8 results on '"Mussotter, Franz"'
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2. Proteomics analysis of dendritic cell activation by contact allergens reveals possible biomarkers regulated by Nrf2
- Author
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Mussotter, Franz, Tomm, Janina Melanie, El Ali, Zeina, Pallardy, Marc, Kerdine-Römer, Saadia, Götz, Mario, von Bergen, Martin, Haase, Andrea, and Luch, Andreas
- Published
- 2016
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3. Application of proteomics and metabolomics in molecular investigations of sensitization to contact allergens
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Mußotter, Franz
- Subjects
proteomics ,contact allergens ,dendritic cells ,allergic contact dermatitis ,metabolomics ,sensitization - Abstract
ACD represents a widespread adaptive immune reaction in the human skin elicited by substances contained in consumable products. Mediating the selection and activation of contact allergen-specific T cells during sensitization, DCs play a central role during the development of ACD. Reliable identification and classification of contact allergens is of high importance to curtail ACD. Analyses of the sensitization to contact allergens on the molecular level may contribute to the advancement of alternative testing methods. In the thesis on hand, proteomics and metabolomics analyses were conducted to investigate the sensitization to contact allergens on the molecular level using DC models. In the first study, gel-based proteomics was applied to identify differentially regulated proteins in nrf2+/+ and nrf2-/- BMDCs after treatment with the contact allergens DNCB, CA, NiSO4 and the irritant SDS. Alterations on the proteome level were identified using 2D-PAGE and ESI-MS/MS. Regulated proteins in nrf2+/+ BMDCs could be mapped to essential processes of BMDC activation including unfolded protein response, cell signaling, protein expression and re-organization of the cytoskeleton. A central finding was that contact allergens induced enzymes of the carbohydrate metabolism indicating metabolic reprogramming in BMDCs. Comparative analysis of nrf2+/+ and nrf2-/- data confirmed Nrf2 targets on the protein level for the first time and identified further putative Nrf2 targets. On the pathway level, differential regulation of oxidative stress response proteins confirmed the induction of the Keap1/Nrf2 pathway by contact allergens. In summary, this study confirmed a central role of Nrf2 during the response of BMDCs treated with contact allergens and led to the identification of promising biomarker candidates for the identification of contact allergens in vitro. In the second study, the response of human THP-1 cells to the contact allergen DNCB and the irritant SDS was investigated using a multi-omics strategy. For the targeted analysis of metabolites LC-MS/MS and FIA-MS/MS was employed. Untargeted quantitative proteomics analysis was based on SILAC and MALDI-TOF-MS/MS. Consistent findings on the metabolome and proteome level indicated metabolic reprogramming in THP 1 cells treated with DNCB. An induction of lipid synthesis was confirmed by the up-regulation of phospholipids and fatty acid synthase, a key enzyme of lipid synthesis. Additionally, proteins involved in protein synthesis and UPR were induced and an interruption of the TCA cycle could be concluded. Biogenic amines and long-chained phospholipids could be proposed as promising biomarker candidates for the activation of THP-1 cells by contact allergens. In summary, this study supported the hypothesis that contact allergens may induce metabolic reprogramming in THP-1 cells. Furthermore, the results confirmed that the analysis of metabolic endpoints may represent a promising approach for the identification of contact allergens in vitro., Allergische