| 1. |
- Ishizaki, Taku, et al.
(author)
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The asparaginyl endopeptidase legumain after experimental stroke.
- 2010
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In: Journal of cerebral blood flow and metabolism : official journal of the International Society of Cerebral Blood Flow and Metabolism. - : SAGE Publications. - 1559-7016. ; Apr 7, s. 1756-1766
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Journal article (peer-reviewed)abstract
- Various proteases in the brain contribute to ischemic brain injury. We investigated the involvement of the asparaginyl endopeptidase legumain after experimental stroke. On the basis of gene array studies and in situ hybridizations, we observed an increase of legumain expression in the peri-infarct area of rats after transient occlusion of the middle cerebral artery (MCAO) for 120 mins with a maximum expression at 24 and 48 h. Immunohistochemical analyses revealed the expression of legumain in Iba1(+) microglial cells and glial fibrillary acidic protein-positive astrocytes of the peri-infarct area in mice after MCAO. Post-stroke recovery was also studied in aged legumain-deficient mice (45 to 58 weeks old). Legumain-deficient mice did not show any differences in physiologic parameters compared with respective littermates before, during MCAO (45 mins), and the subsequent recovery period of 8 days. Moreover, legumain deficiency had no effect on mortality, infarct volume, and the neurologic deficit determined by the rotating pole test, a standardized grip strength test, and the pole test. However, a reduced number of invading CD74(+) cells in the ischemic hemisphere indicates an involvement in post-stroke inflammation. We conclude that legumain is not essential for the functional deficit after MCAO but may be involved in mechanisms of immune cell invasion.Journal of Cerebral Blood Flow & Metabolism advance online publication, 17 March 2010; doi:10.1038/jcbfm.2010.39.
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| 2. |
- Kuric, Enida, et al.
(author)
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Demonstration of Tissue Resident Memory CD8 T Cells in Insulitic Lesions in Adult Patients with Recent-Onset Type 1 Diabetes
- 2017
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In: American Journal of Pathology. - : Elsevier BV. - 0002-9440 .- 1525-2191. ; 187:3, s. 581-588
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Journal article (peer-reviewed)abstract
- Subtypes of CD8(+) T cells in insulitic lesions in biopsy specimens from six subjects with recent-onset type 1 diabetes (T1D) and six nondiabetic matched controls were analyzed using simultaneous multicolor immunofluorescence. Also, insulitic islets based on accumulation of CD3(+) T cells were microdissected with laser-capture microscopy, and gene transcripts associated with inflammation and autoimmunity were analyzed. We found a substantial proportion, 43%, of the CD8(+) T cells in the insulitic lesions to display a tissue resident memory T cell (TRM) (CD8(+)CD69(+)CD103(+)) phenotype in T1D subjects. Most TRM cells were located in the insulitic lesion in the endocrine-exocrine interface. TRM cells were also sporadically found in islets of control subjects. Moreover, gene expression analysis showed a lack of active transcription of genes associated with acute inflammatory or cytotoxic T-cell responses. We present evidence that a substantial proportion of T cells in insulitic lesions of recent-onset T1D patients are TRM cells and not classic cytotoxic CD8(+) T cells. Our findings highlight the need for further analysis of the T cells involved in insulitis to elucidate their role in the etiology of T1D.
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| 3. |
- Kuric, Enida, et al.
(author)
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Dopamine receptor activation increases glial cell line-derived neurotrophic factor in experimental stroke.
