Progress in regulatory effects of thymic stromal lymphopoietin on immune cells / 中华微生物学和免疫学杂志

Thymic stromal lymphopoietin (TSLP), as a pleiotropic cell growth factor, not only participants in the processes of human skin fibrosis, epidermal proliferation and angiogenesis, but also plays a critical role in regulating a variety of immune cells in immune-related diseases (such as respiratory diseases and allergic diseases). TSLP regulates various innate immune cells (such as dendritic cells, mast cells, macrophages, eosinophils, basophils, natural killer T cells and innate lymphocytes) and adaptive immune cells (T lymphocytes and B lymphocytes) mainly through JAK/STAT, NF-κB and other signal pathways mediated by TSLP receptor. This paper summarized the progress in the regulatory roles of TSLP in the proliferation, differentiation and function of various immune cells.

Role of epithelium-derived cytokines interleukin-33, interleukin-25 and thymic stromal lymphopoietin in the pathogenesis of atopic dermatitis / 中华皮肤科杂志

Epidermal barrier defects and immune abnormalities are the main pathophysiological changes in the development of atopic dermatitis (AD) . Skin keratinocytes can release a variety of inflammatory factors and mediators under the treatment with various harmful factors. Three epithelium-derived cytokines interleukin (IL) -33, IL-25 and thymic stromal lymphopoietin are considered to be effective inducers of Th2 immune response in skin or mucosal barrier, which can activate immune cells, cause the secretion of Th2 cytokines, enhance the Th2 immune response, and participate in the occurrence and development of AD. This review focuses on the role of the above 3 epithelium-derived cytokines in the pathogenesis of AD.

The sensitization of thymic stromal lymphopoietin to type 2 lymphocytes in the lung / 国际儿科学杂志

Thymic stromal lymphopoietin(TSLP)can enhance the secretion of type 2 cytokines by type 2 T helper(Th2)cells and type 2 innate iymphocytes(ILC2s)in the lung through direct and indirect action.In terms of indirect effects, TSLP can promote the proliferation and expansion of naive CD4 + T cells by using dendritic cells, and promote the recruitment of Th2 cells to the initial inflammatory site.It can also use stromal mononuclear/macrophages cells to start the acute primary Th2-dependent immune response.In the aspect of direct effect, TSLP can activate ILC2s before the adaptive immune response is initiated, and also can use immature T cells, effector Th2 cells and memory Th2 cells to participate in the adaptive immune response.Therefore, TSLP plays an important role in allergic immune response of lung.

Abnormal thymic stromal lymphopoietin expression in the gastrointestinal mucosa of patients with eosinophilic gastroenteritis / Expressão anormal da linfopoietina estromal tímica na mucosa gastrointestinal depacientes com gastroenterite e osinofílica

Abstract Objective: To investigate the differential expression of the thymic stromal lymphopoietin isoforms, short and long, and discern their biological implications under eosinophilic gastroenteritis. Methods: The expression of thymic stromal lymphopoietin and its two isoforms in tissues was assessed by quantitative RT-PCR in healthy controls (n = 24) and patients with eosinophilic gastroenteritis (n = 17). Results: Thymic stromal lymphopoietin mRNA was significantly reduced in eosinophilic gastroenteritis when compared with healthy controls (p < 0.0001). A significantly lower amount of short thymic stromal lymphopoietin mRNA was observed in eosinophilic gastroenteritis when compared with controls (p < 0.05), while a significantly higher amount of long thymic stromal lymphopoietin mRNA was observed in eosinophilic gastroenteritis when compared with controls (p < 0.05). Peak eosinophilic count is significantly positively correlated with the expression of long thymic stromal lymphopoietin mRNA in the gastrointestinal mucosal of patients with eosinophilic gastroenteritis (r s = 0.623, p < 0.005), while peak eosinophilic count is significantly negatively correlated with the expression of short thymic stromal lymphopoietin mRNA in the gastrointestinal mucosal of patients with eosinophilic gastroenteritis (r s = −0.4474, p < 0.05). Conclusions: Abnormal mucosal thymic stromal lymphopoietin expression may contribute to gastrointestinal mucosa damage in eosinophilic gastroenteritis.

