Publishing type：Research paper (scientific journal)   Publisher：ELSEVIER SCIENCE INC 360 PARK AVE SOUTH, NEW YORK, NY 10010-1710 USA  

DOI： 10.1016/j.jid.2017.12.014

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Publishing type：Research paper (scientific journal)  

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：JOHN LIBBEY EUROTEXT LTD  

Background: In the pathogenesis of atopic dermatitis (AD), skin barrier dysfunction and T-helper (Th) type 2 immune reactions play important roles. Alterations in the stratum spinosum of AD have not been studied in much detail. Objectives: In this study, we investigated the changes of structural proteins and adhesion molecules residing in the stratum spinosum of AD lesional skin, and whether Th2 cytokines including interleukin (IL)-4 and IL-13 alter the expression of these proteins. Materials & Methods: Skin samples were collected from patients with AD and from healthy controls. Normal human epidermal keratinocytes were cultured and differentiated in the presence or absence of either IL-4 or IL-13 in vitro. The expression of keratin 1, keratin 10, desmoglein 1 and desmocollin 1 was examined by immunofluorescence and western blotting. Results: The expression of these proteins was downregulated in AD lesional skin, and IL-4 and IL-13 were shown to suppress their expression. Conclusion: These results suggest that the stratum spinosum is impaired in AD lesional skin, possibly by Th2 cytokines, which may be involved in the pathogenesis of AD.

DOI： 10.1684/ejd.2017.2985

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Publishing type：Research paper (scientific journal)   Publisher：Wiley Online Library  

DOI： 10.1111/sji.12397

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Publishing type：Research paper (scientific journal)   Publisher：Elsevier  

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Authorship：Lead author,　Corresponding author   Language：English   Publishing type：Research paper (scientific journal)   Publisher：NATURE PUBLISHING GROUP  

T helper type 2 (Th2) cytokines, IL-4 and IL-13, attenuate the expression of genes that regulate epidermal cellular structures and the barrier function at the terminal stage of keratinocyte differentiation. However, whether these Th2 cytokines act at earlier stages remains unknown. We investigated the roles of cytokines in expression levels of mRNAs and/or proteins in primary mouse keratinocytes and human keratinocyte HaCaT cells at earlier stages. We showed that IL-4 downregulated the expression levels of Krt1, Krt10, Dsg1, and Dsc1 via IL-4R alpha- and signal transducer and activator of transcription factor 6 (STAT6)-dependent mechanisms in differentiating mouse keratinocytes at early stages. As the expression levels of keratin-1 and -10 in the keratinocytes transiently expressing an active form of STAT6 were not downregulated, STAT6 and other IL-4-induced molecules may synergistically regulate this expression. The restoration of the downregulated expression levels of Krt1 and Krt10 induced by IL-4 with the MEK (mitogen-activated protein kinase (MAPK)/extracellular signal regulated kinase kinase) inhibitor U0126 indicated the involvement of the p44/42 MAPK signaling pathway in the attenuated expression. IL-13 also downregulated the expression of the four genes. Furthermore, IL-4 or IL-13 caused the downregulation of these genes in HaCaT cells and promoted the fragmentation of cell sheets with mechanical stress. Our results showed that IL-4 or IL-13 acted on differentiating keratinocytes in vitro at early stages to attenuate the gene expression.

DOI： 10.1038/jid.2013.503

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Publishing type：Research paper (scientific journal)  

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Language：English   Publisher：AMER ASSOC IMMUNOLOGISTS  

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Publishing type：Research paper (scientific journal)   Publisher：Nature Publishing Group  

DOI： 10.1038/mi.2012.14

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Other Link： http://www.nature.com/articles/mi201214

Language：English   Publishing type：Research paper (scientific journal)   Publisher：OXFORD UNIV PRESS  

