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TH17-immune responses
ABSTRACTS | Inflammation, Immunity and Infection 226
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Effects of culture temperature on functions of monocytes/dendritic cells J Hosoi and E Takai Shiseido Global Innovation Center, Yokohama, Japan Since the skin is exposed directly to external environment, it is affected by environmental climate. Epidermal Langerhans cells which reside in the upper layewr of epidermis are, therfore, affected by einvironmental temperature. The effects of culture temperature on epidermal Langerhans cells were examined by using model cell lines, THP-1 and U937, as well as differentiated cells from CD34-positive umbilical cord blood cells. Gene expression profile of THP-1 cells was compared between RNA from cells culture at 37 C and ones cultured at 33 C. Regulated genes by temperature include growth controling factors, inflammation related factors, hormone receptors, and others. Inhibitors/stimulators are upregulated at 33C and downregulated at 37 C or vice versa ion each category. Proliferation of THP-1 cells and CD34+ cell-derived Langerhans cell-like cells were declined at 33 C. Response to hapten, DNCB, was slightly suppressed by culture at 33 C. These results suggest that Langerhans function differently in situ from the regular culture condition at 37 C.
Effect of different cooking methods on histamine levels in selected foods B Chung1, Y Byun1, J Son1, Y Cho1, H Kim1, H Cho2 and C Park1 1 Dermatology, Kangnam Sacred Heart Hospital, Seoul, Korea (the Republic of) and 2 Dermatology, Chuncheon Sacred Heart Hospital, ChunCheon, Korea (the Republic of) Histamine in food is known to cause food poisoning and allergic reactions. We usually ingest histamine in cooked food, but there are few studies about the influence of cooking method on the histamine level. The purpose of this study was to determine the influence of cooking methods on the concentration of histamine of foods. The foods chosen were those kinds consumed frequently and cooked by various methods. The histamine level of the food was measured using enzyme-linked immunosorbent assay (ELISA). Grilled seafood had higher histamine levels than raw or boiled seafood. For meat, grilling increased the histamine level, while boiling decreased it. For eggs and vegetables, there was not much difference in histamine level according to cooking method. But fermenting vegetables greatly increased histamine levels. And fermented foods didn’t show much difference in histamine level after being boiled. The histamine level in food has changed according to the cooking method used to prepare it. Frying, grilling and fermenting increased histamine level of foods, whereas boiling had little influence or even decreased it. This study could be beneficial to histamine sensitive patients by providing dietary advice for reducing histamine levels in their diets.
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A novel splenic B1 regulatory B cell subset with a unique CD9+CD80+ phenotype suppresses an allergic disease via PI3K-Akt pathway activation T Matsushita1, M Fujimoto2 and K Takehara1 1 Kanazawa University, Kanazawa, Japan and 2 University of Tsukuba, Tsukuba, Japan IL-10-producing regulatory B (B10) cells potently suppress the autoimmune diseases and allergic diseases, such as contact hypersensitivity (CHS). The signaling mechanisms of B10 cell development remains poorly understood. Furthermore, splenic B10 cells share overlapping phenotypic markers with CD1dhiCD5+ B cell and CD1dhiCD21+CD23+ T2-marginal zone (MZ) precursor B cell subsets, but do not exclusively belong to either subset. Therefore, it is necessary to clarify the signaling mechanisms of B10 cell development and identify a unique phenotypic marker for B10 cells. In this study, we performed transcriptome analysis of B10 cells comparing IL-10+ and IL-10- B cells. Microarray analysis revealed the phosphatidylinositol 3-kinase (PI3K)-Akt pathway is important for IL-10 production in B cells. PI3K-Akt pathway inhibitors reduced B10 cells in vitro. Furthermore, B10 cells were significantly increased in B cell-specific phosphatase and tensin homolog (PTEN)-deficient mice, which exhibit aberrant activation of the PI3K-Akt pathway in B cells. The CHS response was significantly diminished in B cell-specific PTEN-deficient mice. Unexpectedly, splenic B10 cells in these mice were found within B1 B cell subset but not within MZ B cell subset, which is similar to CD1dhiCD5+ B cell or T2-MZ precursor B cell subsets. In addition, splenic B10 cells in wild type mice were found within not only MZ B cell subset but also B1 B cell subset, and both MZ B cell and B1 B cell subset inhibited CHS response. These results suggested two major B10 cell subsets designated as “MZ-B10 cells” and “B1-B10 cells”. Microarray analysis also revealed CD9 and CD80 were useful markers for identifying B10 cell subset. In addition, MZ-B10 cells and B1-B10 cells subset were predominantly found within a unique CD9+CD80+ phenotype. Taken together, our findings show the PI3K-Akt pathway in B cell is critical for B10 cell development and CHS response. Furthermore, CD9+CD80+ is a novel phenotype for both MZ-B10 cells and B1-B10 cells.
