Immunostaining of LYVE-1 and Podoplanin (D2-40) in MS brains. (A) D2-40 Immunostaining of control and MS human postmortem brain tissue. D2-40 immunostaining was mainly localized to the inner wall of brain endothelium in normal brain tissue sections and at perivascular inflammatory regions in MS brain tissue sections. Magnification 40×. (B) LYVE-1 immunostaining in control and MS human postmortem brain tissue. LYVE-1 immunostaining was not confined to vasculature in the MS tissue since, neuronal like cells were found to be LYVE-1+ in sample numbers 572, 3422, and 3816. Inset shows the axonal positivity of LYVE-1 in MS brain tissue. Magnification 40×. (C) Double immunofluorescence analysis of LYVE-1 and SMI311 in patient samples 3422 and 572. We observed a strong co-localization of SMI311 (recognizes both neuronal bodies and axons) stained in green (Alex-488) with LYVE-1 stained in red (cy3) in RRMS brain tissue samples. Furthermore, SMI311 was also observed to be co-localized on axons. LYVE-1 staining was primarily confined around the axons indicating oligodendrocyte/myelin positivity. Inset shows a higher magnification of SMI311 co-localization with axonal structures in MS 572 (magnification 100×). (D) LYVE-1 immunofluorescence also showed higher numbers of LYVE-1+ immune cell infiltrates in the MS brain vasculature (572, 3422) compared to controls (4064).