Correlation of serum interleukin-9 and platelet-activating factor levels with total IgE levels, disease severity and disease course in patients with chronic spontaneous urticaria / 中华皮肤科杂志

Objective:To explore the relationship of serum interleukin (IL) -9 and platelet-activating factor (PAF) levels with serum total immunoglobulin E (IgE) levels, disease severity and disease course in patients with chronic spontaneous urticaria (CSU) .Methods:A total of 60 patients with active CSU were collected from Department of Dermatology, the First Affiliated Hospital of Hebei North University from March 2018 to March 2019 (active CSU group), and divided into mild group, moderate group and severe group according to 7-day urticaria activity score (UAS7). After 28-day standard antihistamine therapy, the patients whose condition became stable were included in the stable CSU group. During the same period, 30 health examinees were included in the healthy control group. Peripheral venous blood samples were collected from the subjects in each group, enzyme-linked immunosorbent assay (ELISA) was performed to detect serum levels of IL-9 and PAF, and immunoturbidimetric assay to detect the serum total IgE level. Correlations of serum IL-9 and PAF levels with serum total IgE levels, UAS7 scores and disease courses were analyzed in patients with CSU. One-way analysis of variance was used for comparisons among multiple groups, least significant difference- t test for multiple comparisons, and Pearson correlation analysis for correlation analysis. Results:Totally, 28 males and 32 females were included in the active CSU group, their age ranged from 11 to 68 years (34.68 ± 8.62 years), and the disease duration ranged from 2 months to 7 years (1.42 ± 0.41 years). In the healthy control group, 14 were males and 16 were females, and their age ranged from 10 to 70 years (35.06 ± 7.89 years). According to UAS7, 12, 26, and 22 patients were diagnosed with mild, moderate and severe CSU respectively, and 22 were included in the stable CSU group after standard treatment. The levels of serum IL-9, PAF and total IgE significantly differed among the active CSU group, stable CSU group and healthy control group (IL-9: 144.34 ± 23.19 vs. 109.25 ± 20.77 vs. 107.23 ± 19.23 pg/ml; PAF: 362.45 ± 51.45 vs. 223.18 ± 32.46 vs. 221.23 ± 28.38 pg/ml; total IgE: 168.12 ± 32.48 vs. 24.04 ± 7.04 vs. 21.76 ± 5.95 IU/ml; F = 38.80, 148.38, 499.12, respectively, all P < 0.001), and were significantly higher in the active CSU group than in the stable CSU group and healthy control group (all P < 0.05), while there was no significant difference between the stable CSU group and healthy control group (all P > 0.05). Pearson correlation analysis showed that the serum IL-9 and PAF levels were positively correlated with serum total IgE levels and UAS7 scores (all P < 0.05), but not correlated with the disease course (both P > 0.05) . Conclusion:Serum IL-9 and PAF levels in patients with active CSU were markedly elevated along with the increase in disease severity, and closely correlated with serum total IgE levels.

Progress on the involvement of Th9 cells in the pathogenesis of ulcerative colitis / 中华微生物学和免疫学杂志

Ulcerative colitis (UC) is one of the main types of inflammatory bowel disease (IBD). It is a chronic, nonspecific, and inflammatory disease of the colon that is prone to repeated attacks and requires long-term maintenance treatment. Its clinical features include: diarrhea, weight loss, abdominal pain, fever, blood in the stool and the risk of cancer. It is currently believed that the pathogenesis of UC is due to environmental factors acting on genetically susceptible individuals, causing abnormal activation of the immune system and destruction of the intestinal mucosal barrier, resulting in inflammatory changes of the colonic mucosa, in which T cells and cytokines secreted by them is the important topics in UC pathogenesis research. Th9 cells are a newly discovered subset of T cells. IL-9 secreted by it has been shown to be involved in a variety of autoimmune disease. Now we will review the differentiation of Th9 cells and the mechanisms involved in the pathogenesis of UC.

Changes and clinical significance of serum interleukin-9 and other T helper cell-related cytokines in patients with acute anterior uveitis / 中华实验眼科杂志

Objective:To explore the role of interleukin (IL)-9 and other T helper (Th) cell-related cytokines in the pathogenesis of acute anterior uveitis.Methods:A cross-sectional study was conducted.Thirty-six patients (36 eyes) with acute anterior uveitis who were treated at Gansu Provincial Hospital from May 2018 to May 2019 and 40 matched healthy subjects (40 eyes) who had no eye diseases or systemic diseases in the same period were enrolled as the acute anterior uveitis group and healthy control group, respectively.The disease severity of the subjects in the acute anterior uveitis group was graded and the subjects were divided into mild, moderate and severe groups according to the grading.Serum of all subjects was collected to determine the concentration of serum IL-9, IL-17, transforming growth factor-β 1 (TGF-β 1), interferon-γ (IFN-γ), IL-4, IL-35 and IL-22 by enzyme linked immunosorbent assay (ELISA) method.Spearman rank correlation analysis was used to evaluate the relationship between IL-9 concentration and other Th cell-related cytokines.This study adhered to the Declaration of Helsinki.The study protocol was approved by an Ethics Committee of Gansu Provincial Hospital (No.2019-204). Written informed consent was obtained from each subject prior to any medical examination. Results:The serum levels of IL-9, IFN-γ, IL-4, TGF-β 1, IL-35 and IL-22 in the acute anterior uveitis group were significantly higher than those in the healthy control group, and the differences were statistically significant (all at P<0.05). There was no statistically significant difference in the concentration of IL-17 between the two groups ( U=704.500, P=0.872). The IL-9 concentration of patients with acute anterior uveitis in the mild, moderate and severe groups was 57.24 (47.47, 65.10), 71.68 (67.55, 78.91) and 114.01 (74.78, 139.30) ng/L, respectively, and the overall difference was statistically significant ( Z=8.766, P=0.012), and the IL-9 concentration of the mild group and the moderate group was significantly lower than that of the severe group (both at P<0.05). The concentration of IL-9 in the patients with acute anterior uveitis was positively correlated with the concentration of IL-17, TGF-β 1 and IL-35 ( rs=0.449, 0.517, 0.400; all at P<0.05), and no significant correlations were found between the concentration of IL-9 and the concentration of IFN-γ, IL-4 and IL-22 ( rs=0.293, 0.286, 0.316; all at P>0.05). Conclusions:IL-9 plays a role in promoting the immune inflammatory response in the occurrence and development of acute anterior uveitis, and it is closely related to Th17 and Treg cell-related cytokines (TGF-β 1, IL-35).

