89 results on '"Dunay IR"'
Search Results
2. Microglial sex differences in innate high anxiety and modulatory effects of minocycline.
- Author
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Ugursu B, Sah A, Sartori S, Popp O, Mertins P, Dunay IR, Kettenmann H, Singewald N, and Wolf SA
- Subjects
- Animals, Female, Male, Mice, Brain metabolism, Brain drug effects, Mice, Inbred C57BL, Hippocampus metabolism, Hippocampus drug effects, Disease Models, Animal, Synapses drug effects, Synapses metabolism, Phagocytosis drug effects, Minocycline pharmacology, Microglia metabolism, Microglia drug effects, Anxiety metabolism, Anxiety drug therapy, Sex Characteristics
- Abstract
Microglia modulate synaptic refinement in the central nervous system (CNS). We have previously shown that a mouse model with innate high anxiety-related behavior (HAB) displays higher CD68
+ microglia density in the key regions of anxiety circuits compared to mice with normal anxiety-related behavior (NAB) in males, and that minocycline treatment attenuated the enhanced anxiety of HAB male. Given that a higher prevalence of anxiety is widely reported in females compared to males, little is known concerning sex differences at the cellular level. Herein, we address this by analyzing microglia heterogeneity and function in the HAB and NAB brains of both sexes. Single-cell RNA sequencing revealed ten distinct microglia clusters varied by their frequency and gene expression profile. We report striking sex differences, especially in the major microglia clusters of HABs, indicating a higher expression of genes associated with phagocytosis and synaptic engulfment in the female compared to the male. On a functional level, we show that female HAB microglia engulfed a greater amount of hippocampal vGLUT1+ excitatory synapses compared to the male. We moreover show that female HAB microglia engulfed more synaptosomes compared to the male HAB in vitro. Due to previously reported effects of minocycline on microglia, we finally administered oral minocycline to HABs of both sexes and showed a significant reduction in the engulfment of synapses by female HAB microglia. In parallel to our microglia-specific findings, we further showed an anxiolytic effect of minocycline on female HABs, which is complementary to our previous findings in the male HABs. Our study, therefore, identifies the altered function of synaptic engulfment by microglia as a potential avenue to target and resolve microglia heterogeneity in mice with innate high anxiety., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Author(s). Published by Elsevier Inc. All rights reserved.)- Published
- 2024
- Full Text
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3. Integrity of neural extracellular matrix is required for microglia-mediated synaptic remodeling.
- Author
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Cangalaya C, Sun W, Stoyanov S, Dunay IR, and Dityatev A
- Subjects
- Animals, Mice, Neuronal Plasticity physiology, Neuronal Plasticity drug effects, Complement C3 metabolism, Calreticulin metabolism, Male, Phagocytosis physiology, Phagocytosis drug effects, Mice, Transgenic, Macrophage-1 Antigen metabolism, Microglia metabolism, Microglia drug effects, Extracellular Matrix metabolism, Extracellular Matrix drug effects, Synapses metabolism, Synapses drug effects, Synapses physiology, Complement C1q metabolism, Mice, Inbred C57BL, Chondroitin ABC Lyase pharmacology
- Abstract
Microglia continuously remodel synapses, which are embedded in the extracellular matrix (ECM). However, the mechanisms, which govern this process remain elusive. To investigate the influence of the neural ECM in synaptic remodeling by microglia, we disrupted ECM integrity by injection of chondroitinase ABC (ChABC) into the retrosplenial cortex of healthy adult mice. Using in vivo two-photon microscopy we found that ChABC treatment increased microglial branching complexity and ECM phagocytic capacity and decreased spine elimination rate under basal conditions. Moreover, ECM attenuation largely prevented synaptic remodeling following synaptic stress induced by photodamage of single synaptic elements. These changes were associated with less stable and smaller microglial contacts at the synaptic damage sites, diminished deposition of calreticulin and complement proteins C1q and C3 at synapses and impaired expression of microglial CR3 receptor. Thus, our findings provide novel insights into the function of the neural ECM in deposition of complement proteins and synaptic remodeling by microglia., (© 2024 The Author(s). GLIA published by Wiley Periodicals LLC.)
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- 2024
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4. Microvascular damage, neuroinflammation and extracellular matrix remodeling in Col18a1 knockout mice as a model for early cerebral small vessel disease.
- Author
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Khoshneviszadeh M, Henneicke S, Pirici D, Senthilnathan A, Morton L, Arndt P, Kaushik R, Norman O, Jukkola J, Dunay IR, Seidenbecher C, Heikkinen A, Schreiber S, and Dityatev A
- Subjects
- Animals, Humans, Infant, Mice, Endostatins, Extracellular Matrix metabolism, Heparan Sulfate Proteoglycans metabolism, Mice, Knockout, Cerebral Small Vessel Diseases genetics, Cerebral Small Vessel Diseases metabolism, Collagen Type XVIII genetics, Collagen Type XVIII metabolism, Neuroinflammatory Diseases
- Abstract
Collagen type XVIII (COL18) is an abundant heparan sulfate proteoglycan in vascular basement membranes. Here, we asked (i) if the loss of COL18 would result in blood-brain barrier (BBB) breakdown, pathological alterations of small arteries and capillaries and neuroinflammation as found in cerebral small vessel disease (CSVD) and (ii) if such changes may be associated with remodeling of synapses and neural extracellular matrix (ECM). We found that 5-month-old Col18a1
-/- mice had elevated BBB permeability for mouse IgG in the deep gray matter, and intravascular erythrocyte accumulations were observed brain-wide in capillaries and arterioles. BBB permeability increased with age and affected cortical regions and the hippocampus in 12-month-old Col18a1-/- mice. None of the Col18a1-/- mice displayed hallmarks of advanced CSVD, such as hemorrhages, and did not show perivascular space enlargement. Col18a1 deficiency-induced BBB leakage was accompanied by activation of microglia and astrocytes, a loss of aggrecan in the ECM of perineuronal nets associated with fast-spiking inhibitory interneurons and accumulation of the perisynaptic ECM proteoglycan brevican and the microglial complement protein C1q at excitatory synapses. As the pathway underlying these regulations, we found increased signaling through the TGF-ß1/Smad3/TIMP-3 cascade. We verified the pivotal role of COL18 for small vessel wall structure in CSVD by demonstrating the protein's involvement in vascular remodeling in autopsy brains from patients with cerebral hypertensive arteriopathy. Our study highlights an association between the alterations of perivascular ECM, extracellular proteolysis, and perineuronal/perisynaptic ECM, as a possible substrate of synaptic and cognitive alterations in CSVD., Competing Interests: Declaration of competing interest The authors declare that they have no competing interests., (Copyright © 2024 The Author(s). Published by Elsevier B.V. All rights reserved.)- Published
- 2024
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5. Spatio-temporal dynamics of microglia phenotype in human and murine cSVD: impact of acute and chronic hypertensive states.
- Author
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Morton L, Arndt P, Garza AP, Henneicke S, Mattern H, Gonzalez M, Dityatev A, Yilmazer-Hanke D, Schreiber S, and Dunay IR
- Subjects
- Rats, Humans, Mice, Animals, Microglia metabolism, Rats, Inbred SHR, Phenotype, Hypertension complications, Hypertension pathology, Cerebral Small Vessel Diseases pathology
- Abstract
Vascular risk factors such as chronic hypertension are well-established major modifiable factors for the development of cerebral small vessel disease (cSVD). In the present study, our focus was the investigation of cSVD-related phenotypic changes in microglia in human disease and in the spontaneously hypertensive stroke-prone rat (SHRSP) model of cSVD. Our examination of cortical microglia in human post-mortem cSVD cortical tissue revealed distinct morphological microglial features specific to cSVD. We identified enlarged somata, an increase in the territory occupied by thickened microglial processes, and an expansion in the number of vascular-associated microglia. In parallel, we characterized microglia in a rodent model of hypertensive cSVD along different durations of arterial hypertension, i.e., early chronic and late chronic hypertension. Microglial somata were already enlarged in early hypertension. In contrast, at late-stage chronic hypertension, they further exhibited elongated branches, thickened processes, and a reduced ramification index, mirroring the findings in human cSVD. An unbiased multidimensional flow cytometric analysis revealed phenotypic heterogeneity among microglia cells within the hippocampus and cortex. At early-stage hypertension, hippocampal microglia exhibited upregulated CD11b/c, P2Y12R, CD200R, and CD86 surface expression. Detailed analysis of cell subpopulations revealed a unique microglial subset expressing CD11b/c, CD163, and CD86 exclusively in early hypertension. Notably, even at early-stage hypertension, microglia displayed a higher association with cerebral blood vessels. We identified several profound clusters of microglia expressing distinct marker profiles at late chronic hypertensive states. In summary, our findings demonstrate a higher vulnerability of the hippocampus, stage-specific microglial signatures based on morphological features, and cell surface protein expression in response to chronic arterial hypertension. These results indicate the diversity within microglia sub-populations and implicate the subtle involvement of microglia in cSVD pathogenesis., (© 2023. The Author(s).)
- Published
- 2023
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6. Phytohormones regulate asexual Toxoplasma gondii replication.
- Author
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Wagner T, Bangoura B, Wiedmer S, Daugschies A, and Dunay IR
- Subjects
- Humans, Plant Growth Regulators pharmacology, Abscisic Acid pharmacology, DNA, Toxoplasma, Toxoplasmosis parasitology
- Abstract
The protozoan Toxoplasma gondii (T. gondii) is a zoonotic disease agent causing systemic infection in warm-blooded intermediate hosts including humans. During the acute infection, the parasite infects host cells and multiplies intracellularly in the asexual tachyzoite stage. In this stage of the life cycle, invasion, multiplication, and egress are the most critical events in parasite replication. T. gondii features diverse cell organelles to support these processes, including the apicoplast, an endosymbiont-derived vestigial plastid originating from an alga ancestor. Previous studies have highlighted that phytohormones can modify the calcium-mediated secretion, e.g., of adhesins involved in parasite movement and cell invasion processes. The present study aimed to elucidate the influence of different plant hormones on the replication of asexual tachyzoites in a human foreskin fibroblast (HFF) host cell culture. T. gondii replication was measured by the determination of T. gondii DNA copies via qPCR. Three selected phytohormones, namely abscisic acid (ABA), gibberellic acid (GIBB), and kinetin (KIN) as representatives of different plant hormone groups were tested. Moreover, the influence of typical cell culture media components on the phytohormone effects was assessed. Our results indicate that ABA is able to induce a significant increase of T. gondii DNA copies in a typical supplemented cell culture medium when applied in concentrations of 20 ng/μl or 2 ng/μl, respectively. In contrast, depending on the culture medium composition, GIBB may potentially serve as T. gondii growth inhibitor and may be further investigated as a potential treatment for toxoplasmosis., (© 2023. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)
- Published
- 2023
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7. Astrocytes evoke a robust IRF7-independent type I interferon response upon neurotropic viral infection.
- Author
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Weichert L, Düsedau HP, Fritzsch D, Schreier S, Scharf A, Grashoff M, Cebulski K, Michaelsen-Preusse K, Erck C, Lienenklaus S, Dunay IR, and Kröger A
- Subjects
- Animals, Mice, Antibodies, Astrocytes, Central Nervous System, Interferon Regulatory Factor-7 genetics, Interferon Type I, Encephalitis, Tick-Borne immunology
- Abstract
Background: Type I interferons (IFN-I) are fundamental in controlling viral infections but fatal interferonopathy is restricted in the immune-privileged central nervous system (CNS). In contrast to the well-established role of Interferon Regulatory Factor 7 (IRF7) in the regulation of IFN-I response in the periphery, little is known about the specific function in the CNS., Methods: To investigate the role for IRF7 in antiviral response during neurotropic virus infection, mice deficient for IRF3 and IRF7 were infected systemically with Langat virus (LGTV). Viral burden and IFN-I response was analyzed in the periphery and the CNS by focus formation assay, RT-PCR, immunohistochemistry and in vivo imaging. Microglia and infiltration of CNS-infiltration of immune cells were characterized by flow cytometry., Results: Here, we demonstrate that during infection with the neurotropic Langat virus (LGTV), an attenuated member of the tick-borne encephalitis virus (TBEV) subgroup, neurons do not rely on IRF7 for cell-intrinsic antiviral resistance and IFN-I induction. An increased viral replication in IRF7-deficient mice suggests an indirect antiviral mechanism. Astrocytes rely on IRF7 to establish a cell-autonomous antiviral response. Notably, the loss of IRF7 particularly in astrocytes resulted in a high IFN-I production. Sustained production of IFN-I in astrocytes is independent of an IRF7-mediated positive feedback loop., Conclusion: IFN-I induction in the CNS is profoundly regulated in a cell type-specific fashion., (© 2023. BioMed Central Ltd., part of Springer Nature.)
- Published
- 2023
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8. Plasma concentrations of anti-inflammatory cytokine TGF-β are associated with hippocampal structure related to explicit memory performance in older adults.
- Author
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Raschick M, Richter A, Fischer L, Knopf L, Schult A, Yakupov R, Behnisch G, Guttek K, Düzel E, Dunay IR, Seidenbecher CI, Schraven B, Reinhold D, and Schott BH
- Subjects
- Humans, Aged, Magnetic Resonance Imaging, Neuropsychological Tests, Hippocampus diagnostic imaging, Cognition, Anti-Inflammatory Agents, Cytokines, Transforming Growth Factor beta
- Abstract
Human cognitive abilities, and particularly hippocampus-dependent memory performance typically decline with increasing age. Immunosenescence, the age-related disintegration of the immune system, is increasingly coming into the focus of research as a considerable factor contributing to cognitive decline. In the present study, we investigated potential associations between plasma levels of pro- and anti-inflammatory cytokines and learning and memory performance as well as hippocampal anatomy in young and older adults. Plasma concentrations of the inflammation marker CRP as well as the pro-inflammatory cytokines IL-6 and TNF-α and the anti-inflammatory cytokine TGF-β
1 were measured in 142 healthy adults (57 young, 24.47 ± 4.48 years; 85 older, 63.66 ± 7.32 years) who performed tests of explicit memory (Verbal Learning and Memory Test, VLMT; Wechsler Memory Scale, Logical Memory, WMS) with an additional delayed recall test after 24 h. Hippocampal volumetry and hippocampal subfield segmentation were performed using FreeSurfer, based on T1-weighted and high-resolution T2-weighted MR images. When investigating the relationship between memory performance, hippocampal structure, and plasma cytokine levels, we found that TGF-β1 concentrations were positively correlated with the volumes of the hippocampal CA4-dentate gyrus region in older adults. These volumes were in turn positively associated with better performance in the WMS, particularly in the delayed memory test. Our results support the notion that endogenous anti-inflammatory mechanisms may act as protective factors in neurocognitive aging., (© 2023. The Author(s).)- Published
- 2023
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9. Classifying flow cytometry data using Bayesian analysis helps to distinguish ALS patients from healthy controls.