The cytokine thymic stromal lymphopoietin (TSLP) functions as a regulator of bone marrow B-cell development and a key initiator of allergic inflammation. In the current study, we show that mature B cells, derived from transgenic mice with systemically elevated levels of TSLP (K5-TSLP mice), exhibit markedly enhanced mitogenic responses in vitro and that this enhanced responsiveness leads to polyclonal B-cell activation and development of autoimmune hemolytic anemia in vivo. In contrast, B cells derived from K5-TSLP mice lacking CD4(+) T cells failed to show polyclonal activation. Furthermore, neither mature B-cell activation nor hemolytic anemia occurred in IL-4-deficient K5-TSLP mice. Consistent with these findings, activation of mature B cells occurred independently of B-cell intrinsic TSLP signals. Taken together, our results demonstrate that systemic alterations in TSLP, through induction of IL-4 from CD4(+) T cells and other cell types, functions as an important factor in peripheral B-cell homeostasis and promotion of humoral autoimmunity.

DOI： 10.1093/intimm/dxr113

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Publishing type：Research paper (scientific journal)   Publisher：Nature Publishing Group  

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：ACADEMIC PRESS INC ELSEVIER SCIENCE  

To elucidate whether leukocyte cell derived chemotaxin 2 (LECT2) controls the progression of staphylococcal enterotoxin A (SEA) induced toxicity, we examined the role of LECT2 in a mouse model Almost all the C57BL/6 J (B6) mice survived for 72 h after the injection of 0 1 mu g of SEA and 20 mg of D galactosamme (D GalN) However, the same treatment protocol in LECT2(-/-) mice produced a high lethality (similar to 90%), severe hepatic apoptosis, and massive hepatic and pulmonary hemorrhage, similar to the situation observed in B6 mice treated with 1 0 mu g SEA/D GalN The plasma LECT2 levels in B6 mice treated with 1 0 mu g SEA/D GalN were inversely correlated with the plasma cytokine levels and were associated with prognosis LECT2 administration increased the survival of B6 mice and down regulated TNF alpha and IL 6 These results suggest the involvement of LECT2 in the regulation of fatal SEA induced toxicity in D GalN sensitized mice (C) 2010 Elsevier Inc All rights reserved

DOI： 10.1016/j.clim.2010.08.002

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：Oxford University Press (OUP)  

DOI： 10.1093/intimm/dxq012

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CiNii Books

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：AMER ASSOC IMMUNOLOGISTS  

Thymic stromal lymphopoietin (TSLP) is an IL-7-related cytokine, produced by epithelial cells, that has been linked to atopic dermatitis and asthma; however, it remains unclear how TSLP shapes the adaptive immune response that causes these allergic disorders. In this study, we demonstrate a role for TSLP in a Th2 model of contact hypersensitivity in mice. TSLP is required for the development of Th2-type contact hypersensitivity induced by the hapten FITC in combination with the sensitizing agent dibutyl phthalate. TSLPR-deficient mice exhibited a dramatically reduced response, including markedly reduced local infiltration by eosinophils, Th2 cytokine production, and serum IgE levels, following FITC sensitization and challenge. The reduced response by TSLPR-deficient mice is likely due to decreased frequency and reduced T cell stimulatory function of skin-derived Ag-bearing FITC(+)CD11c(+) dendritic cells in draining lymph nodes following FITC sensitization. These data suggest that skin-derived dendritic cells are direct or indirect targets of TSLP in the development of type 2 immune responses in the skin, where TSLP drives their maturation, accumulation in skin draining lymph nodes, and ability to induce proliferation of naive allergen-specific T cells. The Journal of Immunology, 2010, 184: 2974-2984.

