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Article ; Online: Skin Whole-Mount Immunofluorescent Staining Protocol, 3D Visualization, and Spatial Image Analysis.

Schmidt, Alfonso J / Wright, Graham D / Ronchese, Franca / Price, Kylie M

Current protocols

2023 Volume 3, Issue 6, Page(s) e820

Abstract: The use of polychromatic immunofluorescent staining on whole-mount skin enables cell type characterization and aids in the delineation of the physiological and immunological strategies used by the skin to combat pathogens. Using whole-mount skin for ... ...

Abstract	The use of polychromatic immunofluorescent staining on whole-mount skin enables cell type characterization and aids in the delineation of the physiological and immunological strategies used by the skin to combat pathogens. Using whole-mount skin for polychromatic immunofluorescent staining removes the need for histological sectioning and enables the visualization of anatomical structures and immune cell types in three dimensions. Here we present a detailed protocol for immunostaining with fluorescence-conjugated primary antibodies in whole-mount skin to reveal structural landmarks and specific immune cell types using confocal laser scanning microscopy (CLSM) (Basic Protocol 1). The optimized staining panel reveals structural features such as blood vessels (CD31 antibody) and the lymphatic network (LYVE-1 antibody), in combination with MHCII antibodies for antigen-presenting cells (APCs), CD64 for macrophages and monocytes, CD103 for dendritic epidermal T cells (DETC), and CD326 for Langerhans cells (LC). Basic Protocol 2 describes image visualization pipelines using open-source software (ImageJ/FIJI), enabling four visualization options (z-projections, orthogonal views, 3D visualization, and animation). Basic Protocol 3 describes a quantitative analysis pipeline using CellProfiler to characterize the spatial relationship between cell types using mathematical indices such as Spatial Distribution Index (SDI), Neighborhood Frequency (NF), and Normalized Median Evenness (NME). These protocols will enable researchers to stain, record, analyze, and interpret data from whole-mount skin using commercially available reagents in a CLSM-equipped laboratory and freely available analysis software. © 2023 Wiley Periodicals LLC. Basic Protocol 1: Immunofluorescent staining and imaging for whole-mount mouse skin Basic Protocol 2: File rendering and visualization using FIJI Basic Protocol 3: Spatial image analysis using CellProfiler.
MeSH term(s)	Animals ; Mice ; Imaging, Three-Dimensional/methods ; Skin/diagnostic imaging ; Staining and Labeling ; Coloring Agents ; Microscopy, Confocal/methods
Chemical Substances	Coloring Agents
Language	English
Publishing date	2023-06-19
Publishing country	United States
Document type	Journal Article
ISSN	2691-1299
ISSN (online)	2691-1299
DOI	10.1002/cpz1.820
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Article ; Online: IL-13 in dermal type-2 dendritic cell specialization: From function to therapeutic targeting.

Lamiable, Olivier / Brewerton, Maia / Ronchese, Franca

European journal of immunology

2022 Volume 52, Issue 7, Page(s) 1047–1057

Abstract: Skin functions as a barrier protecting the host against physical, thermal, chemical changes as well as microbial insults. The skin is populated by several immune cell types that are crucial to host defense and to maintain self-tolerance as well as ... ...

Abstract	Skin functions as a barrier protecting the host against physical, thermal, chemical changes as well as microbial insults. The skin is populated by several immune cell types that are crucial to host defense and to maintain self-tolerance as well as equilibrium with beneficial microbiota. Conventional dendritic cells (cDCs) are antigen-presenting cells that patrol the skin and all other nonlymphoid tissues for self or foreign antigens, and then migrate to draining lymph nodes to initiate T-cell responses. This review article describes recent developments on skin cDC specialization, focusing on the role of IL-13, a cytokine essential to allergic immune responses that is also secreted at steady state by type-2 innate lymphoid cells in healthy skin, and is required for dermal cDC differentiation. Furthermore, we contextualize how different therapeutics that block IL-13 signaling and were recently approved for the treatment of atopic dermatitis might affect cDCs in human skin.
MeSH term(s)	Dendritic Cells ; Humans ; Immunity, Innate ; Interleukin-13/metabolism ; Lymphocytes ; Skin/pathology
Chemical Substances	IL13 protein, human ; Interleukin-13
Language	English
Publishing date	2022-06-09
Publishing country	Germany
Document type	Journal Article ; Review ; Research Support, Non-U.S. Gov't
ZDB-ID	120108-6
ISSN	1521-4141 ; 0014-2980
ISSN (online)	1521-4141
ISSN	0014-2980
DOI	10.1002/eji.202149677
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Zs.A 489: Show issues			Location: Je nach Verfügbarkeit (siehe Angabe bei Bestand) bis Jg. 1994: Bestellungen von Artikeln über das Online-Bestellformular Jg. 1995 - 2021: Lesesall (1.OG) ab Jg. 2022: Lesesaal (EG)
Zs.MG 85: Show issues

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Article ; Online: Antigen presentation by dendritic cells and their instruction of CD4+ T helper cell responses.

