myelin-oligodendrocyte-glycoprotein-(35-55) and Infarction--Middle-Cerebral-Artery

Topics: Animals; Chloride-Bicarbonate Antiporters; Disease Models, Animal; Infarction, Middle Cerebral Artery; Macrophages; Mice, Inbred C57BL; Microglia; Myelin-Oligodendrocyte Glycoprotein; Peptide Fragments; Stroke; Sulfate Transporters

Chemoattraction of leukocytes into the brain after induction of middle cerebral artery occlusion (MCAO) increases the lesion size and worsens disease outcome. Our previous studies demonstrated that partial MHC class II constructs can reverse this process. However, the potential application of pMHC to human stroke is limited by the need to rapidly match recipient MHC class II with the β1 domain of the pMHC construct. We designed a novel recombinant protein comprised of the HLA-DRα1 domain linked to MOG-35-55 peptide but lacking the β1 domain found in pMHC and treated MCAO after 4 h reperfusion in humanized DR2 mice. Infarct volumes were quantified after 96 h reperfusion and immune cells from the periphery and CNS were evaluated for expression of CD74 and other cell surface, cytokine and pathway markers. This study demonstrates that four daily treatments with DRα1-MOG-35-55 reduced infarct size by 40 % in the cortex, striatum and hemisphere, inhibited the migration of activated CD11b+CD45high cells from the periphery to the brain and reversed splenic atrophy. Furthermore, DRα1-MOG-35-55 bound to CD74 on monocytes and blocked both binding and downstream signaling of macrophage migration inhibition factor (MIF) that may play a key role in infarct development. The novel DRα1-MOG-35-55 construct is highly therapeutic in experimental stroke and could be given to all patients at least 4 h after stroke onset without the need for tissue typing due to universal expression of DRα1 in humans.

Topics: Animals; Antigens, Differentiation, B-Lymphocyte; Atrophy; Chemotaxis, Leukocyte; Drug Administration Schedule; Drug Evaluation, Preclinical; Gene Expression Profiling; Histocompatibility Antigens Class II; HLA-B15 Antigen; HLA-DRB1 Chains; Humans; Infarction, Middle Cerebral Artery; Male; Mice; Mice, Transgenic; Microglia; Monocytes; Myelin-Oligodendrocyte Glycoprotein; Nerve Tissue Proteins; Peptide Fragments; Protein Structure, Tertiary; Random Allocation; Recombinant Fusion Proteins; Spleen; Tumor Necrosis Factor-alpha

We aimed at validating a plasma biomarker for neuronal damage that can be used in acute and chronic models of neurological diseases.. We investigated two different models, middle cerebral artery occlusion followed by reperfusion and MOG35-55-induced experimental autoimmune encephalomyelitis (EAE). In stroke experiments we measured infarct sizes by magnetic resonance imaging and vital stainings and correlated them with plasma levels of neuron specific enolase (NSE) at different time points after reperfusion. Equally, in EAE experiments, we correlated NSE levels with neurological scores and histopathological damage of axons at different time points. We detected plasma NSE levels by ELISA.. Plasma NSE levels correlated significantly with stroke size, EAE score and histopathological damage in EAE. Investigations into the dynamics of neuronal loss over time correlated well with the dynamics of NSE levels. NSE even predicted the onset of EAE, before clinical signs were recordable.. Plasma NSE is a valid and simple experimental biomarker that allows quantifying the degree of neuronal injury in a non-invasive approach.

Topics: Amyloid beta-Protein Precursor; Animals; Brain; Brain Infarction; Cells, Cultured; Disease Models, Animal; Embryo, Mammalian; Glutamic Acid; Infarction, Middle Cerebral Artery; L-Lactate Dehydrogenase; Magnetic Resonance Imaging; Mice; Mice, Inbred C57BL; Microtubule-Associated Proteins; Multiple Sclerosis; Myelin-Oligodendrocyte Glycoprotein; Neurons; Peptide Fragments; Phosphopyruvate Hydratase; Time Factors

Mucosal tolerance has been used successfully to treat animal models of autoimmune diseases and is being tested in human diseases. In this work we demonstrate the reduction of infarct size following mucosal tolerance by myelin oligodendrocyte glycoprotein (MOG) (35-55) peptide in mouse stroke model. Nasal MOG was most efficacious and reduced ischemic infarct size by 70% at 24 h as well as improving behavior score. Using immunohistological methods and IL-10 -/- mice, we demonstrate the importance of IL-10-producing CD4+ T cells in the reduction of the ischemic infarct volume following middle cerebral artery occlusion (MCAO). Furthermore, adoptive transfer of CD4+ T cells from nasally tolerized mice to untreated mice prior to MCAO surgery significantly decreased stroke size (p<0.001 vs. control), whereas CD4+ T cells from nasally tolerized IL-10-deficient mice had no significant effect. Based on these results, modulation of cerebral inflammation by mucosal tolerance to myelin antigens may have applicability both as prophylactic therapy and treatment following ischemia attacks.

Topics: Analysis of Variance; Animals; Antigens, CD; Behavior, Animal; Brain Infarction; CD4-Positive T-Lymphocytes; Cells, Cultured; Cytokines; Disease Models, Animal; Drug Administration Routes; Drug Administration Schedule; Enzyme-Linked Immunosorbent Assay; Female; Glycoproteins; Immune Tolerance; Immunohistochemistry; Infarction, Middle Cerebral Artery; Interleukin-10; Mice; Mice, Inbred C57BL; Mice, Knockout; Models, Immunological; Myelin-Oligodendrocyte Glycoprotein; Peptide Fragments; Severity of Illness Index; Stroke; Time Factors

Inflammation plays an important role in ischemic stroke and in humans IL-10 may have a beneficial effect in stroke. We mucosally administered myelin oligodendrocyte glycoprotein (MOG) 35-55 peptide to C57BL/6 mice before middle cerebral artery occlusion (MCAO) to induce an anti-inflammatory T cell response directed at CNS myelin. Nasal and oral administration of MOG(35-55) peptide decreased ischemic infarct size at 24 and 72 h after MCAO surgery. Nasal MOG(35-55) peptide was most efficacious and reduced infarct size by 70% at 24 h and by 50% at 72 h (p

Topics: Administration, Intranasal; Administration, Oral; Adoptive Transfer; Animals; Brain Ischemia; CD4-Positive T-Lymphocytes; Cytokines; Disease Models, Animal; Female; Glycoproteins; Immune Tolerance; Immunity, Mucosal; Immunohistochemistry; Infarction, Middle Cerebral Artery; Interleukin-10; Mice; Mice, Inbred C57BL; Mice, Knockout; Myelin-Oligodendrocyte Glycoprotein; Neuroprotective Agents; Peptide Fragments; Vaccines