Two cases of cutaneous vasculitis in cogan’s syndrome: The relationship between the pathophysiology and chlamydia infection Y Kawasaki2, S Kawana3 and T Uehara1 1 Clinical Oncology and Respiratory Medicine, Nippon Medical School, Tama Nagayama Hospital, Tama, Japan, 2 Dermatology, Nippon Medical School, Chiba Hokusoh Hospital, Inzai, Japan and 3 Dermatology, Nippon Medical School, Bunkyo, Japan Cogan’s syndrome (CS), a rare disorder characterized by bilateral sensorineural hearing loss, vestibular symptoms, and inflammatory ocular manifestations with a variable risk of developing into a systemic disease, can be associated with various types of vasculitis in multiple organs. Vasculitis affects large vessels predominantly, while small arteritis and small vessel vasculitis are rare. To date, only four cases of CS with polyarteritis nodosa (PAN) and two with small vessel vasculitis have been histologically confirmed in the English literatures. Recently, we observed two cases of CS with cutaneous vasculitis. One of them simultaneously showed typical symptoms of Takayasu’s arteritis and multiple purpuric skin lesions in the lower extremities with histological findings of small vessel vasculitis of the dermis. The other case showed subcutaneous nodules in the lower extremities and axillae with histological findings of small arteritis in the interlobular area. Both cases were demonstrated to have high IgA- and IgG-antibody titers against Chlamydia trachomatis in the serum, which indicates an active stage of Chlamydia infection. We conclude that chronic Chlamydia infection, such as infectious perihepatitis detected in one of our patient, can possibly be associated with the pathophysiology of vasculitis in CS through immunological mechanisms. Dermatologists should know that CS may develop various skin symptoms caused by cutaneous vasculitis affecting vessels in the dermis and the interlobular area.
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Aryl hydrocarbon receptor deficiency exacerbates immune complex-mediated vascular injury through up-regulation of CD16 expression on macrophages R Nakajima, T Miyagaki, M Kabasawa, S Morimura, M Sugaya and S Sato Department of Dermatology, University of Tokyo Graduate School of Medicine, Tokyo, Japan Aryl hydrocarbon receptor (AhR) is a ligand-activated transcription factor that mediates numerous cellular responses to toxins and plays critical modulatory roles in various immune cells during innate and adaptive immune responses. Recently, a number of studies have examined the functions of AhR in the immune system using mouse models of a wide variety of inflammatory and autoimmune diseases. However, studies on AhR involvement in vasculitis have been limited. Immune complex (IC) deposition causes significant vascular injury associated with various autoimmune diseases. Arthus reaction is a classic experimental model for immune complex-mediated vascular injury, which is characterized by edema, hemorrhage, and neutrophil infiltration. In this study, to assess the function of AhR in ICmediated vascular injury, cutaneous and peritoneal reverse-passive Arthus reaction was examined in mice with heterozygous deficiency of AhR (AhR+/-). In the cutaneous Arthus reaction, dermal edema, which indicates cutaneous inflammation, was more severe in AhR+/- mice than in wild-type (WT) mice. The number of infiltrating neutrophils, mRNA expression levels of CXC chemokine ligand (CXCL) 1, which is a chemoattractant for neutrophils, and interleukin (IL)-6 were also increased in AhR+/- mice. Similarly, in peritoneal Arthus reaction, infiltration of neutrophils in peritoneal lavage was significantly increased in AhR+/- mice compared with WT mice. Peritoneal macrophages from AhR+/- mice expressed higher levels of CD16, a FcgRIII that binds to IC, than those from WT mice. Furthermore, after in vitro IC-treatment, the former produced higher levels of CXCL1 and IL-6 than the latter. These results suggest that AhR deficiency induces up-regulation of CD16 expression and thereby higher production of IL-6 and CXCL1 from macrophages, resulting in increased infiltration of neutrophils, followed by exacerbation of IC-mediated vascular injury.
