No / IL-33 is a member of the IL-1 family of cytokines that is constitutively expressed in healthy skin and was found to be increased in the skin of patients with atopic dermatitis (AD). Because it can be released after tissue damage or physical stress including scratching of the skin,1 it has been classified as an alarmin concerned with alerting the immune system.2 It enhances TH2 responses by inducing IL-5 and IL-13 as well as TH1 responses via upregulation of IFN-γ. Keratinocytes are known producer cells of IL-33 and also express the receptor complex consisting of ST2 and IL-1RAcP on their surface. The aim of this study was to investigate the effect of IL-33 on keratinocytes, skin biopsies, and living skin equivalents with regard to the regulation of the skin barrier molecule filaggrin (FLG).

No / Neuroimmune dysregulation characterizes atopic disease, but its nature and clinical impact remain ill-defined. Induced by stress, the neurotrophin nerve growth factor (NGF) may worsen cutaneous inflammation. We therefore studied the role of NGF in the cutaneous stress response in a mouse model for atopic dermatitis–like allergic dermatitis (AlD). Combining several methods, we found that stress increased cutaneous but not serum or hypothalamic NGF in telogen mice. Microarray analysis showed increased mRNAs of inflammatory and growth factors associated with NGF in the skin. In stress-worsened AlD, NGF-neutralizing antibodies markedly reduced epidermal thickening together with NGF, neurotrophin receptor (tyrosine kinase A and p75 neurotrophin receptor), and transforming growth factor-β expression by keratinocytes but did not alter transepidermal water loss. Moreover, NGF expression by mast cells was reduced; this corresponded to reduced cutaneous tumor necrosis factor-α (TNF-α) mRNA levels but not to changes in mast cell degranulation or in the T helper type 1 (Th1)/Th2 cytokine balance. Also, eosinophils expressed TNF receptor type 2, and we observed reduced eosinophil infiltration after treatment with NGF-neutralizing antibodies. We thus conclude that NGF acts as a local stress mediator in perceived stress and allergy and that increased NGF message contributes to worsening of cutaneous inflammation mainly by enhancing epidermal hyperplasia, pro-allergic cytokine induction, and allergy-characteristic cellular infiltration.

Abstract An excess of atopic allergies has been found in patients with depression, and conversely, increased amounts of depressive symptoms have been reported in patients with atopic disorders. Thus far, however, the findings have mainly been based on clinical samples. In this thesis, the association between atopic disorders and depression was investigated at epidemiological level by using data from the Northern Finland 1966 Birth Cohort. An unselected cohort of 12058 liveborn children was followed prospectively from prenatal stages until 1997. During the 31-year follow-up, 6025 cohort members underwent skin prick tests. Data on lifetime depression diagnoses and atopic conditions were obtained from postal questionnaires and Finnish Hospital Discharge Registers, and the severity of the depressive symptoms was assessed with Hopkins Symptom Checklist-25. Information on the family histories of the atopic disorders was obtained from questionnaires of the 31-year follow-up. Females with positive skin prick test responses and self-reported histories of allergic symptoms exhibited a 2.7-fold probability of developing lifetime depression. The corresponding probability increased in line with the increased severity of depressive symptoms in atopic but not in non-atopic females, ranging from 3.0 to 4.7-fold. Among males, the atopy-depression association was seen only in the highest depression scores, the odds ratio being up to 6.3-fold. While the most severe, hospital-treated manifestations of both disorders were considered, atopic disorders increased the risk of depression 3-fold independently of the subject's gender and sociodemographic characteristics. When investigating the effect of familial atopy on a child's depression, maternal atopy increased the probability of lifetime depression nearly 2-fold in females, and over 4-fold, when a female cohort member's own atopy was also present. At epidemiological level, the presence of atopic conditions seemed to increase the probability of lifetime depression especially in females. Since both atopic disorders and depression are illnesses of major public health importance in Western countries, also the co-morbidity between these disorders should be seriously taken into account in clinical practice. Further investigations are called for in evaluating whether this association is specific to atopic disorders, since increased risks of depression have been noted in connection with many other physical diseases as well.

