1.The immune response of human keratinocytes to Trichophyton rubrum conidia is partially mediated by toll-like receptor-2, 4, dectin-1 and cytokines.
Ying LI ; Jian CHEN ; Miao-Jian WAN ; Wei LAI ; Yue ZHENG ; Mei-Rong LI ; Rong-Zhang CHEN ; Xiao-Xin LI
Journal of Southern Medical University 2011;31(4):678-681
OBJECTIVETo investigate the effects of Trichophyton rubrum exposure on the expressions of toll-like receptor-2 (TLR-2), TLR-4 and dendritic cell associated C-type lectin-1 (Dectin-1) and cytokine secretions in human keratinocytes cell line HaCaT.
METHODSThe mRNA of TLR-2,4, and dectin-1 in the HaCaT co-cultured with the conidia of Trichophyton rubrum conidia for 24 h was measured with real-time PCR. The mean fluorescence intensity and the percentage of cells positive for TLR-2, 4, and dectin-1 was detected during the co-culture using flow cytometry. The cytokine secretion profiles in the cell culture supernatant was analyzed using a cytokine antibody array.
RESULTSThe TLR-2,4, and dectin-1 mRNA expressions, mean fluorescence intensity and percentage of positive cells for TLR-2,4, and dectin-1 all increased in HaCaT cells in response to Trichophyton rubrum conidia exposure. The results of cytokine antibody array demonstrated obviously increased secretions of IL-8, I-309, IFN-γ, IL-6, and IL-13 in the culture supernatant of HaCaT cells in response to Trichophyton rubrum exposure.
CONCLUSIONThe immune responses and immunological recognition of human keratinocytes to Trichophyton rubrum conidia are partially mediated by up-regulating the expressions of TLR-2, TLR-4 and dectin-1 and secretions of multiple cytokines.
Cell Line ; Chemokine CCL1 ; secretion ; Coculture Techniques ; Humans ; Interferon-gamma ; secretion ; Interleukin-13 ; secretion ; Interleukin-6 ; secretion ; Interleukin-8 ; secretion ; Keratinocytes ; metabolism ; Lectins, C-Type ; metabolism ; Toll-Like Receptor 2 ; metabolism ; Toll-Like Receptor 4 ; metabolism ; Trichophyton ; Tumor Necrosis Factor-alpha ; secretion
2.Molecular mechanism of interleukin-13-induced mucus hypersecretion in rat airway.
De-peng JIANG ; Victor P KOLOSOV ; Juliy M PERELMAN ; Xiang-dong ZHOU
Journal of Southern Medical University 2011;31(1):73-76
OBJECTIVETo investigate the effect of interleukin-13 (IL-13) on mucus secretion in vivo and the possible mechanism.
METHODSThe SD rats were randomly divided into control group, IL-13 group and IL-13 plus SP600125 group. The phosphorylation of extracellular signal-regulated kinase 1/2 (ERK1/2), c-Jun N-terminal kinase 1/2 (JNK1/2) and the level of MUC5AC in the lung tissues were examined using Western blotting. RT-PCR was performed to examine the mRNA level of STAT4 and STAT6, and electrophoretic mobility shift assays (EMSA) was used to detect the DNA-binding activities of Forkhead box a2 (FOXA2) and activator protein-1 (AP-1).
RESULTSIL-13 caused a significant increase in MUC5AC and p-JNK1/2 expression, but did not affect the phosphorylation of ERK1/2. The expression of MUC5AC was attenuated after treatment with SP600125. A significant increase in STAT6 was observed in IL-13 group compared with that in the control group, whereas the expression of STAT4 mRNA was not significantly affected. The DNA-binding activity of FOXA2 was down-regulated after IL-13 exposure, which did not affect the DNA-binding activity of AP-1.
CONCLUSIONIL-13 down-regulates mucus secretion via STAT6-FOXA2 pathway in vitro.
