Journal: Acta Neuropathologica

Article Title: Oxidative tissue injury in multiple sclerosis is only partly reflected in experimental disease models

doi: 10.1007/s00401-014-1263-5

Figure Lengend Snippet: Primary antibodies and antigen retrieval procedures

Article Snippet: Ox8 , Mouse (mAb) , Anti-rat CD8 , 1:400 , 0 , Seralab; MAS041.

Techniques: Binding Assay, Virus

Journal: Acta Neuropathologica

Article Title: Oxidative tissue injury in multiple sclerosis is only partly reflected in experimental disease models

doi: 10.1007/s00401-014-1263-5

Figure Lengend Snippet: Quantification of markers for inflammation and oxidative injury in different experimental models for CNS inflammation and multiple sclerosis

Article Snippet: Ox8 , Mouse (mAb) , Anti-rat CD8 , 1:400 , 0 , Seralab; MAS041.

Techniques: Injection, Virus

Journal: Acta Neuropathologica

Article Title: Oxidative tissue injury in multiple sclerosis is only partly reflected in experimental disease models

doi: 10.1007/s00401-014-1263-5

Figure Lengend Snippet: Oxidative burst, oxidative injury and iron accumulation in CD8 + T cell-mediated EAE, in innate immunity-driven inflammation and in toxic cuprizone-mediated demyelination. a – f Brain inflammation induced by CD8 + T cells. Perivascular inflammatory infiltrates were associated with massive microglial activation in the adjacent tissue ( a ; Iba-1). Perivascular inflammatory cells and to lower degree also the surrounding microglia highly expressed p22phox ( b ), while iNOS expression was minor and restricted to a few perivascular cells ( c ). The immunohistochemistry for oxidised phospholipids (E06; d ) was negative. Iron deposition was lacking in most lesions ( e ) but occurred in few lesions within perivascular macrophages ( f ). g – m Inflammatory demyelinating lesions induced by local injection of LPS into the spinal cord dorsal column. In the early inflammatory stage (1 day after LPS injection), p22phox + granulocytes ( g ) and iNOS + macrophages ( h ) were present in the lesions. At the peak of active demyelination (9 days after LPS injection), lesions were densely infiltrated by macrophages and showed microglial activation at the lesion edge ( i ; Iba-1, asterisk ). Macrophages in these lesions and, to a lower degree, microglia at the lesion edges showed expression of p22phox ( j ). Despite a very strong expression of p22phox in active lesions ( j ), the expression of iNOS was very low ( k ). There was a weak reactivity for oxidised phospholipids in the lesions ( l ; E06). Individual perivascular macrophages were iron positive ( m ). n – s Actively demyelinating lesion in the corpus callosum induced by cuprizone diet. Extensive loss of myelin was seen in the corpus callosum ( n ), which was densely packed with macrophages and microglia ( o ), although their majority did not express p22phox ( p ) and iNOS only very weakly ( q ). Additionally, there was no evidence for oxidised phospholipids ( r ; E06) or iron deposition ( s ) in the lesions. Scale bar 50 μm

Article Snippet: Ox8 , Mouse (mAb) , Anti-rat CD8 , 1:400 , 0 , Seralab; MAS041.

Techniques: Activation Assay, Expressing, Immunohistochemistry, Injection

Journal: Acta Neuropathologica

Article Title: Oxidative tissue injury in multiple sclerosis is only partly reflected in experimental disease models

doi: 10.1007/s00401-014-1263-5

Figure Lengend Snippet: Oxidative burst, oxidative injury and iron accumulation in inflammatory demyelinating lesions in MHV-JHM coronavirus-induced encephalomyelitis. a – i Basic characterisation of inflammatory demyelinating lesions in the spinal cord of an infected Lewis rat with demyelinating encephalomyelitis after infection with MHV-JHM coronavirus. There was extensive plaque-like demyelination ( a ; MBP) with relative preservation of axons ( b and f ; e NAWM; Bodian silver impregnation). Virus antigen was present in the periplaque white and grey matter ( c ; immunohistochemistry for nucleocapsid protein N) in neurons ( g ) as well as in glia cells in the white matter ( h ). Active demyelination was associated with profound T cell-mediated inflammation ( d ), the majority of inflammatory cells being CD8 + T cells ( blue ; i ) and ED1 + macrophages ( brown ; i ). j – n Determination of microglial activation in MHV-JHM coronavirus-induced lesions. Activated microglia formed microglial nodules in the NAWM reactive for Iba-1 ( j ) and p22phox ( l ). The edge of an active lesion is shown with immunocytochemistry for macrophages/microglia (Iba-1; k ), for myelin (MBP; m ) and for expression of NADPH oxidase (p22phox; n ). Active demyelination was associated with profound microglial activation ( k ; Iba-1). Their numbers increased in the periplaque white matter (PPWM) towards the lesion edge ( k ; left side ) showing the highest density at the zone of initial demyelination ( k , middle ) and decreasing towards the centre of the lesion, where many cells exhibited a macrophage phenotype ( k ; right side ). Immunohistochemistry for p22phox in serial-cut adjacent sections from the lesion ( n ) showed a staining pattern similar to that of Iba-1. iNOS expression was sparse in the lesions ( o ). p – u Accumulation of oxidative damage and iron in MHV-JHM coronavirus-induced lesions. Lesions showed abundant reactivity for oxidised phospholipids ( p , q , s E06) in myelin ( p ), in cells with apoptotic nuclei ( q ), in macrophage granules ( q ) and in axonal spheroids ( s ). In addition, nuclei within the lesions contained oxidised DNA (8OHdH reactivity; r ). Most lesions did not show iron deposition ( t ), although iron-positive perivascular macrophages accumulated in individual lesions ( u ). Scale bar 50 μm except for a – d = 0.5 mm

Article Snippet: Ox8 , Mouse (mAb) , Anti-rat CD8 , 1:400 , 0 , Seralab; MAS041.

Techniques: Infection, Preserving, Virus, Immunohistochemistry, Activation Assay, Immunocytochemistry, Expressing, Staining