INTRODUÇÃO: A dermatite atópica (DA) é uma doença cutânea inflamatória, acompanhada por prurido intenso e xerose cutânea. A etiopatogenia da DA é multifatorial, envolvendo fatores genéticos, ambientais e imunológicos, dentre outros. OBJETIVOS: Avaliar a influência das enterotoxinas A e B do Staphylococcus aureus (SEA e SEB) na resposta mediada por células Th17 e Th22 nos indivíduos adultos com DA. MÉTODOS: Foram selecionados 38 pacientes adultos com DA e um grupo controle com 40 indivíduos adultos, pareados por idade e gênero Os métodos utilizados foram: 1) ELISA: dosagem dos níveis séricos de IL-6, IL-17, IL-22 e IL-12p40/IL-23 e em sobrenadantes de culturas de células mononucleares do sangue periférico (PBMC) estimuladas com SEA e SEB; 2) Imuno-histoquímica: análise da expressão de IL-17 em fragmentos de pele; 3) Citometria de fluxo: a) análise das citocinas circulantes em amostras de soro: IL-2, IL-4, IL-5, IL-6, IL-10, TNF, IL-17A e IFN-y b)avaliação das células T CD4+ mono e polifuncionais secretoras de IL-17, IL-22, TNF, IFN-y, MIP-1beta, e expressão do marcador de ativação celular CD38; c) células Th22 e Tc22 estimuladas com SEA e SEB. RESULTADOS: 1) Através do ELISA, a secreção de IL-22 sérica e em PBMC induzidas por SEA e SEB foi significativamente mais elevada, quando comparada ao grupo controle; 2) houve aumento na expressão de IL-17 em amostras de pele de doentes de DA através da imuno-histoquímica; 3) Através da citometria de fluxo, foram detectados: a) níveis séricos de IL-2, 5, 6, 10, 17A e IFN-y elevados no grupo com DA em relação aos controles; houve diferença significativa nos níveis circulantes de IL-17A nos pacientes com DA moderada e grave; b) na avaliação monofuncional das células T CD4+ sob estímulo de SEA/SEB, houve redução da expressão das citocinas IFN-y, IL-17A, IL-22 ou TNF na DA, quando comparadas ao grupo controle; na análise polifuncional das células T CD4+/CD8+, ocorreu redução da resposta na DA em relação aos controles; nos pacientes atópicos encontramos aumento da resposta em situação basal na dependência de CD38, e redução na resposta frente a SEA/SEB na ausência de CD38; c) encontramos resposta reduzida das células Th22, e elevada de células Tc22 frente aos estímulos SEA e SEB, nos pacientes com DA. CONCLUSÕES: O estudo corrobora o papel patogênico das enterotoxinas estafilocócicas na DA. A ativação crônica com superantígenos estafilocócicos pode contribuir com a alta frequência de células T CD4+ CD38+ polifuncionais, e com a resposta polifuncional anérgica, mediadas por células T CD38- / BACKGROUND: Atopic dermatitis (AD) is an inflammatory skin disease with intense itching and xerosis. AD pathogenesis is multifactorial, involving genetic, environmental, and immunological factors, among others. OBJECTIVES: To evaluate the influence of enterotoxins A and B from Staphylococcus aureus (SEA and SEB) in Th17 and Th22 cell response in adults with AD. METHODS: We evaluated 38 adult patients with AD, and a control group of 40 adults, age and gender matched. Assays: 1) ELISA: evaluation of IL-6, IL-17, IL-12p40/IL-23 and IL-22 serum levels and in supernatants of mononuclear cell cultures from peripheral blood (PBMC), stimulated with SEA/SEB; 2) Immunohistochemistry: analysis of IL-17 expression in skin specimens; 3) Flow cytometry: a) analysis of circulating cytokines in serum samples: IL-2, IL-4, IL-5, IL-6, IL-10, TNF, IL-17A and IFN-y b) evaluation of mono and polyfunctional TCD4+ cells that secrete IL-17, IL-22, TNF, IFN-y, MIP-1beta, and expression of the activation marker CD38; c) analysis of Tc22 and Th22 cells stimulated with SEA and SEB. RESULTS: 1) Secretion of IL-22 in the serum and from supernatants of cell cultures from PBMC, stimulated with SEA and SEB were higher in AD patients, when compared to the control group by ELISA; 2) there was an increase of IL-17 expression in skin samples by immunohistochemistry; 3) Flow cytometry showed: a) elevated serum levels of IL-2, 5, 6, 10, 17A and IFN-y in AD, when compared to controls; there was a significant difference in circulating levels of IL-17A in patients with moderate and severe disease; b) monofunctional evaluation of T CD4+ cells under SEA/SEB stimuli showed reduced expression of IFN-y, IL-17A, IL-22 or TNF cytokines in AD, compared to controls; the same was observed for polyfunctional CD4+/CD8+ T cells analysis, exhibiting a diminished response in AD. In atopic patients under basal conditions, there was an augmented CD38- dependent response and reduced pattern to SEA/SEB in the absence of CD38; c) finally, we observed a reduced response of Th22 cells and enhanced Tc22 cells under SEA/SEB stimuli in patients with AD. CONCLUSIONS: This study corroborates the pathogenic role of staphylococcal enterotoxins in AD. Chronic activation with staphylococcal superantigens may contribute to the high frequency of polyfunctional CD4 +CD38+ T cells and with the anergic polyfunctional response mediated by T CD38- T cells