Kontaktdermatitis (ACD) stellt eine weitverbreitete, adaptive Immunreaktion der menschlichen Haut dar, welche durch Substanzen in verbrauchernahen Produkten ausgelöst werden kann. Dendritische Zellen (DC) spielen eine entscheidende Rolle während der Entstehung von ACD, indem sie die Selektion und Aktivierung kontaktallergen-spezifischer T-Zellen während der Sensibilisierung vermitteln. Die zuverlässige Identifizierung und Klassifizierung von Kontaktallergenen spielt eine wichtige Rolle bei der Eindämmung von ACD. Analysen der Sensibilisierung gegen Kontaktallergene auf molekularer Ebene können zur Verbesserung alternativer Testmethoden beitragen. In der vorliegenden Arbeit wurden proteomische und metabolomische Analysen an verschiedenen Modellen für DC durchgeführt, um die Sensibilisierung gegen Kontaktallergene auf der molekularen Ebene zu untersuchen. In der ersten Studie der Arbeit wurden murine, aus dem Knochenmark gewonnene dendritische Zellen (Bone Marrowed Derived Dendritic Cells, BMDC) mit den Kontaktallergenen 2,4-Dinitrochlorbenzol (DNCB), Zimtaldehyd (CA) und Nickel(II)-sulfat (NiSO4) sowie dem Irritanz Natriumdodecylsulfat (SDS) behandelt. Veränderungen auf Proteom-Ebene wurden mittels 2-dimensionaler Polyacrylamidgelelektrophorese (2D-PAGE) und Elektrosprayionisation-gekoppelter Tandem-Massenspektrometrie (ESI-MS/MS) identifiziert. Regulierte Proteine in wildtypischen BMDC (nrf2+/+) konnten essentiellen Prozessen der Aktivierung von BMDC zugeordnet werden, einschließlich der Antwort auf ungefaltete Proteine, Signaltransduktion, Expression von Proteinen und Reorganisation des Cytoskeletts. Ein zentrales Ergebnis war, dass Kontaktallergene Enzyme des Stoffwechsels von Kohlenhydraten induzierten, was eine metabolische Reprogrammierung von BMDC anzeigte. Die vergleichende Analyse von Daten aus nrf2+/+ und BMDC mit Nrf2-Knockout (nrf2-/-) bestätigte vorhergesagte Nrf2-Zielmoleküle zum ersten Mal auf der Proteinebene und konnte weitere Kandidaten für Nrf2-Zielmoleküle identifizieren. Auf der Signalweg-Ebene bestätigte die differentielle Regulierung von Proteinen der oxidativen Stressantwort die Aktivierung des Keap1/Nrf2-Signalweges durch Kontaktallergene. Zusammengefasst bestätigte diese Studie eine zentrale Rolle für Nrf2 während der Antwort von BMDC auf die Behandlung mit Kontaktallergenen und führte zur Identifizierung von vielversprechenden Biomarker-Kandidaten für die Identifizierung von Kontaktallergenen in vitro. In der zweiten Studie wurde die zelluläre Antwort humaner THP-1 Zellen nach Behandlung mit dem Kontaktallergen DNCB oder dem Irritanz SDS mit Hilfe von Metabolomics und Proteomics analysiert. Für eine gezielte Metabolit-Analyse wurden LC-MS/MS und FIA-MS/MS angewendet. Die ungezielte Protein-Analyse basierte auf SILAC und MALDI-TOF-MS/MS. Konsistente Ergebnisse auf Metabolom- und Proteom-Ebene zeigte eine metabolische Reprogrammierung der mit DNCB behandelten THP-1 Zellen an. Eine Aktivierung der Lipid-Synthese wurde durch die Hochregulierung von Phospholipiden und der Fettsäure-Synthase, einem zentralen Enzym der Lipid-Synthese, bestätigt. Zusätzlich wurden Proteine induziert, die an Protein-Synthese und der Antwort auf ungefaltete Proteine beteiligt sind. Außerdem konnte auf eine Beeinträchtigung des Citratzyklus geschlossen werden. Langkettige Phospholipide sowie die biogenen Amine Taurin und Spermin konnten als vielversprechende Biomarker-Kandidaten für die Aktivierung von THP-1 Zellen durch Kontaktallergene vorgeschlagen werden. Zusammengefasst unterstützt diese Studie die Hypothese, dass Kontaktallergene metabolische Reprogrammierung von THP-1 Zellen auslösen können. Außerdem bestätigten die Ergebnisse, dass Analysen metabolischer Endpunkte ein vielversprechender Ansatz für die Identifizierung von Kontaktallergenen in vitro sein können.