Change and clinical significance of T helper 9 cells in previously untreated patients with chronic hepatitis C / 临床肝胆病杂志

ObjectiveTo investigate the changes of T helper 9 (Th9) cells, interleukin-9 (IL-9), and related transcription factors in previously untreated patients with chronic hepatitis C, as well as their association with clinical indices. MethodsA total of 29 previously untreated patients with chronic hepatitis C who attended Hainan Provincial People’s Hospital from December 2018 to July 2019 were enrolled, and 15 healthy individuals were enrolled as healthy controls. The patients with chronic hepatitis C received sofosbuvir/velpatasvir antiviral therapy for 12 weeks, and then plasma and peripheral mononuclear cells (PBMCs) were isolated. Flow cytometry was used to measure the percentage of CD3+CD4+IL-9+ Th9 cells in PBMCs; ELISA was used to measure the plasma level of IL-9; quantitative real-time PCR was used to measure the relative mRNA expression of IL-9 and the transcription factors PU.1 and Foxo1 in PBMCs. The t-test or the paired t-test was used for comparison between two groups, and a Pearson correlation analysis was used to investigate correlation. ResultsCompared with the healthy controls, the previously untreated chronic hepatitis C patients had significantly lower percentage of peripheral Th9 cells (0.92%±0.14% vs 1.14%±0.21%, t=4.31, P＜0.001) and plasma IL-9 level (248.2±66.97 pg/ml vs 309.02±88.48 pg/ml, t=2.63, P=0.012). The previously untreated chronic hepatitis C patients had significantly lower relative mRNA expression of IL-9 and PU.1 than the healthy controls (t=20.67 and 23.21, both P＜0.001), while there was no significant difference in the relative mRNA expression of Foxo1 between the previously untreated chronic hepatitis C patients and the healthy controls (P＞0.05). In the previously untreated chronic hepatitis C patients, the percentage of peripheral Th9 cells, IL-9 level, and mRNA expression of IL-9 and PU.1 were negatively correlated with HCV RNA (r=-0.46, -0.38, -0.52, and -0.41, all P＜0.05), but they were not correlated with the level of alanine aminotransferase (all P＞0.05). Sofosbuvir/velpatasvir antiviral therapy achieved virologic response in 29 chronic hepatitis C patients, and the percentage of peripheral Th9 cells and the mRNA expression of PU.1 after antiviral therapy were significantly higher than those at baseline (t=2.20 and 6.52, both P＜0.05), while there were no significant changes in the plasma level of IL-9 and the relative mRNA expression of IL-9 from baseline to after treatment (both P＞0.05). ConclusionChronic hepatitis C virus infection may suppress the activation of Th9 cells, suggesting that Th9 cells might be involved in the chronicity of hepatitis C virus infection.

Regulatory Effect of Modified Erchentang on Levels of Lymphokines Relating to Th1,Th2 and Th9 Cells and Expressions of IL-4R1/IL-13RA1 in Bronchioles of Rats with Chronic Obstructive Pulmonary Disease / 中国实验方剂学杂志

Objective:To study the effect of modified Erchentang on levels of interleukin-12 (IL-12), interferon-γ (IFN-γ), interleukin-9 (IL-9), interleukin-4 (IL-4) and interleukin-13 (IL-13) in plasma and bronchoalveolar lavage fluid (BALF) of all rats, as well as expressions of interleukin-4 (IL-4) receptor (IL-4R1) and interleukin-13 (IL-13) receptor (IL-13RA1) in bronchioles tissue of rats with chronic obstructive pulmonary disease (COPD). Method:Fifty SD rats were randomly divided into 5 groups, namely normal group, model group, and low, middle and high-dose modified Erchentang groups (5, 10, 20 g·kg-1), with 10 rats in each group. COPD in rat was prepared by using cigarette smoke combined with dripping lipopolysaccharide (LPS) in trachea. After the modeling, normal and model groups were given normal saline solution through intragastric (ig) administration, while other groups were given corresponding herbal drugs (5, 10, 20 g·kg-1) intragastrically (ig) for 14 days. The levels of IL-12, IFN-γ, IL-9, IL-4 and IL-13 in plasma and BALF were detected by Enzyme-linked immunosorbent assay (ELISA) method， and immunohistochemistry (IHC) method was used to detect the expressions of IL-4R1 and IL-13RA1 in bronchioles tissue of all of the groups. Result:Compared with the normal group, the levels of IL-12 and IFN-γ were decreased significantly (P<0.01), but the levels of IL-9, IL-4 and IL-13 in plasma and BALF were significantly increased (P<0.01), and the expressions of IL-4R1 and IL-13RA1 in bronchioles tissue were increased significantly (P<0.01) in model group. Compared with the model group, the levels of IL-12 and IFN-γ were increased significantly, while the levels of IL-9, IL-4 and IL-13 in plasma and BALF were decreased significantly (P<0.01), and the expressions of IL-4R1 and IL-13RA1 in bronchioles tissue were decreased significantly (P<0.01) in modified Erchentang groups (10, 20 g·kg-1). Conclusion:Modified Erchentang has effects in resisting inflammatory and protecting tissue structure of bronchioles. Its mechanism may be correlated with increasing the levels of IL-12, IFN-γ and reducing the levels of IL-9, IL-4 and IL-13 in plasma and BALF, and inhibiting the expressions of IL-4R1 and IL-13RA1 in bronchioles tissue.