Sy, Hing Yee. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2011. / Includes bibliographical references (leaves lxxiv-xciv). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstract also in Chinese; appendixes I-III in Chinese.

A resposta proliferativa das células mononucleares do sangue periférico dos adultos com dermatite atópica mostrou-se diminuída após estímulos com mitógenos (enterotoxinas estafilocócicas A e B, "pokeweed" e fitohemaglutinina) e antígenos (toxóide tetânico e Candida albicans). A correlação positiva dos níveis séricos de interleucina-18, IgE e escore de gravidade da doença sugerem que esta citocina seria um marcador de atividade da dermatite atópica do adulto / A reduced proliferative response of peripheral blood mononuclear cells in adults with atopic dermatitis was detected when stimuli with mitogens (staphylococcal enterotoxins A and B, phytohemaglutinin, pokeweed), and with antigens (tetanus toxoid and Candida albicans) were performed. A positive correlation of interleukin-18, IgE levels and severity scores of the disease suggest that this cytokine could be a marker of disease activity in adult atopic dermatitis

A resposta proliferativa das células mononucleares do sangue periférico dos adultos com dermatite atópica mostrou-se diminuída após estímulos com mitógenos (enterotoxinas estafilocócicas A e B, "pokeweed" e fitohemaglutinina) e antígenos (toxóide tetânico e Candida albicans). A correlação positiva dos níveis séricos de interleucina-18, IgE e escore de gravidade da doença sugerem que esta citocina seria um marcador de atividade da dermatite atópica do adulto / A reduced proliferative response of peripheral blood mononuclear cells in adults with atopic dermatitis was detected when stimuli with mitogens (staphylococcal enterotoxins A and B, phytohemaglutinin, pokeweed), and with antigens (tetanus toxoid and Candida albicans) were performed. A positive correlation of interleukin-18, IgE levels and severity scores of the disease suggest that this cytokine could be a marker of disease activity in adult atopic dermatitis

