In order to confirm upregulation of expression of IFN-inducible GTP-binding protein (IIGP), we performed immunostaining in skin sections of K14-Cre-IKK2fl/fl mice at day 7. in: Fluorescent stainings were analyzed using a Leica TCS SP2 upright confocal laser-scanning microscope (Leica Microsystems) at excitation wavelengths of 488 nm and 543 nm. Open up mouse by pulling skin with both hands to each side of mouse. White dotted lines indicate the position of the epidermal basement membrane. 1994. 17, 31, 32), and T cells have been shown to play a crucial role in the development of psoriasis-like skin changes in mouse models (for example, see ref. 2009 May;41(5):963-8. doi: 10.1016/j.biocel.2008.10.022. Scale bar: 50 μm. Scale bars: 100 μm (H&E); 40 μm (immunostaining). Mice deficient for CD18 have been reported previously to be defective for granulocyte migration into areas of organ inflammation (23). Targeted deletion of IFN-γ receptor…, Figure 8. We conclude that the psoriasis-like skin condition in K14-Cre-IKK2fl/fl mice, like human psoriasis, responds to anti-TNF treatment. Google Scholar, Find articles by Clodronate was a gift of Roche Diagnostics GmbH. Previous histological and immunohistological analysis of the skin of K14-Cre-IKK2fl/fl mice showed massive accumulation of macrophages (F4/80-positive) and granulocytes (GR-1–positive) as well as an increase in the numbers of mast cells in the skin of these mice. | 1997. We also showed that these immune cells expressed TNF (18). Localization and functions of macrophage and monocyte subpopulations. Skin graft survival and antidonor rat humoral responses were quantified. 848 pp. in: Improvement of psoriasis-like skin disease in K14-Cre-IKK2, Characterization of macrophages and dendritic cells in the skin of K14-Cre-IKK2. Mast cells and macrophages in early relapsing psoriasis. At P7 or P8 mice were sacrificed, and skin samples from the upper and lower back were fixed in 4% paraformaldehyde overnight or embedded in Tissue-Tek (Sakura) for cryosectioning. Wang H, Peters T, Sindrilaru A, Scharffetter-Kochanek K. J Invest Dermatol. National Center for Biotechnology Information, Unable to load your collection due to an error, Unable to load your delegates due to an error, Light microscopic and confocal images of paraffin-embedded skin sections (H&E, K14, K10, and loricrin [Lor]) and confocal images of cryostat skin sections (CD3, GR-1, F4/80) obtained at P7 from K14-Cre-IKK2, Confocal images of immunostainings of skin of untreated (, Light microscopic images of paraffin-embedded skin sections (top 3 panels) and confocal images of cryostat skin sections (bottom 6 panels) from control mice and K14-Cre-IKK2, Immunostainings of paraffin-embedded skin sections with antibodies against the epidermal differentiation markers K14, K10, loricrin, and filaggrin (Fil) or of frozen skin with antibodies against the immune cell markers GR-1 for granulocytes and CD3 for T lymphocytes. Pathogenic function of IL-1 beta in psoriasiform skin lesions of flaky skin (fsn/fsn) mice. We have previously shown in our wound paradigm that the Thepen, T., et al. By permitting the emptying of the niche occupied in the skin dermis and intestinal lamina propria by CD64 + myeloid cells, the CD64 dtr mouse model showed that all the mo-DC and macrophage subsets that were present in adult skin and intestine before DT treatment can be readily reconstituted from circulating Ly-6C high monocytes. In K14-Cre-IKK2fl/fl mice we have observed a similar phenomenon: spindle-shaped macrophages that partly expressed CD14 accumulated directly beneath the epidermal basement membrane. (L and M) F4/80-positive epithelium-lining macrophages in a developing lesion at P4 (L) and in a fully developed lesion at P7 (M). Mast cell numbers were as follows (no. 2001. Since macrophages have been identified previously as a major source of TNF in skin (40), they are also likely to produce pathogenic TNF in the skin of K14-Cre-IKK2fl/fl mice. A subset of macrophages located along the basement membrane (“lining cells”) is a characteristic histopathological feature of psoriasis. Front Cell Dev Biol. JCI TNF-mediated inflammatory skin disease in mice with epidermis-specific deletion of IKK2. Inhibition of granulocyte migration into the skin therefore does not appear to be a promising therapeutic target in psoriasis. Rupec, R. Inflammatory skin diseases, T cells and immune surveillance. A polygenic mouse model of psoriasiform skin disease in CD18-deficient mice. Pathogenic role for skin macrophages in a mouse model of keratinocyte-induced psoriasis-like skin inflammation Athanasios Stratis, 1 Manolis Pasparakis, 2,3 Rudolf