Effect of 1, 25-dihydroxy vitamin D3 on food allergy in mice / 中华实用儿科临床杂志

Objective@#To investigate the effects of 1, 25-dihydroxy vitamin D3[1, 25(OH)2D3] on food allergy(FA) in mice and its mechanism.@*Methods@#A total of 40 BALB/c mice were randomly divided into 5 groups, 8 in each group, including control group (group C) and FA model group (FA group), according to the dose of 1, 25(OH)2D3 intervention, the mice of the FA group were divided into FA0 group (0), FAl group [10 μg/(kg·d)], FAm group [50 μg/(kg·d)] and FAh group[100 μg/(kg·d)]. Egg albumin was used to establish a food allergy model, with different doses of 1, 25(OH)2D3 for gastric intervention, and the control group was replaced by 9 g/L saline.The serum levels of ovalbumin-immunoglobulin E(OVA-IgE), interleukin(IL)-9 and IL-17 of mice were measured by using enzyme linked immunosorbent assay after the last excitation, and HE staining and histopathological examination were carried out in the small intestine of mice.@*Results@#Compared with group C, FA0 group and FAh group small intestinal mucosa in mice had different degrees of damage, partial peeling off, structure disorder, villi epithelial cell focal falls peeling off, necrosis, lamina propria edema, congestion, a large number of inflammatory cells infiltration, low but the FAl group and FAm group had light mucosa damage, intestinal epithelial basically intact, with integrity, no congestion, edema, and inflammatory cells infiltration to a lesser degree.The mean concentrations of serum IgE, IL-9 and IL-17 in different groups were statistically significant (F=40.770, 9.530, 5.624, all P<0.05). Compared with the FA0 group [(41.87±3.19) ng/L], the OVA-IgE of the FAl group [(22.71±4.77) ng/L] and the FAm group [(16.34±2.81) ng/L] were significantly reduced (t=5.533, 11.835, all P<0.01), but there was no significant difference between the FAh group [(36.29±6.52) ng/L] (t=1.673, P>0.05). Compared with the FA0 group [(161.77±50.44) ng/L], the IL-9 levels of the FAl group [(94.29±18.79) ng/L] and the FAm group[(84.45±30.88) ng/L] were significantly lower (t=3.267, 3.366, all P<0.01), while that of the FAh group [(36.29±6.52) ng/L] was not significantly lower (t=0.777, P>0.05). Compared with FA0 group [(81.55±29.37) ng/L], IL-17 levels of FAh group [(133.58±47.05) ng/L] was significantly increased (t=2.653, P<0.05), while IL-17 level of FAl group [(79.41±25.15) ng/L] and FAm group [(58.81±26.00) ng/L] were lower than that of FA0 group [(81.55±29.37) ng/L], but the difference was not statistically significant (t=0.154, 1.640, all P>0.05).@*Conclusions@#Low, medium dose of 1, 25(OH)2D3 supplements can inhibit mice food allergies, but high doses of 1, 25(OH)2D3 improve performance in mice food allergies, and 1, 25(OH)2D3′s influence on the secretion of IL-9 is one that influences mechanism of food allergy.

Effect of 1,25-dihydroxy vitamin D3 on food allergy in mice / 中华实用儿科临床杂志

Objective To investigate the effects of 1,25-dihydroxy vitamin D3 [1,25 (OH)2D3] on food allergy(FA) in mice and its mechanism.Methods A total of 40 BALB/c mice were randomly divided into 5 groups,8 in each group,including control group (group C) and FA model group (FA group),according to the dose of 1,25 (OH)2 D3 intervention,the mice of the FA group were divided into FA0 group (0),FA1 group [10 μg/(kg · d)],FAm group [50 μg/(kg · d)] and FAh group[100 μg/(kg · d)].Egg albumin was used to establish a food allergy model,with different doses of 1,25 (OH)2D3 for gastric intervention,and the control group was replaced by 9 g/L saline.The serum levels of ovalbumin-immunoglobulin E(OVA-IgE),interleukin (IL)-9 and IL-17 of mice were measured by using enzyme linked immunosorbent assay after the last excitation,and HE staining and histopathological examination were carried out in the small intestine of mice.Results Compared with group C,FAo group and FAh group small intestinal mucosa in mice had different degrees of damage,partial peeling off,structure disorder,villi epithelial cell focal falls peeling off,necrosis,lamina propria edema,congestion,a large number of inflammatory cells infiltration,low but the FA1 group and FAm group had light mucosa damage,intestinal epithelial basically intact,with integrity,no congestion,edema,and inflammatory cells infiltration to a lesser degree.The mean concentrations of serum IgE,IL-9 and IL-17 in different groups were statistically significant (F =40.770,9.530,5.624,all P ＜ 0.05).Compared with the FA0 group [(41.87 ±3.19) ng/L],the OVA-IgE of the FA1 group [(22.71 ±4.77) ng/L] and the FAm group [(16.34 ±2.81) ng/L] were significantly reduced (t =5.533,1 1.835,all P ＜ 0.01),but there was no significant difference between the FAh group [(36.29 ± 6.52) ng/L] (t =1.673,P ＞ 0.05).Compared with the FA0 group [(161.77 ±50.44) ng/L],the IL-9 levels of the FA1 group [(94.29 ± 18.79) ng/L] and the FAm group [(84.45 ± 30.88) ng/L] were significantly lower (t =3.267,3.366,all P ＜ 0.01),while that of the FAh group [(36.29 ±6.52) ng/L] was not significantly lower (t =0.777,P ＞0.05).Compared with FA0 group [(81.55 ±29.37) ng/L],IL-17 levels of FAh group [(133.58 ± 47.05) ng/L] was significantly increased (t =2.653,P ＜0.05),while IL-17 level of FA1 group [(79.41 ± 25.15) ng/L] and FAm group [(58.81 ± 26.00) ng/L] were lower than that of FAo group [(81.55 ±29.37) ng/L],but the difference was not statistically significant (t =0.154,1.640,all P ＞ 0.05).Conclusions Low,medium dose of 1,25 (OH) 2 D3 supplements can inhibit mice food allergies,but high doses of 1,25 (OH)2 D3 improve performance in mice food allergies,and 1,25 (OH)2 D3's influence on the secretion of IL-9 is one that influences mechanism of food allergy.