- Author
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Räuber S, Nelke C, Schroeter CB, Barman S, Pawlitzki M, Ingwersen J, Akgün K, Günther R, Garza AP, Marggraf M, Dunay IR, Schreiber S, Vielhaber S, Ziemssen T, Melzer N, Ruck T, Meuth SG, and Herty M
- Subjects
- Bayes Theorem, Algorithms, Humans, Models, Theoretical, Male, Female, Adult, Middle Aged, Aged, Aged, 80 and over, Flow Cytometry classification, Flow Cytometry methods, Amyotrophic Lateral Sclerosis diagnosis
- Abstract
Introduction: Given its wide availability and cost-effectiveness, multidimensional flow cytometry (mFC) became a core method in the field of immunology allowing for the analysis of a broad range of individual cells providing insights into cell subset composition, cellular behavior, and cell-to-cell interactions. Formerly, the analysis of mFC data solely relied on manual gating strategies. With the advent of novel computational approaches, (semi-)automated gating strategies and analysis tools complemented manual approaches., Methods: Using Bayesian network analysis, we developed a mathematical model for the dependencies of different obtained mFC markers. The algorithm creates a Bayesian network that is a HC tree when including raw, ungated mFC data of a randomly selected healthy control cohort (HC). The HC tree is used to classify whether the observed marker distribution (either patients with amyotrophic lateral sclerosis (ALS) or HC) is predicted. The relative number of cells where the probability q is equal to zero is calculated reflecting the similarity in the marker distribution between a randomly chosen mFC file (ALS or HC) and the HC tree., Results: Including peripheral blood mFC data from 68 ALS and 35 HC, the algorithm could correctly identify 64/68 ALS cases. Tuning of parameters revealed that the combination of 7 markers, 200 bins, and 20 patients achieved the highest AUC on a significance level of p < 0.0001. The markers CD4 and CD38 showed the highest zero probability. We successfully validated our approach by including a second, independent ALS and HC cohort (55 ALS and 30 HC). In this case, all ALS were correctly identified and side scatter and CD20 yielded the highest zero probability. Finally, both datasets were analyzed by the commercially available algorithm 'Citrus', which indicated superior ability of Bayesian network analysis when including raw, ungated mFC data., Discussion: Bayesian network analysis might present a novel approach for classifying mFC data, which does not rely on reduction techniques, thus, allowing to retain information on the entire dataset. Future studies will have to assess the performance when discriminating clinically relevant differential diagnoses to evaluate the complementary diagnostic benefit of Bayesian network analysis to the clinical routine workup., Competing Interests: SR received travel reimbursements from Merck Healthcare Germany GmbH and Alexion Pharmaceuticals. She served on a scientific advisory board from Merck Healthcare Germany GmbH and her research was supported by Novartis and the Stiftung zur Förderung junger Neurowissenschaftler. CS received travel reimbursements from Merck Healthcare Germany GmbH and Alexion Pharmaceuticals. She served on a scientific advisory board from Merck Healthcare Germany GmbH. MP received honoraria for lecturing from Argenx, Biogen, Bayer, Novartis, Hexal, Sanofi and Merck. He received research funding from Biogen. KA received personal compensation for consulting services and speaker honoraria from Roche, Novartis, Merck, Sanofi, Teva, BMS, Biogen, and Celgene. SM received honoraria for lecturing and travel expenses for attending meetings from Almirall, Amicus Therapeutics Germany, Bayer HealthCare, Biogen, Celgene, Diamed, Genzyme, MedDay Pharmaceuticals, Merck Serono, Novartis, Novo Nordisk, ONO Pharma, Roche, Sanofi-Aventis, Chugai Pharma, QuintilesIMS, and Teva. His research is funded by the German Ministry for Education and Research BMBF, Bundesinstitut für Risikobewertung BfR, Deutsche Forschungsgemeinschaft DFG, Else Kröner Fresenius Foundation, Gemeinsamer Bundesausschuss G-BA, German Academic Exchange Service, Hertie Foundation, Interdisciplinary Center for Clinical Studies IZKF Muenster, German Foundation Neurology and by Alexion, Almirall, Amicus Therapeutics Germany, Biogen, Diamed, Fresenius Medical Care, Genzyme, HERZ Burgdorf, Merck Serono, Novartis, ONO Pharma, Roche, and Teva. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Räuber, Nelke, Schroeter, Barman, Pawlitzki, Ingwersen, Akgün, Günther, Garza, Marggraf, Dunay, Schreiber, Vielhaber, Ziemssen, Melzer, Ruck, Meuth and Herty.)
- Published
- 2023
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10. Deletion of p75 NTR rescues the synaptic but not the inflammatory status in the brain of a mouse model for Alzheimer's disease.
- Author
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Demuth H, Hosseini S, Düsedeau HP, Dunay IR, Korte M, and Zagrebelsky M
- Abstract
Introduction: Alzheimer's disease (AD), is characterized by a gradual cognitive decline associated with the accumulation of Amyloid beta (Aβ)-oligomers, progressive neuronal degeneration and chronic neuroinflammation. Among the receptors shown to bind and possibly transduce the toxic effects of Aβ-oligomers is the p75 neurotrophin receptor (p75
NTR ). Interestingly, p75NTR mediates several crucial processes in the nervous system, including neuronal survival and apoptosis, maintenance of the neuronal architecture, and plasticity. Furthermore, p75NTR is also expressed in microglia, the resident immune cells of the brain, where it is markedly increased under pathological conditions. These observations indicate p75NTR as a potential candidate for mediating Aβ-induced toxic effects at the interface between the nervous and the immune system, thereby potentially participating in the crosstalk between these two systems., Methods: Here we used APP/PS1 transgenic mice (APP/PS1tg) and compared the Aβ-induced alterations in neuronal function, chronic inflammation as well as their cognitive consequences between 10 months old APP/PS1tg and APP/PS1tg x p75NTRexonIV knockout mice., Results: Electrophysiological recordings show that a loss of p75NTR rescues the impairment in long-term potentiation at the Schaffer collaterals in the hippocampus of APP/PS1tg mice. Interestingly, however loss of p75NTR does not influence the severity of neuroinflammation, microglia activation or the decline in spatial learning and memory processes observed in APP/PS1tg mice., Conclusion: Together these results indicate that while a deletion of p75NTR rescues the synaptic defect and the impairment in synaptic plasticity, it does not affect the progression of the neuroinflammation and the cognitive decline in a mouse model for AD., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Demuth, Hosseini, Düsedeau, Dunay, Korte and Zagrebelsky.)- Published
- 2023
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11. Brain Vascular Health in ALS Is Mediated through Motor Cortex Microvascular Integrity.
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Schreiber S, Bernal J, Arndt P, Schreiber F, Müller P, Morton L, Braun-Dullaeus RC, Valdés-Hernández MDC, Duarte R, Wardlaw JM, Meuth SG, Mietzner G, Vielhaber S, Dunay IR, Dityatev A, Jandke S, and Mattern H
- Subjects
- Humans, Vascular Endothelial Growth Factor A metabolism, Motor Neurons pathology, Blood-Brain Barrier pathology, Amyotrophic Lateral Sclerosis metabolism, Motor Cortex metabolism
- Abstract
Brain vascular health appears to be critical for preventing the development of amyotrophic lateral sclerosis (ALS) and slowing its progression. ALS patients often demonstrate cardiovascular risk factors and commonly suffer from cerebrovascular disease, with evidence of pathological alterations in their small cerebral blood vessels. Impaired vascular brain health has detrimental effects on motor neurons: vascular endothelial growth factor levels are lowered in ALS, which can compromise endothelial cell formation and the integrity of the blood-brain barrier. Increased turnover of neurovascular unit cells precedes their senescence, which, together with pericyte alterations, further fosters the failure of toxic metabolite removal. We here provide a comprehensive overview of the pathogenesis of impaired brain vascular health in ALS and how novel magnetic resonance imaging techniques can aid its detection. In particular, we discuss vascular patterns of blood supply to the motor cortex with the number of branches from the anterior and middle cerebral arteries acting as a novel marker of resistance and resilience against downstream effects of vascular risk and events in ALS. We outline how certain interventions adapted to patient needs and capabilities have the potential to mechanistically target the brain microvasculature towards favorable motor cortex blood supply patterns. Through this strategy, we aim to guide novel approaches to ALS management and a better understanding of ALS pathophysiology.
- Published
- 2023
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12. Initial and ongoing tobacco smoking elicits vascular damage and distinct inflammatory response linked to neurodegeneration.
- Author
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Garza AP, Morton L, Pállinger É, Buzás EI, Schreiber S, Schott BH, and Dunay IR
- Abstract
Tobacco smoking is strongly linked to vascular damage contributing to the development of hypertension, atherosclerosis, as well as increasing the risk for neurodegeneration. Still, the involvement of the innate immune system in the development of vascular damage upon chronic tobacco use before the onset of clinical symptoms is not fully characterized. Our data provide evidence that a single acute exposure to tobacco elicits the secretion of extracellular vesicles expressing CD105 and CD49e from endothelial cells, granting further recognition of early preclinical biomarkers of vascular damage. Furthermore, we investigated the effects of smoking on the immune system of healthy asymptomatic chronic smokers compared to never-smokers, focusing on the innate immune system. Our data reveal a distinct immune landscape representative for early stages of vascular damage in clinically asymptomatic chronic smokers, before tobacco smoking related diseases develop. These results indicate a dysregulated immuno-vascular axis in chronic tobacco smokers that are otherwise considered as healthy individuals. The distinct alterations are characterized by increased CD36 expression by the blood monocyte subsets, neutrophilia and increased plasma IL-18 and reduced levels of IL-33, IL-10 and IL-8. Additionally, reduced levels of circulating BDNF and elevated sTREM2, which are associated with neurodegeneration, suggest a considerable impact of tobacco smoking on CNS function in clinically healthy individuals. These findings provide profound insight into the initial and ongoing effects of tobacco smoking and the potential vascular damage contributing to neurodegenerative disorders, specifically cerebrovascular dysfunction and dementia., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (© 2023 The Authors.)
- Published
- 2023
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13. IFN-γ-mediated neuronal defense mechanism targets Toxoplasma.
- Author
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Figueiredo CA and Dunay IR
- Subjects
- Interferon-gamma, Neurons, Immunity, Cellular, Defense Mechanisms, Toxoplasma
- Abstract
Toxoplasma gondii encysts preferentially within neurons in the central nervous system, establishing lifelong persistence. Despite recent discoveries, this neuronal preference was thought, in part, to be secondary to a lack of neuronal cell-autonomous immunity. By showing that neurons can mount interferon-gamma (IFN-γ)-mediated cell-autonomous anti-T. gondii defenses, Chandrasekaran et al. have challenged long held assumptions., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2022 Elsevier Ltd. All rights reserved.)
- Published
- 2022
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14. The neuropeptide PACAP alleviates T. gondii infection-induced neuroinflammation and neuronal impairment.
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Figueiredo CA, Düsedau HP, Steffen J, Ehrentraut S, Dunay MP, Toth G, Reglödi D, Heimesaat MM, and Dunay IR
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- Humans, Pituitary Adenylate Cyclase-Activating Polypeptide pharmacology, Pituitary Adenylate Cyclase-Activating Polypeptide therapeutic use, Pituitary Adenylate Cyclase-Activating Polypeptide metabolism, Neuroinflammatory Diseases, Nerve Growth Factors, Inflammation drug therapy, Inflammation Mediators, Toxoplasmosis complications, Toxoplasmosis drug therapy, Toxoplasma
- Abstract
Background: Cerebral infection with the protozoan Toxoplasma gondii (T. gondii) is responsible for inflammation of the central nervous system (CNS) contributing to subtle neuronal alterations. Albeit essential for brain parasite control, continuous microglia activation and recruitment of peripheral immune cells entail distinct neuronal impairment upon infection-induced neuroinflammation. PACAP is an endogenous neuropeptide known to inhibit inflammation and promote neuronal survival. Since PACAP is actively transported into the CNS, we aimed to assess the impact of PACAP on the T. gondii-induced neuroinflammation and subsequent effects on neuronal homeostasis., Methods: Exogenous PACAP was administered intraperitoneally in the chronic stage of T. gondii infection, and brains were isolated for histopathological analysis and determination of pathogen levels. Immune cells from the brain, blood, and spleen were analyzed by flow cytometry, and the further production of inflammatory mediators was investigated by intracellular protein staining as well as expression levels by RT-qPCR. Neuronal and synaptic alterations were assessed on the transcriptional and protein level, focusing on neurotrophins, neurotrophin-receptors and signature synaptic markers., Results: Here, we reveal that PACAP administration reduced the inflammatory foci and the number of apoptotic cells in the brain parenchyma and restrained the activation of microglia and recruitment of monocytes. The neuropeptide reduced the expression of inflammatory mediators such as IFN-γ, IL-6, iNOS, and IL-1β. Moreover, PACAP diminished IFN-γ production by recruited CD4
+ T cells in the CNS. Importantly, PACAP promoted neuronal health via increased expression of the neurotrophin BDNF and reduction of p75NTR , a receptor related to neuronal cell death. In addition, PACAP administration was associated with increased expression of transporters involved in glutamatergic and GABAergic signaling that are particularly affected during cerebral toxoplasmosis., Conclusions: Together, our findings unravel the beneficial effects of exogenous PACAP treatment upon infection-induced neuroinflammation, highlighting the potential implication of neuropeptides to promote neuronal survival and minimize synaptic prejudice., (© 2022. The Author(s).)- Published
- 2022
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15. Persisting Microbiota and Neuronal Imbalance Following T. gondii Infection Reliant on the Infection Route.
