlation between the results from in vitro and in vivo infections should not be discarded since both were induced by promastigotes of Leishmania spp. that turn into amastigotes inside of the macrophages after a few hours.…”

Section: Resultsmentioning

confidence: 99%

“…Interestingly, Old World cutaneous leishmaniasis can heal spontaneously but may also respond well to pentavalent antimonials . However, treatment of New World cutaneous leishmaniasis requires systemic treatment with pentavalent antimonials and is sometimes combined with antibiotics due to the development of parasite resistance against antimonials. − Indeed, previous reports from our group have demonstrated that disease progression and infectivity caused by L. amazonensis is more severe than L. major . − …”

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confidence: 99%

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Tandem Mass Tag Proteomic Analysis of in Vitro and in Vivo Models of Cutaneous Leishmaniasis Reveals Parasite-Specific and Nonspecific Modulation of Proteins in the Host

et al. 2019

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Cutaneous leishmaniasis, the most common form of leishmaniasis, is endemic in several regions of the world, and if not treated properly, it can cause disfiguring scars on the skin. Leishmania spp. infection causes an inflammatory response in its host, and it modulates the host metabolism differently depending on the Leishmania species. Since Leishmania spp. has begun to develop resistance against current therapies, we believe efforts to identify new possibilities for treatment are critical for future control of the disease. Proteomics approaches such as isobaric labeling yield accurate relative quantification of protein abundances and, when combined with chemometrics/statistical analysis, provide robust information about protein modulation across biological conditions. Using a mass spectrometry-based proteomics approach and tandem mass tag labeling, we have investigated protein modulation in murine macrophages (in vitro model) and skin biopsies after exposure to Leishmania spp. (in vivo murine model). Infections induced by L. amazonensis (endemic in the New World) and L. major (endemic in the Old World) were compared to an inflammation model to search for Leishmania-specific and nonspecific protein modulation in the host. After protein extracts obtained from in vitro and in vivo experiments were digested, the resulting peptides were labeled with isobaric tags and analyzed by liquid chromatography-MS (LC-MS). Several proteins that were found to be changed upon infection with Leishmania spp. provide interesting candidates for further investigation into disease mechanism and development of possible immunotherapies.

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confidence: 99%

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Tandem Mass Tag Proteomic Analysis of in Vitro and in Vivo Models of Cutaneous Leishmaniasis Reveals Parasite-Specific and Nonspecific Modulation of Proteins in the Host

et al. 2019

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“…Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI TOF MS) is a powerful analytical technique widely used for analysis of proteins in a broad range of biological samples which vary from lysed cells extracts [1][2][3][4][5][6][7][8][9][10] to tissue sections [11][12][13][14][15][16][17][18]. Whole cell MALDI fingerprinting is based on the mass spectral analysis of a whole cell without additional preparatory steps such as fractionation or extraction.…”

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confidence: 99%

Whole Cell MALDI Fingerprinting Is a Robust Tool for Differential Profiling of Two-Component Mammalian Cell Mixtures

Petukhova

Young

Wang³

et al. 2018

J. Am. Soc. Mass Spectrom.

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MALDI fingerprinting was first described two decades ago as a technique to identify microbial cell lines. Microbial fingerprinting has since evolved into an automated platform for microorganism identification and classification which is now routinely used in clinical and environmental sectors. The extension of fingerprinting to mammalian cells has yet to progress partly due to compartmentalization of eukaryotic cells and overall higher cellular complexity. A number of publications on mammalian whole cell fingerprinting suggests that the method could be useful for classification of different cell types, cell states, and monitoring cell differentiation. We report the optimization of MALDI fingerprinting workflow parameters for mammalian cells and its application for differential profiling of mammalian cell lines and two component cell line mixtures. Murine fallopian tube cells and high-grade ovarian carcinoma cell lines and their mixtures are used as model mammalian cell lines. Two-component cell mixtures serve to determine the method's feasibility for complex biological samples as the ability to detect cancer cells in a mixed cell population. The level of detection of cancer cells in the two-component mixture by principle component analysis (PCA) starts to deteriorate at 5% but with application of a different statistical approach, Wilcoxon rank sum test, the level of detection was determined to be 1%. The ability to differentiate heterogeneous cell mixtures will help further extend whole cell MALDI fingerprinting to complex biological systems.

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“…L. major is another causative agent of cutaneous leishmaniasis that is exclusively found in the eastern hemisphere, northern Africa, the Middle East, northwestern China, and Northwestern India . Using a murine infection model combining traditional histopathology and matrix assisted laser desorption ionization (MALDI) imaging, our group previously demonstrated that L. amazonensis leads to more aggressive infection than L. major . However, since in situ top-down proteomics technologies are/were still under development at the time, we were unable to identify the proteins involved in the pathogenesis of CL.…”

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Comparative Proteomic Analysis of Murine Cutaneous Lesions Induced by Leishmania amazonensis or Leishmania major

et al. 2019

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Cutaneous leishmaniasisis is the most common clinical form of leishmaniasis and one of the most relevant neglected diseases. It is known that the progress of the disease is species specific and the host's immune response plays an important role in its outcome. However, the pathways that lead to parasite clearance or survival remain unknown. In this work, skin tissue from mice experimentally infected with L. amazonensis, one of the causative agents of cutaneous leishmaniasis in the Amazon region, L. major, another causative agent of cutaneous leishmaniasis in Africa, the Middle East, China, and India, or lipopolysaccharides from Escherichia coli as an inflammation model were investigated using label-free proteomics to unveil Leishmaniaspecific protein alterations. Proteomics is a powerful tool to investigate host−pathogen relationships to address biological questions. In this work, proteins from mice skin biopsies were identified and quantified using nano-LC coupled with tandem mass spectrometry analyses. Integrated Proteomics Pipeline was used for peptide/protein identification and quantification. Western blot was used for validation of protein quantification by mass spectrometry, and protein pathways were predicted using Ingenuity Pathway Analysis. In this proteomics study, several proteins were pointed out as hypothetical targets to guide future studies on Leishmania-specific modulation of proteins in the host. We identified hundreds of exclusively modulated proteins after Leishmania spp. infection and 17 proteins that were differentially modulated in the host after L. amazonensis or L. major infection.

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Murine cutaneous leishmaniasis investigated by MALDI mass spectrometry imaging

Cited by 10 publications

References 39 publications

Tandem Mass Tag Proteomic Analysis of in Vitro and in Vivo Models of Cutaneous Leishmaniasis Reveals Parasite-Specific and Nonspecific Modulation of Proteins in the Host

Tandem Mass Tag Proteomic Analysis of in Vitro and in Vivo Models of Cutaneous Leishmaniasis Reveals Parasite-Specific and Nonspecific Modulation of Proteins in the Host

Whole Cell MALDI Fingerprinting Is a Robust Tool for Differential Profiling of Two-Component Mammalian Cell Mixtures

Comparative Proteomic Analysis of Murine Cutaneous Lesions Induced by Leishmania amazonensis or Leishmania major

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