A. Rupec, 4 Doreen Markur, 1 Karin Hartmann, 1 Karin Scharffetter-Kochanek, 5 Thorsten Peters, 5 Nico van Rooijen, 6 Thomas Krieg, 1 and Ingo Haase 1 Boyman, O., et al. In order to evaluate contributions of other immune cell populations to the skin disease, we selectively eliminated macrophages and granulocytes from the skin of mice with epidermis-specific deletion of IKK2 (K14-Cre-IKK2fl/fl mice). The liposome-mediated macrophage ‘suicide’ technique. In addition, the inflammatory cell infiltrate was less dense compared with control liposome–injected mice. Macrophages in the dermis of K14-Cre-IKK2fl/fl mice stained positive for F4/80, CD11b, and CD206 and partly positive for CD16/32 (Figure 2). Google Scholar, Find articles by Phone: 49-221-478-86360; Fax: 49-221-478-5949; E-mail: USA.gov. Nuclei are shown in red. 1Department of Dermatology and Center for Molecular Medicine, University of Cologne (CMMC), Cologne, Germany. Int J Mol Sci. Hemalaun was used for counterstaining. Pantelyushin S, Haak S, Ingold B, Kulig P, Heppner FL, Navarini AA, Becher B. J Clin Invest. 1997. Google Scholar, Find articles by Scharffetter-Kochanek, K., et al. The term psoriasis designates a heterogeneous group of chronic inflammatory disorders that are characterized by a typical morphological and histopathological pattern of skin abnormalities. Analysis of expression of K14, K10, and filaggrin showed no major differences among K14-Cre-IKK2fl/fl mice, CD18+/– K14-Cre-IKK2fl/fl mice, and CD18–/– K14-Cre-IKK2fl/fl mice (Figure 6). Targeting NF-kappa B with a natural triterpenoid alleviates skin inflammation in a mouse model of psoriasis. Ghoreschi, K., et al. | Indicated markers are stained green, and nuclei are stained red. Schön, M., et al. Elimination of granulocytes from the…, Figure 7. Spontaneous skin ulceration and defective T cell function in CD18 null mice. They play critical roles in homeostasis and host defense in each tissue. While the source of TNF in K14-Cre-IKK2fl/fl mice remains unclear at the moment, epidermal keratinocytes are not likely to be responsible for the release of pathogenic TNF, as suggested by our previous studies (18) and by skin grafting experiments from TNF-deficient mice with targeted deletion of RelA/c-Rel onto Rag–/– mice, in which grafts developed a hyperproliferative, inflammatory phenotype in the absence of donor keratinocyte–derived TNF (39). Transgenic expression of the human amphiregulin gene induces a psoriasis-like phenotype. Immune response in mice that lack the interferon-gamma receptor. Note relative increase in numbers of neutrophils (stabs and polymorphonuclear granulocytes) and relative decrease in numbers of lymphocytes. Response of psoriasis to a lymphocyte-selective toxin (DAB389IL-2) suggests a primary immune, but not keratinocyte, pathogenic basis. FASEB J. (K) Double staining with antibodies against CD83 (green) and CD80 (red). Since elimination of skin macrophages prevented both the accumulation of granulocytes and T cells and the development of the psoriasis-like skin condition, it was unclear whether the improvement of the skin disease was a direct consequence of macrophage depletion or the result of inhibited granulocyte and T cell migration or expansion. Skin phenotypes that met these criteria were consequently termed psoriasis or psoriasiform or psoriasis-like skin disease (reviewed in ref. a Skin sections were stained with anti-CD68 (red) to visualize macrophages in expanded and non-expanded skin at day 7, day 14 and day 35.b Normal skin sections stained with anti-CD68.c The total number of macrophages in the skin was quantified on 3 sections of 5–6 rats … Our present study revealed evidence for the activity of IFN-γ–dependent gene regulatory pathways, which are also known to be essential for classical macrophage activation, during the initial stages of disease development. By combining CD64 and CCR2 staining, we successfully identified each of these cell types and studied their origin, transcriptomic … Sano, S., et al. Scale bar: 40 μm. | Research ArticleDermatology Syed SN, Raue R, Weigert A, von Knethen A, Brüne B. For immunostainings, indicated markers are stained green, and nuclei are stained red. 2004. IL-12 is produced by macrophages and can stimulate the production of IFN-γ in an autocrine fashion (42); IL- 23 is known to be produced by keratinocytes in psoriatic lesions (43, 44) and can directly activate macrophages (45). | 1995. In situ localization of CD83-positive dendritic cells in psoriatic lesions. mice does not suppress inflammation. Claassen, I., Van Rooijen, N., Claassen, E. 1990. (B and C) Light microscopic images of chloroacetate