DOI： 10.4049/jimmunol.0803478

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：AMER ASSOC IMMUNOLOGISTS  

The epithelial-derived cytokine thymic stromal lymphopoietin (TSLP) is sufficient to induce asthma or atopic dermatitis-like phenotypes when selectively overexpressed in transgenic mice, or when driven by topical application of vitamin D3 or low-calcemic analogues. Although T and B cells have been reported to be dispensable for the TSLP-induced inflammation in these models, little is known about the downstream pathways or additional cell types involved in the inflammatory response driven by TSLP. To characterize the downstream effects of TSLP in vivo, we examined the effects of exogenous administration of TSLP protein to wild-type and genetically deficient mice. TSLP induced a systemic Th2 inflammatory response characterized by increased circulating IgE and IgG1 as well as increased draining lymph node size and cellularity, Th2 cytokine production in draining lymph node cultures, inflammatory cell infiltrates, epithelial hyperplasia, subcuticular fibrosis, and up-regulated Th2 cytokine and chemokine messages in the skin. Responses to TSLP in various genetically deficient mice demonstrated T cells and eosinophils were required, whereas mast cells and TNF-alpha were dispensable. TSLP-induced responses were significantly, but not completely reduced in IL-41 and IL-13-deficient mice. These results shed light on the pathways and cell types involved in TSLP-induced inflammation.

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Publishing type：Research paper (scientific journal)  

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：NATURE PUBLISHING GROUP  

This protocol describes methods to identify, purify and culture CD1d restricted invariant natural killer T (iNKT) cells from mouse tissue or human blood samples. The methods for identification and purification of iNKT cells are based on the interaction between iNKT cell receptor and its ligand. The iNKT cell receptor is composed of the invariant V alpha 14J alpha 18/V beta 8.2 in mice or V alpha 24J alpha 18/V beta 11 in humans and is expressed only on iNKT cells but not on conventional T cells. The iNKT cell antigen receptor in both species recognizes alpha-galactosylceramide (alpha-GalCer) presented by the MHC class I-like CD1d. Thus, alpha-GalCer-loaded CD1d dimer can be used for analysis and purification by fluorescence-activated cell sorting (FACS). Isolation of 1 x 10(6) purified iNKT cells from mouse thymus, spleen or liver requires 5-6 mice and takes 1-2 h for mononuclear cell preparation from mouse tissues, 1.5 h for enrichment by magnetic beads and 4 h for detection and purification of the iNKT cells by FACS. In the case of isolation of human peripheral blood mononuclear cells (PBMCs) from whole blood, it takes 2 h and requires 5 ml of blood to obtain 5 x 10(6) PBMCs, which contain 500-25,000 iNKT cells.

DOI： 10.1038/nprot.2007.515

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：NATURE PUBLISHING GROUP  

The cytokine thymic stromal lymphopoietin ( TSLP) drives immature B cell development in vitro and may regulate T helper type 2 responses. Here we analyzed the involvement of TSLP in B cell development in vivo with a doxycycline-inducible, keratin 5 driven transgene encoding TSLP (K5-TSLP). K5-TSLP-transgenic mice given doxycycline showed an influx of immature B cells into the periphery, with population expansion of follicular mature B cells, near-complete loss of marginal zone and marginal zone precursor B cells, and 'preferential' population expansion of peritoneal B-1b B cells. These changes promoted cryoglobulin production and immune complex-mediated renal disease. Identical events occurred in mice without T cells, in alternative TSLP-transgenic models and in K5-TSLP-transgenic mice with undetectable systemic TSLP. These observations suggest that signals mediating localized TSLP expression may modulate systemic B cell development and promote humoral autoimmunity.

DOI： 10.1038/ni1452

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：AMER ASSOC IMMUNOLOGISTS  

The cytokine thymic stromal lymphopoietin (TSLP) has been implicated in the development and progression of allergic inflammation in both humans and mice. Although the underlying mechanism is not known, TSLP-stimulated dendritic cells have been shown to prime human CD4(+) T cells into Th2 cytokine-producing cells. However, its direct effect on CD4(+) T cells has not been extensively investigated. In this study, we show that TSLP can drive Th2 differentiation in the absence of exogenous IL-4 and APCs. IL-4 blockade inhibited TSLP-mediated Th2 differentiation, demonstrating that IL-4 is involved in this process. Further analysis has shown that TSLP-induced Th2 differentiation is dependent on Stat6 and independent of IL-2 and that TSLP treatment leads to immediate, direct 11-4 gene transcription. Taken together, these data demonstrate that TSLP is directly involved in Th2-mediated responses via the induction of IL-4 production.