S200 Journal of Investigative Dermatology (2016), Volume 136
Free fatty acids sensitize dermal cells to amplify TH1/TH17-immune responses D Herbert1, A Lorz1, K Stelzner1, Y Popkova2, J Simon1 and A Saalbach1 1 Dermatology, Allergology and Venerology, Universal Hospital Leipzig, Leipzig, Germany and 2 Institute of Medical Physics and Biophysics, Leipzig, Germany Obesity is associated with body fat gain and impaired glucose metabolism. Here, we identified both body fat gain in obesity and impaired glucose metabolism as two independent risk factors for increased levels of serum free fatty acids (FFAs). Furthermore, obese subjects show increased and/or delayed resolution of inflammation observed in various chronic inflammatory diseases such as psoriasis. Thus, we hypothesized a possible impact of FFAs on immune cell function relevant for the pathogenesis of chronic inflammation. Surprisingly, stimulation of human monocyte-derived and mouse bone marrow-derived dendritic cells (DCs) with FFAs, such as palmitic acid (PA) and oleic acid (OA), did not affect the pro-inflammatory immune response of these immune cells. In contrast, pre-incubation with PA and OA sensitizes DCs resulting in augmented secretion of TH1/TH17-instructive cytokines upon pro-inflammatory stimulation with LPS. Similar results according to a sensitization of dermal cells, such as keratinocytes and fibroblasts, to danger signals could be shown in this study by incubation with FFAs. The relevance of this observation was confirmed in vivo when obesity in mice worsened a TH1/TH17-driven psoriasis-like skin inflammation. A strong correlation of the amount of total FFA, PA and OA in serum and the severity of skin inflammation point to a critical role of FFAs in obesity mediating exacerbation of skin inflammation. Our data suggest that increased levels of FFAs might be a pre-disposing factor promoting a TH1/TH17mediated inflammation such as psoriasis in response to an inflammatory danger signal.
227
Effects of culture temperature on functions of monocytes/dendritic cells J Hosoi and E Takai Shiseido Global Innovation Center, Yokohama, Japan Since the skin is exposed directly to external environment, it is affected by environmental climate. Epidermal Langerhans cells which reside in the upper layewr of epidermis are, therfore, affected by einvironmental temperature. The effects of culture temperature on epidermal Langerhans cells were examined by using model cell lines, THP-1 and U937, as well as differentiated cells from CD34-positive umbilical cord blood cells. Gene expression profile of THP-1 cells was compared between RNA from cells culture at 37 C and ones cultured at 33 C. Regulated genes by temperature include growth controling factors, inflammation related factors, hormone receptors, and others. Inhibitors/stimulators are upregulated at 33C and downregulated at 37 C or vice versa ion each category. Proliferation of THP-1 cells and CD34+ cell-derived Langerhans cell-like cells were declined at 33 C. Response to hapten, DNCB, was slightly suppressed by culture at 33 C. These results suggest that Langerhans function differently in situ from the regular culture condition at 37 C.
Effect of different cooking methods on histamine levels in selected foods B Chung1, Y Byun1, J Son1, Y Cho1, H Kim1, H Cho2 and C Park1 1 Dermatology, Kangnam Sacred Heart Hospital, Seoul, Korea (the Republic of) and 2 Dermatology, Chuncheon Sacred Heart Hospital, ChunCheon, Korea (the Republic of) Histamine in food is known to cause food poisoning and allergic reactions. We usually ingest histamine in cooked food, but there are few studies about the influence of cooking method on the histamine level. The purpose of this study was to determine the influence of cooking methods on the concentration of histamine of foods. The foods chosen were those kinds consumed frequently and cooked by various methods. The histamine level of the food was measured using enzyme-linked immunosorbent assay (ELISA). Grilled seafood had higher histamine levels than raw or boiled seafood. For meat, grilling increased the histamine level, while boiling decreased it. For eggs and vegetables, there was not much difference in histamine level according to cooking method. But fermenting vegetables greatly increased histamine levels. And fermented foods didn’t show much difference in histamine level after being boiled. The histamine level in food has changed according to the cooking method used to prepare it. Frying, grilling and fermenting increased histamine level of foods, whereas boiling had little influence or even decreased it. This study could be beneficial to histamine sensitive patients by providing dietary advice for reducing histamine levels in their diets.