- 2013
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In: Experimental Neurology. - : Elsevier BV. - 0014-4886. ; 247:May,9, s. 202-208
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Journal article (peer-reviewed)abstract
- Treatment with levodopa enhances functional recovery after experimental stroke but its mechanisms of action are elusive. Reactive astrocytes in the ischemic hemisphere are involved in mechanisms promoting recovery and also express dopamine 1 (D1) and dopamine 2 (D2) receptors. Here we investigated if the activation of astrocytic dopamine receptors (D1 and D2) regulates the expression of glial cell line-derived neurotrophic factor (GDNF) after combined in vitro hypoxia/aglycemia (H/A) and studied the expression of GDNF in the ischemic brain after treatment with levodopa/benserazide following transient occlusion of the middle cerebral artery (tMCAO) in the rat. Twenty-four hours after H/A, GDNF levels were upregulated in exposed astrocytes compared to normoxic control cultures and further elevated by the addition of the selective D1 receptor agonist (R)-(+)-SKF-38393 hydrochloride while D1 receptor antagonism by R(+)-SCH-23390 hydrochloride significantly reduced GDNF. No effect on GDNF levels was observed by the application of the D2 receptor agonist R(-)-2,10,11-trihydroxy-N-propyl-noraporphine hydrobromide hydrate or S-(-)-eticlopride hydrochloride (D2 receptor antagonist). After tMCAO, GDNF was upregulated in D1 expressing reactive astrocytes in the peri-infarct area. In addition, treatment with levodopa/benserazide significantly increased GDNF levels in the infarct core and peri-infarct area after tMCAO without affecting the expression of glial fibrillar acidic protein (GFAP), an intermediate filament and marker of reactive gliosis. After stroke, GDNF levels increase in the ischemic hemisphere in rats treated with levodopa, implicating GDNF in the mechanisms of tissue reorganization and plasticity and in l-DOPA enhanced recovery of lost brain function. Our results support levodopa treatment as a potential recovery enhancing therapy in stroke patients.
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| 4. |
- Kuric, Enida, et al.
(author)
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Dynamics of major histocompatibility complex class II-positive cells in the postischemic brain - influence of levodopa treatment
- 2014
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In: Journal of Neuroinflammation. - : Springer Science and Business Media LLC. - 1742-2094. ; 11
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Journal article (peer-reviewed)abstract
- Background: Cerebral ischemia activates both the innate and the adaptive immune response, the latter being activated within days after the stroke onset and triggered by the recognition of foreign antigens. Methods: In this study we have investigated the phenotype of antigen presenting cells and the levels of associated major histocompatibility complex class II (MHC II) molecules in the postischemic brain after transient occlusion of the middle cerebral artery (tMCAO) followed by levodopa/benserazide treatment. Male Sprague Dawley rats were subjected to tMCAO for 105 minutes and received levodopa (20 mg/kg)/benserazide (15 mg/kg) for 5 days starting on day 2 after tMCAO. Thereafter, immune cells were isolated from the ischemic and contralateral hemisphere and analyzed by flow cytometry. Complementarily, the spatiotemporal profile of MHC II-positive (MHC II+) cells was studied in the ischemic brain during the first 30 days after tMCAO; protein levels of MHC II and the levels of inflammation associated cytokines were determined in the ischemic hemisphere. Results: We found that microglia/macrophages represent the main MHC II expressing cell in the postischemic brain one week after tMCAO. No differences in absolute cell numbers were found between levodopa/benserazide and vehicle-treated animals. In contrast, MHC II protein levels were significant downregulated in the ischemic infarct core by levodopa/benserazide treatment. This reduction was accompanied by reduced levels of IFN-gamma, TNF-alpha and IL-4 in the ischemic hemisphere. In the contralateral hemisphere, we exclusively detected MHC II+ cells in the corpus callosum. Interestingly, the number of cells was increased by treatment with levodopa/benserazide independent from the infarct size 14 days after tMCAO. Conclusions: Results suggest that dopamine signaling is involved in the adaptive immune response after stroke and involves microglia/macrophages.
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| 5. |
- Kuric, Enida
(author)
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Effects of Dopamine Signaling on Recovery and Inflammation after Ischemic Stroke
- 2014
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Doctoral thesis (other academic/artistic)abstract
- Ischemic stroke, resulting from occlusion of a brain artery is the most common type of stroke with a prevalence of 15 million people and the leading cause of long-term disability worldwide. Importantly, partial spontaneous recovery of lost neurological functions occurs mainly during the first months after stroke onset, albeit to a limited extent. Recovery has been attributed to mechanisms of compensation but also by activation of endogenous repair processes. The area surrounding the infarct has been extensively studied and revealed functional and structural changes, however, the exact mechanisms underlying functional recovery have not been elucidated. The focus of the present study has been to investigate if levodopa/benserazide treatment, a common medication used as symptomatic treatment in Parkinson’s disease, affects processes enhancing recovery after stroke. Importantly levodopa/benserazide treatment also enhanced motor recovery and motor learning in preliminary stroke trials. In a reverse translational approach we have demonstrated an improved functional recovery by levodopa/benserazide treatment in a rat stroke model. We identified astrocytes and immune cells to respond to the treatment and to be involved in recovery enhancing effects. Among the processes affected by dopamine signaling were increased levels of the astrocyte derived neurotrophic factor, glial cell line-derived neurotrophic factor. In addition, we also demonstrated for the first time that dopamine signaling modulates inflammatory processes in the postischemic brain and periphery involving T-cells. Overall this investigation presents novel data regarding the role of dopamine signaling in the postischemic brain of relevance for recovery after stroke.