- Author
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French T, Steffen J, Glas A, Osbelt L, Strowig T, Schott BH, Schüler T, and Dunay IR
- Subjects
- Animals, Cytokines, Inflammation, Mice, Mice, Inbred C57BL, Neurons, Microbiota, Toxoplasma
- Abstract
Toxoplasma gondii is a highly successful parasite capable of infecting all warm-blooded animals. The natural way of infection in intermediate hosts is the oral ingestion of parasite-contaminated water or food. In murine experimental models, oral infection ( p.o. ) of mice with T. gondii is applied to investigate mucosal and peripheral immune cell dynamics, whereas intraperitoneal infection ( i.p. ) is frequently used to study peripheral inflammation as well as immune cell - neuronal interaction in the central nervous system (CNS). However, the two infection routes have not yet been systematically compared along the course of infection. Here, C57BL/6 mice were infected p.o. or i.p. with a low dose of T. gondii cysts, and the acute and chronic stages of infection were compared. A more severe course of infection was detected following i.p. challenge, characterized by an increased weight loss and marked expression of proinflammatory cytokines particularly in the CNS during the chronic stage. The elevated proinflammatory cytokine expression in the ileum was more prominent after p.o. challenge that continued following the acute phase in both i.p. or p.o. infected mice. This resulted in sustained microbial dysbiosis, especially after p.o. challenge, highlighted by increased abundance of pathobionts from the phyla proteobacteria and a reduction of beneficial commensal species. Further, we revealed that in the CNS of i.p. infected mice CD4 and CD8 T cells displayed higher IFNγ production in the chronic stage. This corresponded with an increased expression of C1q and CD68 in the CNS and reduced expression of genes involved in neuronal signal transmission. Neuroinflammation-associated synaptic alterations, especially PSD-95, VGLUT, and EAAT2 expression, were more pronounced in the cortex upon i.p. infection highlighting the profound interplay between peripheral inflammation and CNS homeostasis., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 French, Steffen, Glas, Osbelt, Strowig, Schott, Schüler and Dunay.)
- Published
- 2022
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16. Acute alcohol consumption increases systemic endotoxin bioactivity for days in healthy volunteers-with reduced intestinal barrier loss in female.
- Author
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Sturm R, Haag F, Janicova A, Xu B, Vollrath JT, Bundkirchen K, Dunay IR, Neunaber C, Marzi I, and Relja B
- Subjects
- Alcohol Drinking adverse effects, Biomarkers, Blood Alcohol Content, Endotoxins, Female, Healthy Volunteers, Humans, Syndecan-1, Young Adult, Alcoholic Intoxication, Lipopolysaccharide Receptors
- Abstract
Objective: Trauma is the most common cause of death among young adults. Alcohol intoxication plays a significant role as a cause of accidents and as a potent immunomodulator of the post-traumatic response to tissue injury. Polytraumatized patients are frequently at risk to developing infectious complications, which may be aggravated by alcohol-induced immunosuppression. Systemic levels of integral proteins of the gastrointestinal tract such as syndecan-1 or intestinal fatty acid binding proteins (FABP-I) reflect the intestinal barrier function. The exact impact of acute alcohol intoxication on the barrier function and endotoxin bioactivity have not been clarified yet., Methods: 22 healthy volunteers received a precisely defined amount of alcohol (whiskey-cola) every 20 min over a period of 4 h to reach the calculated blood alcohol concentration (BAC) of 1‰. Blood samples were taken before alcohol drinking as a control, and after 2, 4, 6, 24 and 48 h after beginning with alcohol consumption. In addition, urine samples were collected. Intestinal permeability was determined by serum and urine values of FABP-I, syndecan-1, and soluble (s)CD14 as a marker for the endotoxin translocation via the intestinal barrier by ELISA. BAC was determined., Results: Systemic FABP-I was significantly reduced 2 h after the onset of alcohol drinking, and remained decreased after 4 h. However, at 6 h, FABP-I significantly elevated compared to previous measurements as well as to controls (p < 0.05). Systemic sCD14 was significantly elevated after 6, 24 and 48 h after the onset of alcohol consumption (p < 0.05). Systemic FABP-I at 2 h after drinking significantly correlated with the sCD14 concentration after 24 h indicating an enhanced systemic LPS bioactivity. Women showed significantly lower levels of syndecan-1 in serum and urine and urine for all time points until 6 h and lower FABP-I in the serum after 2 h., Conclusions: Even relative low amounts of alcohol affect the immune system of healthy volunteers, although these changes appear minor in women. A potential damage to the intestinal barrier and presumed enhanced systemic endotoxin bioactivity after acute alcohol consumption is proposed, which represents a continuous immunological challenge for the organism and should be considered for the following days after drinking., (© 2021. The Author(s).)
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- 2022
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17. IFNAR signaling in fibroblastic reticular cells can modulate CD8 + memory fate decision.
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Knop L, Spanier J, Larsen PK, Witte A, Bank U, Dunay IR, Kalinke U, and Schüler T
- Subjects
- Animals, CD8-Positive T-Lymphocytes, Fibroblasts, Mice, Mice, Inbred C57BL, Vaccines, Subunit, Cancer Vaccines, Vesicular Stomatitis metabolism, Vesicular Stomatitis pathology
- Abstract
CD8
+ memory T cells (TM ) are crucial for long-term protection from infections and cancer. Multiple cell types and cytokines are involved in the regulation of CD8+ T cell responses and subsequent TM formation. Besides their direct antiviral effects, type I interferons (IFN-I) modulate CD8+ T cell immunity via their action on several immune cell subsets. However, it is largely unclear how nonimmune cells are involved in this multicellular network modulating CD8+ TM formation. Fibroblastic reticular cells (FRCs) form the 3D scaffold of secondary lymphoid organs, express the IFN-I receptor (IFNAR), and modulate adaptive immune responses. However, it is unclear whether and how early IFNAR signals in lymph node (LN) FRCs affect CD8+ TM differentiation. Using peptide vaccination and viral infection, we studied CD8+ TM differentiation in mice with an FRC-specific IFNAR deletion (FRCΔIFNAR ). We show here that the differentiation of CD8+ TCR-transgenic T cells into central memory cells (TCM ) is enhanced in peptide-vaccinated FRCΔIFNAR mice. Conversely, vesicular stomatitis virus infection of FRCΔIFNAR mice is associated with impaired TCM formation and the accumulation of vesicular stomatitis virus specific double-positive CD127hi KLRG-1hi effector memory T cells. In summary, we provide evidence for a context-dependent contribution of FRC-specific IFNAR signaling to CD8+ TM differentiation., (© 2022 The Authors. European Journal of Immunology published by Wiley-VCH GmbH.)- Published
- 2022
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18. Type 1 innate lymphoid cells regulate the onset of Toxoplasma gondii-induced neuroinflammation.
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Steffen J, Ehrentraut S, Bank U, Biswas A, Figueiredo CA, Hölsken O, Düsedau HP, Dovhan V, Knop L, Thode J, Romero-Suárez S, Duarte CI, Gigley J, Romagnani C, Diefenbach A, Klose CSN, Schüler T, and Dunay IR
- Subjects
- Animals, Immunity, Innate, Killer Cells, Natural, Mice, Neuroinflammatory Diseases, Toxoplasma, Toxoplasmosis
- Abstract
Cerebral infections are restrained by a complex interplay of tissue-resident and recruited peripheral immune cells. Whether innate lymphoid cells (ILCs) are involved in the orchestration of the neuroinflammatory dynamics is not fully understood. Here, we demonstrate that ILCs accumulate in the cerebral parenchyma, the choroid plexus, and the meninges in the onset of cerebral Toxoplasma gondii infection. Antibody-mediated depletion of conventional natural killer (cNK) cells and ILC1s in the early stage of infection results in diminished cytokine and chemokine expression and increased cerebral parasite burden. Using cNK- and ILC1-deficient murine models, we demonstrate that exclusively the lack of ILC1s affects cerebral immune responses. In summary, our results provide evidence that ILC1s are an early source of IFN-γ and TNF in response to cerebral T. gondii infection, thereby inducing host defense factors and initiating the development of a neuroinflammatory response., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2022 The Authors. Published by Elsevier Inc. All rights reserved.)
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- 2022
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19. Immune response and pathogen invasion at the choroid plexus in the onset of cerebral toxoplasmosis.
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Figueiredo CA, Steffen J, Morton L, Arumugam S, Liesenfeld O, Deli MA, Kröger A, Schüler T, and Dunay IR
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- Blood-Brain Barrier metabolism, Endothelial Cells, Humans, Immunity, Choroid Plexus metabolism, Toxoplasmosis, Cerebral metabolism
- Abstract
Background: Toxoplasma gondii (T. gondii) is a highly successful parasite being able to cross all biological barriers of the body, finally reaching the central nervous system (CNS). Previous studies have highlighted the critical involvement of the blood-brain barrier (BBB) during T. gondii invasion and development of subsequent neuroinflammation. Still, the potential contribution of the choroid plexus (CP), the main structure forming the blood-cerebrospinal fluid (CSF) barrier (BCSFB) have not been addressed., Methods: To investigate T. gondii invasion at the onset of neuroinflammation, the CP and brain microvessels (BMV) were isolated and analyzed for parasite burden. Additionally, immuno-stained brain sections and three-dimensional whole mount preparations were evaluated for parasite localization and morphological alterations. Activation of choroidal and brain endothelial cells were characterized by flow cytometry. To evaluate the impact of early immune responses on CP and BMV, expression levels of inflammatory mediators, tight junctions (TJ) and matrix metalloproteinases (MMPs) were quantified. Additionally, FITC-dextran was applied to determine infection-related changes in BCSFB permeability. Finally, the response of primary CP epithelial cells to T. gondii parasites was tested in vitro., Results: Here we revealed that endothelial cells in the CP are initially infected by T. gondii, and become activated prior to BBB endothelial cells indicated by MHCII upregulation. Additionally, CP elicited early local immune response with upregulation of IFN-γ, TNF, IL-6, host-defence factors as well as swift expression of CXCL9 chemokine, when compared to the BMV. Consequently, we uncovered distinct TJ disturbances of claudins, associated with upregulation of MMP-8 and MMP-13 expression in infected CP in vivo, which was confirmed by in vitro infection of primary CP epithelial cells. Notably, we detected early barrier damage and functional loss by increased BCSFB permeability to FITC-dextran in vivo, which was extended over the infection course., Conclusions: Altogether, our data reveal a close interaction between T. gondii infection at the CP and the impairment of the BCSFB function indicating that infection-related neuroinflammation is initiated in the CP., (© 2022. The Author(s).)
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- 2022
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20. Influenza A Virus (H1N1) Infection Induces Microglial Activation and Temporal Dysbalance in Glutamatergic Synaptic Transmission.
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Düsedau HP, Steffen J, Figueiredo CA, Boehme JD, Schultz K, Erck C, Korte M, Faber-Zuschratter H, Smalla KH, Dieterich D, Kröger A, Bruder D, and Dunay IR
- Subjects
- Animals, Brain pathology, Chemokines, Cytokines, Gene Expression, Humans, Inflammation virology, Influenza, Human virology, Mice, Orthomyxoviridae Infections virology, Excitatory Amino Acid Agents pharmacology, Influenza A Virus, H1N1 Subtype immunology, Influenza A virus genetics, Microglia metabolism, Synaptic Transmission physiology
- Abstract
Influenza A virus (IAV) causes respiratory tract disease and is responsible for seasonal and reoccurring epidemics affecting all age groups. Next to typical disease symptoms, such as fever and fatigue, IAV infection has been associated with behavioral alterations presumably contributing to the development of major depression. Previous experiments using IAV/H1N1 infection models have shown impaired hippocampal neuronal morphology and cognitive abilities, but the underlying pathways have not been fully described. In this study, we demonstrate that infection with a low-dose non-neurotrophic H1N1 strain of IAV causes ample peripheral immune response followed by a temporary blood-brain barrier disturbance. Although histological examination did not reveal obvious pathological processes in the brains of IAV-infected mice, detailed multidimensional flow cytometric characterization of immune cells uncovered subtle alterations in the activation status of microglial cells. More specifically, we detected an altered expression pattern of major histocompatibility complex classes I and II, CD80, and F4/80 accompanied by elevated mRNA levels of CD36, CD68, C1QA, and C3, suggesting evolved synaptic pruning. To closer evaluate how these profound changes affect synaptic balance, we established a highly sensitive multiplex flow cytometry-based approach called flow synaptometry. The introduction of this novel technique enabled us to simultaneously quantify the abundance of pre- and postsynapses from distinct brain regions. Our data reveal a significant reduction of VGLUT1 in excitatory presynaptic terminals in the cortex and hippocampus, identifying a subtle dysbalance in glutamatergic synapse transmission upon H1N1 infection in mice. In conclusion, our results highlight the consequences of systemic IAV-triggered inflammation on the central nervous system and the induction and progression of neuronal alterations. IMPORTANCE Influenza A virus (IAV) causes mainly respiratory tract disease with fever and fatigue but is also associated with behavioral alterations in humans. Here, we demonstrate that infection with a low-dose non-neurotrophic H1N1 strain of IAV causes peripheral immune response followed by a temporary blood-brain barrier disturbance. Characterization of immune cells uncovered subtle alterations in the activation status of microglia cells that might reshape neuronal synapses. We established a highly sensitive multiplex flow cytometry-based approach called flow synaptometry to more closely study the synapses. Thus, we detected a specific dysbalance in glutamatergic synapse transmission upon H1N1 infection in mice. In conclusion, our results highlight the consequences of systemic IAV-triggered inflammation on the central nervous system and the induction and progression of neuronal alterations.
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- 2021
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21. Enhanced Susceptibility of ADAP-Deficient Mice to Listeria monocytogenes Infection Is Associated With an Altered Phagocyte Phenotype and Function.
