title: Antimicrobial peptide β-defensin-1 expression is upregulated in Alzheimer's brain.
abstract: BACKGROUND
  The human β-defensins (hBDs) are a highly conserved family of cationic antimicrobial and immunomodulatory peptides expressed primarily by epithelial cells in response to invasion by bacteria, fungi and some viruses. To date, the most studied members of this family of peptides are hBD-1, -2, and -3. Expression of hBD-1 and -2 has been demonstrated previously in cultured microglia and astrocytes of both mouse and human brain. Unlike inducible hBD-2 and -3, hBD-1 is constitutively expressed and is not generally upregulated by proinflammatory factors. In this study, we investigated whether hBDs, as active components of the innate immune response, are affected by pathological events in the Alzheimer's disease (AD) brain. We assessed the expression of hBD-1, -2, and -3 in tissue obtained at autopsy from AD and age-matched control brains.
  
  
  METHODS
  Fixed and frozen choroid plexus and the CA1 region of the hippocampus were obtained at autopsy from individuals diagnosed with AD, or from age-matched control brains without diagnosed neurodegenerative disease. Histopathologically diagnosed AD brain tissue was obtained for our study. Immunocytochemical analysis was performed using affinity purified polyclonal antibodies directed against hBD-1, -2, or -3. TaqMan gene expression assays were used to quantify the mRNA of hBD-1, -2, and -3 in the choroid plexus and hippocampus. Immunocytochemical detection of iron deposits was achieved using a modified Perl's stain for redox-active iron. In vitro experiments were performed on human primary oral epithelial cells to model the human choroid plexus epithelial response to ferric chloride. Cells were then exposed to ferric chloride added to selected wells at 0, 1, or 10 mM concentrations for 24 h at 37°C. Total mRNA was isolated to quantify hBD-1 mRNA expression by RTqPCR.
  
  
  RESULTS
  hBD-1 peptide is apparent in astrocytes of the AD hippocampus and hippocampal neurons, notably within granulovacuolar degeneration structures (GVD). A higher level of hBD-1 was also seen in the choroid plexus of AD brain in comparison to age-matched control tissue. Increased expression of hBD-1 mRNA was observed only in the choroid plexus of the AD brain when compared to expression level in age-matched control brain. Redox-active iron was also elevated in the AD choroid plexus and in vitro addition of Fe⁺³Cl₃ to cultured epithelial cells induced hBD-1 mRNA expression.
  
  
  CONCLUSIONS
  Our findings suggest interplay between hBD-1 and neuroimmunological responses in AD, marked by microglial and astrocytic activation, and increased expression of the peptide within the choroid plexus and accumulation within GVD. As a constitutively expressed component of the innate immune system, we propose that hBD-1 may be of considerable importance early in the disease process. We also demonstrate that increased iron deposition in AD may contribute to the elevated expression of hBD-1 within the choroid plexus. These findings represent a potentially important etiological aspect of Alzheimer's disease neuropathology not previously reported.
authors.affiliation: Department of Biological Sciences, School of Dental Medicine, Case Western Reserve University, 2124 Cornell Rd,, Cleveland, OH 44106, USA. wmw5@case.edu.
authors.forename: Wesley M
authors.initials: WM
authors.lastname: Williams
authors.forename: Sandy
authors.initials: S
authors.lastname: Torres
authors.forename: Sandra L
authors.initials: SL
authors.lastname: Siedlak
authors.forename: Rudy J
authors.initials: RJ
authors.lastname: Castellani
authors.forename: George
authors.initials: G
authors.lastname: Perry
authors.forename: Mark A
authors.initials: MA
authors.lastname: Smith
authors.forename: Xiongwei
authors.initials: X
authors.lastname: Zhu
mesh_terms: D000368:Aged; D000369:Aged, 80 and over; D000544:Alzheimer Disease / Q000378:metabolism*; D015415:Biomarkers / Q000032:analysis*; D002831:Choroid Plexus / Q000378:metabolism*; D005260:Female; D006624:Hippocampus / Q000378:metabolism*; D006801:Humans; D007150:Immunohistochemistry; D007501:Iron / Q000378:metabolism; D008297:Male; D020411:Oligonucleotide Array Sequence Analysis; D010084:Oxidation-Reduction; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015854:Up-Regulation; D023083:beta-Defensins / Q000096:biosynthesis*
chemical_list: D015415:Biomarkers; C406407:DEFB1 protein, human; D023083:beta-Defensins; D007501:Iron