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| 6. |
- Kuric, Enida, et al.
(author)
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No Evidence for Presence of Mucosal-Associated Invariant T Cells in the Insulitic Lesions in Patients Recently Diagnosed with Type 1 Diabetes
- 2018
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In: American Journal of Pathology. - : Elsevier BV. - 0002-9440 .- 1525-2191. ; 188:8, s. 1744-1748
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Journal article (peer-reviewed)abstract
- Mucosal-associated invariant T (MAIT) cells are innate T cells that recognize bacteria-infected cells and are thought to play a role in autoimmune diseases. Translocation of duodenal bacteria and viruses to the pancreas through the pancreatic duct has been hypothesized to initiate an innate inflammatory response that could contribute to the development of type 1 diabetes, a process that could involve MAIT cells. In this study, we used immunohistochemistry and quantitative PCR to search for evidence of MAIT cells in the insulitic lesions in the pancreas of human patients recently diagnosed with type 1 diabetes. Only a few scattered MAIT cells were found within the exocrine parenchyma in all pancreatic samples, but no MAIT cells were found in association to the islets. Also, only low gene expression levels of the MAIT T-cell receptor V alpha 7.2-3 alpha 33 were found in the pancreas of patients with type 1 diabetes, in similar Levels as that in nondiabetic organ donors used as control. The absence of MAIT cells shown in insulitic lesions in humans questions the direct cytotoxic role of these cells in beta-cell destruction.
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| 7. |
- Kuric, Enida, et al.
(author)
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Reduction of rat brain CD8(+) T-cells by levodopa/benserazide treatment after experimental stroke.
- 2014
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In: European Journal of Neuroscience. - : Wiley. - 1460-9568 .- 0953-816X. ; 40:2, s. 2463-2470
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Journal article (peer-reviewed)abstract
- The activation of inflammatory cascades in the ischemic hemisphere impairs mechanisms of tissue reorganization with consequences for recovery of lost neurological function. Recruitment of T-cell populations to the post-ischemic brain occurs and represents a significant part of the inflammatory response. This study was conducted to investigate if treatment with levodopa, potentially acting as an immunomodulator, affects the T-cell accumulation in the post-ischemic brain. Male Sprague-Dawley rats were subjected to transient occlusion of the middle cerebral artery (tMCAO) for 105 min followed by levodopa/benserazide treatment (20 mg/kg/15 mg/kg) for 5 days initiated on day 2 post-stroke. One week after tMCAO, T-cell populations were analysed from brains, and levels of interleukin (IL)-1β, chemokine (C-X-C motif) ligand 1, IL-4, IL-5, interferon gamma and IL-13 were analysed. After levodopa/benserazide treatment, we found a significant reduction of cytotoxic T-cells (CD3(+) CD8(+) ) in the ischemic hemisphere together with reduced levels of T-cell-associated cytokine IL-5, while other T-cell populations (CD3(+) , CD3(+) CD4(+) , CD3(+) CD4(+) CD25(+) ) were unchanged compared with vehicle-treated rats. Moreover, a reduced number of cells was associated with reduced levels of intercellular adhesion molecule 1, expressed in endothelial cells, in the infarct core of levodopa/benserazide-treated animals. Together, we provide the first evidence that dopamine can act as a potential immunomodulator by attenuating inflammation in the post-ischemic brain.
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| 8. |
- Kuric, Enida, et al.
