title: A specific anti-Aire antibody reveals aire expression is restricted to medullary thymic epithelial cells and not expressed in periphery.
abstract: Autoimmune polyendocrinopathy-candidiasis-ectodermal dystrophy is an autoimmune disorder caused by mutations in the autoimmune regulator gene AIRE. We examined the expression of Aire in different organs (thymus, spleen, and lymph nodes) in C57BL/6 mice, using a novel rat mAb, specific for murine Aire. Using flow cytometry, directly fluorochrome-labeled mAb revealed Aire expression in a rare thymic cellular subset that was CD45(-), expressed low levels of Ly51, and was high for MHC-II and EpCam. This subset also expressed a specific pattern of costimulatory molecules, including CD40, CD80, and PD-L1. Immunohistochemical analysis revealed that Aire(+) cells were specifically localized to the thymus or, more precisely, to the cortico-medulla junction and medulla, correlating with the site of negative selection. Although in agreement with previous studies, low levels of Aire mRNA was detected in all dendritic cell subtypes however lacZ staining, immunohistochemistry and flow cytometry failed to detect Aire protein. At a cellular level, Aire was expressed in perinuclear speckles within the nucleus. This report provides the first detailed analysis of Aire protein expression, highlighting the precise location at both the tissue and cellular level.
authors.affiliation: Division of Molecular and Medicine, Walter and Eliza Hall Institute of Medical Research, University of Melbourne, Parkville, Victoria, Australia. hubert@wehi.edu.au
authors.forename: François-Xavier
authors.initials: FX
authors.lastname: Hubert
authors.forename: Sarah A
authors.initials: SA
authors.lastname: Kinkel
authors.forename: Kylie E
authors.initials: KE
authors.lastname: Webster
authors.forename: Ping
authors.initials: P
authors.lastname: Cannon
authors.forename: Pauline E
authors.initials: PE
authors.lastname: Crewther
authors.forename: Anna I
authors.initials: AI
authors.lastname: Proeitto
authors.forename: Li
authors.initials: L
authors.lastname: Wu
authors.forename: William R
authors.initials: WR
authors.lastname: Heath
authors.forename: Hamish S
authors.initials: HS
authors.lastname: Scott
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000032:analysis* / Q000096:biosynthesis; D000918:Antibody Specificity; D000937:Antigen-Antibody Reactions; D002467:Cell Nucleus / Q000276:immunology / Q000378:metabolism; D004847:Epithelial Cells / Q000276:immunology* / Q000378:metabolism*; D016130:Immunophenotyping; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D008822:Mice, Transgenic; D008969:Molecular Sequence Data; D045663:Quantum Dots; D051381:Rats; D013950:Thymus Gland / Q000166:cytology* / Q000276:immunology* / Q000378:metabolism; D014157:Transcription Factors / Q000096:biosynthesis* / Q000172:deficiency / Q000235:genetics / Q000276:immunology*; D000096927:AIRE Protein
chemical_list: D000911:Antibodies, Monoclonal; D014157:Transcription Factors; D000096927:AIRE Protein