228
229
A novel splenic B1 regulatory B cell subset with a unique CD9+CD80+ phenotype suppresses an allergic disease via PI3K-Akt pathway activation T Matsushita1, M Fujimoto2 and K Takehara1 1 Kanazawa University, Kanazawa, Japan and 2 University of Tsukuba, Tsukuba, Japan IL-10-producing regulatory B (B10) cells potently suppress the autoimmune diseases and allergic diseases, such as contact hypersensitivity (CHS). The signaling mechanisms of B10 cell development remains poorly understood. Furthermore, splenic B10 cells share overlapping phenotypic markers with CD1dhiCD5+ B cell and CD1dhiCD21+CD23+ T2-marginal zone (MZ) precursor B cell subsets, but do not exclusively belong to either subset. Therefore, it is necessary to clarify the signaling mechanisms of B10 cell development and identify a unique phenotypic marker for B10 cells. In this study, we performed transcriptome analysis of B10 cells comparing IL-10+ and IL-10- B cells. Microarray analysis revealed the phosphatidylinositol 3-kinase (PI3K)-Akt pathway is important for IL-10 production in B cells. PI3K-Akt pathway inhibitors reduced B10 cells in vitro. Furthermore, B10 cells were significantly increased in B cell-specific phosphatase and tensin homolog (PTEN)-deficient mice, which exhibit aberrant activation of the PI3K-Akt pathway in B cells. The CHS response was significantly diminished in B cell-specific PTEN-deficient mice. Unexpectedly, splenic B10 cells in these mice were found within B1 B cell subset but not within MZ B cell subset, which is similar to CD1dhiCD5+ B cell or T2-MZ precursor B cell subsets. In addition, splenic B10 cells in wild type mice were found within not only MZ B cell subset but also B1 B cell subset, and both MZ B cell and B1 B cell subset inhibited CHS response. These results suggested two major B10 cell subsets designated as “MZ-B10 cells” and “B1-B10 cells”. Microarray analysis also revealed CD9 and CD80 were useful markers for identifying B10 cell subset. In addition, MZ-B10 cells and B1-B10 cells subset were predominantly found within a unique CD9+CD80+ phenotype. Taken together, our findings show the PI3K-Akt pathway in B cell is critical for B10 cell development and CHS response. Furthermore, CD9+CD80+ is a novel phenotype for both MZ-B10 cells and B1-B10 cells.
Two cases of cutaneous vasculitis in cogan’s syndrome: The relationship between the pathophysiology and chlamydia infection Y Kawasaki2, S Kawana3 and T Uehara1 1 Clinical Oncology and Respiratory Medicine, Nippon Medical School, Tama Nagayama Hospital, Tama, Japan, 2 Dermatology, Nippon Medical School, Chiba Hokusoh Hospital, Inzai, Japan and 3 Dermatology, Nippon Medical School, Bunkyo, Japan Cogan’s syndrome (CS), a rare disorder characterized by bilateral sensorineural hearing loss, vestibular symptoms, and inflammatory ocular manifestations with a variable risk of developing into a systemic disease, can be associated with various types of vasculitis in multiple organs. Vasculitis affects large vessels predominantly, while small arteritis and small vessel vasculitis are rare. To date, only four cases of CS with polyarteritis nodosa (PAN) and two with small vessel vasculitis have been histologically confirmed in the English literatures. Recently, we observed two cases of CS with cutaneous vasculitis. One of them simultaneously showed typical symptoms of Takayasu’s arteritis and multiple purpuric skin lesions in the lower extremities with histological findings of small vessel vasculitis of the dermis. The other case showed subcutaneous nodules in the lower extremities and axillae with histological findings of small arteritis in the interlobular area. Both cases were demonstrated to have high IgA- and IgG-antibody titers against Chlamydia trachomatis in the serum, which indicates an active stage of Chlamydia infection. We conclude that chronic Chlamydia infection, such as infectious perihepatitis detected in one of our patient, can possibly be associated with the pathophysiology of vasculitis in CS through immunological mechanisms. Dermatologists should know that CS may develop various skin symptoms caused by cutaneous vasculitis affecting vessels in the dermis and the interlobular area.