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Böning MAL, Parzmair GP, Jeron A, Düsedau HP, Kershaw O, Xu B, Relja B, Schlüter D, Dunay IR, Reinhold A, Schraven B, and Bruder D
- Subjects
- Adaptor Proteins, Signal Transducing metabolism, Animals, Disease Models, Animal, Disease Susceptibility, Female, Immunity, Listeriosis metabolism, Listeriosis microbiology, Liver metabolism, Male, Mice, Mice, Knockout, Phagocytes metabolism, Phenotype, Spleen metabolism, Adaptor Proteins, Signal Transducing genetics, Listeria monocytogenes immunology, Listeriosis immunology, Phagocytes immunology
- Abstract
The adhesion and degranulation-promoting adaptor protein (ADAP) serves as a multifunctional scaffold and is involved in the formation of immune signaling complexes. To date, only limited data exist regarding the role of ADAP in pathogen-specific immunity during in vivo infection, and its contribution in phagocyte-mediated antibacterial immunity remains elusive. Here, we show that mice lacking ADAP (ADAPko) are highly susceptible to the infection with the intracellular pathogen Listeria monocytogenes ( Lm ) by showing enhanced immunopathology in infected tissues together with increased morbidity, mortality, and excessive infiltration of neutrophils and monocytes. Despite high phagocyte numbers in the spleen and liver, ADAPko mice only inefficiently controlled pathogen growth, hinting at a functional impairment of infection-primed phagocytes in the ADAP-deficient host. Flow cytometric analysis of hallmark pro-inflammatory mediators and unbiased whole genome transcriptional profiling of neutrophils and inflammatory monocytes uncovered broad molecular alterations in the inflammatory program in both phagocyte subsets following their activation in the ADAP-deficient host. Strikingly, ex vivo phagocytosis assay revealed impaired phagocytic capacity of neutrophils derived from Lm -infected ADAPko mice. Together, our data suggest that an alternative priming of phagocytes in ADAP-deficient mice during Lm infection induces marked alterations in the inflammatory profile of neutrophils and inflammatory monocytes that contribute to enhanced immunopathology while limiting their capacity to eliminate the pathogen and to prevent the fatal outcome of the infection., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Böning, Parzmair, Jeron, Düsedau, Kershaw, Xu, Relja, Schlüter, Dunay, Reinhold, Schraven and Bruder.)
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- 2021
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22. Robustly High Hippocampal BDNF levels under Acute Stress in Mice Lacking the Full-length p75 Neurotrophin Receptor.
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Schott BH, Kronenberg G, Schmidt U, Düsedau HP, Ehrentraut S, Geisel O, von Bohlen Und Halbach O, Gass P, Dunay IR, and Hellweg R
- Subjects
- Animals, Hippocampus metabolism, Mice, Receptors, Nerve Growth Factor genetics, Receptors, Nerve Growth Factor metabolism, Signal Transduction, Brain-Derived Neurotrophic Factor genetics, Brain-Derived Neurotrophic Factor metabolism, Receptor, Nerve Growth Factor metabolism
- Abstract
Background: Brain-derived neurotrophic factor (BDNF) exerts its effects on neural plasticity via 2 distinct receptor types, the tyrosine kinase TrkB and the p75 neurotrophin receptor (p75NTR). The latter can promote inflammation and cell death while TrkB is critically involved in plasticity and memory, particularly in the hippocampus. Acute and chronic stress have been associated with suppression of hippocampal BDNF expression and impaired hippocampal plasticity. We hypothesized that p75NTR might be involved in the hippocampal stress response, in particular in stress-induced BDNF suppression, which might be accompanied by increased neuroinflammation., Method: We assessed hippocampal BDNF protein concentrations in wild-type mice compared that in mice lacking the long form of the p75NTR (p75NTR
ExIII-/- ) with or without prior exposure to a 1-hour restraint stress challenge. Hippocampal BDNF concentrations were measured using an optimized ELISA. Furthermore, whole-brain mRNA expression of pro-inflammatory interleukin-6 ( Il6 ) was assessed with RT-PCR., Results: Deletion of full-length p75NTR was associated with higher hippocampal BDNF protein concentration in the stress condition, suggesting persistently high hippocampal BDNF levels in p75NTR-deficient mice, even under stress. Stress elicited increased whole-brain Il6 mRNA expression irrespective of genotype; however, p75NTRExIII-/- mice showed elevated baseline Il6 expression and thus a lower relative increase., Conclusions: Our results provide evidence for a role of p75NTR signaling in the regulation of hippocampal BDNF levels, particularly under stress. Furthermore, p75NTR signaling modulates baseline but not stress-related Il6 gene expression in mice. Our findings implicate p75NTR signaling as a potential pathomechanism in BDNF-dependent modulation of risk for neuropsychiatric disorders., Competing Interests: The authors declare that they have no conflict of interest., (Thieme. All rights reserved.)- Published
- 2021
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23. Toxoplasma gondii infection and its implications within the central nervous system.
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Matta SK, Rinkenberger N, Dunay IR, and Sibley LD
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- Animals, Central Nervous System Infections immunology, Central Nervous System Infections parasitology, Central Nervous System Infections pathology, Chronic Disease, Humans, Interferons immunology, Toxoplasma growth & development, Toxoplasma pathogenicity, Toxoplasmosis drug therapy, Toxoplasmosis immunology, Virulence, Toxoplasma physiology, Toxoplasmosis parasitology
- Abstract
Toxoplasma gondii is a parasite that infects a wide range of animals and causes zoonotic infections in humans. Although it normally only results in mild illness in healthy individuals, toxoplasmosis is a common opportunistic infection with high mortality in individuals who are immunocompromised, most commonly due to reactivation of infection in the central nervous system. In the acute phase of infection, interferon-dependent immune responses control rapid parasite expansion and mitigate acute disease symptoms. However, after dissemination the parasite differentiates into semi-dormant cysts that form within muscle cells and neurons, where they persist for life in the infected host. Control of infection in the central nervous system, a compartment of immune privilege, relies on modified immune responses that aim to balance infection control while limiting potential damage due to inflammation. In response to the activation of interferon-mediated pathways, the parasite deploys an array of effector proteins to escape immune clearance and ensure latent survival. Although these pathways are best studied in the laboratory mouse, emerging evidence points to unique mechanisms of control in human toxoplasmosis. In this Review, we explore some of these recent findings that extend our understanding for proliferation, establishment and control of toxoplasmosis in humans.
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- 2021
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24. Acute Alcohol Intoxication Modulates Monocyte Subsets and Their Functions in a Time-Dependent Manner in Healthy Volunteers.
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Janicova A, Haag F, Xu B, Garza AP, Dunay IR, Neunaber C, Nowak AJ, Cavalli P, Marzi I, Sturm R, and Relja B
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- Adolescent, Adult, Biomarkers, Healthy Volunteers, Humans, Immunophenotyping, Interleukin-1beta metabolism, Middle Aged, Time Factors, Toll-Like Receptor 4 metabolism, Young Adult, Alcoholic Intoxication immunology, Alcoholic Intoxication metabolism, Cell Plasticity immunology, Monocytes immunology, Monocytes metabolism
- Abstract
Background: Excessive alcohol intake is associated with adverse immune response-related effects, however, acute and chronic abuse differently modulate monocyte activation. In this study, we have evaluated the phenotypic and functional changes of monocytes in acutely intoxicated healthy volunteers (HV)., Methods: Twenty-two HV consumed individually adjusted amounts of alcoholic beverages until reaching a blood alcohol level of 1‰ after 4h (T4). Peripheral blood was withdrawn before and 2h (T2), 4h (T4), 6h (T6), 24h (T24), and 48h (T48) after starting the experiment and stained for CD14, CD16 and TLR4. CD14
bright CD16- , CD14bright CD16+ and CD14dim CD16+ monocyte subsets and their TLR4 expression were analyzed by flow cytometry. Inflammasome activation via caspase-1 in CD14+ monocytes was measured upon an ex vivo in vitro LPS stimulation. Systemic IL-1β and adhesion capacity of isolated CD14+ monocytes upon LPS stimulation were evaluated., Results: The percentage of CD14+ monocyte did not change following alcohol intoxication, whereas CD14bright CD16- monocyte subset significantly increased at T2 and T24, CD14bright CD16+ at T2, T4 and T6 and CD14dim CD16+ at T4 and T6. The relative fraction of TLR4 expressing CD14+ monocytes as well as the density of TLR4 surface presentation increased at T2 and decreased at T48 significantly. TLR4+ CD14+ monocytes were significantly enhanced in all subsets at T2. TLR4 expression significantly decreased in CD14bright CD16- at T48, in CD14bright CD16+ at T24 and T48, increased in CD14dim CD16+ at T2. IL-1β release upon LPS stimulation decreased at T48, correlating with TLR4 receptor expression. Alcohol downregulated inflammasome activation following ex vivo in vitro stimulation with LPS between T2 and T48 vs . T0. The adhesion capacity of CD14+ monocytes decreased from T2 with significance at T4, T6 and T48. Following LPS administration, a significant reduction of adhesion was observed at T4 and T6., Conclusions: Alcohol intoxication immediately redistributes monocyte subsets toward the pro-inflammatory phenotype with their subsequent differentiation into the anti-inflammatory phenotype. This is paralleled by a significant functional depression, suggesting an alcohol-induced time-dependent hyporesponsiveness of monocytes to pathogenic triggers., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Janicova, Haag, Xu, Garza, Dunay, Neunaber, Nowak, Cavalli, Marzi, Sturm and Relja.)- Published
- 2021
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25. Lymphotoxin β Receptor: a Crucial Role in Innate and Adaptive Immune Responses against Toxoplasma gondii.
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Tersteegen A, Sorg UR, Virgen-Slane R, Helle M, Petzsch P, Dunay IR, Köhrer K, Degrandi D, Ware CF, and Pfeffer K
- Subjects
- Animals, Disease Models, Animal, Lymphotoxin beta Receptor genetics, Mice, Mice, Knockout, Toxoplasmosis parasitology, Adaptive Immunity, Host-Parasite Interactions immunology, Immunity, Innate, Lymphotoxin beta Receptor metabolism, Toxoplasma immunology, Toxoplasmosis immunology, Toxoplasmosis metabolism
- Abstract
The lymphotoxin β receptor (LTβR) plays an essential role in the initiation of immune responses to intracellular pathogens. In mice, the LTβR is crucial for surviving acute toxoplasmosis; however, until now, a functional analysis was largely incomplete. Here, we demonstrate that the LTβR is a key regulator required for the intricate balance of adaptive immune responses. Toxoplasma gondii -infected LTβR-deficient (LTβR
-/- ) mice show globally altered interferon-γ (IFN-γ) regulation, reduced IFN-γ-controlled host effector molecule expression, impaired T cell functionality, and an absent anti-parasite-specific IgG response, resulting in a severe loss of immune control of the parasites. Reconstitution of LTβR-/- mice with toxoplasma immune serum significantly prolongs survival following T. gondii infection. Notably, analysis of RNA-seq data clearly indicates a specific effect of T. gondii infection on the B cell response and isotype switching. This study uncovers the decisive role of the LTβR in cytokine regulation and adaptive immune responses to control T. gondii ., (Copyright © 2021 Tersteegen et al.)- Published
- 2021
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26. The Immunoproteasome Subunits LMP2, LMP7 and MECL-1 Are Crucial Along the Induction of Cerebral Toxoplasmosis.
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French T, Israel N, Düsedau HP, Tersteegen A, Steffen J, Cammann C, Topfstedt E, Dieterich D, Schüler T, Seifert U, and Dunay IR
- Subjects
- Animals, Apoptosis, Biomarkers, Cytokines metabolism, Disease Models, Animal, Disease Susceptibility, Mice, Signal Transduction, Toxoplasma, Cysteine Endopeptidases metabolism, Immunomodulation, Proteasome Endopeptidase Complex metabolism, Toxoplasmosis, Cerebral etiology, Toxoplasmosis, Cerebral metabolism
- Abstract
Cell survival and function critically relies on the fine-tuned balance of protein synthesis and degradation. In the steady state, the standard proteasome is sufficient to maintain this proteostasis. However, upon inflammation, the sharp increase in protein production requires additional mechanisms to limit protein-associated cellular stress. Under inflammatory conditions and the release of interferons, the immunoproteasome (IP) is induced to support protein processing and recycling. In antigen-presenting cells constitutively expressing IPs, inflammation-related mechanisms contribute to the formation of MHC class I/II-peptide complexes, which are required for the induction of T cell responses. The control of Toxoplasma gondii infection relies on Interferon-γ (IFNγ)-related T cell responses. Whether and how the IP affects the course of anti-parasitic T cell responses along the infection as well as inflammation of the central nervous system is still unknown. To answer this question we used triple knockout (TKO) mice lacking the 3 catalytic subunits of the immunoproteasome (β1i/LMP2, β2i/MECL-1 and β5i/LMP7). Here we show that the numbers of dendritic cells, monocytes and CD8
+ T cells were reduced in Toxoplasma gondii -infected TKO mice. Furthermore, impaired IFNγ, TNF and iNOS production was accompanied by dysregulated chemokine expression and altered immune cell recruitment to the brain. T cell differentiation was altered, apoptosis rates of microglia and monocytes were elevated and STAT3 downstream signaling was diminished. Consequently, anti-parasitic immune responses were impaired in TKO mice leading to elevated T. gondii burden and prolonged neuroinflammation. In summary we provide evidence for a critical role of the IP subunits β1i/LMP2, β2i/MECL-1 and β5i/LMP7 for the control of cerebral Toxoplasma gondii infection and subsequent neuroinflammation., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 French, Israel, Düsedau, Tersteegen, Steffen, Cammann, Topfstedt, Dieterich, Schüler, Seifert and Dunay.)- Published
- 2021
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27. Alterations of Phagocytic Activity and Capacity in Granulocytes and Monocytes Depend on the Pathogen Strain in Porcine Polytrauma.