Resumo Objetivo: Investigar a expressão diferencial das isoformas da linfopoietina estromal tímica, curta e longa, e discernir suas implicações biológicas na gastroenterite eosinofílica. Métodos: Avaliamos a expressão das isoformas da linfopoietina estromal tímica e suas duas isoformas através da técnica RT-PCR quantitativa em tecidos de controles saudáveis (n = 24) e pacientes com gastroenterite eosinofílica (n = 17). Resultados: Demonstramos que o RNAm das isoformas da linfopoietina estromal tímica estava significativamente reduzido na gastroenterite eosinofílica em comparação com os controles saudáveis (p < 0,0001). Também descobrimos uma quantidade significativamente menor de RNAm das isoformas da linfopoietina estromal tímica curta na gastroenterite eosinofílica em comparação com os controles (p < 0,05) e uma quantidade significativamente maior de RNAm das isoformas da linfopoietina estromal tímica longa na gastroenterite eosinofílica em comparação com os controles (p < 0,05). O pico da contagem eosinofílica está correlacionado positiva e significativamente com a expressão do RNAm das isoformas da linfopoietina estromal tímica longa na mucosa gastrointestinal de pacientes com gastroenterite eosinofílica (rs = 0,623, p < 0,005), enquanto o pico de contagem eosinofílica está negativa e significativamente correlacionado com a expressão do RNAm das isoformas da linfopoietina estromal tímica curta na mucosa gastrointestinal de pacientes com gastroenterite eosinofílica (rs = -0,4474, p < 0,05). Conclusões: A expressão anormal das isoformas da linfopoietina estromal tímica na mucosa pode contribuir para o dano da mucosa gastrointestinal na gastroenterite eosinofílica.

The inhibition of 18β-sodium glycyrrhetinic acid on thymic stromal lymphopoietin expression in the nasal mucosa of allergic rhinitis rats / 中华耳鼻咽喉头颈外科杂志

Objective@#To explore the effect of 18β-sodium glycyrrhetinic acid on thymic stromal lymphopoietin (TSLP) in nasal mucosa of allergic rhinitis (AR) rats.@*Methods@#One hundred Wistar rats,half male and half female,were randomly divided into 5 groups by random number table method: control group, AR model group,budesonide group,18β-sodium glycyrrhetinic acid at dose of 20 mg/kg and 40 mg/kg groups, with 20 rats in each group. AR animal models were established by ovalbumin (OVA) sensitization in the other four experimental groups. After successful modeling, budesonide and 18β-sodium glycyrrhetinic acid were given in each group,and the detection time points were 2 weeks and 4 weeks. The distribution of TSLP in rat nasal mucosa was detected by immunohistochemistry,and the expression of TSLP in rat nasal mucosa was determined by Western blot at the protein level. The expression of TSLP-mRNA in rat nasal mucosa was detected and compared by real-time fluorescence quantitative PCR (RT-PCR) at mRNA level. The concentrations of IL-4 and OVA-sIgE in rat serum were measured and compared by ELISA. One-way analysis of variance and the least significant difference method were used for the comparison among groups, LSD t test was used for the comparison between the two groups,and the difference was statistically significant (P<0.05).@*Results@#Immunohistochemistry confirmed existence of TSLP in rat nasal mucosa, especially in epithelial cells,endothelial cells and epithelial cilia. Western blot and RT-PCR suggested that the expression of TSLP and TSLP-mRNA in nasal mucosa of AR model group was significantly higher than that of control group (2 weeks TSLP: 1.795 9±0.131 4 vs 0.990 5±0.164 2, 4 weeks TSLP: 1.809 7±0.253 4 vs 0.870 3±0.124 4; 2 weeks TSLP-mRNA:4.582 9±0.697 7 vs 1.108 7±0.081 1, 4 weeks TSLP-mRNA:4.814 4±0.662 8 vs 1.001 0±0.155 3; all P<0.05). After 2 weeks and 4 weeks of drug intervention,the expression of TSLP and TSLP-mRNA was inhibited in nasal mucosa of budesonide group,18β-sodium sodium glycyrrhetinic acid at dose of 20 mg/kg and 40 mg/kg group,which was significantly different from that of AR model group (2 weeks TSLP: (0.897 8±0.081 8)/(1.072 1±0.113 6)/(1.396 6±0.133 9) vs 1.795 9±0.131 4; 4 weeks TSLP: (1.191 0±0.161 3)/(1.141 0±0.152 3)/(1.200 5±0.189 6) vs 1.809 7±0.253 4; 2 weeks TSLP-mRNA: (1.175 6±0.100 9)/(1.254 4±0.078 2)/(2.037 2±0.559 2) vs 4.582 9±0.697 7; 4 weeks TSLP-mRNA: (1.158 3±0.104 3)/(1.224 0±0.034 0)/(1.275 2±0.099 6) vs 4.814 4±0.662 8; all P<0.05), and not significantly different from control group. With the inhibition of TSLP, the concentrations of IL-4 and OVA-sIgE in rat serum were also decreased.@*Conclusion@#18β-sodium glycyrrhetinic acid has obvious inhibitory effect on TSLP in nasal mucosa of AR rats, which can control Th2 type immune inflammatory reaction.