INTRODUÇÃO: A dermatite atópica (DA) é uma doença cutânea inflamatória, acompanhada por prurido intenso e xerose cutânea. A etiopatogenia da DA é multifatorial, envolvendo fatores genéticos, ambientais e imunológicos, dentre outros. OBJETIVOS: Avaliar a influência das enterotoxinas A e B do Staphylococcus aureus (SEA e SEB) na resposta mediada por células Th17 e Th22 nos indivíduos adultos com DA. MÉTODOS: Foram selecionados 38 pacientes adultos com DA e um grupo controle com 40 indivíduos adultos, pareados por idade e gênero Os métodos utilizados foram: 1) ELISA: dosagem dos níveis séricos de IL-6, IL-17, IL-22 e IL-12p40/IL-23 e em sobrenadantes de culturas de células mononucleares do sangue periférico (PBMC) estimuladas com SEA e SEB; 2) Imuno-histoquímica: análise da expressão de IL-17 em fragmentos de pele; 3) Citometria de fluxo: a) análise das citocinas circulantes em amostras de soro: IL-2, IL-4, IL-5, IL-6, IL-10, TNF, IL-17A e IFN-y b)avaliação das células T CD4+ mono e polifuncionais secretoras de IL-17, IL-22, TNF, IFN-y, MIP-1beta, e expressão do marcador de ativação celular CD38; c) células Th22 e Tc22 estimuladas com SEA e SEB. RESULTADOS: 1) Através do ELISA, a secreção de IL-22 sérica e em PBMC induzidas por SEA e SEB foi significativamente mais elevada, quando comparada ao grupo controle; 2) houve aumento na expressão de IL-17 em amostras de pele de doentes de DA através da imuno-histoquímica; 3) Através da citometria de fluxo, foram detectados: a) níveis séricos de IL-2, 5, 6, 10, 17A e IFN-y elevados no grupo com DA em relação aos controles; houve diferença significativa nos níveis circulantes de IL-17A nos pacientes com DA moderada e grave; b) na avaliação monofuncional das células T CD4+ sob estímulo de SEA/SEB, houve redução da expressão das citocinas IFN-y, IL-17A, IL-22 ou TNF na DA, quando comparadas ao grupo controle; na análise polifuncional das células T CD4+/CD8+, ocorreu redução da resposta na DA em relação aos controles; nos pacientes atópicos encontramos aumento da resposta em situação basal na dependência de CD38, e redução na resposta frente a SEA/SEB na ausência de CD38; c) encontramos resposta reduzida das células Th22, e elevada de células Tc22 frente aos estímulos SEA e SEB, nos pacientes com DA. CONCLUSÕES: O estudo corrobora o papel patogênico das enterotoxinas estafilocócicas na DA. A ativação crônica com superantígenos estafilocócicos pode contribuir com a alta frequência de células T CD4+ CD38+ polifuncionais, e com a resposta polifuncional anérgica, mediadas por células T CD38- / BACKGROUND: Atopic dermatitis (AD) is an inflammatory skin disease with intense itching and xerosis. AD pathogenesis is multifactorial, involving genetic, environmental, and immunological factors, among others. OBJECTIVES: To evaluate the influence of enterotoxins A and B from Staphylococcus aureus (SEA and SEB) in Th17 and Th22 cell response in adults with AD. METHODS: We evaluated 38 adult patients with AD, and a control group of 40 adults, age and gender matched. Assays: 1) ELISA: evaluation of IL-6, IL-17, IL-12p40/IL-23 and IL-22 serum levels and in supernatants of mononuclear cell cultures from peripheral blood (PBMC), stimulated with SEA/SEB; 2) Immunohistochemistry: analysis of IL-17 expression in skin specimens; 3) Flow cytometry: a) analysis of circulating cytokines in serum samples: IL-2, IL-4, IL-5, IL-6, IL-10, TNF, IL-17A and IFN-y b) evaluation of mono and polyfunctional TCD4+ cells that secrete IL-17, IL-22, TNF, IFN-y, MIP-1beta, and expression of the activation marker CD38; c) analysis of Tc22 and Th22 cells stimulated with SEA and SEB. RESULTS: 1) Secretion of IL-22 in the serum and from supernatants of cell cultures from PBMC, stimulated with SEA and SEB were higher in AD patients, when compared to the control group by ELISA; 2) there was an increase of IL-17 expression in skin samples by immunohistochemistry; 3) Flow cytometry showed: a) elevated serum levels of IL-2, 5, 6, 10, 17A and IFN-y in AD, when compared to controls; there was a significant difference in circulating levels of IL-17A in patients with moderate and severe disease; b) monofunctional evaluation of T CD4+ cells under SEA/SEB stimuli showed reduced expression of IFN-y, IL-17A, IL-22 or TNF cytokines in AD, compared to controls; the same was observed for polyfunctional CD4+/CD8+ T cells analysis, exhibiting a diminished response in AD. In atopic patients under basal conditions, there was an augmented CD38- dependent response and reduced pattern to SEA/SEB in the absence of CD38; c) finally, we observed a reduced response of Th22 cells and enhanced Tc22 cells under SEA/SEB stimuli in patients with AD. CONCLUSIONS: This study corroborates the pathogenic role of staphylococcal enterotoxins in AD. Chronic activation with staphylococcal superantigens may contribute to the high frequency of polyfunctional CD4 +CD38+ T cells and with the anergic polyfunctional response mediated by T CD38- T cells