- Author
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Vollrath JT, Klingebiel F, Bläsius FM, Greven J, Bolierakis E, Janicova A, Dunay IR, Hildebrand F, Marzi I, and Relja B
- Abstract
Background: Polytraumatized patients undergo a strong immunological stress upon insult. Phagocytes (granulocytes and monocytes) play a substantial role in immunological defense against bacteria, fungi and yeast, and in the clearance of cellular debris after tissue injury. We have reported a reduced monocytes phagocytic activity early after porcine polytrauma before. However, it is unknown if both phagocyte types undergo those functional alterations, and if there is a pathogen-specific phagocytic behavior. We characterized the phagocytic activity and capacity of granulocytes and monocytes after polytrauma. Methods: Eight pigs ( Sus scrofa ) underwent polytrauma consisting of lung contusion, liver laceration, tibial fracture and hemorrhagic shock with fluid resuscitation and fracture fixation with external fixator. Intensive care treatment including mechanical ventilation for 72 h followed. Phagocytic activity and capacity were investigated using an in vitro ex vivo whole blood stimulation phagocytosis assays before trauma, after surgery, 24, 48, and 72 h after trauma. Blood samples were stimulated with Phorbol-12-myristate-13-acetate and incubated with FITC-labeled E. coli, S. aureus or S. cerevisiae for phagocytosis assessment by flow cytometry. Results: Early polytrauma-induced significant increase of granulocytes and monocytes declined to baseline values within 24 h. Percentage of E. coli -phagocytizing granulocytes significantly decreased after polytrauma and during further intensive care treatment, while their capacity significantly increased. Interestingly, both granulocytic phagocytic activity and capacity of S. aureus significantly decreased after trauma, although a recovery was observed after 24 h and yet was followed by another decrease. The percentage of S. cerevisiae -phagocytizing granulocytes significantly increased after 24 h, while their impaired capacity after surgery and 72 h later was detected. Monocytic E. coli -phagocytizing percentage did not change, while their capacity increased after 24-72 h. After a significant decrease in S. aureus -phagocytizing monocytes after surgery, a significant increase after 24 and 48 h was observed without capacity alterations. No significant changes in S. cerevisiae -phagocytizing monocytes occurred, but their capacity dropped 48 and 72 h. Conclusion: Phagocytic activity and capacity of granulocytes and monocytes follow a different pattern and significantly change within 72 h after polytrauma. Both phagocytic activity and capacity show significantly different alterations depending on the pathogen strain, thus potentially indicating at certain and possibly more relevant infection causes after polytrauma., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Vollrath, Klingebiel, Bläsius, Greven, Bolierakis, Janicova, Dunay, Hildebrand, Marzi and Relja.)
- Published
- 2021
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28. Directional mast cell degranulation of tumor necrosis factor into blood vessels primes neutrophil extravasation.
- Author
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Dudeck J, Kotrba J, Immler R, Hoffmann A, Voss M, Alexaki VI, Morton L, Jahn SR, Katsoulis-Dimitriou K, Winzer S, Kollias G, Fischer T, Nedospasov SA, Dunay IR, Chavakis T, Müller AJ, Schraven B, Sperandio M, and Dudeck A
- Subjects
- Animals, Blood Circulation, Cell Degranulation, Cells, Cultured, Immune System Diseases, Leukocyte Disorders, Mice, Mice, Inbred C57BL, Mice, Knockout, Neutrophil Activation, Receptors, Tumor Necrosis Factor, Type I metabolism, Secretory Vesicles metabolism, Tumor Necrosis Factor-alpha genetics, Blood Vessels immunology, Dermatitis, Contact immunology, Inflammation immunology, Mast Cells immunology, Neutrophils immunology, Skin pathology, Tumor Necrosis Factor-alpha metabolism
- Abstract
Tissue resident mast cells (MCs) rapidly initiate neutrophil infiltration upon inflammatory insult, yet the molecular mechanism is still unknown. Here, we demonstrated that MC-derived tumor necrosis factor (TNF) was crucial for neutrophil extravasation to sites of contact hypersensitivity-induced skin inflammation by promoting intraluminal crawling. MC-derived TNF directly primed circulating neutrophils via TNF receptor-1 (TNFR1) while being dispensable for endothelial cell activation. The MC-derived TNF was infused into the bloodstream by directional degranulation of perivascular MCs that were part of the vascular unit with access to the vessel lumen. Consistently, intravenous administration of MC granules boosted neutrophil extravasation. Pronounced and rapid intravascular MC degranulation was also observed upon IgE crosslinking or LPs challenge indicating a universal MC potential. Consequently, the directional MC degranulation of pro-inflammatory mediators into the bloodstream may represent an important target for therapeutic approaches aimed at dampening cytokine storm syndromes or shock symptoms, or intentionally pushing immune defense., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2020 Elsevier Inc. All rights reserved.)
- Published
- 2021
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29. Club Cell Protein 16 Attenuates CD16 bright CD62 dim Immunosuppressive Neutrophils in Damaged Tissue upon Posttraumatic Sepsis-Induced Lung Injury.
- Author
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Becker N, Störmann P, Janicova A, Köhler K, Horst K, Dunay IR, Neunaber C, Marzi I, Vollrath JT, and Relja B
- Subjects
- Acute Lung Injury pathology, Animals, Biomarkers, Disease Models, Animal, Immunohistochemistry, Immunophenotyping, Male, Mice, Neutrophil Infiltration immunology, Neutrophils pathology, Sepsis etiology, Thoracic Injuries complications, Uteroglobin metabolism, Acute Lung Injury etiology, Acute Lung Injury metabolism, Neutrophils immunology, Neutrophils metabolism, Receptors, IgG metabolism, Selectins metabolism, Sepsis complications, Uteroglobin genetics
- Abstract
Background: Recently, identification of immunosuppressive polymorphonuclear leukocytes (PMNL) that were traditionally described as proinflammatory cells emerged in the field of posttraumatic immunity. To understand their local and remote distribution after trauma, PMNL-subsets and the impact of immunomodulatory Club Cell protein (CC)16 that correlates with pulmonary complications were assessed., Methods: C57BL/6N mice were divided into three groups, receiving isolated blunt chest trauma (TxT), undergoing TxT followed by cecal ligation and puncture (CLP, TxT + CLP) after 24 h, or sham undergoing analgosedation ( n = 18/group). Further, each group was subdivided into three groups receiving either no treatment (ctrl) or intratracheal neutralization of CC16 by application of anti-CC16-antibody or application of an unspecific IgG control antibody ( n = 6/group). Treatment was set at the time point after TxT. Analyses followed 6 h post-CLP. PMNL were characterized via expression of CD11b, CD16, CD45, CD62L, and Ly6G by flow cytometry in bone marrow (BM), blood, spleen, lung, liver, and bronchoalveolar and peritoneal lavage fluid (BALF and PL). Apoptosis was assessed by activated (cleaved) caspase-3. Results from untreated ctrl and IgG-treated mice were statistically comparable between all corresponding sham, TxT, and TxT + CLP groups., Results: Immature (CD16
dim CD62Lbright ) PMNL increased significantly in BM, circulation, and spleen after TxT vs . sham and were significantly attenuated in the lungs, BALF, PL, and liver. Classical-shaped (CD16bright CD62Lbright ) PMNL increased after TxT vs . sham in peripheral tissue and were significantly attenuated in circulation, proposing a trauma-induced migration of mature or peripheral differentiation of circulating immature PMNL. Immunosuppressive (CD16bright CD62Ldim ) PMNL decreased significantly in the lungs and spleen, while they systemically increased after TxT vs . sham. CLP in the TxT + CLP group reduced immunosuppressive PMNL in PL and increased their circulatory rate vs . isolated TxT, showing local reduction in affected tissue and their increase in nonaffected tissue. CC16 neutralization enhanced the fraction of immunosuppressive PMNL following TxT vs . sham and decreased caspase-3 in the lungs post-CLP in the TxT + CLP group, while apoptotic cells in the liver diminished post-TxT. Posttraumatic CC16 neutralization promotes the subset of immunosuppressive PMNL and antagonizes their posttraumatic distribution., Conclusion: Since CC16 affects both the distribution of PMNL subsets and apoptosis in tissues after trauma, it may constitute as a novel target to beneficially shape the posttraumatic tissue microenvironment and homeostasis to improving outcomes., Competing Interests: The authors state that they have no conflict of interest., (Copyright © 2021 Nils Becker et al.)- Published
- 2021
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30. Attenuation of the extracellular matrix restores microglial activity during the early stage of amyloidosis.
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Stoyanov S, Sun W, Düsedau HP, Cangalaya C, Choi I, Mirzapourdelavar H, Baidoe-Ansah D, Kaushik R, Neumann J, Dunay IR, and Dityatev A
- Subjects
- Alzheimer Disease, Amyloid beta-Peptides, Animals, Disease Models, Animal, Extracellular Matrix, Mice, Mice, Transgenic, Microglia, Plaque, Amyloid, Amyloidosis
- Abstract
In the advanced stages of Alzheimer's disease (AD), microglia are transformed to an activated phenotype with thickened and retracted processes, migrate to the site of amyloid-beta (Aβ) plaques, and proliferate. In the early stages of AD, it is still poorly understood whether the microglial function is altered and which factors may regulate these changes. Here, we focused on studying microglia in the retrosplenial cortex (RSC) in 3- to 4-month-old 5xFAD mice as a transgenic mouse model of AD. At this age, there are neither Aβ plaques, nor activation of microglia, nor dysregulation in the expression of genes encoding major extracellular matrix (ECM) molecules or extracellular proteases in the RSC. Still, histochemical evaluation of the fine structure of neural ECM revealed increased levels of Wisteria floribunda agglutinin labeling in holes of perineuronal nets and changes in the perimeter of ECM barriers around the holes in 5xFAD mice. Two-photon vital microscopy demonstrated normal morphology and resting motility of microglia but strongly diminished number of microglial cells that migrated to the photolesion site in 5xFAD mice. Enzymatic digestion of ECM by chondroitinase ABC (ChABC) ameliorated this defect. Accordingly, the characterization of cell surface markers by flow cytometry demonstrated altered expression of microglial CD45. Moreover, ChABC treatment reduced the invasion of myeloid-derived mononuclear cells into the RSC of 5xFAD mice. Hence, the migration of both microglia and myeloid cells is altered during the early stages of amyloidosis and can be restored at least partially by the attenuation of the ECM., (© 2020 The Authors. GLIA published by Wiley Periodicals LLC.)
- Published
- 2021
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31. NK Cells Negatively Regulate CD8 T Cells to Promote Immune Exhaustion and Chronic Toxoplasma gondii Infection.
- Author
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Ivanova DL, Krempels R, Denton SL, Fettel KD, Saltz GM, Rach D, Fatima R, Mundhenke T, Materi J, Dunay IR, and Gigley JP
- Subjects
- Animals, CD8-Positive T-Lymphocytes, Killer Cells, Natural, Mice, Spleen, Toxoplasma, Toxoplasmosis
- Abstract
NK cells regulate CD4+ and CD8+ T cells in acute viral infection, vaccination, and the tumor microenvironment. NK cells also become exhausted in chronic activation settings. The mechanisms causing these ILC responses and their impact on adaptive immunity are unclear. CD8+ T cell exhaustion develops during chronic Toxoplasma gondii ( T. gondii ) infection resulting in parasite reactivation and death. How chronic T. gondii infection impacts the NK cell compartment is not known. We demonstrate that NK cells do not exhibit hallmarks of exhaustion. Their numbers are stable and they do not express high PD1 or LAG3. NK cell depletion with anti-NK1.1 is therapeutic and rescues chronic T. gondii infected mice from CD8+ T cell exhaustion dependent death, increases survival after lethal secondary challenge and alters cyst burdens in brain. Anti-NK1.1 treatment increased polyfunctional CD8+ T cell responses in spleen and brain and reduced CD8+ T cell apoptosis in spleen. Chronic T. gondii infection promotes the development of a modified NK cell compartment, which does not exhibit normal NK cell characteristics. NK cells are Ly49 and TRAIL negative and are enriched for expression of CD94/NKG2A and KLRG1. These NK cells are found in both spleen and brain. They do not produce IFNγ, are IL-10 negative, do not increase PDL1 expression, but do increase CD107a on their surface. Based on the NK cell receptor phenotype we observed NKp46 and CD94-NKG2A cognate ligands were measured. Activating NKp46 (NCR1-ligand) ligand increased and NKG2A ligand Qa-1b expression was reduced on CD8+ T cells. Blockade of NKp46 rescued the chronically infected mice from death and reduced the number of NKG2A+ cells. Immunization with a single dose non-persistent 100% protective T. gondii vaccination did not induce this cell population in the spleen, suggesting persistent infection is essential for their development. We hypothesize chronic T. gondii infection induces an NKp46 dependent modified NK cell population that reduces functional CD8+ T cells to promote persistent parasite infection in the brain. NK cell targeted therapies could enhance immunity in people with chronic infections, chronic inflammation and cancer., (Copyright © 2020 Ivanova, Krempels, Denton, Fettel, Saltz, Rach, Fatima, Mundhenke, Materi, Dunay and Gigley.)
- Published
- 2020
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32. Short Exposure to Ethanol Diminishes Caspase-1 and ASC Activation in Human HepG2 Cells In Vitro.
