title: Signalling protein complexes isolated from primary human skin-resident T cells can be analysed by Multiplex IP-FCM.
abstract: Studying signal transduction in skin-resident T cells (sr-T cells) can be limited by the small size of clinical biopsies. Here, we isolated sr-T cells from clinical samples and analysed signalling protein complexes by multiplex immunoprecipitation detected by flow cytometry (mIP-FCM). In samples from two independent donors, antigenic stimulation induced signalling proteins to join shared complexes that were observed in seven pairwise combinations among five proteins. This demonstrates that sr-T cells isolated from small clinical samples provide sufficient material for mIP-FCM-based analysis of signalling-induced protein complexes. We propose that this strategy may be useful for gaining improved mechanistic insight of sr-T cell signal transduction associated with dermatological disease.
authors.affiliation: Department of Immunology, Mayo Clinic College of Medicine, Rochester, MN, USA.
authors.forename: Stephen E P
authors.initials: SE
authors.lastname: Smith
authors.forename: Steven C
authors.initials: SC
authors.lastname: Neier
authors.forename: Tessa R
authors.initials: TR
authors.lastname: Davis
authors.forename: Mark R
authors.initials: MR
authors.lastname: Pittelkow
authors.forename: Diana
authors.initials: D
authors.lastname: Gil
authors.forename: Adam G
authors.initials: AG
authors.lastname: Schrum
mesh_terms: D005434:Flow Cytometry; D006801:Humans; D047468:Immunoprecipitation / Q000379:methods*; D046912:Multiprotein Complexes / Q000032:analysis*; D015398:Signal Transduction; D012867:Skin / Q000737:chemistry* / Q000166:cytology / Q000276:immunology; D013601:T-Lymphocytes / Q000737:chemistry*
chemical_list: D046912:Multiprotein Complexes
keywords: T cell antigen receptor; multiplex technology; protein-protein interaction; signal transduction; skin-resident T cell