title: The skin immune atlas: three-dimensional analysis of cutaneous leukocyte subsets by multiphoton microscopy.
abstract: Site-specific differences in skin response to pathogens and in the course of cutaneous inflammatory diseases are well appreciated. The composition and localization of cutaneous leukocytes has been studied extensively using histology and flow cytometry. However, the precise three-dimensional (3D) distribution of distinct immune cell subsets within skin at different body sites requires visualization of intact living skin. We used intravital multiphoton microscopy in transgenic reporter mice in combination with quantitative flow cytometry to generate a 3D immune cell atlas of mouse skin. The 3D location of innate and adaptive immune cells and site-specific differences in the densities of macrophages, T cells, and mast cells at four defined sites (ear, back, footpad, and tail) is presented. The combinatorial approach further demonstrates an as yet unreported age-dependent expansion of dermal gamma-delta T cells. Localization of dermal immune cells relative to anatomical structures was also determined. Although dendritic cells were dispersed homogeneously within the dermis, mast cells preferentially localized to the perivascular space. Finally, we show the functional relevance of site-specific mast cell disparities using the passive cutaneous anaphylaxis model. These approaches are applicable to assessing immune cell variations and potential functional consequences in the setting of infection, as well as the pathogenesis of inflammatory skin conditions.
authors.affiliation: Centenary Institute, Newtown, New South Wales, Australia; Discipline of Dermatology, The University of Sydney, Camperdown, New South Wales, Australia; Department of Dermatology, Royal Prince Alfred Hospital, Camperdown, New South Wales, Australia. Electronic address: p.tong@centenary.org.au.
authors.forename: Philip L
authors.initials: PL
authors.lastname: Tong
authors.affiliation: Centenary Institute, Newtown, New South Wales, Australia; Sydney Medical School, The University of Sydney, Camperdown, New South Wales, Australia.
authors.forename: Ben
authors.initials: B
authors.lastname: Roediger
authors.affiliation: Centre for Cancer Biology, University of South Australia and SA Pathology, Adelaide, South Australia, Australia.
authors.forename: Natasha
authors.initials: N
authors.lastname: Kolesnikoff
authors.affiliation: Centenary Institute, Newtown, New South Wales, Australia; Sydney Medical School, The University of Sydney, Camperdown, New South Wales, Australia.
authors.forename: Maté
authors.initials: M
authors.lastname: Biro
authors.affiliation: Centenary Institute, Newtown, New South Wales, Australia.
authors.forename: Szun S
authors.initials: SS
authors.lastname: Tay
authors.affiliation: Centenary Institute, Newtown, New South Wales, Australia; Sydney Medical School, The University of Sydney, Camperdown, New South Wales, Australia.
authors.forename: Rohit
authors.initials: R
authors.lastname: Jain
authors.affiliation: Centenary Institute, Newtown, New South Wales, Australia.
authors.forename: Lisa E
authors.initials: LE
authors.lastname: Shaw
authors.affiliation: Sydney Medical School, The University of Sydney, Camperdown, New South Wales, Australia.
authors.forename: Michele A
authors.initials: MA
authors.lastname: Grimbaldeston
authors.affiliation: Centenary Institute, Newtown, New South Wales, Australia; Discipline of Dermatology, The University of Sydney, Camperdown, New South Wales, Australia; Department of Dermatology, Royal Prince Alfred Hospital, Camperdown, New South Wales, Australia. Electronic address: w.weninger@centenary.org.au.
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Weninger
mesh_terms: D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics; D003713:Dendritic Cells / Q000473:pathology; D003872:Dermatitis / Q000473:pathology*; D020405:Dermis / Q000098:blood supply / Q000473:pathology; D046169:Dermoscopy / Q000379:methods*; D004431:Ear, External; D004817:Epidermis / Q000473:pathology; D005260:Female; D005434:Flow Cytometry; D049452:Green Fluorescent Proteins / Q000235:genetics; D021621:Imaging, Three-Dimensional / Q000379:methods*; D007962:Leukocytes / Q000473:pathology*; D008164:Luminescent Proteins / Q000235:genetics; D016131:Lymphocyte Subsets / Q000473:pathology; D008264:Macrophages / Q000473:pathology; D008297:Male; D008407:Mast Cells / Q000473:pathology; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D036641:Microscopy, Fluorescence, Multiphoton / Q000379:methods*; D012867:Skin / Q000098:blood supply / Q000473:pathology*
chemical_list: D001426:Bacterial Proteins; D008164:Luminescent Proteins; C054562:yellow fluorescent protein, Bacteria; D049452:Green Fluorescent Proteins