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Hörauf JA, Kany S, Janicova A, Xu B, Vrdoljak T, Sturm R, Dunay IR, Martin L, and Relja B
- Subjects
- Adamantane analogs & derivatives, Adamantane pharmacology, Amino Acid Chloromethyl Ketones pharmacology, Aminoquinolines pharmacology, Hep G2 Cells, Humans, Inflammasomes drug effects, Inflammasomes metabolism, Lipopolysaccharides pharmacology, Liver drug effects, Liver metabolism, Reactive Oxygen Species metabolism, Vanadates pharmacology, Caspase 1 metabolism, Ethanol pharmacology
- Abstract
This paper discusses how the assembly of pro-caspase-1 and apoptosis-associated speck-like protein containing a caspase-recruitment domain (ASC) in macromolecular protein complexes, inflammasomes, activates caspase-1. The present study investigates the molecular mechanisms of inflammasome activation in HepG2 cells and examines how short exposures to ethanol (EtOH) affect inflammasome activation. HepG2 cells were treated with lipopolysaccharide (LPS), ATP or nigericin (NIG) in a two-step model. After LPS priming, ATP or NIG were added. As inhibitors, sodium orthovanadate (general inhibitor of tyrosine phosphatases), AC-YVAD-CMK (caspase-1 inhibitor) or AZ10606120 (purinergic receptor P2X7R inhibitor) were applied after LPS priming. To monitor the inflammasome activation, the caspase-1 activity, ASC speck formation, reactive oxygen species (ROS) production and cell death were analyzed. To elucidate the mechanistical approach of EtOH to the inflammasome assembly, the cells were treated with EtOH either under simultaneous LPS administration or concurrently with ATP or NIG application. The co-stimulation with LPS and ATP induced a significant ASC speck formation, caspase-1 activation, cell death and ROS generation. The inhibition of the ATP-dependent purinoreceptor P2X7 decreased the caspase-1 activation, whereas sodium orthovanadate significantly induced caspase-1. Additional treatment with EtOH reversed the LPS and ATP-induced caspase-1 activation, ASC speck formation and ROS production. The ASC speck formation and caspase-1 induction require a two-step signaling with LPS and ATP in HepG2 cells. Inflammasome activation may depend on P2X7. The molecular pathway of an acute effect of EtOH on inflammasomes may involve a reduction in ROS generation, which in turn may increase the activity of tyrosine phosphatases., Competing Interests: The authors declare no conflict of interest.
- Published
- 2020
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33. Regulatory T Cells Modulate CD4 Proliferation after Severe Trauma via IL-10.
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Sturm R, Xanthopoulos L, Heftrig D, Oppermann E, Vrdoljak T, Dunay IR, Marzi I, and Relja B
- Abstract
Objective: Severely injured patients frequently develop an immunological imbalance following the traumatic insult, which might result in infectious complications evoked by a persisting immunosuppression. Regulatory T cells (Tregs) maintain the immune homeostasis by suppressing proinflammatory responses, however, their functionality after trauma is unclear. Here, we characterized the role of Tregs in regulating the proliferation of CD4
+ lymphocytes in traumatized patients (TP)., Methods: Peripheral blood was obtained daily from 29 severely injured TP (Injury Severity Score, ISS ≥16) for ten days following admission to the emergency department (ED). Ten healthy volunteers (HV) served as controls. The frequency and activity of Tregs were assessed by flow cytometry. Proliferation of CD4+ cells was analyzed either in presence or absence of Tregs, or after blocking of either IL-10 or IL-10R1., Results: The frequencies of CD4+ CD25high and CD4+ CD25+ CD127- Tregs were significantly decreased immediately upon admission of TP to the ED and during the following 10 post-injury days. Compared with HV CD4+ T cell proliferation in TP increased significantly upon their admission and on the following days. As expected, CD4+ CD25+ CD127- Tregs reduced the proliferation of CD4+ cells in HV, nevertheless, CD4+ proliferation in TP was increased by Tregs. Neutralization of IL-10 as well as blocking the IL-10R1 increased further CD4+ T cell proliferation in Tregs-depleted cultures, thereby confirming an IL-10-mediated mechanism of IL-10-regulated CD4+ T cell proliferation. Neutralization of IL-10 in TP decreased CD4+ T cell proliferation in Tregs-depleted cultures, whereas blocking of the IL-10R1 receptor had no significant effects., Conclusions: The frequency of Tregs in the CD4+ T lymphocyte population is reduced after trauma; however, their inductiveness is increased. The mechanisms of deregulated influence of Tregs on CD4+ T cell proliferation are mediated via IL-10 but not via the IL-10R1.- Published
- 2020
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34. c-FLIP is crucial for IL-7/IL-15-dependent NKp46 + ILC development and protection from intestinal inflammation in mice.
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Bank U, Deiser K, Plaza-Sirvent C, Osbelt L, Witte A, Knop L, Labrenz R, Jänsch R, Richter F, Biswas A, Zenclussen AC, Vivier E, Romagnani C, Kühl AA, Dunay IR, Strowig T, Schmitz I, and Schüler T
- Subjects
- Animals, Antigens, Ly genetics, Apoptosis physiology, B-Lymphocytes immunology, CASP8 and FADD-Like Apoptosis Regulating Protein genetics, Cells, Cultured, Colitis chemically induced, Colitis pathology, Killer Cells, Natural immunology, Mice, Mice, Knockout, Natural Cytotoxicity Triggering Receptor 1 genetics, Phosphorylation, T-Lymphocytes immunology, Antigens, Ly metabolism, CASP8 and FADD-Like Apoptosis Regulating Protein metabolism, Colitis prevention & control, Interleukin-15 metabolism, Interleukin-7 metabolism, Natural Cytotoxicity Triggering Receptor 1 metabolism, STAT5 Transcription Factor metabolism
- Abstract
NKp46
+ innate lymphoid cells (ILC) modulate tissue homeostasis and anti-microbial immune responses. ILC development and function are regulated by cytokines such as Interleukin (IL)-7 and IL-15. However, the ILC-intrinsic pathways translating cytokine signals into developmental programs are largely unknown. Here we show that the anti-apoptotic molecule cellular FLICE-like inhibitory protein (c-FLIP) is crucial for the generation of IL-7/IL-15-dependent NKp46+ ILC1, including conventional natural killer (cNK) cells, and ILC3. Cytokine-induced phosphorylation of signal transducer and activator of transcription 5 (STAT5) precedes up-regulation of c-FLIP, which protects developing NKp46+ ILC from TNF-induced apoptosis. NKp46+ ILC-specific inactivation of c-FLIP leads to the loss of all IL-7/IL-15-dependent NKp46+ ILC, thereby inducing early-onset chronic colitis and subsequently microbial dysbiosis; meanwhile, the depletion of cNK, but not NKp46+ ILC1/3, aggravates experimental colitis. In summary, our data demonstrate a non-redundant function of c-FLIP for the generation of NKp46+ ILC, which protect T/B lymphocyte-sufficient mice from intestinal inflammation.- Published
- 2020
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35. Protection of Batf3-deficient mice from experimental cerebral malaria correlates with impaired cytotoxic T-cell responses and immune regulation.
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Kuehlwein JM, Borsche M, Korir PJ, Risch F, Mueller AK, Hübner MP, Hildner K, Hoerauf A, Dunay IR, and Schumak B
- Subjects
- Animals, Basic-Leucine Zipper Transcription Factors genetics, Blood-Brain Barrier immunology, Blood-Brain Barrier parasitology, Brain metabolism, Brain parasitology, Cells, Cultured, Dendritic Cells metabolism, Dendritic Cells parasitology, Disease Models, Animal, Female, Granzymes immunology, Granzymes metabolism, Host-Parasite Interactions, Interferon-gamma immunology, Interferon-gamma metabolism, Interleukin-10 immunology, Interleukin-10 metabolism, Malaria, Cerebral immunology, Malaria, Cerebral metabolism, Malaria, Cerebral parasitology, Mice, Inbred C57BL, Mice, Knockout, Plasmodium berghei immunology, Repressor Proteins genetics, Spleen immunology, Spleen metabolism, Spleen parasitology, T-Lymphocytes, Cytotoxic metabolism, T-Lymphocytes, Cytotoxic parasitology, Basic-Leucine Zipper Transcription Factors deficiency, Brain immunology, Dendritic Cells immunology, Malaria, Cerebral prevention & control, Plasmodium berghei pathogenicity, Repressor Proteins deficiency, T-Lymphocytes, Cytotoxic immunology
- Abstract
Excessive inflammatory immune responses during infections with Plasmodium parasites are responsible for severe complications such as cerebral malaria (CM) that can be studied experimentally in mice. Dendritic cells (DCs) activate cytotoxic CD8
+ T-cells and initiate immune responses against the parasites. Batf3-/- mice lack a DC subset, which efficiently induces strong CD8 T-cell responses by cross-presentation of exogenous antigens. Here we show that Batf3-/- mice infected with Plasmodium berghei ANKA (PbA) were protected from experimental CM (ECM), characterized by a stable blood-brain barrier (BBB) and significantly less infiltrated peripheral immune cells in the brain. Importantly, the absence of ECM in Batf3-/- mice correlated with attenuated responses of cytotoxic T-cells, as their parasite-specific lytic activity as well as the production of interferon gamma and granzyme B were significantly decreased. Remarkably, spleens of ECM-protected Batf3-/- mice had elevated levels of regulatory immune cells and interleukin 10. Thus, protection from ECM in PbA-infected Batf3-/- mice was associated with the absence of strong CD8+ T-cell activity and induction of immunoregulatory mediators and cells., (© 2019 The Authors. Immunology Published by John Wiley & Sons Ltd.)- Published
- 2020
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36. The Central Nervous System Contains ILC1s That Differ From NK Cells in the Response to Inflammation.
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Romero-Suárez S, Del Rio Serrato A, Bueno RJ, Brunotte-Strecker D, Stehle C, Figueiredo CA, Hertwig L, Dunay IR, Romagnani C, and Infante-Duarte C
- Subjects
- Animals, Antigens, Differentiation genetics, Antigens, Differentiation immunology, Brain pathology, Cell Movement genetics, Cytokines genetics, Cytokines immunology, Encephalomyelitis, Autoimmune, Experimental genetics, Encephalomyelitis, Autoimmune, Experimental pathology, Female, Inflammation genetics, Inflammation immunology, Inflammation pathology, Killer Cells, Natural pathology, Mice, Mice, Transgenic, Brain immunology, Cell Movement immunology, Encephalomyelitis, Autoimmune, Experimental immunology, Killer Cells, Natural immunology
- Abstract
Innate lymphoid cells (ILCs) are tissue resident cells with organ-specific properties. Here, we show that the central nervous system (CNS) encompasses ILCs. In particular, CD3
- NK1.1+ cells present in the murine CNS comprise natural killer (NK) cells, ILC1s, intermediate ILC1s (intILC1s) and ex-ILC3s. We investigated the properties of CNS-ILC1s in comparison with CNS-NK cells during steady state and experimental autoimmune encephalomyelitis (EAE). ILC1s characteristically express CXCR3, CXCR6, DNAM-1, TRAIL, and CD200R and display heightened TNF-α production upon stimulation. In addition, ILC1s express perforin and are able to degranulate, although in a lesser extent than NK cells. Within the CNS compartments, ILC1s are enriched in the choroid plexus where very few NK cells are present, and also reside in the brain parenchyma and meninges. During EAE, ILC1s maintain stable IFN-γ and TNF-α levels while in NK cells the production of these cytokines increases as EAE progresses. Moreover, the amount of ILC1s and intILC1s increase in the parenchyma during EAE, but in contrast to NK cells, they show no signs of local proliferation. The upregulation in the inflamed brain of chemokines involved in ILC1 migration, such as CXCL9, CXCL10, and CXCL16 may lead to a recruitment of ILC1s from meninges or choroid plexus into the brain parenchyma. In sum, CNS-ILC1 phenotype, distribution and moderate inflammatory response during EAE suggest that they may act as gatekeepers involved in the control of neuroinflammation., (Copyright © 2019 Romero-Suárez, Del Rio Serrato, Bueno, Brunotte-Strecker, Stehle, Figueiredo, Hertwig, Dunay, Romagnani and Infante-Duarte.)- Published
- 2019
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37. Neuronal impairment following chronic Toxoplasma gondii infection is aggravated by intestinal nematode challenge in an IFN-γ-dependent manner.
- Author
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French T, Düsedau HP, Steffen J, Biswas A, Ahmed N, Hartmann S, Schüler T, Schott BH, and Dunay IR
- Subjects
- Animals, Brain parasitology, Coinfection, Macrophage Activation physiology, Mice, Microglia parasitology, Nematospiroides dubius, Neurons parasitology, Synapses metabolism, Synapses parasitology, Toxoplasma, Brain metabolism, Interferon-gamma metabolism, Microglia metabolism, Neurons metabolism, Strongylida Infections metabolism, Toxoplasmosis metabolism
- Abstract
Background: It has become increasingly evident that the immune and nervous systems are closely intertwined, relying on one another during regular homeostatic conditions. Prolonged states of imbalance between neural and immune homeostasis, such as chronic neuroinflammation, are associated with a higher risk for neural damage. Toxoplasma gondii is a highly successful neurotropic parasite causing persistent subclinical neuroinflammation, which is associated with psychiatric and neurodegenerative disorders. Little is known, however, by what means neuroinflammation and the associated neural impairment can be modulated by peripheral inflammatory processes., Methods: Expression of immune and synapse-associated genes was assessed via quantitative real-time PCR to investigate how T. gondii infection-induced chronic neuroinflammation and associated neuronal alterations can be reshaped by a subsequent acute intestinal nematode co-infection. Immune cell subsets were characterized via flow cytometry in the brain of infected mice. Sulfadiazine and interferon-γ-neutralizing antibody were applied to subdue neuroinflammation., Results: Neuroinflammation induced by T. gondii infection of mice was associated with increased microglia activation, recruitment of immune cells into the brain exhibiting Th1 effector functions, and enhanced production of Th1 and pro-inflammatory molecules (IFN-γ, iNOS, IL-12, TNF, IL-6, and IL-1β) following co-infection with Heligmosomoides polygyrus. The accelerated cerebral Th1 immune response resulted in enhanced T. gondii removal but exacerbated the inflammation-related decrease of synapse-associated gene expression. Synaptic proteins EAAT2 and GABA
A α1, which are involved in the excitation/inhibition balance in the CNS, were affected in particular. These synaptic alterations were partially recovered by reducing neuroinflammation indirectly via antiparasitic treatment and especially by application of IFN-γ-neutralizing antibody. Impaired iNOS expression following IFN-γ neutralization directly affected EAAT2 and GABAA α1 signaling, thus contributing to the microglial regulation of neurons. Besides, reduced CD36, TREM2, and C1qa gene expression points toward inflammation induced synaptic pruning as a fundamental mechanism., Conclusion: Our results suggest that neuroimmune responses following chronic T. gondii infection can be modulated by acute enteric nematode co-infection. While consecutive co-infection promotes parasite elimination in the CNS, it also adversely affects gene expression of synaptic proteins, via an IFN-γ-dependent manner.- Published
- 2019
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38. Immunomodulatory Effects of the Neuropeptide Pituitary Adenylate Cyclase-Activating Polypeptide in Acute Toxoplasmosis .