Protease-Activated Receptors 2-Antagonist Suppresses Asthma by Inhibiting Reactive Oxygen Species-Thymic Stromal Lymphopoietin Inflammation and Epithelial Tight Junction Degradation

PURPOSE: Protease-activated receptor 2 (PAR2) reportedly triggers the immune response in allergic asthma. We aimed to investigate the mechanism on allergic inflammation mediated by PAR2. METHODS: Human lung epithelial cells (A549 cells) were used for in vitro, and the German cockroach extract (GCE)-induced mouse model was developed for in vivo studies. RESULTS: In A549 cells, the levels of reactive oxygen species (ROS) and thymic stromal lymphopoietin (TSLP) were significantly increased by GCE treatment, but were suppressed by PAR2-antagonist (PAR2-ant) or N-acetylcysteine (NAC) treatment. Claudin-1 was degraded by GCE, and was restored by PAR2-ant or NAC in the cells. In the mouse model, the clinical appearance including bronchial hyperresponsiveness, bronchoalveolar lavage fluid analysis and total immunoglobulin E were significantly suppressed by PAR2-ant or NAC. Moreover, TSLP levels in the lung were suppressed by the same treatments in the lung. Claudin-1 was also degraded by GCE, and was restored by PAR2-ant or NAC. CONCLUSIONS: ROS generation and epidermal tight junction degradation are triggered by protease, followed by the induction of TSLP in allergic asthma. Our findings could suggest that PAR2-ant or anti-oxidants could be considered for allergic diseases as preventive alternatives.

Auxo action and mechanism of tgf-β1 up-regulating smad3 phosphorylation on the expression of thymic stromal lymphopoietin in human bronchial Epithelia / 解放军医学杂志

[Abstract] Objective To discuss the effect and the corresponding mechanism of transforming growth factor β1 (TGF-β1) in promoting the bronchial epithelia synthesis and the expression of thymic stromal lymphopoietin (TSLP), so seek out a potential therapeutic target for asthma. Methods Human bronchial epithelia cells (HBEc) were cultured in vitro, and then divided into 0h group, 3h group, 6h group, 12h group, 24h group and 48h group to evaluate the effect of TGF-β1 stimulation in different time points; and divided into 0ng/ml group, 0.1ng/ml group, 1ng/ml group and 10ng/ml group to evaluate the effect of TGF-β1 stimulation in different concentrations. SB431542, a TGF-β1 antagonist, was used to block the effect of TGF-β1, HBEc were divided into negative control group, TGF-β1 group (1ng/ml TGF-β1) and TGF-β1+SB431542 group (1ng/ml TGF-β1+10μmol/L SB431542). Western blotting was performed to detect the protein expression level of TSLP, p-Smad3 and Smad3, while qRT-PCR was performed to determine the mRNA transcription level of TSLP. Concentrations of TSLP in HBEc culture supernatants were measured by ELISA. Results As the co-culture time with TGF-β1 prolonged, the expression of TSLP in HBEc increased. The relative expression of TSLP protein was significantly higher in 24h group (0.803±0.022) than in 0h group (0.350±0.032, P<0.05), and the relative expression of TSLP mRNA also increased (4.957±0.391 vs. 1.002±0.086, P<0.05). The levels of TSLP mRNA transcription and protein expression were significantly higher in 1ng/ml TGF-β1 group (7.954±2.004; 1.522±0.003) than in 0ng/ml TGF-β1 group (1.008±0.152; 0.758±0.014, P<0.05). The concentrations of TSLP in HBEc culture supernatants were markedly higher in 1ng/ml TGF-β1 group than in 0ng/ml TGF-β1 group (160.157±7.050 vs. 138.817±1.940, P<0.05). The ratio of p-Smad3/Smad3 declined obviously in TGF-β1+SB431542 group than in TGF-β1 group (0.808±0.063 vs. 1.116±0.049, P<0.05). Meanwhile, the relative expression of TSLP protein was significantly lower in TGF-β1+SB431542 group than in TGF-β1 group (1.016±0.030 vs. 1.186±0.045, P<0.05). Conclusion TGF-β1 may induce the expression of TSLP in HBEc by up-regulating Smad3 phosphorylation, which may be a novel method in curing asthma.