Expression and significance of peripheral T helper type 9 cells and their related cytokines in patients with alopecia areata / 中华皮肤科杂志

Objective To investigate the relationship of the expression of peripheral T helper type 9 (Th9) cells and their related cytokines with the severity and clinical course of alopecia areata,and to explore their significance in the occurrence of alopecia areata.Methods From May to December in 2017,74 outpatients with alopecia areata enrolled from Department of Dermatology,Huashan Hospital,Fudan University served as the alopecia areata group,and 57 health checkup examinees in Huashan Hospital served as the control group.Peripheral blood samples were obtained from the patients and controls.Enzyme-linked immunosorbent assay (ELISA) was performed to detect the serum levels of interleukin (IL)-9,IL-4,transforming growth factor (TGF)-β1 and interferon (IFN)-γ,flow cytometry to determine the proportion of Th9 cells (CD4+IL-9+ T helper cells) in peripheral blood mononuclear cells (PBMC),and real-time fluorescence-based quantitative PCR to measure the mRNA expression of IL-9 and PU.1 in the PBMCs.Data were recorded in Microsoft Excel 2017 software,and statistically analyzed with SPSS17.0 software using two-sample t test and Spearman rank correlation analysis.Statistical charts were drawn with Graphpad prism 6 software.Results Compared with the control group,the alopecia areata group showed significantly decreased serum level of IL-9 (190.40 ± 12.33 ng/L vs.288.10 ± 17.38 ng/L,t =4.71,P ＜ 0.01),but significantly increased serum levels of TGF-β1 (6 191.00 ± 355.50 ng/L vs.4 026.00 ± 258.00 ng/L,t =4.41,P ＜ 0.05) and IFN-γ(15.71 ± 3.00 ng/L vs.8.79 ± 0.60 ng/L,t =2.001,P ＜ 0.05).However,there was no significant difference in the serum level of IL-4 between the alopecia areata group and control group (P ＞ 0.05).The serum level of IFN-γ was significantly lower in the patients with severe alopecia areata than in the patients with mild alopecia areata (P =0.02),and the serum level of IFN-γ in the patients with alopecia areata was negatively correlated with the severity of alopecia tool (SALT) score (ru =-0.298,P =0.010).There were no significant differences in the serum levels of IL-9,IL-4 and TGF-β1 between the patients with severe alopecia areata and those with mild alopecia areata (all P ＞ 0.05).The serum levels of IL-9,IL-4,TGF-β1 and IFN-γdid not differ between the patients with active alopecia areata and those with stable alopecia areata,as well as between the patients with clinical course of ＜ 6 months and those with clinical course of ＞ 6 months (P ＞ 0.05).The alopecia areata group showed significantly decreased proportion of Th9 cells in the PBMCs (t =2.04,P =0.045) and mRNA expression of IL-9 and PU.1 (t =2.12,2.178,both P ＜ 0.05) compared with the control group.Condusion The serum level of IL-9 and proportion of peripheral blood Th9 cells both decrease in patients with alopecia areata,and Th9 cells and their related cytokines may be involved in the occurrence of alopecia areata.

Ectopically Expressed Membrane-bound Form of IL-9 Exerts Immune-stimulatory Effect on CT26 Colon Carcinoma Cells

IL-9 is a known T cell growth factor with pleiotropic immunological functions, especially in parasite infection and colitis. However, its role in tumor growth is controversial. In this study, we generated tumor clones expressing the membrane-bound form of IL-9 (MB-IL-9) and investigated their influences on immune system. MB-IL-9 tumor clones showed reduced tumorigenicity but shortened survival accompanied with severe body weight loss in mice. MB-IL-9 expression on tumor cells had no effect on cell proliferation or major histocompatibility complex class I expression in vitro. MB-IL-9 tumor clones were effective in amplifying CD4⁺ and CD8⁺ T cells and increasing cytotoxic activity against CT26 cells in vivo. We also observed a prominent reduction in body weights and survival period of mice injected intraperitoneally with MB-IL-9 clones compared with control groups. Ratios of IL-17 to interferon (IFN)-γ in serum level and tumor mass were higher in mice implanted with MB-IL-9 tumor clones than those observed in mice implanted with control cells. These results indicate that the ectopic expression of the MB-IL-9 on tumor cells exerts an immune-stimulatory effect with toxicity. To exploit its benefits as a tumor vaccine, a strategy to control the toxicity of MB-IL-9 tumor clones should be developed.