- Author
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Figueiredo CA, Düsedau HP, Steffen J, Gupta N, Dunay MP, Toth GK, Reglodi D, Heimesaat MM, and Dunay IR
- Subjects
- Animals, Antigens, Ly, Brain-Derived Neurotrophic Factor metabolism, Chemokines metabolism, Cytokines metabolism, Disease Models, Animal, Female, Immunity, Innate, Inflammation, Interleukin-10, Macrophages, Mice, Mice, Inbred C57BL, Nerve Growth Factors, Receptors, Vasoactive Intestinal Peptide, Type II, Receptors, Vasoactive Intestinal Polypeptide, Type I, Toxoplasma, Up-Regulation, Host-Pathogen Interactions immunology, Immunomodulation, Neuropeptides immunology, Pituitary Adenylate Cyclase-Activating Polypeptide administration & dosage, Pituitary Adenylate Cyclase-Activating Polypeptide immunology, Toxoplasmosis immunology
- Abstract
Pituitary Adenylate Cyclase-Activating Polypeptide (PACAP) is an endogenous neuropeptide with distinct functions including the regulation of inflammatory processes. PACAP is able to modify the immune response by directly regulating macrophages and monocytes inhibiting the production of inflammatory cytokines, chemokines and free radicals. Here, we analyzed the effect of exogenous PACAP on peripheral immune cell subsets upon acute infection with the parasite Toxoplasma gondii (T. gondii) . PACAP administration was followed by diminished innate immune cell recruitment to the peritoneal cavity of T. gondii -infected mice. PACAP did not directly interfere with parasite replication, instead, indirectly reduced parasite burden in mononuclear cell populations by enhancing their phagocytic capacity. Although proinflammatory cytokine levels were attenuated in the periphery upon PACAP treatment, interleukin (IL)-10 and Transforming growth factor beta (TGF-β) remained stable. While PACAP modulated VPAC1 and VPAC2 receptors in immune cells upon binding, it also increased their expression of brain-derived neurotrophic factor (BDNF). In addition, the expression of p75 neurotrophin receptor (p75
NTR ) on Ly6Chi inflammatory monocytes was diminished upon PACAP administration. Our findings highlight the immunomodulatory effect of PACAP on peripheral immune cell subsets during acute Toxoplasmosis , providing new insights about host-pathogen interaction and the effects of neuropeptides during inflammation.- Published
- 2019
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39. Comprehensive Kinetic Survey of Intestinal, Extra-Intestinal and Systemic Sequelae of Murine Ileitis Following Peroral Low-Dose Toxoplasma gondii Infection.
- Author
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Heimesaat MM, Dunay IR, and Bereswill S
- Subjects
- Animal Structures parasitology, Animal Structures pathology, Animals, Disease Models, Animal, Dysbiosis, Gastrointestinal Microbiome, Histocytochemistry, Ileitis complications, Ileitis parasitology, Ileum parasitology, Ileum pathology, Intestinal Mucosa parasitology, Intestinal Mucosa pathology, Longitudinal Studies, Mice, Inbred C57BL, Parasitemia parasitology, Parasitemia pathology, Toxoplasmosis, Animal parasitology, Ileitis pathology, Toxoplasma growth & development, Toxoplasmosis, Animal pathology
- Abstract
We have recently shown that following peroral low-dose Toxoplasma gondii infection susceptible mice develop subacute ileitis within 10 days. Data regarding long-term intestinal and extra-intestinal sequelae of infection are scarce, however. We therefore challenged conventional C57BL/6 mice with one cyst of T. gondii ME49 strain by gavage and performed a comprehensive immunopathological survey 10, 36, and 57 days later. As early as 10 days post-infection, mice were suffering from subacute ileitis as indicated by mild-to-moderate histopathological changes of the ileal mucosa. Furthermore, numbers of apoptotic and proliferating/regenerating epithelial cells as well as of T and B lymphocytes in the mucosa and lamina propria of the ileum were highest at day 10 post-infection, but declined thereafter, and were accompanied by enhanced pro-inflammatory mediator secretion in ileum, colon and mesenteric lymph nodes that was most pronounced during the early phase of infection. In addition, subacute ileitis was accompanied by distinct shifts in the commensal gut microbiota composition in the small intestines. Remarkably, immunopathological sequelae of T. gondii infection were not restricted to the intestines, but could also be observed in extra-intestinal tissues including the liver, kidneys, lungs, heart and strikingly, in systemic compartments that were most prominent at day 10 post-infection. We conclude that the here provided long-term kinetic survey of immunopathological sequalae following peroral low-dose T. gondii infection provides valuable corner stones for a better understanding of the complex interactions within the triangle relationship of (parasitic) pathogens, the host immunity and the commensal gut microbiota during intestinal inflammation. The low-dose T. gondii infection model may be applied as valuable gut inflammation model in future pre-clinical studies in order to test potential treatment options for intestinal inflammatory conditions in humans.
- Published
- 2019
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40. Pituitary Adenylate Cyclase-Activating Polypeptide-A Neuropeptide as Novel Treatment Option for Subacute Ileitis in Mice Harboring a Human Gut Microbiota.
- Author
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Bereswill S, Escher U, Grunau A, Kühl AA, Dunay IR, Tamas A, Reglodi D, and Heimesaat MM
- Subjects
- Animals, Female, Humans, Interleukin-6 immunology, Mice, T-Lymphocytes immunology, T-Lymphocytes pathology, Toxoplasma immunology, Toxoplasmosis immunology, Tumor Necrosis Factor-alpha immunology, Fecal Microbiota Transplantation, Gastrointestinal Microbiome immunology, Ileitis immunology, Ileitis pathology, Ileitis therapy, Inflammatory Bowel Diseases immunology, Inflammatory Bowel Diseases pathology, Inflammatory Bowel Diseases therapy, Pituitary Adenylate Cyclase-Activating Polypeptide immunology
- Abstract
The neuropeptide Pituitary adenylate cyclase-activating polypeptide (PACAP) is well-known for its important functions in immunity and inflammation. Data regarding anti-inflammatory properties of PACAP in the intestinal tract are limited, however. In our present preclinical intervention study we addressed whether PACAP treatment could alleviate experimental subacute ileitis mimicking human gut microbiota conditions. Therefore, secondary abioitic mice were subjected to human fecal microbiota transplantation (FMT) and perorally infected with low-dose Toxoplasma gondii to induce subacute ileitis on day 0. From day 3 until day 8 post-infection, mice were either treated with synthetic PACAP38 or placebo. At day 9 post-infection, placebo, but not PACAP treated mice exhibited overt macroscopic sequelae of intestinal immunopathology. PACAP treatment further resulted in less distinct apoptotic responses in ileal and colonic epithelia that were accompanied by lower T cell numbers in the mucosa and lamina propria and less secretion of pro-inflammatory cytokines in intestinal ex vivo biopsies. Notably, ileitis-associated gut microbiota shifts were less distinct in PACAP as compared to placebo treated mice. Inflammation-ameliorating effects of PACAP were not restricted to the intestines, but could also be observed in extra-intestinal including systemic compartments as indicated by lower apoptotic cell counts and less pro-inflammatory cytokine secretion in liver and lungs taken from PACAP treated as compared to placebo control mice, which also held true for markedly lower serum TNF and IL-6 concentrations in the former as compared to the latter. Our preclinical intervention study provides strong evidence that synthetic PACAP alleviates subacute ileitis and extra-intestinal including systemic sequelae of T cell-driven immunopathology. These findings further support PACAP as a novel treatment option for intestinal inflammation including inflammatory bowel diseases (IBD).
- Published
- 2019
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41. Innate Lymphoid Cells in Protection, Pathology, and Adaptive Immunity During Apicomplexan Infection.
- Author
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Ivanova DL, Denton SL, Fettel KD, Sondgeroth KS, Munoz Gutierrez J, Bangoura B, Dunay IR, and Gigley JP
- Subjects
- Animals, Biomarkers, Cell Plasticity immunology, Cytokines metabolism, Host-Parasite Interactions, Humans, Immunophenotyping, Inflammation Mediators metabolism, Plasmodium immunology, Adaptive Immunity, Apicomplexa immunology, Immunity, Innate, Lymphocyte Subsets immunology, Lymphocyte Subsets metabolism, Protozoan Infections immunology, Protozoan Infections parasitology
- Abstract
Apicomplexans are a diverse and complex group of protozoan pathogens including Toxoplasma gondii, Plasmodium spp., Cryptosporidium spp., Eimeria spp., and Babesia spp. They infect a wide variety of hosts and are a major health threat to humans and other animals. Innate immunity provides early control and also regulates the development of adaptive immune responses important for controlling these pathogens. Innate immune responses also contribute to immunopathology associated with these infections. Natural killer (NK) cells have been for a long time known to be potent first line effector cells in helping control protozoan infection. They provide control by producing IL-12 dependent IFNγ and killing infected cells and parasites via their cytotoxic response. Results from more recent studies indicate that NK cells could provide additional effector functions such as IL-10 and IL-17 and might have diverse roles in immunity to these pathogens. These early studies based their conclusions on the identification of NK cells to be CD3-, CD49b+, NK1.1+, and/or NKp46+ and the common accepted paradigm at that time that NK cells were one of the only lymphoid derived innate immune cells present. New discoveries have lead to major advances in understanding that NK cells are only one of several populations of innate immune cells of lymphoid origin. Common lymphoid progenitor derived innate immune cells are now known as innate lymphoid cells (ILC) and comprise three different groups, group 1, group 2, and group 3 ILC. They are a functionally heterogeneous and plastic cell population and are important effector cells in disease and tissue homeostasis. Very little is known about each of these different types of ILCs in parasitic infection. Therefore, we will review what is known about NK cells in innate immune responses during different protozoan infections. We will discuss what immune responses attributed to NK cells might be reconsidered as ILC1, 2, or 3 population responses. We will then discuss how different ILCs may impact immunopathology and adaptive immune responses to these parasites.
- Published
- 2019
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42. Interferon-γ Receptor Signaling in Dendritic Cells Restrains Spontaneous Proliferation of CD4 + T Cells in Chronic Lymphopenic Mice.
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Knop L, Frommer C, Stoycheva D, Deiser K, Kalinke U, Blankenstein T, Kammertoens T, Dunay IR, and Schüler T
- Subjects
- Animals, Cell Proliferation, Chronic Disease, Interleukin-6 immunology, Mice, Transgenic, Receptors, Interferon genetics, Signal Transduction, Interferon gamma Receptor, CD4-Positive T-Lymphocytes immunology, Dendritic Cells immunology, Lymphopenia immunology, Receptors, Interferon immunology
- Abstract
In lymphopenic mice, T cells become activated and undergo lymphopenia-induced proliferation (LIP). However, not all T cells are equally sensitive to lymphopenia. Several lymphopenia-insensitive T cell clones were described and their non-responsiveness was mainly attributed to clone-specific properties. Here, we provide evidence for an additional, host-dependent mechanism restraining LIP of lymphopenia-insensitive CD4
+ T cells. We show that such cells undergo LIP in lymphopenic mice lacking IFN-γ receptor (IFN-γR) expression, a process, which is promoted by the autocrine action of T cell-derived IFN-γ. Additionally, LIP of lymphopenia-insensitive CD4+ T cells requires an intact microflora and is accompanied by the massive accumulation of IL-6 and dendritic cells (DCs). Consistent with these results, IL-6 neutralization and the DC-specific restoration of IFN-γR expression are both sufficient to restrict LIP. Hence, the insensitivity of CD4+ T cells to lymphopenia relies on cell-intrinsic properties and a complex interplay between the commensal microflora, IL-6, IFN-γR+ DCs, and T cell-derived IFN-γ.- Published
- 2019
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43. p75 NTR regulates brain mononuclear cell function and neuronal structure in Toxoplasma infection-induced neuroinflammation.
- Author
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Düsedau HP, Kleveman J, Figueiredo CA, Biswas A, Steffen J, Kliche S, Haak S, Zagrebelsky M, Korte M, and Dunay IR
- Subjects
- Animals, Female, Immunity, Innate physiology, Inflammation immunology, Inflammation pathology, Leukocytes, Mononuclear pathology, Mice, Mice, Inbred C57BL, Mice, Knockout, Mice, Transgenic, Toxoplasmosis pathology, Leukocytes, Mononuclear physiology, Neurons physiology, Receptor, Nerve Growth Factor physiology, Toxoplasma, Toxoplasmosis immunology
- Abstract
Neurotrophins mediate neuronal growth, differentiation, and survival via tropomyosin receptor kinase (Trk) or p75 neurotrophin receptor (p75
NTR ) signaling. The p75NTR is not exclusively expressed by neurons but also by certain immune cells, implying a role for neurotrophin signaling in the immune system. In this study, we investigated the effect of p75NTR on innate immune cell behavior and on neuronal morphology upon chronic Toxoplasma gondii (T. gondii) infection-induced neuroinflammation. Characterization of the immune cells in the periphery and central nervous system (CNS) revealed that innate immune cell subsets in the brain upregulated p75NTR upon infection in wild-type mice. Although cell recruitment and phagocytic capacity of p75NTRexonIV knockout (p75-/- ) mice were not impaired, the activation status of resident microglia and recruited myeloid cell subsets was altered. Importantly, recruited mononuclear cells in brains of infected p75-/- mice upregulated the production of the cytokines interleukin (IL)-10, IL-6 as well as IL-1α. Protein levels of proBDNF, known to negatively influence neuronal morphology by binding p75NTR , were highly increased upon chronic infection in the brain of wild-type and p75-/- mice. Moreover, upon infection the activated immune cells contributed to the proBDNF release. Notably, the neuroinflammation-induced changes in spine density were rescued in the p75-/- mice. In conclusion, these findings indicate that neurotrophin signaling via the p75NTR affects innate immune cell behavior, thus, influencing the structural plasticity of neurons under inflammatory conditions., (© 2018 The Authors. Glia published by Wiley Periodicals, Inc.)- Published
- 2019
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44. Treatment of Toxoplasmosis: Historical Perspective, Animal Models, and Current Clinical Practice.
