Journal: Experimental neurology

Article Title: Efficacy of vitamin D in treating multiple sclerosis-like neuroinflammation depends on developmental stage.

doi: 10.1016/j.expneurol.2013.08.002

Figure Lengend Snippet: Fig. 4. Antibody extravasation and complement activation in the spinal cord of juvenile/adolescent rats affected by inflammation/demyelination evidenced the humoral immunity. Group-representative neuropathology of spinal cord in juvenile/adolescent rats with EAE fed either with vitamin D supplemented (AI–DI), regular (AII–DII) or deprived (AIII–DIII) diet (IHC was performed on the same tissue as histopathology and IHC presented in Fig. 3). Analysis of the Ig isotypes IgG (AI–AIII) and IgM (BI–BIII) revealed the lowest immunoglobulin reactivity in the vitamin D supplemented group (AI, BI respectively) compared to the other two diet regimens (AII, BII and AIII, BIII, respectively; magnification ×100). Staining for C3 (CI–CIII; magnification ×100) revealed massive depositions upon regular diet (CII) and vitamin D deprived diet (CIII) matching the demyelinated areas. C3 deposition in the treated animal (CI) is much reduced and mainly restricted to the perivascular area (arrows). C5b-9 was detected in the lesions of rats that received regular diet (DII) and vitamin D deprived diet (DIII). Virtually no C5b-9 depositions in the lesions of the vitamin D supplemented rat (DI). Staining revealed also the presence of granulocytes in the lesions of rats fed with vitamin D deprived (DIII) and the regular diet (DII), whereas the infiltrate in the vitamin D supplemented animals is mainly composed of mononuclear cells (DI).

Article Snippet: In adjacent serial sections IHC was performed by using antibodies against the following targets: lysosomes of rat activatedmacrophages andmicroglia cells (ED1,mouse anti-ratmonoclonal antibody, AbD Serotec, Germany; 1:1000), rat T lymphocytes (W3/13, mouse anti-rat, Abcam, UK, 1:100) and anti-amyloid precursor protein (APP, mouse anti-rat, Millipore Chemikon, 1:500), complement components (mouse monoclonal antiC1q antibody (Abcam UK, 1:100), mouse monoclonal complement C3 antibody (Novus Biologicals, UK, 1:400), rabbit polyclonal anti-C5b-9 antibody (Bioss, USA, 1:500)) and immunoglobulins (goat anti-rat IgG (biotinylated, GE Healthcare, 1:200) and goat anti-rat IgM antibody (biotinylated, Novus Biologicals, UK, 1:1000)).

Techniques: Activation Assay, Histopathology, Staining