Expression of interleukin-9 in colon cancer tissues and its clinical significance / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the expression of interleukin-9 (IL-9) in colon cancer tissues and its clinical significance.</p><p><b>METHODS</b>Immunohistochenmistry and qRT-PCR were used to detect the expressions of IL-9 protein and mRNA in 92 colon cancer tissues and paired adjacent normal tissues. The correlation of IL-9 expressions with the clinicopathological features and prognosis of the patients was analyzed.</p><p><b>RESULTS</b>IL-9 protein and mRNA expressions were significantly higher in adjacent normal tissues than in the colon cancer tissues ( < 0.001). In colon cancer patients, IL-9 expression was significantly correlated with TNM stage (=0.013), Ducks stage (=0.025) and lymph node metastasis (=0.004) but not with gender, age, tumor size, differentiation or hepatic metastasis ( > 0.05). The survival time of colon cancer patients with positive IL-9 expression was significantly longer than that of patients negative for IL-9 expression (=0.015).</p><p><b>CONCLUSIONS</b>IL-9 expression is lowered in colon cancer tissues compoved with in the adjacent normal tissues. IL-9 expression is negatively correlated with TNM staging, Ducks staging and lymph node metastasis but positively with good prognosis, suggesting its important role in the tumor microenvironment of colon cancer.</p>

Effects of overexpression tumor necrosis factor-related ligand-1A on T helper 9 cells in chronic experimental colitis / 中华消化杂志

Objective To investigate the effects of tumor necrosis factor-related ligand-1A(TL1A)on activation of T helper 9(Th9)cells of colonic tissues in chronic experimental colitis mice.Methods The chronic experimental colitis mice model was established with drinking dextran sulfate sodium salt(DSS).A total of 32 lymphocytes TL1A highly expressed mice and wild type(WT)mice were divided into WT control group, transgene control group,WT modeling group and transgene modeling group.The mice of control groups were administrated with distilled water. The mice of modeling groups received 3% DSS in drinking water discontinuously.The mice were sacrificed on 29 days after modeling.Body mass was measured,length of colon was recorded,scores of gross colon and the disease activity index(DAI)were calculated.The colonic morphological changes were observed by hematoxylin-eosin(H-E)staining.The lamina propria mononuclear cells(LPMC)were isolated and the number of Th9 cells was tested by flow cytometry.The levels of interleukin-9(IL-9)in serum and LPMC were detected by enzyme-linked immunosorbent assay(ELISA).The expressions of IL-9 protein and mRNA of the colonic tissues were measured by Western blotting and real-time polymerase chain reaction(PCR),respectively.T test and single factor analysis of variance were performed for statistical analysis.Results The percentage of body mass loss of WT modeling group was lower than that of transgene modeling group(16.2% ± 1.0% vs 18.9% ± 1.2%),and the difference was statistically significant(t=4.90, P<0.05).The scores of gross colon,DAI and pathology of transgene modeling group were all higher than those of WT modeling group(2.80 ± 0.64 vs 1.60 ± 0.31,2.55 ± 0.20 vs 1.58 ± 0.17,and 11.85 ± 0.86 vs 9.50 ± 0.79),and the differences were statistically significant(t=4.77,10.45 and 5.69,all P<0.05).The number of LPMC in transgene modeling group was higher than that of WT modeling group(3.70×106± 0.28×106vs 2.65×106± 0.32 × 106)and the difference was statistically significant(t= 6.98,P< 0.05).The percentage of Th9 in total CD4+T cells of LPMC in colonic tissues of transgene modeling group was higher than that of WT modeling group(0.54% ± 0.04% vs 0.23% ± 0.03%),and the difference was statistically significant(t= 17.54,P< 0.05).The serum IL-9 level of transgene modeling group was higher than that of WT modeling group((170.23 ± 5.69)pg/mL vs(150.62 ± 6.45)pg/mL),and the difference was statistically significant(t= 6.50,P< 0.05).The level of IL-9 secreted by LMPC of transgene modeling group was higher than that of WT modeling group((265.21 ± 8.76)pg/mL vs (237.58 ± 10.24)pg/mL),and the difference was statistically significant(t= 5.80,P< 0.05).The expressions of IL-9 protein and mRNA of transgene modeling group were higher than those of WT modeling group(1.31 ± 0.09 vs 1.18 ± 0.03,and 8.26 ± 1.13 vs 2.25 ± 0.29,respectively),and the differences were statistically significant(t=3.88 and 14.57,both P< 0.05).Conclusion TL1A high expression in lymphocytes can promote Th9 cells differentiation and IL-9 secretion which involved in the genesis of chronic experimental colitis.

Helper T cell Th9 and its cytokine interleukin-9 in peripheral blood of patients with Parkinson's disease / 中华神经科杂志

Objective To investigate the proportion of helper T cell subset Th 9 and the expression of its cytokine interleukin-9(IL-9)in Parkinson's disease(PD)and the clinical significance.Methods Seventy-two patients diagnosed with PD between January 2016 and June 2017 and 20 healthy volunteers in the same period were selected.The PD patients were staged according to the Hoehn-Yahr(H-Y)staging method,21 in stage one,19 in stage two,18 in stage three,11 in stage four,and three in stage five.The proportion of Th9 subset in peripheral blood of PD patients and healthy volunteers was measured by flow cytometry.The expression of IL-9 in peripheral blood of PD patients and healthy volunteers was measured by enzyme-linked immunosorbent assay.Results The proportion of Th9 cells in peripheral blood of PD patients(1.27%±0.34%)was significantly higher than that of healthy volunteers(0.61%±0.11%,t=8.530,P<0.05),and the higher stage of PD,the higher proportion of Th9,suggesting that the proportion of Th9 was related to the PD staging.IL-9 was also highly expressed in PD patients((16.04 ±2.94) pg/ml)and had statistically significant difference compared with healthy volunteers((7.53 ±0.70)pg/ml;t=12.781,P<0.05).IL-9 was similar to Th9, the higher stage of PD, the higher expression of IL-9. Conclusion The proportion of Th9 cells and the expression of IL-9 in peripheral blood of patients with PD increased significantly,having a significant relationship with the H-Y staging of PD.