Bamboo salt suppresses skin inflammation in mice with 2, 4-dinitrofluorobenzene-induced atopic dermatitis / 中国天然药物

Bamboo salt (BS) is a traditional Korean food, and has been reported to have anti-cancer, anti-inflammatory, and anti-metastatic effects. However, the anti-atopic dermatitis (AD) activity of BS has not been described yet. In the present study, we examined the preventive effect of BS on AD. The effect of oral administration of BS was tested in a 2, 4-dinitrofluorobenzene (DNFB)-induced AD animal model, by histological analysis, enzyme-linked immunosorbent assay, reverse transcription-polymerase chain reaction, caspase-1 assay, and Western blotting analysis. BS administration reduced the total clinical severity and scratching frequencies, compared with the AD group. In the serum of DNFB-induced AD mice, the levels of IgE, histamine, thymic stromal lymphopoietin (TSLP), interleukin (IL)-5, and IL-13 were significantly reduced by BS treatment. BS significantly reduced the protein and mRNA expression of TSLP, IL-6, and tumor necrosis factor-α in the AD skin lesions. BS markedly reduced the infiltration of inflammatory cells. Furthermore, the activation of caspase-1 was reduced by BS in the AD skin lesions. Our results suggested that BS should be considered as a candidate treatment for allergic inflammatory diseases including AD.

The Role of Thymic Stromal Lymphopoietin (TSLP) in Glomerulonephritis

Thymic stromal lymphopoietin (TSLP) is an interleukin-7-like cytokine that is an important trigger and initiator of many allergic diseases. TSLP promotes a T-helper type 2 (Th2) cytokine response that can be pathological. A relationship is formed both at the induction phase of the Th2 response through polarization of dendritic cells to drive Th2 cell differentiation and at the effector phase of the response, by promoting the expansion of activated T cells and their secretion of Th2 cytokines and TSLP. In transgenic mice with TSLP overexpression, it has been reported that TSLP leads to the development of mixed cryoglobulinemic membranoproliferative glomerulonephritis. In addition, TSLP can play an important role in the pathogenesis of IgA nephropathy and systemic lupus erythematosus-related nephritis. From our knowledge of the role of TSLP in the kidney, further studies including the discovery of new therapies need to be considered based on the relationship between TSLP and glomerulonephritis.

Catechin remits the inflammation in allergic asthma mice by suppressing NF-κB-TSLP signal pathway / 中国药理学通报

Aim To explore the mechanism of catechin in inhibiting Th2-driven inflammation responses of allergic asthma mice.Methods Female BALB/c mice were chosen to establish an allergic asthma model by sensitized and excited with OVA.Then,the administered group was intervened with catechin by series of dose (75,150,300 mg · kg-1).The proportion of peripheral leukocyte cells was analysed,and pathological changes in lung tissues were observed by HE dyeing.The levels of TSLP,IL-5,IL-13 in lung tissues and OVA specific IgE in mouse serum were measured using ELISA kit.NF-κB p65,IκB protein expression levels in lung tissues were investigated using Western blot,and NF-κB p65 nuclear translocation was detected with immunofluorescent technique.Results Catechin could not only effectively reduce the proportion of peripheral leukocyte cells,but relieve the levels of TSLP,Th2 inflammatory factor IL-5 and IL-13,and the activation of NF-κB signal pathway in lung tissues of allergic asthma mice.Conclusions Catechin can effectively relieve Th2 inflammation in allergic asthma mice induced by allergen OVA.The possible mechanism may be that it could reduce expression of TSLP by inhibiting the NF-κB signal.

Effect of Xiaoqinglong Decoction on the expression of TSLP in airway epithelial cells of asthmatic model mice / 重庆医学

Objective To study the effect of Xiaoqinglong Decoction on the expression of TSLP in airway epithelial cells of asthmatic model mice.Methods Thirty healthy female BALB/C mice were divided into the blank control group,asthma model group and Xiaoqinglong Decoction group,10 cases in each group.The each group adopted corresponding intervention measures. Then the bronchial and lung tissues were taken.The expression level of bronchial TSLP was detected by immunofluorescence,and the expression level of TSLP mRNA and protein in lung tissue was detected by qRT-PCR and Western Blot.Results The mouse bronchial epithelial cell membrane TSLP staining was strongly positive in the asthma model group,which was weakly positive in the Xiaoqinglong Decoction group and negative in the blank control group.Compared with the asthma model group,mice lung tissue TSLP mRNA expression level in the Xiaoqinglong Decoction group and blank control group was decreased(P<0.05),the Xiaoqin-glong Decoction group was slightly higher than the blank control group,but the inter-group difference was not statistically signifi-cant(P>0.05).The expression level of lung tissue TSLP protein in the Xiaoqinglong Decoction group and blank control group was lower than that in the asthma model group(P<0.05),the Xiaoqinglong Decoction group was slightly higher than the blank control group,but the inter-group difference was not statistically significant(P>0.05).Conclusion Xiaoqinglong decoction has obvious inhibiting effect on the expression of TSLP in airway epithelial cells of asthmatic model mice.