(author)
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Reversal of stroke induced lymphocytopenia by levodopa/benserazide treatment.
- 2014
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In: Journal of Neuroimmunology. - : Elsevier BV. - 1872-8421 .- 0165-5728. ; 269:1-2, s. 94-97
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Journal article (peer-reviewed)abstract
- Evidences exist that dopamine is involved in T-cell homeostasis. Here, we investigated if daily treatment with levodopa (20mg/kg)/benserazide (15mg/kg) regulates the number of T-cells in the blood of Sprague Dawley rats subjected to transient occlusion of the middle cerebral artery (tMCAO). T-cells and associated inflammatory cytokines were analyzed in the blood one week after tMCAO. Treatment with levodopa/benserazide completely recovered T-helper cell depletion in animals subjected to tMCAO whereas T-cell associated cytokines remained unaffected after stroke, however, cytokines were downregulated in levodopa treated sham operated animals. Together, we demonstrate that dopamine has beneficial effects on poststroke immunodepression.
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| 9. |
- Lundberg, Marcus, et al.
(author)
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Expression of Interferon-Stimulated Genes in Insulitic Pancreatic Islets of Patients Recently Diagnosed With Type 1 Diabetes
- 2016
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In: Diabetes. - : American Diabetes Association. - 0012-1797 .- 1939-327X. ; 65:10, s. 3104-3110
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Journal article (peer-reviewed)abstract
- A primary insult to the pancreatic islets of Langerhans, leading to the activation of innate immunity, has been suggested as an important step in the inflammatory process in type 1 diabetes (T1D). The aim of this study was to examine whether interferon (IFN)-stimulated genes (ISGs) are overexpressed in human T1D islets affected with insulitis. By using laser capture microdissection and a quantitative PCR array, 23 of 84 examined ISGs were found to be overexpressed by at least fivefold in insulitic islets from living patients with recent-onset T1D, participating in the Diabetes Virus Detection (DiViD) study, compared with islets from organ donors without diabetes. Most of the overexpressed ISGs, including GBP1, TLR3, OAS1, EIF2AK2, HLA-E, IFI6, and STAT1, showed higher expression in the islet core compared with the peri-islet area containing the surrounding immune cells. In contrast, the T-cell attractant chemokine CXCL10 showed an almost 10-fold higher expression in the peri-islet area than in the islet, possibly partly explaining the localization of T cells mainly to this region. In conclusion, insulitic islets from recent-onset T1D subjects show overexpression of ISGs, with an expression pattern similar to that seen in islets infected with virus or exposed to IFN-gamma/interleukin-1 beta or IFN-alpha.
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| 10. |
- Ruscher, Karsten, et al.
(author)
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Effects of chronic clozapine administration on apolipoprotein D levels and on functional recovery following experimental stroke.
- 2010
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In: Brain Research. - : Elsevier BV. - 1872-6240 .- 0006-8993. ; 1321, s. 152-163
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Journal article (peer-reviewed)abstract
- Elevated brain levels of apolipoprotein D (ApoD) correlate with improved neurological recovery after experimental stroke. Hence, a pharmacological induction of ApoD in the postischemic brain could be beneficial for recovery after stroke. Here we investigated the effect of Clozapine, a compound that increases the expression of ApoD, in two rat models of experimental stroke. Rats were subjected to permanent occlusion of the middle cerebral artery (pMCAO) and treated with Clozapine (i.p. 10mg/kg body weight) or saline for 8 or 28days starting on the second day after MCAO. ApoD levels increased by 35% in the peri-infarct area after 10 and 30days after pMCAO, mainly in neuron-specific nuclear protein (NeuN) positive neurons and glial fibrillary acidic protein (GFAP) positive astrocytes. Clozapine did not affect the neurological deficit assessed by the rotating pole test and a grip strength test at 7d, 14d, 21d, and 28days after pMCAO. Functional outcome and the infarct size were similar in rats subjected to transient MCAO and injected with Clozapine (i.p. 10mg/kg body weight) or saline for 26days starting on the second day after tMCAO. We conclude that Clozapine affects cellular processes involved in peri-infarct tissue reorganization, but does not affect functional recovery after MCAO.
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