The adaptor protein Shb has been implicated in the signaling of several tyrosine kinase receptors and previous studies have suggested a role for Shb in the signal transduction of T cells. Shb associates with the T cell receptor (TCR) and partakes in the signal propagation of activated T lymphocytes. In order to explore Shb’s influence on TCR signaling in vivo, T cell development and function was studied in a Shb knockout mouse. The loss of Shb led to aberrant TCR signaling in both thymocytes and peripheral CD4+ TH cells, with elevated basal phosphorylation of key components in the signal cascade. Shb was found to be dispensable for thymocyte development, but its absence resulted in a TH2 bias in in vitro stimulated peripheral CD4+ TH cells. As imbalances in TH2 responses are linked to allergic diseases, we further explored Shb’s role in immune regulation in a mouse model of atopic dermatitis. Shb knockout mice exhibit more aggravated signs of atopic dermatitis, including increased immune cell recruitment to the affected areas and elevated mRNA levels of typical TH2 cytokines. The effect of Shb on hematopoiesis in general was determined by examining populations of long-term hematopoietic stem cells (LT-HSCs) and hematopoietic progenitor cells in bone marrow of Shb knockout and wild type mice. Shb deficient bone marrow was found to contain significantly fewer relative numbers of LT-HSCs due to a proliferative defect. The reduced cell cycle activity of Shb LT-HSCs could further be linked to an abnormal regulation of the focal adhesion kinase/Rac1/p21-activated kinase pathway. Since alterations in LT-HSC proliferative abilities may have implications for leukemia development, BCR-Abl induced myeloid neoplasia was investigated in the absence of Shb. Shb deficiency confers a more aggressive progression of BCR-Abl induced myeloid neoplasia characterized by an increased peripheral blood neutrophilia and a deregulated cytokine profile. In addition, focal adhesion kinase and STAT3 signaling is hyperactivated in Shb knockout leukemic cells. In conclusion, Shb appears to be a multifunctional signaling mediator that controls several responses in hematopoietic cells, under homeostatic as well as disease conditions.

ABSTRACT Introduction and Rationale: Atopic Dermatitis (AD) is a common skin condition, characterized by markedly pruritic eczematous lesions, that most often presents in childhood. The majority of children diagnosed with AD will have mild disease and will first present with symptoms to a primary care provider (PCP), however approximately 85% of pediatricians only provide limited initial care followed by a referral to dermatology (Eichenfield et al., 2015). While there are specialty care based treatment guidelines for childhood AD, there are no guidelines available that specifically address primary care management of childhood AD. Purpose and Objective: The primary purpose of this DNP project is to develop an evidence-based clinical practice guideline (CPG) for pediatric PCPs. The secondary purpose is to develop a corresponding atopic dermatitis action plan (ADAP) to be used by children and parents. The objective is to equip PCPs to better manage children with AD in the primary care setting and to guide patients and parents in the importance of daily control measures and in the individualized treatment plan prescribed by the PCP. Methods: The Appraisal of Guidelines for Research & Evaluation II (AGREE II) framework and Social Cognitive Theory (SCT) serve as the theoretical frameworks for CPG and ADAP development. The American Academy of Pediatrics (AAP) process for evidence based policy setting is used as a model for key action statement development. Results: Evaluation of the CPG was completed using the AGREE II tool, a reliable and validated tool for evaluating CPGs. Five of the six domains evaluated, yielded combined scores of at least 90%, with one domain a combined score of 63%. The overall standard deviation was 0.58, indicating an overall low level of user discrepancy Additions and revisions were made based on the results of the AGREE II evaluation scores with specific emphasis on the lowest scoring domain. Conclusion: This DNP Project identified the need for a CPG specific to pediatric primary care. A CPG with accompanying ADAP was developed and evaluated using the AGREE II tool. The CPG was found to meet the recommended standards and recommended for use in pediatric primary care.