TSLP Polymorphisms in Atopic Dermatitis and Atopic March in Koreans

BACKGROUND: Atopic march (AM) is the progression from atopic dermatitis (AD) to allergic rhinitis and asthma. The development of AD is as high as 20% in children worldwide and continues to increase. AD seems to be caused by both genetic and environmental factors. Recently, polymorphisms of the thymic stromal lymphopoietin (TSLP) gene associated with allergic disorders were reported in ethnic groups from various countries. OBJECTIVE: Identification of TSLP polymorphisms in Koreans with AD or AM. METHODS: Whole-exome sequencing was performed in 20 AD and 20 AM patients. RESULTS: Nine single nucleotide polymorphisms (SNPs) of TSLP were detected (rs191607411, rs3806933, rs2289276, rs2289277, rs2289278, rs139817258, rs11466749, rs11466750, rs10073816). These SNPs have been correlated with susceptibility to allergic diseases in ethnic groups from China, Japan, Turkey, and Costa Rica in previous studies. Remarkably, one of 20 patients in the AD group lacked all SNPs, compared to six of 20 patients in the AM group. Odds ratios showed that Korean patients without the nine TSLP variants had an 8.14 times higher risk of moving from AD to AM. Two haplotype blocks were validated in 60 AD and 59 AM patients using Sanger sequencing. The haplotype blocks (rs3806933 and rs2289276) and (rs11466749 and rs11466750) were in high linkage disequilibrium, respectively (D′=0.97, D′=1). CONCLUSION: The increase of major allele frequency of respective nine TSLP variants may enhance the risk of AM. These data will contribute to improved genetic surveillance system in the early diagnosis technology of allergic disease.

Potential Role of Interleukin-25/Interleukin-33/Thymic Stromal Lymphopoietin-Fibrocyte Axis in the Pathogenesis of Allergic Airway Diseases / 中华医学杂志(英文版)

Objective@#Allergic airway diseases (AADs) are a group of heterogeneous disease mediated by T-helper type 2 (Th2) immune response and characterized with airway inflammation and remodeling, including allergic asthma, allergic rhinitis, and chronic rhinosinusitis with allergic background. This review aimed to discuss the abnormal epithelial-mesenchymal crosstalk in the pathogenesis of AADs.@*Data Sources@#Articles referred in this review were collected from the database of PubMed published in English up to January 2018.@*Study Selection@#We had done a literature search using the following terms "allergic airway disease OR asthma OR allergic rhinitis OR chronic sinusitis AND IL-25 OR IL-33 OR thymic stromal lymphopoietin OR fibrocyte". Related original or review articles were included and carefully analyzed.@*Results@#It is now believed that abnormal epithelial-mesenchymal crosstalk underlies the pathogenesis of AADs. However, the key regulatory factors and molecular events involved in this process still remain unclear. Epithelium-derived triple cytokines, including interleukin (IL)-25, IL-33, and thymic stromal lymphopoietin (TSLP), are shown to act on various target cells and promote the Th2 immune response. Circulating fibrocyte is an important mesenchymal cell that can mediate tissue remodeling. We previously found that IL-25-circulating fibrocyte axis was significantly upregulated in patients with asthma, which may greatly contribute to asthmatic airway inflammation and remodeling.@*Conclusions@#In view of the redundancy of cytokines and "united airway" theory, we propose a new concept that IL-25/IL-33/TSLP-fibrocyte axis may play a vital role in the abnormal epithelial-mesenchymal crosstalk in some endotypes of AADs. This novel idea will guide potential new intervention schema for the common treatment of AADs sharing common pathogenesis in the future.

Bamboo salt suppresses skin inflammation in mice with 2, 4-dinitrofluorobenzene-induced atopic dermatitis / 中国天然药物

Bamboo salt (BS) is a traditional Korean food, and has been reported to have anti-cancer, anti-inflammatory, and anti-metastatic effects. However, the anti-atopic dermatitis (AD) activity of BS has not been described yet. In the present study, we examined the preventive effect of BS on AD. The effect of oral administration of BS was tested in a 2, 4-dinitrofluorobenzene (DNFB)-induced AD animal model, by histological analysis, enzyme-linked immunosorbent assay, reverse transcription-polymerase chain reaction, caspase-1 assay, and Western blotting analysis. BS administration reduced the total clinical severity and scratching frequencies, compared with the AD group. In the serum of DNFB-induced AD mice, the levels of IgE, histamine, thymic stromal lymphopoietin (TSLP), interleukin (IL)-5, and IL-13 were significantly reduced by BS treatment. BS significantly reduced the protein and mRNA expression of TSLP, IL-6, and tumor necrosis factor-α in the AD skin lesions. BS markedly reduced the infiltration of inflammatory cells. Furthermore, the activation of caspase-1 was reduced by BS in the AD skin lesions. Our results suggested that BS should be considered as a candidate treatment for allergic inflammatory diseases including AD.