Animals ; Bronchi ; secretion ; Female ; Hepatocyte Nuclear Factor 3-beta ; genetics ; metabolism ; Interleukin-13 ; pharmacology ; Male ; Mucin 5AC ; metabolism ; Mucus ; secretion ; RNA, Messenger ; genetics ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; STAT6 Transcription Factor ; genetics ; metabolism ; Signal Transduction ; drug effects
3.Effect of polyI: C on secretion of thymic stromal lymphopoietin and airway inflammation in mice with respiratory syncytial virus-induced asthma exacerbation.
Hu XIA ; Hua-peng YU ; Li-min LUO ; Shao-xi CAI
Journal of Southern Medical University 2011;31(3):434-437
OBJECTIVETo investigate the effects of polyinosinic-polycytidylic acid (polyI:C) on the production of thymic stromal lymphopoietin (TSLP) and airway inflammation in mice with exacerbated asthma induced by respiratory syncytial virus (RSV).
METHODSThirty-two female BALB/c mice were randomly divided into 4 groups, namely the PBS control group, OVA group, OVA/RSV group, and OVA/RSV/polyI:C group. In the latter 3 groups, the mice were sensitized by OVA and stimulated with nebulized OVA. RSV was inoculated into the nasal cavity of the sensitized mice and polyI:C (1 mg/kg) was intramuscularly administered. The airway response to metacholine was examined, and the serum levels of IL-4, IL-5, IL-13, and IFN-γ and TSLP in the supernatants of bronchoalveolar lavage fluid (BALF) were detected using ELISA. The total BALF cells, eosinophils, lymphocytes and neutrophils were counted. The lung specimens were collected to observe the inflammation with HE staining, and immunohistochemistry was employed to determine TSLP production in the airway epithelial cells.
RESULTSThe mice in RSV/OVA/polyI:C group showed a significantly lower airway responsiveness to metacholine than those in OVA/RSV group (P<0.01). Compared with OVA/RSV group, RSV/OVA/polyI:C group showed significantly lower serum levels of IL-4, IL-5, IL-13 and TSLP in BALF (P<0.05), with also lower total BALF cells, eosinophils and lymphocytes (P<0.05) and lessened infiltration of the airway inflammatory cells. Immunohistochemistry of TSLP also demonstrated a lower production of TSLP in the airway epithelial cells in RSV/OVA/polyI:C group than in OVA/RSV group.
CONCLUSIONSpolyI:C can inhibit the increase in TSLP production in the airway epithelial cells after RSV infection and relieve airway inflammation in mice with RSV-induced asthma exacerbation.
Animals ; Asthma ; blood ; metabolism ; virology ; Bronchoalveolar Lavage Fluid ; Cytokines ; secretion ; Female ; Inflammation ; pathology ; Interleukin-13 ; blood ; Interleukin-4 ; blood ; Interleukin-5 ; blood ; Mice ; Mice, Inbred BALB C ; Poly I-C ; pharmacology ; Respiratory Syncytial Virus Infections ; blood ; metabolism ; Respiratory Syncytial Viruses
4.Protease-Activated Receptor 2 Is Involved in Th2 Responses against Trichinella spiralis Infection.
Mi Kyung PARK ; Min Kyoung CHO ; Shin Ae KANG ; Hye Kyung PARK ; Yun Seong KIM ; Ki Uk KIM ; Soon Cheol AHN ; Dong Hee KIM ; Hak Sun YU
The Korean Journal of Parasitology 2011;49(3):235-243
In order to get a better understanding of the role of protease-activated receptor 2 (PAR2) in type 2 helper T (Th2) cell responses against Trichinella spiralis infection, we analyzed Th2 responses in T. spiralis-infected PAR2 knockout (KO) mice. The levels of the Th2 cell-secreted cytokines, IL-4, IL-5, and IL-13 were markedly reduced in the PAR2 KO mice as compared to the wild type mice following infection with T. spiralis. The serum levels of parasite-specific IgE increased significantly in the wild type mice as the result of T. spiralis infection, but this level was not significantly increased in PAR2 KO mice. The expression level of thymic stromal lymphopoietin, IL-25, and eotaxin gene (the genes were recently known as Th2 response initiators) of mouse intestinal epithelial cells were increased as the result of treatment with T. spiralis excretory-secretory proteins. However, the expression of these chemokine genes was inhibited by protease inhibitor treatments. In conclusion, PAR2 might involve in Th2 responses against T. spiralis infection.