Hilligan, Kerry L / Ronchese, Franca

Cellular & molecular immunology

2020 Volume 17, Issue 6, Page(s) 587–599

Abstract: Dendritic cells are powerful antigen-presenting cells that are essential for the priming of T cell responses. In addition to providing T-cell-receptor ligands and co-stimulatory molecules for naive T cell activation and expansion, dendritic cells are ... ...

Abstract	Dendritic cells are powerful antigen-presenting cells that are essential for the priming of T cell responses. In addition to providing T-cell-receptor ligands and co-stimulatory molecules for naive T cell activation and expansion, dendritic cells are thought to also provide signals for the differentiation of CD4+ T cells into effector T cell populations. The mechanisms by which dendritic cells are able to adapt and respond to the great variety of infectious stimuli they are confronted with, and prime an appropriate CD4+ T cell response, are only partly understood. It is known that in the steady-state dendritic cells are highly heterogenous both in phenotype and transcriptional profile, and that this variability is dependent on developmental lineage, maturation stage, and the tissue environment in which dendritic cells are located. Exposure to infectious agents interfaces with this pre-existing heterogeneity by providing ligands for pattern-recognition and toll-like receptors that are variably expressed on different dendritic cell subsets, and elicit production of cytokines and chemokines to support innate cell activation and drive T cell differentiation. Here we review current information on dendritic cell biology, their heterogeneity, and the properties of different dendritic cell subsets. We then consider the signals required for the development of different types of Th immune responses, and the cellular and molecular evidence implicating different subsets of dendritic cells in providing such signals. We outline how dendritic cell subsets tailor their response according to the infectious agent, and how such transcriptional plasticity enables them to drive different types of immune responses.
MeSH term(s)	Animals ; Antigen Presentation/immunology ; Cell Differentiation/immunology ; Dendritic Cells/immunology ; Humans ; Models, Biological ; T-Lymphocytes, Helper-Inducer/immunology ; Transcription, Genetic
Language	English
Publishing date	2020-05-20
Publishing country	China
Document type	Journal Article ; Research Support, Non-U.S. Gov't ; Review
ZDB-ID	2435097-7
ISSN	2042-0226 ; 1672-7681
ISSN (online)	2042-0226
ISSN	1672-7681
DOI	10.1038/s41423-020-0465-0
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Zs.A 6681: Show issues

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Article ; Online: A guideline for the appropriate recognition of shared resource laboratories in publication.

Ferrer-Font, Laura / Schmidt, Alfonso / Ronchese, Franca / Price, Kylie M

Cytometry. Part A : the journal of the International Society for Analytical Cytology

2022 Volume 103, Issue 3, Page(s) 193–197

Abstract: The issue of what level of contribution warrants authorship, determining a fair order of authors and when and whom to acknowledge in publications is often a cause of debate, and in some instances, has also been a focus of conflict at certain institutions. ...

Abstract	The issue of what level of contribution warrants authorship, determining a fair order of authors and when and whom to acknowledge in publications is often a cause of debate, and in some instances, has also been a focus of conflict at certain institutions. Shared resource laboratories (SRLs) play a fundamental role in supporting publications, and SRL staff scientists can contribute to numerous areas such as experimental design, sample preparation, data acquisition, data analysis and manuscript drafting and review. However, SRL staff scientists are often unfairly omitted from the author list. To avoid SRLs and SRL staff scientist contributions going unnoticed, the authors have formulated a set of guidelines to aid in the conceptualization and recognition of the technical and intellectual contributions of SRLs. As a better understanding of the role SRL staff scientists play in the achievement of the scientific lead's experimental aims will foster a positive feedback loop, where acknowledgements can lead to more support and funding for SRLs and more engaged SRL staff capable of supporting discoveries and technological innovations that underpin major advancements in the field of life sciences.
MeSH term(s)	Humans ; Laboratories ; Authorship ; Research Design
Language	English
Publishing date	2022-12-21
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2099868-5
ISSN	1552-4930 ; 0196-4763 ; 1552-4922
ISSN (online)	1552-4930
ISSN	0196-4763 ; 1552-4922
DOI	10.1002/cyto.a.24713
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Zs.A 1688: Show issues			Location: Je nach Verfügbarkeit (siehe Angabe bei Bestand) bis Jg. 1994: Bestellungen von Artikeln über das Online-Bestellformular Jg. 1995 - 2021: Lesesall (1.OG) ab Jg. 2022: Lesesaal (EG)
Zs.MG 68: Show issues

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Article ; Online: Dendritic cells and the skin environment.