Dynamic analysis of thymic stromal lymphopoietin and its downstream factors in lung tissue of young rats infected with respiratory syncytial virus / 中华实用儿科临床杂志

Objective To study the expressions of thymic stromal lymphopoietin (TSLP),GATA-3 transcription factor and serum interleukin (IL)-4 and IL-13 in lung tissue of young rats infected with respiratory syncytial virus (RSV) at different time points,and to analyze the expression level of TSLP and its downstream related factors.Methods Sixty healthy Wistar rats were randomly divided into normal group and model group.Under the ether anesthesia,RSV Long virus solution was inoculated into the nasal cavity of the model group for 3 days,once a day,and the virus infection model was established.The normal conditions of the 2 groups were observed and recorded on the 3rd,5th and 7th day after the model was established.The expression levels of RSV mRNA,TSLP and its downstream factors were analyzed by real-time polymerase chain reaction and enzyme-linked immunosorbent assay.Results (1)Compared with the normal group,the model group was in poor condition and progressively increased.(2) After the RSV infection,the virus load in the model group reached the peak on the 5th day,which was 4.58 ±0.21.(3)The expression levels of TSLP at the corresponding time points were 0.94 ± 0.26,1.27 ± 0.10 and 1.27 ± 0.01,respectively,which were significantly higher than those in the normal group on the 5th,7th day (t =-11.409,-57.482,all P ＜0.05).(4) The expression of GATA3 at the each time point in the model group were 1.16 ± 0.07,2.61 ± 0.41 and 0.64 ± 0.06,raspectively,which were higher than those in the normal group,and the expression level of GATA3 was statistically significant (t =-26.899,-10.637,-15.187,all P ＜ 0.05).(5) Compared with the normal group,the expressions of IL-4 and IL-13 in the model group were higher than those in the normal group,and the expressions level of IL-4 and IL-13 in the model group showed an increasing trend.Conclusion RSV infected rats can up-regulate the dynamic response of Th2 cells by increasing the expression of TSLP,GATA3 transcription factor and IL-4 and IL-13.

Inflammatory-promoting roles of thymic stromal lymphopoietin and interleukin-4 in allergic conjunctivitis mice / 中华实验眼科杂志

Background Researches showed that thymic stromal lymphopoietin (TSLP) is an interleukin-17-like inflammatory factor and plays important roles in the pathogenesis and development of allergic diseases.However,the study whether TSLP plays roles in allergic conjunctivitis is rare.Objective This study was to investigate the expression change of TSLP and IL-4 in ocular surface tissue and cervical lymph node in the mice with allergic conjunctivitis induced by artemisia annua,a common plant in China,and to explore the role of TSLP and IL-4 in the pathogenesis and development of allergic conjunctivitis.Methods Twenty female BALB/c mice aged 6-8 weeks were randomized into normal control group and model group.Antigen solution was prepared using 400 μl extracting solution of artemisia annua pollen with 400 μl antigen solvent.The mouse models of allergic conjunctivitis were established by injection of 50 μl antigen solution in footpad followed by ocular topical administration of extracting solution once a day from day 10 to 12 after injection,and no any intervention was given in the normal control group.The mice were sacrificed and eyeballs were obtained 13 days after modeling,and corneal epithelium,conjunctiva and cervical lymph nodes were harvested for the detection of TSLP mRNA and IL-4 mRNA by real-time PCR.Immunochemistry was employed to assay the expression of TSLP and IL-4 proteins in corneal,conjunctival and subconjunctival tissues.Results Ocular inflammatory signs appeared 0.5 hours after ocular topical administration of extracting solution,including eyelid edema,conjunctival congestion,tears,scratching eyelids,etc.The symptoms lasted for 6-24 hours,with the model successful rate 80％.Real-time PCR indicated that the expression of IL-4 was absent in corneal epithelium in both model group and normal control group.The relative expression levels of TSLP mRNA in the corneal epithelium,conjunctiva and cervical lymph nodes in the model group were more (1.63±0.20)times,(2.71±0.48) times and (1.48 ±0.05) times than the normal control group,showing significant differences between the two groups (t =4.44,14.16,5.01,all at P＜0.05).Compared with the normal control group,the relative expression levels of IL-4 mRNA in the model group increased (2.94±0.39) times and (1.74±0.09) times,with significant differences between the two groups (t =9.92,14.54,both at P＜0.05).Immunochemistry assay showed that the expression of TSLP protein in the corneal and conjunctival epithelial cells were enhanced in the model group compared with the normal control group.In addition,an intensive expression of IL-4 protein was seen in subconjunctival tissue in the model group,while the expression of IL-4 was absent in the normal control group.Conclusions TSLP is mainly expressed in the cornea,conjunctiva and cervical lymph nodes of the mice with allergic conjunctivitis,suggesting that TSLP promotes the inflammatory process of cornea and conjunctiva;IL-4 is mainly expressed in conjunctiva,showing IL-4 participates in conjunctival inflammatory process.TSLP and IL-4 play synergistic roles in promoting the inflammatory process of ocular surface in the mice with allergic conjunctivitis,which may be new therapeutic targets.