Animals
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Antibodies, Helminth/blood
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Chemokine CCL11/biosynthesis
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Cytokines/biosynthesis
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Female
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Gene Expression Profiling
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Immunoglobulin E/blood
;
Interleukin-13/secretion
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Interleukin-4/secretion
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Interleukin-5/secretion
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Interleukins/biosynthesis
;
Mice
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Mice, Inbred C57BL
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Mice, Knockout
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Receptor, PAR-2/*metabolism
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Th2 Cells/*immunology
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Trichinella spiralis/*immunology
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Trichinellosis/*immunology
5.Effect of dexamethasone on expression of AGR2 protein in asthmatic mice.
Min ZHOU ; Hui-long CHEN ; Sheng CHENG ; Li MEI ; Hui-lan ZHANG ; Min XIE ; Wei-ning XIONG ; Yong-jian XU
Journal of Huazhong University of Science and Technology (Medical Sciences) 2013;33(1):33-36
This study examined the expression of the anterior gradient-2 (AGR2) protein and Muc5ac protein in the lung tissues of asthmatic mice and the effect of dexamethasone, with an attempt to explore the role of AGR2 in the over-secretion of mucus in the airway. Eighteen BALB/c mice were divided into asthma group, control group and dexamethasone group. In dexamethasone group, dexamethasone was intraperitoneally administered. Expression of AGR2 protein and Muc5ac protein in the murine lung tissues was immunohistochemically detected. IL-13 level was determined in the bronchoalveolar lavage fluid (BALF) by ELISA. The results exhibited that the expression of AGR2 protein in asthma group (0.522±0.041) was significantly higher than that in normal controls (0.361±0.047) (P<0.01) and bore a positive linear relationship to the expression of Muc5ac protein (r=0.873, P<0.05) and IL-13 level (r=0.828, P<0.05). Expression of AGR2 protein in the dexamethasone group (0.456±0.049) was significantly lower than that in the asthma group. It was concluded that: (1) the expression of AGR2 protein was significantly higher in asthmatic mice as compared with their normal counterparts; (2) the expression was obviously related to the expression of Muc5ac protein and IL-13; (3) dexamethasone could down-regulate the expression of AGR2 protein. Our findings suggested that AGR2 might be involved in the over-secretion of mucus in the airway in asthma.
Animals
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Anti-Inflammatory Agents
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pharmacology
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Asthma
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drug therapy
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metabolism
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Dexamethasone
;
pharmacology
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Female
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Interleukin-13
;
metabolism
;
Lung
;
drug effects
;
metabolism
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Mice
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Mice, Inbred BALB C
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Mucin 5AC
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metabolism
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Mucoproteins
;
metabolism
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Mucus
;
secretion
;
Treatment Outcome
6.Effects of glucocorticoid on IL-13-induced Muc5ac expression in airways of mice.