Ronchese, Franca / Hilligan, Kerry L / Mayer, Johannes U

Current opinion in immunology

2020 Volume 64, Page(s) 56–62

Abstract: The skin is inhabited by several immune cell populations that serve as a first line of defence against pathogen invasion. Amongst these populations are dendritic cells, which play an essential sentinel function by taking up antigen or infectious agents ... ...

Abstract	The skin is inhabited by several immune cell populations that serve as a first line of defence against pathogen invasion. Amongst these populations are dendritic cells, which play an essential sentinel function by taking up antigen or infectious agents and transporting them to the lymph node for T cell recognition and the priming of immune responses. In this review, we briefly summarise recent advances showing how skin dendritic cells are connected to a network of epithelial and stromal cells, which provide structural support, growth factors, spatial cues, contact with the external environment and the skin microbiome, and favour interactions with other immune cells. We propose that this network creates a unique skin environment that may condition dendritic cell phenotype and function.
MeSH term(s)	Dendritic Cells ; Humans ; Langerhans Cells ; Lymph Nodes ; Microbiota ; Skin
Language	English
Publishing date	2020-05-05
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't ; Review
ZDB-ID	1035767-1
ISSN	1879-0372 ; 0952-7915
ISSN (online)	1879-0372
ISSN	0952-7915
DOI	10.1016/j.coi.2020.03.006
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Zs.A 2773: Show issues			Location: Je nach Verfügbarkeit (siehe Angabe bei Bestand) bis Jg. 1994: Bestellungen von Artikeln über das Online-Bestellformular Jg. 1995 - 2021: Lesesall (2.OG) ab Jg. 2022: Lesesaal (EG)
Zs.MG 13: Show issues

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Article ; Online: Dendritic cells in Th2 immune responses and allergic sensitization.

Lamiable, Olivier / Mayer, Johannes U / Munoz-Erazo, Luis / Ronchese, Franca

Immunology and cell biology

2020 Volume 98, Issue 10, Page(s) 807–818

Abstract: Allergic responses are characterized by the activation of a specific subset of effector ... ...

Abstract	Allergic responses are characterized by the activation of a specific subset of effector CD4
MeSH term(s)	Allergens ; Dendritic Cells/immunology ; Humans ; Hypersensitivity ; Immunity ; Th2 Cells/immunology
Chemical Substances	Allergens
Language	English
Publishing date	2020-09-10
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't ; Review
ZDB-ID	284057-1
ISSN	1440-1711 ; 0818-9641
ISSN (online)	1440-1711
ISSN	0818-9641
DOI	10.1111/imcb.12387
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Uc I Zs.175: Show issues

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Article ; Online: Comment on: Repositioning T

Jankovic, Dragana / Ciucci, Thomas / Coffman, Robert L / Coquet, Jonathan M / Le Gros, Graham / Mosmann, Tim R / Sher, Alan / Ronchese, Franca

Nature immunology

2022 Volume 23, Issue 4, Page(s) 501–502

MeSH term(s)	Cytokines ; Lymphocyte Activation ; Th1 Cells ; Th2 Cells
Chemical Substances	Cytokines
Language	English
Publishing date	2022-02-21
Publishing country	United States
Document type	Letter ; Comment
ZDB-ID	2016987-5
ISSN	1529-2916 ; 1529-2908
ISSN (online)	1529-2916
ISSN	1529-2908
DOI	10.1038/s41590-022-01144-y
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Zs.A 5294: Show issues			Location: Je nach Verfügbarkeit (siehe Angabe bei Bestand) bis Jg. 1994: Bestellungen von Artikeln über das Online-Bestellformular Jg. 1995 - 2021: Lesesall (2.OG) ab Jg. 2022: Lesesaal (EG)
Zs.MG 42: Show issues

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Article ; Online: BCL6 deletion in CD4 T cells does not affect Th2 effector mediated immunity in the skin.