230
231
Aryl hydrocarbon receptor deficiency exacerbates immune complex-mediated vascular injury through up-regulation of CD16 expression on macrophages R Nakajima, T Miyagaki, M Kabasawa, S Morimura, M Sugaya and S Sato Department of Dermatology, University of Tokyo Graduate School of Medicine, Tokyo, Japan Aryl hydrocarbon receptor (AhR) is a ligand-activated transcription factor that mediates numerous cellular responses to toxins and plays critical modulatory roles in various immune cells during innate and adaptive immune responses. Recently, a number of studies have examined the functions of AhR in the immune system using mouse models of a wide variety of inflammatory and autoimmune diseases. However, studies on AhR involvement in vasculitis have been limited. Immune complex (IC) deposition causes significant vascular injury associated with various autoimmune diseases. Arthus reaction is a classic experimental model for immune complex-mediated vascular injury, which is characterized by edema, hemorrhage, and neutrophil infiltration. In this study, to assess the function of AhR in ICmediated vascular injury, cutaneous and peritoneal reverse-passive Arthus reaction was examined in mice with heterozygous deficiency of AhR (AhR+/-). In the cutaneous Arthus reaction, dermal edema, which indicates cutaneous inflammation, was more severe in AhR+/- mice than in wild-type (WT) mice. The number of infiltrating neutrophils, mRNA expression levels of CXC chemokine ligand (CXCL) 1, which is a chemoattractant for neutrophils, and interleukin (IL)-6 were also increased in AhR+/- mice. Similarly, in peritoneal Arthus reaction, infiltration of neutrophils in peritoneal lavage was significantly increased in AhR+/- mice compared with WT mice. Peritoneal macrophages from AhR+/- mice expressed higher levels of CD16, a FcgRIII that binds to IC, than those from WT mice. Furthermore, after in vitro IC-treatment, the former produced higher levels of CXCL1 and IL-6 than the latter. These results suggest that AhR deficiency induces up-regulation of CD16 expression and thereby higher production of IL-6 and CXCL1 from macrophages, resulting in increased infiltration of neutrophils, followed by exacerbation of IC-mediated vascular injury.
S200 Journal of Investigative Dermatology (2016), Volume 136
Free fatty acids sensitize dermal cells to amplify TH1/TH17-immune responses D Herbert1, A Lorz1, K Stelzner1, Y Popkova2, J Simon1 and A Saalbach1 1 Dermatology, Allergology and Venerology, Universal Hospital Leipzig, Leipzig, Germany and 2 Institute of Medical Physics and Biophysics, Leipzig, Germany Obesity is associated with body fat gain and impaired glucose metabolism. Here, we identified both body fat gain in obesity and impaired glucose metabolism as two independent risk factors for increased levels of serum free fatty acids (FFAs). Furthermore, obese subjects show increased and/or delayed resolution of inflammation observed in various chronic inflammatory diseases such as psoriasis. Thus, we hypothesized a possible impact of FFAs on immune cell function relevant for the pathogenesis of chronic inflammation. Surprisingly, stimulation of human monocyte-derived and mouse bone marrow-derived dendritic cells (DCs) with FFAs, such as palmitic acid (PA) and oleic acid (OA), did not affect the pro-inflammatory immune response of these immune cells. In contrast, pre-incubation with PA and OA sensitizes DCs resulting in augmented secretion of TH1/TH17-instructive cytokines upon pro-inflammatory stimulation with LPS. Similar results according to a sensitization of dermal cells, such as keratinocytes and fibroblasts, to danger signals could be shown in this study by incubation with FFAs. The relevance of this observation was confirmed in vivo when obesity in mice worsened a TH1/TH17-driven psoriasis-like skin inflammation. A strong correlation of the amount of total FFA, PA and OA in serum and the severity of skin inflammation point to a critical role of FFAs in obesity mediating exacerbation of skin inflammation. Our data suggest that increased levels of FFAs might be a pre-disposing factor promoting a TH1/TH17mediated inflammation such as psoriasis in response to an inflammatory danger signal.