Alternaria Induces Production of Thymic Stromal Lymphopoietin in Nasal Fibroblasts Through Toll-like Receptor 2

PURPOSE: Chronic rhinosinusitis with nasal polyps is a chronic inflammatory disease with markedly increased eosinophils, Th2-type lymphocytes, fibroblasts, and goblet cells. Fungi are commonly associated with airway inflammatory diseases, and thymic stromal lymphopoietin (TSLP) is important in the development of Th2 inflammatory responses. The aim of this study was to investigate the interaction between airborne fungi and nasal fibroblasts in TSLP mRNA and protein expression. METHODS: Inferior turbinate and nasal polyp fibroblasts were stimulated with Alternaria and Aspergillus, respectively, for 48 hours, and TSLP mRNA and protein expressions were measured. The reverse transcriptase polymerase chain reaction was performed for the Toll-like receptor (TLR) mRNA expression of the nasal fibroblasts. To determine the role of TLR in the induction of TSLP, the fibroblasts were transfected with siRNA against TLR2 and TLR5. RESULTS: Alternaria induced TSLP mRNA and protein expression in both inferior turbinate and nasal polyp fibroblasts. The nasal polyp fibroblasts responded more strongly to the fungi. TLR2 and TLR5 mRNA expressions were significantly increased with fungal stimulation and TSLP production was significantly inhibited by siRNA against TLR2. CONCLUSIONS: The results of this study show that TSLP expression could be induced in nasal fibroblasts by exposure to Alternaria and that TLR2 may be involved in the process. The promotion of TSLP production in nasal fibroblasts by airborne fungi may facilitate the development or exacerbation of Th2-type nasal inflammation, especially in CRS with nasal polyps.

On optimum conditons for establishment of calcipotriol-induced mouse atopic dermatitis model / 中国药理学通报

Aim To investigate the method of establishing the atopic dermatitis mice model induced by calcipotriol ( MC903 ) . Methods Induction dose exploratory experiment: since d 0, 0.33,1,3 nmol MC903 was smeared on the right ear of BALB/c mice in each dose group respectively , for 7 consecutive days . The ear swelling degree of the mice was observed every day and the bilateral ears thicknesses were measured .The materials were drawn and analyzed in d 3 and d 7.Induction days exploratory experiment: 2 nmol MC903 was smeared on the right ear of BALB/c mice, for 14 consecutive days .The ear swelling degree of mice was observed every day and the bilateral ears thicknesses were measured .The histopathological examination of the right ear was conducted in 3 d, 7 d, 11 d and 15 d respectively .The tissue homogenate of the mice right ear was prepared .The ex-pressions of thymic stromal lymphopoietin (TSLP),IL-33, IL-4 and IFN-γin the homogenate , CD40 +,CD86 +,the DC surface markers and ILC2 contents in the peripheral lymph nodes were detected.Results ①1,3 nmol MC903 induced ear swelling in mice was significantly increased in d 3, the levels of TSLP and IL-4 were significantly increased .The level of IL-33 in 3 nmol dose group was increased significantly in d 7.② The right ear swelling of 2 nmol MC903 induced atopic dermatitis mice was significant , the ear thickness was increased gradually and reached the peak in d 14.The histopathological examination of the mice ear tissue showed in d 7, the right ear tissue of the mice was swelling and red , the capillary vessels were dilated and the infiltration of inflammatory cells was obvious .The ear in-flammatory symptoms maintained and gradually aggravated for 15 days.Compared with the normal mice , MC903 increased the TSLP level in the right ear tissue homogenate significantly in d 3 and then decreased gradually .The levels of IL-4 and IL-33 were increased significantly in d 7 and then decreased gradually .The levels of ILC2, CD40 +, CD86 + in the peripheral lymph nodes were increased in d 7 and d 15.Conclusion The atopic derma-titis mice model can be successfully established using 2 nmol MC903 induced mice for 7 days.Appropriate testing point of TSLP is d 3.Appropriate testing point of IL-33 and IL-4 is d 7.