Anti-Interleukin-9 Antibody Increases the Effect of Allergen-Specific Immunotherapy in Murine Allergic Rhinitis

PURPOSE: Interleukin (IL)-9 induces allergic responses; however, the roles of anti-IL-9 antibody in the induction of tolerance remain unclear. This study investigated the effects of anti-IL-9 antibody on oral tolerance (OT) in a mouse model of allergic rhinitis (AR). METHODS: BALB/c mice were divided into 4 groups: the control, AR, OT, and OT with anti-IL-9 antibody (OT+IL9AB) groups. Ovalbumin (OVA) was used for sensitization and challenge. Mice in the OT and OT+IL9AB groups were fed OVA for immunotherapy. During immunotherapy, OT+IL9AB mice were injected with anti-IL-9 antibody. Allergic symptoms, tissue eosinophil counts, and serum OVA-specific immunoglobulin E (IgE) were measured. The mRNA expressions of cytokines and transcription factors of T cells of nasal mucosa were determined by real-time polymerase chain reaction (PCR). The protein levels of GATA3, ROR-γt, and Foxp3 in nasal mucosa were determined by Western blot. CD4⁺CD25⁺Foxp3⁺ T cells in the spleen were analyzed by flow cytometry. RESULTS: Administration of anti-IL-9 antibody decreased allergic symptoms, OVA-specific IgE levels, and eosinophil counts. In addition, it inhibited T-helper (Th) 2 responses, but had no effect on Th1 responses. Protein levels of ROR-γt and mRNA levels of PU.1 and ROR-γt were reduced by anti-IL-9 antibody. Anti-IL-9 antibody increased Foxp3 and IL-10 mRNA expression, Foxp3 protein, and induction of CD4⁺CD25⁺Foxp3⁺ T cells. CONCLUSIONS: Anti-IL-9 antibody decreased allergic inflammation through suppression of Th2 and Th17 cells. Anti-IL-9 antibody enhanced the tolerogenic effects of regulatory T cells. These results suggest that anti-IL-9 antibody might represent a potential therapeutic agent for allergen immunotherapy in patients with uncontrolled allergic airway disease.

Effects of gene silence on immune function of colonic dendritic cells in rats with irritable bowel syndrome visceral hypersensitivity / 中华消化杂志

Objective To explore the role and mechanism of protein disulfide isomerase A3 (PDIA3) in abnormal colonic mucosa immune response of irritable bowel syndrome (IBS) rats with visceral hypersensitivity.Methods A total of 48 SD rats were divided into blank control group (n=12),empty virus group (IBS-1) (n=12),PDIA3 silencing group (IBS-2) (n=12) and model control group(IBS-3) (n=12).The expression of CD103 in ileocecus colonic tissues was detected by immunohistochemistry.Dendritic cells (D C) of mesenteric lymph node were isolated by flow cytometry.CD4+/CD8+ T cells of spleen were separated by immune-magnetic beads sorting technique.DC and CD4+/CD8+ T cells were co-cultured.The proliferation ability of lymphocytes promoted by DC was measured by methyl thiazolyl tetrazolium (MTT).The secretion levels of interlcukin (IL) 4 and IL-9 of CD4+/CD8+ T cells stimulated by DC were determined by enzyme linked immunosorbent assay (ELISA) method.Independent sample t-test was performed for statistical analysis.Results The cell number of CD103 positive DC of rats colon of blank control group,IBS-3 group was 6.25±1.14 and 10.83± 1.03(t=10.07,P＜0.05);that of IBS-2 group was 7.42 ± 0.90,and compared with that of IBS-3 group,the difference was statistically significant (t=-9.25,P ＜ 0.05).The MTT value of CD4+ T cells proliferation stimulated by DC of blank control group and IBm3 group was 0.54±0.01 and 0.60±0.01 (t=3.373,P＜0.05);that of IBS-2 group was 0.53±0.01,and compared with that of IBS-3 group,the difference was statistically significant (t =-3.139,P ＜ 0.05).The MTT value of CD8+ T cells proliferation stimulated by DC was 0.52±0.01 and 0.59±0.00(t=3.539,P＜0.01);that of IBS-2 group was 0.54±0.01,and compared with that of IBS-3 group,the difference was statistically significant (t=-3.183,P＜0.05).The level of IL-4 secreted by CD4+T cells promoted by DC of blank control group and IBS-3 group was 10.24±0.09 and 16.61±1.00 (t=3.222,P＜0.05);that of IBS-2 group was 11.75±0.54,and compared with that of IBS-3 group,the difference was statistically significant (t=-3.539,P＜0.01).The level of IL-9 secreted by CD4+T cells stimulated by DC of blank control group and IBS-3 group was 15.86±10.19 and 43.51±11.32 (t=4.529,P＜0.05);that of IBS-2 group was 29.05±2.09,and compared with that of IBS-3 group,the difference was statistically significant (t=-6.841,P＜0.01).The level of IL-4 secreted by CD8+T cells promoted by DC of blank control group and IBS-3 group was 7.35±0.12 and 13.91±0.57 (t=19.557,P＜0.01);that of IBS-2 group was 8.63± 0.24,and compared with that of IBS-3 group,the difference was statistically significant (t =-14.782,P＜0.01).The level of IL-9 secreted by CD8+T cells stimulated by DC of blank control group and IBS-3 group was 29.12±5.14 and 60.70±11.02 (t=4.122,P＜0.05);that of IBS-2 group was 37.17±2.65,and compared with that of IBS-3 group,the difference was statistically significant (t=-3.255,P＜ 0.05).Conclusions Protein disulfide isomerase A3 may mediate the process of DC activating T lymphocyte levels of related cytokines,cause abnormal immune response of colonic mucosa and promote the genesis of IBS visceral hypersensitivity.

