Figure 2.

Cellular composition, relative expression of osteopontin (OPN) and serum calcium binding protein B (S100B), and BRAFV600E expression in central nervous system Langerhans cell histiocytosis (CNS-LCH) lesions and LCH with neurodegenerative disease (LCH-ND). (A) Immuno-histochemical analysis of CD207, VE-1 (identifies BRAF-V600E protein expression), OPN, and CD3 in representative tissue sections obtained from indicated biopsy specimens (original magnification 340). (B) Steady-state protein expression of OPN and S100B in representative LCH lesions and glioblastoma brain tumor biopsies. The blots were stripped and reprobed with glyceraldehyde 3-phosphate dehydrogenase (GADPH) to confirm equivalent loading. (C) Relative messenger RNA expression of the indicated transcripts (SPP1/OPN, blue; CD207, orange; CD1a, gray; CD3, black) as determined by quantitative polymerase chain reaction (qPCR) for LCH lesions from bone, LCH lesions from lungs, LCH with pituitary involvement (Pit), and LCH with early onset neurodegeneration (ND). Data were normalized to adjusted GAPDH expression and analyzed using the ΔCt method. *Complementary DNA level below the limit of detection by qPCR. (D) qPCR for BRAFV600E from genomic DNA (gray), and relative OPN (SPP1) expression (blue) from biopsies from various regions of whole brain autopsy from a patient with advanced LCH-ND. (E) Magnetic resonance images (T2 FLAIR) from the same patient obtained 2 years before death from progression of LCH-ND. The anatomic areas corresponding to the biopsy sections from qPCR are indicated.