Jianbo LIU ; Zhenxiang ZHANG ; Yongjian XU ; Lihua XING ; Huilan ZHANG
Journal of Huazhong University of Science and Technology (Medical Sciences) 2004;24(6):575-577
To study the effects of glucocorticoid on the IL-13-induced Muc5ac expression in airways of mice, and investigate its role in mucus secretion of airways, 24 pathogen-free BALB/c mice were randomly divided into 3 groups. IL-13 group received an nasal instillation of 100 microg of recombinant murine IL-13 solution on days 1, 3 and 5. In dexamethasone group, dexamethasone (0.5 mg/kg) was administered intraperitoneally 24 h before and 1 h before the first instillation of IL-13 and on 4 consecutive days (day 0 to day 5, 6 consecutive days in total), while control group was not treated with IL-13 or dexamethasone. Bronchoalveolar lavage fluid (BALF) was collected and eosinophils were counted, and expression of Muc5ac mRNA and protein in lungs were detected by reverse transcription-polymerase chain reaction (RT-PCR) technology and immunohistochemical assay respectively. Our results showed that the number of mice, with positve Muc5ac protein expression, expression of Muc5ac mRNA and eosinophils in BALF after IL-13 treatment were all significantly higher than that of control group (all P<0.01). Despite eosinophils reduced (P<0.01), the number of mice with positive Muc5ac protein expression, expression of Muc5ac mRNA afterdexamethasone treatment didn't decreas significantly as compared with that of IL-13 group. It is concluded that IL-13 can up-regulate the expression of Muc5ac mRNA and protein, which may play a pivotal role in the mucus overproduction of airways. Dexamethasone can suppress IL-13-induced eosinophilic infiltration in lung but can't inhibit the mucus overproduction.
Animals
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Bronchoalveolar Lavage Fluid
;
cytology
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Dexamethasone
;
pharmacology
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Interleukin-13
;
pharmacology
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Male
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Mice
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Mice, Inbred BALB C
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Mucin 5AC
;
Mucins
;
biosynthesis
;
genetics
;
Mucus
;
secretion
;
RNA, Messenger
;
biosynthesis
;
genetics
;
Random Allocation
;
Respiratory System
;
metabolism
7.Effects of glucocorticoid on IL-13-induced Muc5ac expression in airways of mice.
Jianbo, LIU ; Zhenxiang, ZHANG ; Yongjian, XU ; Lihua, XING ; Huilan, ZHANG
Journal of Huazhong University of Science and Technology (Medical Sciences) 2004;24(6):575-7
To study the effects of glucocorticoid on the IL-13-induced Muc5ac expression in airways of mice, and investigate its role in mucus secretion of airways, 24 pathogen-free BALB/c mice were randomly divided into 3 groups. IL-13 group received an nasal instillation of 100 microg of recombinant murine IL-13 solution on days 1, 3 and 5. In dexamethasone group, dexamethasone (0.5 mg/kg) was administered intraperitoneally 24 h before and 1 h before the first instillation of IL-13 and on 4 consecutive days (day 0 to day 5, 6 consecutive days in total), while control group was not treated with IL-13 or dexamethasone. Bronchoalveolar lavage fluid (BALF) was collected and eosinophils were counted, and expression of Muc5ac mRNA and protein in lungs were detected by reverse transcription-polymerase chain reaction (RT-PCR) technology and immunohistochemical assay respectively. Our results showed that the number of mice, with positve Muc5ac protein expression, expression of Muc5ac mRNA and eosinophils in BALF after IL-13 treatment were all significantly higher than that of control group (all P<0.01). Despite eosinophils reduced (P<0.01), the number of mice with positive Muc5ac protein expression, expression of Muc5ac mRNA afterdexamethasone treatment didn't decreas significantly as compared with that of IL-13 group. It is concluded that IL-13 can up-regulate the expression of Muc5ac mRNA and protein, which may play a pivotal role in the mucus overproduction of airways. Dexamethasone can suppress IL-13-induced eosinophilic infiltration in lung but can't inhibit the mucus overproduction.
Bronchoalveolar Lavage Fluid/cytology
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Dexamethasone/*pharmacology
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Interleukin-13/*pharmacology
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Mice, Inbred BALB C
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Mucins/*biosynthesis
;
Mucins/genetics
;
Mucus/secretion
;
RNA, Messenger/biosynthesis
;
RNA, Messenger/genetics
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Random Allocation
;
Respiratory System/*metabolism