Expression of IL-9 and its receptor mRNA in nasal mucosa of allergic rhinitis mice / 中国耳鼻咽喉头颈外科

OBJECTIVE To investigate the expression and roles of IL-9 and its receptor (IL-9R) mRNA in the nasal mucosa of ovalbumin-induced allergic rhinitis (AR) mice. METHODS Sixteen Balb/c mice were selected and randomly divided into two groups with 8 mice in each group. Mice were used for establishing the animal model of AR with ovalbumin sensitization as AR group, at the same time, the physiological saline as the control group. In each group, 4 mice were randomly taken from each group, and the pathological examination showed that the model was successful. The nasal mucosa was taken from the remaining 4 mice in each group and then to detect IL-9 and IL-9R mRNA in nasal mucosa of the two groups by using real-time quantitative PCR. RESULTS The expression of IL-9 and IL-9R mRNA could be detected in both the control and AR groups. The expression levels of IL-9 and IL-9R mRNA in the nasal mucosa of the AR group were both higher than that in the control group (P<0.05). The expression level of IL-9 mRNA was positively correlated with the expression of IL-9R mRNA in the nasal mucosa of mice (r =0.857, P<0.05). CONCLUSION IL-9 and its receptor IL-9R are involved in the development of AR, and play an important role in the development of allergic rhinitis. It provides a new perspective for the further understanding of the pathogenesis of AR and provides a new clue for the treatment of allergic rhinitis.

Expressions of interleukin-9 and the transcription factor PU.1 in peripheral blood of patients with chronic urticaria and their significance / 中华皮肤科杂志

Objective To investigate the relationship of interleukin-9 (IL-9) and the transcription factor PU.1 with chronic urticaria.Methods Thirty patients with chronic urticaria (patient group) and 30 healthy volunteers (control group) were included in this study.Venous blood samples were collected from these subjects.Enzyme-linked immunosorbent assay (ELISA) was performed to detect serum levels of IgE,flow cytometry (FCM) to determine the percentage of CD4+IL-9+ T cells,real-time fluorescence-based quantitative PCR (RTFQ-PCR) to measure the mRNA expressions of IL-9 and PU.1 in peripheral blood mononuclear cells (PBMCs).Measurement data were compared between the two groups by independent-samples t test,and relationship among these parameters was assessed by Pearson correlation analysis.Results The expressions of IL-9 and PU.1 mRNAs in patients with chronic urticaria were significantly higher than those in healthy controls (4.44 ± 1.90 vs.3.20 ± 1.78,t =2.60,P＜ 0.05; 3.26 ± 1.59 vs.2.34 ± 1.47,t =2.34,P ＜ 0.05).Compared with the control group,the patient group showed increased percentage of CD4+IL-9+ T cells in peripheral blood (0.63％ ± 0.44％ vs.0.22％ ± 0.12％,t =5.04,P ＜ 0.01) and serum IgE levels ((82.04 ± 31.72) vs.(60.74 ± 28.26) IU/ml,t =2.75,P ＜ 0.01).The percentage of CD4+IL-9+ T cells and levels of IL-9 and PU.1 mRNAs in peripheral blood were all positively correlated with serum levels of IgE in these patients (r =0.596,0.767,0.746,respectively,all P ＜ 0.01).Conclusion T helper type 9 (Th9) cells and IL-9 may take part in the occurrence of chronic urticaria.

Influence of Azithromycin on peripheral blood helper T lymphocyte 9 cells and helper T lymphocyte 17 cells in ;children with bronchial asthma / 中华实用儿科临床杂志

Objective To observe the influence of Azithromycin on helper T lymphocyte 9 cells ( Th9 ) and Th17 in peripheral blood of children with bronchial asthma,and to investigate the immunomodulating effect of Azithro-mycin. Methods Twenty-six asthmatic children were selected as the experimental group,and 17 healthy children as a control group. Peripheral blood mononuclear cells(PBMC)were isolated from venous blood by density gradient cen-tri-fugation under aseptic conditions. PBMC were split by phytohemagglutinin( PHA) in vitro. Different concentrations of Azithromycin (0,0. 1,1. 0,10. 0 mg/L)were added into the cultures in the experimental group. The control group was not interfered with azithromycin. The supematant was collected after 72 h. The levels of interleukin ( IL)-4, IL-9,IL-17 and IL-23 in the supematant were determined by enzyme-linked immunosorbent assay(ELISA). SPSS 17. 0 software was used to analyze data. Results (1) The levels of IL-4,IL-9,IL-17 and IL-23 produced from PBMC of the experimental group were significantly higher than those in the control group(t=9. 210,3. 040,8. 965,2. 796,P0. 05). The levels of IL-17 and IL-23 of Azithromycin 10 mg/L were lower than those in Azithromycin 0 mg/L and 0. 1 mg/L(P0. 05). (3)IL-9 level was negatively correlated with IL-4 level in the experimental group(r=-0. 255,P>0. 05). The levels of IL-23 and IL-17 secreted by Th17 in asthmatic chil-dren had correlation. The secretion of IL-17 levels rose with the secretion of IL-23 rise(r=0. 64,P<0. 05). Conclu-sions The function of Th9 and Th17 in asthmatic children was enhanced;Azithromycin could restrain the function of Th9 and Th17 at higher tissue concentrations (10. 0 mg/L),the former was unrelated to changes in the secretion of IL-9 levels,and the latter with the secretion of IL-23 levels decreased close. Azithromycin can play a role of immune modulators in higher concentrations,inflammatory cytokines are inhibited in the asthmatic children,and Azithromycin relieves airway inflammation.