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- 2018
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4. Immunoproteomic identification and characterization of Ni2+-regulated proteins implicates Ni2+ in the induction of monocyte cell death
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Jakob, Annika, primary, Mussotter, Franz, additional, Ohnesorge, Stefanie, additional, Dietz, Lisa, additional, Pardo, Julian, additional, Haidl, Ian D, additional, and Thierse, Hermann-Josef, additional
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- 2017
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5. Application of proteomics in the elucidation of chemical-mediated allergic contact dermatitis
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Höper, Tessa, primary, Mussotter, Franz, additional, Haase, Andrea, additional, Luch, Andreas, additional, and Tralau, Tewes, additional
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- 2017
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6. A proteomics approach to elucidate the role of Nrf2 in primary bone-marrow-derived dendritic cells of mice upon activation by contact allergens
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Mussotter, Franz, primary, Haase, Andrea, additional, Tomm, Janina Melanie, additional, Ali, Zeina El, additional, Kerdine-Römer, Saadia, additional, Pallardy, Marc, additional, von Bergen, Martin, additional, and Luch, Andreas, additional
- Published
- 2014
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7. Application of proteomics in the elucidation of chemical-mediated allergic contact dermatitis
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HöperBoth authors contributed equally., Tessa, Mussotter, Franz, Haase, Andrea, Luch, Andreas, and Tralau, Tewes
- Abstract
Allergic contact dermatitis (ACD) is a widespread hypersensitivity reaction of the skin. The cellular mechanisms underlying its development are complex and involve close interaction of different cell types of the immune system. It is this very complexity which has long prevented straightforward replacement of the corresponding regulatory in vivotests. Recent efforts have already resulted in the development of several in vitrotesting alternatives that address key steps of ACD. Yet identification of suitable biomarkers is still a subject of intense research. Search strategies for the latter encompass transcriptomics, proteomics as well as metabolomics approaches. The scope of this review shall be the application and use of proteomics in the context of ACD. This includes highlighting relevant aspects of the molecular and cellular mechanisms underlying ACD, the exploitation of these mechanisms for testing and biomarkers (e.g., in the context of the OECD's adverse outcome pathway initiative) as well as an outlook on emerging proteome targets, for example during the allergen-induced activation of dendritic cells (DCs).
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- 2017
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8. Immunoproteomic identification and characterization of Ni 2+ -regulated proteins implicates Ni 2+ in the induction of monocyte cell death.
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Jakob A, Mussotter F, Ohnesorge S, Dietz L, Pardo J, Haidl ID, and Thierse HJ
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- Apoptosis drug effects, Caspase 3 metabolism, Caspase 7 metabolism, Cell Nucleus drug effects, Cell Nucleus metabolism, Cells, Cultured, Dendritic Cells drug effects, Dendritic Cells metabolism, Humans, Inflammation metabolism, Monocytes metabolism, Proteomics methods, T-Lymphocytes drug effects, T-Lymphocytes metabolism, Cell Death drug effects, Monocytes drug effects, Nickel pharmacology, Proteome metabolism
- Abstract
Nickel allergy is the most common cause of allergic reactions worldwide, with cutaneous and systemic effects potentially affecting multiple organs. Monocytes are precursors of not only macrophages but also dendritic cells, the most potent activators of nickel hypersensitivity. Monocytes are themselves important antigen-presenting cells, capable of nickel-specific T-cell activation in vivo and in vitro, in addition to being important for immediate innate immune inflammation. To elucidate early Ni
2+ -dependent inflammatory molecular mechanisms in human monocytes, a Ni2+ -specific proteomic approach was applied. Quantitative two-dimensional (2D) differential gel electrophoresis and Delta2D software analyses coupled with matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) revealed that Ni2+ significantly regulated 56 protein species, of which 36 were analyzed by MALDI-MS. Bioinformatics analyses of all identified proteins resulted in Ni2+ -associated functional annotation clusters, such as cell death, metal ion binding, and cytoskeletal remodeling. The involvement of Ni2+ in the induction of monocyte cell death, but not T-cell death, was observed at Ni2+ concentrations at or above 250 μM. Examination of caspase activity during Ni2+ -mediated cell death revealed monocytic cell death independent of caspase-3 and -7 activity. However, confocal microscopy analysis demonstrated Ni2+ -triggered cytoskeletal remodeling and nuclear condensation, characteristic of cellular apoptosis. Thus, Ni2+ -specific peripheral blood mononuclear cell stimulation suggests monocytic cell death at Ni2+ concentrations at or above 250 μM, and monocytic effects on immune regulation at lower Ni2+ concentrations.- Published
- 2017
- Full Text
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