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Publishing type：Research paper (scientific journal)   Publisher：Nature Publishing Group  

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Authorship：Lead author   Publishing type：Research paper (scientific journal)   Publisher：Rockefeller University Press  

The cytokine thymic stromal lymphopoietin (TSLP) has recently been implicated in the pathogenesis of atopic dermatitis (AD) and other allergic diseases in humans. To further characterize its role in this disease process, transgenic mice were generated that express a keratinocyte-specific, tetracycline-inducible TSLP transgene. Skin-specific overexpression of TSLP resulted in an AD-like phenotype, with the development of eczematous lesions containing inflammatory dermal cellular infiltrates, a dramatic increase in Th2 CD4+ T cells expressing cutaneous homing receptors, and elevated serum levels of IgE. These transgenic mice demonstrate that TSLP can initiate a cascade of allergic inflammation in the skin and provide a valuable animal model for future study of this common disease.

DOI： 10.1084/jem.20041503

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Publishing type：Research paper (scientific journal)   Publisher：Am Assoc Immnol  

DOI： 10.4049/jimmunol.173.8.4967

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Authorship：Lead author   Publishing type：Research paper (scientific journal)   Publisher：Elsevier  

DOI： 10.1016/s1074-7613(03)00210-3

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Publishing type：Research paper (scientific journal)   Publisher：ASBMB  

DOI： 10.1074/jbc.m205876200

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：OXFORD UNIV PRESS  

In this study we investigate the stage at which developing T cells in the thymus acquire the ability to differentiate into T(h)1 and T(h)2 cells. We addressed this question by using sorted heat-stable antigen (HSA)(+) and HSA(-) CD4 single-positive (SP) thymocytes prepared from ovalbumin-specific TCRalphabeta transgenic mice and an in vitro T(h)1/T(h)2 differentiation culture system. HSA(-) CD4 SP thymocytes show nearly full functional capacity to differentiate into either T(h)1 or T(h)2 cells. A dramatic difference was observed, however, between HSA(+) and HSA(-) CD4 SP thymocytes in the efficiency for T(h)1 cell differentiation. TCR function of HSA(+) CD4 SP thymocytes appeared to be fully developed because antigen-induced proliferation and IL-2 production were essentially equivalent to that of HSA(-) CD4 SP thymocytes. However, the levels in IL-12 receptor (IL-12R) beta2 chain expression following anti-TCR stimulation were dramatically low in the HSA(+) CD4 SP thymocytes. Decreased IL-12-induced STAT4 phosphorylation was also observed. Moreover, IL-12-dependent transcriptional up-regulation of T-bet and STAT4 was deficient in the HSA(+) CD4 SP thymocytes. Thus, the poor capacity of HSA(+) CD4 SP thymocytes to proceed to T(h)1 cell differentiation appears to be at least partly due to underdeveloped capacity in IL-12R expression and function.

DOI： 10.1093/intimm/dxf057

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：ACADEMIC PRESS LTD  

A monoclonal antibody (MAb; TCL-BE8) for carp peripheral blood leucocytes (PBL) was produced, and its reactivity was analysed by electron microscopy and flow cytometry. Electron microscopy using immunogold labelling showed that this MAb reacted with neutrophils and monocytes. The antibody reacted with 98% of the cells in a fraction of granulocytes separated by flow cytometry. The antigen detected by this was MAb defined as a membrane protein of Mr of 112 kDa. Using a magnetic separator, cells reactive with this antibody were separated from leucocytes with a density of 1.08 g ml(-1) or 1.08-1.09 g ml(-1). MAb-positive cells in the PBL with a density of 1.08 g ml(-1) consisted of a mixture of neutrophils and monocytes, whilst in PBL with a density of 1.09 g ml(-1) they consisted of only neutrophils. The MAb-negative fractions contained mainly lymphocytes and thrombocytes. The MAb-positive cells showed strong phagocytosis, which is a characteristic function of neutrophils and monocytes, but this was absent from the MAb-negative fraction. Purification of monocytes and neutrophils, or isolation of neutrophils, can be achieved by using this MAb allowing more effective analysis of the carp leucocyte functions. (C) 1998 Academic Press Limited.

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