Presence and the significance of Th9 cells in patients with hashimoto thyroiditis / 中华微生物学和免疫学杂志

Objective To investigate the presence and significance of Th9 cells and the related transcription factor ( PU.1 ) and cytokine ( IL-9 ) in peripheral blood of patents with hashimoto thyroiditis ( HT) .Methods Thirty patients with HT and thirty age/gender matched healthy subjects were recruited in this study.The peripheral blood and serum samples were collected from each subject.The percentages of Th9 cells and the transcriptional levels of PU.1 in peripheral blood mononuclear cells ( PBMCs) were meas-ured by flow cytometry analysis and real-time RT-PCR.The concentrations of IL-9, the functions of thyroid and the titers of thyroid-specific autoantibodies ( TPOAb and TgAb) in serum samples were detected by using enzyme-linked immunosorbent assay ( ELISA ) and electrochemiluminescence immunoassay analysis ( ECLIA) .Results Compared with healthy subjects, the percentages of Th9 cells and the expression of PU.1 at mRNA level in PBMCs and the concentrations of IL-9 in serum samples were all significantly in-creased in patients with HT [(1.49±0.68)%vs (0.87±0.24)%], 4.91±2.14 vs 1.66±0.52, (26.90± 7.74) pg/ml vs (16.71±5.87) pg/ml, all P<0.01).Serum concentrations of IL-9 were positively correla-ted with the percentages of Th9 cells (r=0.419, P=0.021).Moreover, the percentages of Th9 cells were positively correlated with the titers of TPOAb and TgAb in serum samples (r=0.394, P=0.032;r=0.457, P=0.011) of patients with HT.Conclusion The levels of Th9 cells and the related cytokine IL-9 were in-creased in the peripheral blood of patients with HT.A positive correlation was found between the percentage of Th9 cells and the titers of thyroid-specific autoantibodies.This study indicated that Th9 cells might be in-volved in the pathogenesis of autoimmune damage in thyroid.

Th9 cells in peripheral blood of adult patients with primary immune thrombocytopenia and its clinical significance / 第二军医大学学报

Objective To observe Th9 cells and IL-9 level in adult patients with primary immune thrombocytopenia (ITP), and to discuss their potential roles in the pathogenesis of ITP. Methods A total of 25 newly diagnosed ITP patients and 25 sex- and age-matched healthy controls were enrolled in the present study. The percentage of Th9 cells in the peripheral blood samples of the two groups were detected by flow cytometry, expressions of IL-9, TGF-ß, PU. 1 and IRF4 mRNA were analyzed by real time-RCR, and IL-9 protein level was examined by ELISA. The platelet count was recorded by sysmex XE-2100. Results Compared with the healthy controls, the ratio of Th9 cells was significantly increased in ITP patients(P30×109/L(P30 × 109/L(P<0.05). Compared with healthy controls, the mRNA expressions of IL-9, TGF-ß, PU.1 and IRF4 were raised significantly in ITP patients(P<0.05). The ratio of Th9 cells and IL-9 protein level were negatively correlated with PLT in ITP patients(r= -0.428 1, P = 0.032 8; r= -0.537 5, P = 0.005 6, respectively). Furthermore, follow-up study of 11 ITP patients found that both Th9 cells and IL-9 protein levels took a declining tendency after effective treatment(P<0.05). Conclusion Abnormal activation of Th9/IL-9 may participate in the occurrence and development of ITP disease, which provides new clues for further understanding of ITP pathogenesis and selecting potential therapeutic targets in immune therapy of ITP.

Expression and Clinical Significance of Interleukin-9 in Patients with Primary Biliary Cirrhosis / 胃肠病学

Background:Primary biliary cirrhosis( PBC ) is an autoimmune liver disease,the cause of the disease remain incompletely understood. In addition to genetic and environmental factors,autoantibodies,multiple immunocytes and cytokines are considered to be involved in the development of PBC. Recent studies indicated that interleukin-9(IL-9)had pleiotropic functions in inflammatory regulation in allergic and autoimmune diseases. Aims:To investigate the expression and clinical significance of IL-9 in patients with PBC. Methods:A total of 30 specimens of peripheral blood and 20 specimens of liver tissue from PBC patients were collected. Ten specimens of peripheral blood from healthy subjects and 4 specimens of normal liver tissue were served as controls. Level of serum IL-9 was determined by ELISA,and expression of IL-9 in liver tissue was detected by immunohistochemistry. Correlations between IL-9 and serum biochemical indicators, immune indicators and histologic stage of liver tissue were analyzed. Results:Level of serum IL-9 was significantly increased in PBC patients than in normal control group(P<0. 05),and was positively correlated with level of serum IgG (r=0. 681,P<0. 01). Amount of IL-9 positive cells in liver tissue was significantly increased in PBC patients than in normal control group(P <0. 01),and was positively correlated with histologic stage of liver tissue(rs =0. 465,P <0. 05). Conclusions:Expression of IL-9 is significantly increased in peripheral blood and liver tissue in patients with PBC and is positively correlated with level of serum IgG and histologic stage of liver tissue,which suggests an important role of IL-9 in the pathogenesis of PBC.