Chandler, Jodie / Prout, Melanie / Old, Sam / Morgan, Cynthia / Ronchese, Franca / Benoist, Christophe / Le Gros, Graham

Immunology and cell biology

2022 Volume 100, Issue 10, Page(s) 791–804

Abstract: Recent studies propose that T follicular helper (Tfh) cells possess a high degree of functional plasticity in addition to their well-defined roles in mediating interleukin-4-dependent switching of germinal center B cells to the production of ... ...

Abstract	Recent studies propose that T follicular helper (Tfh) cells possess a high degree of functional plasticity in addition to their well-defined roles in mediating interleukin-4-dependent switching of germinal center B cells to the production of immunoglobulin (Ig)G1 and IgE antibodies. In particular Tfh cells have been proposed to be an essential stage in Th2 effector cell development that are able to contribute to innate type 2 responses. We used CD4-cre targeted deletion of BCL6 to identify the contribution Tfh cells make to tissue Th2 effector responses in models of atopic skin disease and lung immunity to parasites. Ablation of Tfh cells did not impair the development or recruitment of Th2 effector subsets to the skin and did not alter the transcriptional expression profile or functional activities of the resulting tissue resident Th2 effector cells. However, the accumulation of Th2 effector cells in lung Th2 responses was partially affected by BCL6 deficiency. These data indicate that the development of Th2 effector cells does not require a BCL6 dependent step, implying Tfh and Th2 effector populations follow separate developmental trajectories and Tfh cells do not contribute to type 2 responses in the skin.
MeSH term(s)	CD4-Positive T-Lymphocytes ; T-Lymphocytes, Helper-Inducer ; Cell Differentiation ; Germinal Center ; B-Lymphocytes ; Proto-Oncogene Proteins c-bcl-6/genetics
Chemical Substances	Proto-Oncogene Proteins c-bcl-6
Language	English
Publishing date	2022-10-18
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	284057-1
ISSN	1440-1711 ; 0818-9641
ISSN (online)	1440-1711
ISSN	0818-9641
DOI	10.1111/imcb.12589
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Article ; Online: Fetal dendritic cells give mum a break.

Connor, Lisa M / Lamiable, Olivier / Ronchese, Franca

Immunology and cell biology

2017 Volume 95, Issue 7, Page(s) 575–576

MeSH term(s)	Dendrites ; Dendritic Cells ; Fetus ; Humans
Language	English
Publishing date	2017-07-18
Publishing country	United States
Document type	Journal Article ; Comment
ZDB-ID	284057-1
ISSN	1440-1711 ; 0818-9641
ISSN (online)	1440-1711
ISSN	0818-9641
DOI	10.1038/icb.2017.46
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Article: IL-4 Is a Key Requirement for IL-4- and IL-4/IL-13-Expressing CD4 Th2 Subsets in Lung and Skin.

Prout, Melanie Sarah / Kyle, Ryan L / Ronchese, Franca / Le Gros, Graham

Frontiers in immunology

2018 Volume 9, Page(s) 1211

Abstract: Although IL-4 is long associated with CD4 Th2 immune responses, its role in Th2 subset development in non-lymphoid tissues is less clear. We sought to better define IL-4's role in CD4 Th2 responses by using transgenic mice that express a dual IL-4 AmCyan/ ...

Abstract	Although IL-4 is long associated with CD4 Th2 immune responses, its role in Th2 subset development in non-lymphoid tissues is less clear. We sought to better define IL-4's role in CD4 Th2 responses by using transgenic mice that express a dual IL-4 AmCyan/IL-13 DsRed (IL-4AC/IL-13DR) fluorescent reporter on an IL-4-sufficient or IL-4-deficient background. Using primary Th2 immune response models against house dust mite or
MeSH term(s)	Allergens/immunology ; Animals ; Biomarkers ; Gene Expression ; Genotype ; Immunization ; Immunophenotyping ; Interleukin-13/genetics ; Interleukin-13/metabolism ; Interleukin-4/genetics ; Interleukin-4/metabolism ; Lung/immunology ; Lung/metabolism ; Lymph Nodes/immunology ; Lymph Nodes/metabolism ; Mice ; Mice, Knockout ; Mice, Transgenic ; Models, Biological ; Pyroglyphidae/immunology ; Skin/immunology ; Skin/metabolism ; T-Lymphocyte Subsets/immunology ; T-Lymphocyte Subsets/metabolism ; Th2 Cells/immunology ; Th2 Cells/metabolism
Chemical Substances	Allergens ; Biomarkers ; Interleukin-13 ; Interleukin-4 (207137-56-2)
Language	English
Publishing date	2018-06-01
Publishing country	Switzerland
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2606827-8
ISSN	1664-3224
ISSN	1664-3224
DOI	10.3389/fimmu.2018.01211
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