esterase–stained, paraffin-embedded skin sections from CD18+/– K14-Cre-IKK2fl/fl mice (B) and CD18–/– K14-Cre-IKK2fl/fl mice (C). Mease, P.J., et al. Affymetrix Murine Genome U74Av2 arrays and GeneChip test 3 arrays were used. Chloracetate esterase staining for detection of neutrophile granulocytes was performed on deparaffinized sections using PBS with 0.8% hexazonium and 2% of a solution containing 90 mg naphthol-AS- | In the skin, the lack of markers permitting the unambiguous identification of macrophages and of conventional and monocyte-derived dendritic cells (DCs) complicates understanding of their contribution to skin integrity and to immune responses. Depletion of antigen-presenting cells by clodronate liposomes reverses the psoriatic skin phenotype in KC-Tie2 mice. Phosphatidylcholine (LIPOID E PC) was obtained from Lipoid. Adhesion is disrupted by deletion of CD18, the β2 integrin chain. Inflammatory and immune cell function in psoriasis: II. Adenoviral gene transfer of a single-chain IL-23 induces psoriatic arthritis-like symptoms in NOD mice. These results show that mast cells are not depleted by clodronate liposomes and that their numbers correlate with the severity of the phenotype. We therefore asked whether IFN-γ receptor–mediated signals are required for the development of the skin disease. Top 3 panel are stained with H&E. Trepicchio, W.L., Dorner, A.J. Figure 3. Address correspondence to: Ingo Haase, Department of Dermatology and Center for Molecular Medicine, University of Cologne (CMMC), Joseph-Stelzmann-Strasse 9, 50924 Cologne, Germany. Yolk sac-derived primitive macrophages are generated in two waves from ‘early’ and ‘late’ erythro-myeloid progenitors (EMPs) in the mouse embryo at embryonic age 7.5 and 8.25 (E7.5 and E8.25). Granulocytes are stained red (arrow). Psoriasis is a common skin disease, the pathogenesis of which has not yet been resolved. Upon establishment of the blood circulation, these primitive macrophages are the first to seed the fetal tissues. Cells. Previous studies in psoriatic human skin have described a subset of skin macrophages that migrate to the dermal/epidermal border where they communicate with epidermal keratinocytes, terming these cells epithelium-lining macrophages (24, 34). 2004. White dotted line indicates the position of the epidermal basement membrane. RNA was extracted from the skin of 1 K14-Cre-IKK2fl/fl mouse and 1 control mouse each at P2 and P3 and subjected to Affymetrix gene chip analysis. Macrophage S1PR1 Signaling Alters Angiogenesis and Lymphangiogenesis During Skin Inflammation. Mice were backcrossed to the C57BL/6 genetic background for at least 5 generations. We carried out immunostainings for differentiation specific markers in order to analyze the degree of disturbance of terminal keratinocyte differentiation. Sphingosine Kinases are Involved in Macrophage NLRP3 Inflammasome Transcriptional Induction. (D) Results of counting of chloroacetate esterase–positive granulocytes in skin sections from 8 CD18–/– K14-Cre-IKK2fl/fl mice and 6 CD18+/+ or CD18+/– K14-Cre-IKK2fl/fl mice. Cook, P.W., Brown, J.R., Cornell, K.A., Pittelkow, M.R. Compared with wild-type mice, CX3CR1-deficient mice showed fewer numbers of skin tumors with a lower incidence. Increased oxidative damage to fibroblasts in skin with and without lesions in psoriasis. 2000. TAMs express immune checkpoint modulators [e.g., B7 family, B7-homolog family including programmed death ligand 1 (PD-L1)] (3) that directly suppress activated T cells. Etanercept as monotherapy in patients with psoriasis. Although the migration of granulocytes into the skin and the formation of microabscesses are prominent features of psoriasis, their absence from the skin does not lead to clearance or major improvement of the psoriasis-like skin disease in K14-Cre-IKK2fl/fl mice. One of the most important immune cells involved in wound healing is the macrophage, which exhibits different immunological functions in the skin, including phagocytosis and antigen-presentation. in: Another group of K14-Cre-IKK2fl/fl mice and control mice was left untreated (>10 litters), and a third was treated with human IgG at 20 μg/d (1 litter containing 2 K14-Cre-IKK2fl/fl mice and 8 control mice). Macrophages and monocytes are distributed in various tissues such as the CNS, skin, lung, spleen, liver, bone, blood and serosal cavities. Philadelphia, Pennsylvania, USA. Figure 4. The protocol for detection of deleted IFN-γ receptor alleles was obtained from The Jackson Laboratory. CD80 was expressed by few cells in the deeper dermis, which also stained positive for CD83 (Figure 2K). 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