Effects of narrow-band ultraviolet B on the expressions of thymic stromal lymphopoietin, interleukin-25 and their receptors in peripheral blood of patients with atopic dermatitis / 中华皮肤科杂志

Objective To explore the effects of narrow-band ultraviolet B (NB-UVB)on the serum levels of thymic stromal lymphopoietin (TSLP)and interleukin-25 (IL-25), as well as on the expressions of TSLP receptor (TSLPR)and IL-25 receptor (IL-25R)mRNAs in peripheral blood mononuclear cells (PBMCs)from patients with atopic dermatitis(AD). Methods A total of 40 patients with AD and 30 healthy volunteers were enrolled in this study. All the patients were treated with NB-UVB at 0.3 - 2.5 J/cm2 thrice a week for 12 consecutive weeks. Venous blood samples were obtained from these patients before and after the treatment as well as from these healthy controls. Double-antibody sandwich enzyme-linked immunosorbent assay (ELISA)was performed to detect serum levels of TSLP and IL-25, and reverse transcription PCR(RT-PCR)to determine the mRNA expression levels of TSLPR and IL-25R in PBMCs from these subjects. The scoring atopic dermatitis (SCORAD)system developed by the European Task Force on Atopic Dermatitis was used to estimate the severity of AD, and visual analogue scale (VAS)to evaluate the degree of itch. Statistical analysis was carried out by the two-independent-sample t-test for intergroup comparisons and paired t-test for comparisons between pre- and post-treatment samples from these patients. Results After the treatment with NB-UVB, the total response rate reached 75%(30/40)in these patients, with a significant decrease in SCORAD score from 55.26 ± 10.88 before the treatment to 20.36 ± 5.12 after the treatment (t = 10.29, P 0.05). Conclusions TSLP and IL-25 may play important roles in the development of AD, and NB-UVB may treat AD by downregulating the expressions of them and their receptors.

Ag85B regulates myeloid dendritic cell maturation and suppresses expres-sion of TSLPR and OX40L mediated by TSLP in vitro / 中国病理生理杂志

AIM:To investigate the maturation of mice immature myeloid dendritic cells (mDCs) induced by antigen(Ag)85B of mycobacterium tuberculosis, and the expression of TSLPR and OX40L mediated by TSLP in vitro. METHODS:Recombinant mouse GM-CSF ( rmGM-CSF) and rmIL-4 were used to induce bone marrow precursor cells of C57BL/6 mice to differentiate into immature mDCs in vitro.mDCs were identified followed by purification using CD 11c binding magnetic beads .The morphological characteristic of mDCs was observed under inverted phase-contrast microscope and scanning electron microscope .The surface phenotypes of mDCs were determined by flow cytometry .To obtain the opti-mal concentrations of Ag85B and TSLP, immature mDCs were cultured with different concentrations of Ag 85B or TSLP at 0 (control group), 50, 100 and 200 μg/L for 24 h, and the expression of cell surface molecules CD 80, CD86, TSLPR and OX40L was detected by flow cytometry.In addition, the expression of TSLPR and OX40L in Ag85B and TSLP-co-stimula-ted mDCs was determined by flow cytometry .RESULTS:After 7 d of culture in vitro, the cells showed irregular dendritic protrusions under the inverted-phase contrast microscope , and had wrinkles and dendritic splits under scanning electron mi-croscope , conformed to the morphological characteristics of immature mDCs .The mDCs cells expressed higher level of spe-cific marker CD11c, lower level of co-stimulatory molecules CD80 and CD86, which conformed to the phenotype of imma-ture mDCs.The CD80 +and CD86 +cell ratios of mDCs displayed significant increases in 50, 100 and 200μg/L Ag85B or TSLP groups compared with control group (P<0.05).The ratios of TSLPR +and OX40L+cells did not differ among dif-ferent concentrations of Ag 85B groups.The ratios of TSLPR +and OX40L+cells were significantly increased in 100 μg/L and 200μg/L TSLP groups compared with control group and 50μg/L TSLP group (P<0.05).Under the circumstance of optimal Ag85B or TSLP treatment concentration at 200 μg/L, there was significantly decreased in TSLPR and OX 40L cell ratio of mDCs in Ag85B group or Ag85B combined with TSLP group when compared with TSLP group (P<0.05), and no significant difference among Ag85B group, Ag85B combined with TSLP group and control group was observed .CONCLU-SION: Ag85B enhances mDCs maturation by up-regulating the expression of co-stimulatory molecules CD80 and CD86, and inhibit the expression of pro-inflammatory specific molecules TSLPR and OX40L on TSLP-activated mDCs, indicating that Ag85B modifies the development of asthmatic airway inflammation through the pathway of TSLP -activated mDCs.