- Author
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Dunay IR, Gajurel K, Dhakal R, Liesenfeld O, and Montoya JG
- Subjects
- Animals, Drug Discovery trends, Humans, Models, Animal, Antiprotozoal Agents therapeutic use, Toxoplasmosis drug therapy
- Abstract
Primary Toxoplasma gondii infection is usually subclinical, but cervical lymphadenopathy or ocular disease can be present in some patients. Active infection is characterized by tachyzoites, while tissue cysts characterize latent disease. Infection in the fetus and in immunocompromised patients can cause devastating disease. The combination of pyrimethamine and sulfadiazine (pyr-sulf), targeting the active stage of the infection, is the current gold standard for treating toxoplasmosis, but failure rates remain significant. Although other regimens are available, including pyrimethamine in combination with clindamycin, atovaquone, clarithromycin, or azithromycin or monotherapy with trimethoprim-sulfamethoxazole (TMP-SMX) or atovaquone, none have been found to be superior to pyr-sulf, and no regimen is active against the latent stage of the infection. Furthermore, the efficacy of these regimens against ocular disease remains uncertain. In multiple studies, systematic screening for Toxoplasma infection during gestation, followed by treatment with spiramycin for acute maternal infections and with pyr-sulf for those with established fetal infection, has been shown to be effective at preventing vertical transmission and minimizing the severity of congenital toxoplasmosis (CT). Despite significant progress in treating human disease, there is a strong impetus to develop novel therapeutics for both the acute and latent forms of the infection. Here we present an overview of toxoplasmosis treatment in humans and in animal models. Additional research is needed to identify novel drugs by use of innovative high-throughput screening technologies and to improve experimental models to reflect human disease. Such advances will pave the way for lead candidates to be tested in thoroughly designed clinical trials in defined patient populations., (Copyright © 2018 American Society for Microbiology.)
- Published
- 2018
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45. Chronic Toxoplasma infection is associated with distinct alterations in the synaptic protein composition.
- Author
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Lang D, Schott BH, van Ham M, Morton L, Kulikovskaja L, Herrera-Molina R, Pielot R, Klawonn F, Montag D, Jänsch L, Gundelfinger ED, Smalla KH, and Dunay IR
- Subjects
- Animals, Antiprotozoal Agents pharmacology, Chronic Disease, Cytokines genetics, Cytokines metabolism, Disease Models, Animal, Gene Expression Regulation drug effects, Glutamic Acid metabolism, Mass Spectrometry, Membrane Proteins genetics, Membrane Proteins metabolism, Meta-Analysis as Topic, Mice, Nerve Tissue Proteins genetics, Nerve Tissue Proteins metabolism, Proteomics, RNA, Messenger metabolism, Sulfadiazine pharmacology, Synapses pathology, Synaptosomes drug effects, Tandem Mass Spectrometry, Toxoplasma pathogenicity, Brain metabolism, Gene Expression Regulation physiology, Synapses metabolism, Synaptosomes metabolism, Toxoplasmosis, Animal pathology
- Abstract
Background: Chronic infection with the neurotropic parasite Toxoplasma gondii has been implicated in the risk for several neuropsychiatric disorders. The mechanisms, by which the parasite may alter neural function and behavior of the host, are not yet understood completely., Methods: Here, a novel proteomic approach using mass spectrometry was employed to investigate the alterations in synaptic protein composition in a murine model of chronic toxoplasmosis. In a candidate-based strategy, immunoblot analysis and immunohistochemistry were applied to investigate the expression levels of key synaptic proteins in glutamatergic signaling., Results: A comparison of the synaptosomal protein composition revealed distinct changes upon infection, with multiple proteins such as EAAT2, Shank3, AMPA receptor, and NMDA receptor subunits being downregulated, whereas inflammation-related proteins showed an upregulation. Treatment with the antiparasitic agent sulfadiazine strongly reduced tachyzoite levels and diminished neuroinflammatory mediators. However, in both conditions, a significant number of latent cysts persisted in the brain. Conversely, infection-related alterations of key synaptic protein levels could be partly reversed by the treatment., Conclusion: These results provide evidence for profound changes especially in synaptic protein composition in T. gondii-infected mice with a downregulation of pivotal components of glutamatergic neurotransmission. Our results suggest that the detected synaptic alterations are a consequence of the distinct neuroinflammatory milieu caused by the neurotropic parasite.
- Published
- 2018
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46. Anti-inflammatory Effects of the Octapeptide NAP in Human Microbiota-Associated Mice Suffering from Subacute Ileitis.
- Author
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Escher U, Giladi E, Dunay IR, Bereswill S, Gozes I, and Heimesaat MM
- Abstract
The octapeptide NAP is well known for its neuroprotective properties. We here investigated whether NAP treatment could alleviate pro-inflammatory immune responses during experimental subacute ileitis. To address this, mice with a human gut microbiota were perorally infected with one cyst of Toxoplasma gondii (day 0) and subjected to intraperitoneal synthetic NAP treatment from day 1 until day 8 postinfection (p.i.). Whereas placebo (PLC) control animals displayed subacute ileitis at day 9 p.i., NAP-treated mice exhibited less pronounced pro-inflammatory immune responses as indicated by lower numbers of intestinal mucosal T and B lymphocytes and lower interferon (IFN)- γ concentrations in mesenteric lymph nodes. The NAP-induced anti-inflammatory effects were not restricted to the intestinal tract but could also be observed in extra-intestinal including systemic compartments, given that pro-inflammatory cytokines were lower in liver, kidney, and lung following NAP as compared to PLC application, whereas at day 9 p.i., colonic and serum interleukin (IL)-10 concentrations were higher in the former as compared to the latter. Remarkably, probiotic commensal bifidobacterial loads were higher in the ileal lumen of NAP as compared to PLC-treated mice with ileitis. Our findings thus further support that NAP might be regarded as future treatment option directed against intestinal inflammation.
- Published
- 2018
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47. Group 1 innate lymphoid cells in Toxoplasma gondii infection.
- Author
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Dunay IR and Diefenbach A
- Abstract
Innate lymphoid cells (ILCs) are a group of lymphocytes that carry out important functions in immunity to infections and in organ homeostasis at epithelial barrier surfaces. ILCs are innate immune cells that provide an early source of cytokines to initiate immune responses against pathogens. Cytotoxic ILCs (i.e. conventional (c)NK cells) and several subsets of helper-like ILCs are the major branches of the ILC family. Conventional NK cells and group 1 ILCs share several characteristics such as surface receptors and the ability to produce IFN-γ upon activation, but they differ in their developmental paths and in their dependence on specific transcription factors. Infection of mice with the intracellular parasite Toxoplasma gondii is followed by a strong Th1-mediated immune response. Previous studies indicate that NK1.1
+ cells contribute to the production of IFN-γ and TNF and cytotoxicity during acute T. gondii infection. Upon oral infection, the parasite infects intestinal enterocytes, and within the lamina propria, innate immune responses lead to initial parasite control although the infection disseminates widely and persists long-term in immune privileged sites despite adaptive immunity. Upon parasite entry into the small intestine, during the acute stage, ILC1 produce high levels of IFN-γ and TNF protecting barrier surfaces, thus essentially contributing to early parasite control. We will discuss here the role of innate lymphocytes during T. gondii infection in the context of the only recently appreciated diversity of ILC subsets., (© 2018 John Wiley & Sons Ltd.)- Published
- 2018
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48. Toxoplasma Gondii Exposure and Neurological Disorders: An Age- and Gender-Matched Case-Control Pilot Study.
- Author
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Alvarado-Esquivel C, Rico-Almochantaf YDR, Hernández-Tinoco J, Quiñones-Canales G, Sánchez-Anguiano LF, Torres-González J, Schott B, Liesenfeld O, and Dunay IR
- Abstract
Little is known about the association of Toxoplasma gondii infection and neurological disorders. We performed a case-control study with 344 patients with neurological diseases and 344 neurologically healthy age- and gender-matched subjects. Sera of participants were analyzed for anti- T . gondii IgG and IgM antibodies using commercially available immunoassays. Anti- T . gondii IgG antibodies were detected in 25 (7.3%) cases and in 35 (10.2%) controls (odds ratio [OR] = 0.69; 95% confidence interval [CI]: 0.40-1.18; P = 0.17). Anti- T . gondii IgM antibodies were found in 5 (14.3%) of the 25 IgG seropositive cases and in 13 (37.1°%) of the 35 IgG seropositive controls (P = 0.15). Anti- T . gondii IgG antibodies were found in 8 (3.8%) of 213 female cases and in 23 (10.8%) of 213 female controls (OR = 0.32; 95% CI: 0.14-0.73; P = 0.005); and in 17 (13.0%) of 131 male cases and in 12 (9.2%) of 131 male controls ( P = 0.32). No direct association between IgG seropositivity and specific neurological disorders was detected. We found no support for a role of latent T. gondii infection in the risk for neurological disorders in this setting. With respect to specific neurological disorders, further studies using larger patient cohorts will be required.
- Published
- 2017
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49. Toxoplasma Co-infection Prevents Th2 Differentiation and Leads to a Helminth-Specific Th1 Response.
- Author
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Ahmed N, French T, Rausch S, Kühl A, Hemminger K, Dunay IR, Steinfelder S, and Hartmann S
- Subjects
- Animals, Antibodies, Helminth blood, Antibodies, Protozoan blood, Cell Differentiation, Coinfection pathology, Cytokines metabolism, Eosinophilia, Mice, Nematospiroides dubius immunology, Strongylida Infections pathology, Toxoplasma immunology, Toxoplasmosis, Animal pathology, Coinfection immunology, Strongylida Infections complications, Strongylida Infections immunology, Th1 Cells immunology, Th2 Cells immunology, Toxoplasmosis, Animal complications, Toxoplasmosis, Animal immunology
- Abstract
Nematode infections, in particular gastrointestinal nematodes, are widespread and co-infections with other parasites and pathogens are frequently encountered in humans and animals. To decipher the immunological effects of a widespread protozoan infection on the anti-helminth immune response we studied a co-infection with the enteric nematode Heligmosomoides polygyrus in mice previously infected with Toxoplasma gondii . Protective immune responses against nematodes are dependent on parasite-specific Th2 responses associated with IL-4, IL-5, IL-13, IgE, and IgG1 antibodies. In contrast, Toxoplasma gondii infection elicits a strong and protective Th1 immune response characterized by IFN-γ, IL-12, and IgG2a antibodies. Co-infected animals displayed significantly higher worm fecundity although worm burden remained unchanged. In line with this, the Th2 response to H. polygyrus in co-infected animals showed a profound reduction of IL-4, IL-5, IL-13, and GATA-3 expressing T cells. Co-infection also resulted in the lack of eosinophilia and reduced expression of the Th2 effector molecule RELM-β in intestinal tissue. In contrast, the Th1 response to the protozoan parasite was not diminished and parasitemia of T. gondii was unaffected by concurrent helminth infection. Importantly, H. polygyrus specific restimulation of splenocytes revealed H. polygyrus -reactive CD4
+ T cells that produce a significant amount of IFN-γ in co-infected animals. This was not observed in animals infected with the nematode alone. Increased levels of H. polygyrus -specific IgG2a antibodies in co-infected mice mirrored this finding. This study suggests that polarization rather than priming of naive CD4+ T cells is disturbed in mice previously infected with T. gondii . In conclusion, a previous T. gondii infection limits a helminth-specific Th2 immune response while promoting a shift toward a Th1-type immune response.- Published
- 2017
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50. Behavior of Neutrophil Granulocytes during Toxoplasma gondii Infection in the Central Nervous System.
- Author
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Biswas A, French T, Düsedau HP, Mueller N, Riek-Burchardt M, Dudeck A, Bank U, Schüler T, and Dunay IR
- Subjects
- Animals, Brain immunology, Brain parasitology, Brain pathology, Central Nervous System parasitology, Cytokines metabolism, Disease Models, Animal, Granulocytes metabolism, Host-Parasite Interactions immunology, Inflammation immunology, Interferon-gamma metabolism, Mice, Mice, Inbred C57BL, Microglia immunology, Microglia metabolism, Monocytes immunology, Neutrophil Infiltration, Neutrophils metabolism, Reactive Oxygen Species isolation & purification, Receptors, Chemokine blood, Toxoplasmosis, Cerebral parasitology, Toxoplasmosis, Cerebral pathology, Central Nervous System immunology, Granulocytes immunology, Neutrophils immunology, Toxoplasma immunology, Toxoplasmosis immunology, Toxoplasmosis, Cerebral immunology
- Abstract
Cerebral toxoplasmosis is characterized by activation of brain resident cells and recruitment of specific immune cell subsets from the periphery to the central nervous system (CNS). Our studies revealed that the rapidly invaded Ly6G
+ neutrophil granulocytes are an early non-lymphoid source of interferon-gamma (IFN-γ), the cytokine known to be the major mediator of host resistance to Toxoplasma gondii ( T. gondii ). Upon selective depletion of Ly6G+ neutrophils, we detected reduced IFN-γ production and increased parasite burden in the CNS. Ablation of Ly6G+ cells resulted in diminished recruitment of Ly6Chi monocytes into the CNS, indicating a pronounced interplay. Additionally, we identified infiltrated Ly6G+ neutrophils to be a heterogeneous population. The Ly6G+ CD62-Lhi CXCR4+ subset released cathelicidin-related antimicrobial peptide (CRAMP), which can promote monocyte dynamics. On the other hand, the Ly6G+ CD62-Llo CXCR4+ subset produced IFN-γ to establish early inflammatory response. Collectively, our findings revealed that the recruited Ly6G+ CXCR4+ neutrophil granulocytes display a heterogeneity in the CNS with a repertoire of effector functions crucial in parasite control and immune regulation upon experimental cerebral toxoplasmosis.- Published
- 2017
- Full Text
- View/download PDF
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