[1]范文婷,钟世民,胡琦,等.色氨酸代谢物调控Th17/Treg分化在小鼠哮喘变应原特异性免疫治疗中的作用及机制研究[J].第三军医大学学报,2018,40(08):658-665.
 FAN Wenting,ZHONG Shimin,HU Qi,et al.Tryptophan metabolites regulate Th17/Treg differentiation to alleviate airway inflammation in asthmatic mice receiving allergen-specific immunotherapy[J].J Third Mil Med Univ,2018,40(08):658-665.
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色氨酸代谢物调控Th17/Treg分化在小鼠哮喘变应原特异性免疫治疗中的作用及机制研究(/HTML )
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《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
40卷
期数:
2018年第08期
页码:
658-665
栏目:
基础医学
出版日期:
2018-04-30

文章信息/Info

Title:
Tryptophan metabolites regulate Th17/Treg differentiation to alleviate airway inflammation in asthmatic mice receiving allergen-specific immunotherapy
作者:
范文婷钟世民胡琦曾靖廖伟汪金玉
陆军军医大学(第三军医大学)第一附属医院儿科
Author(s):
FAN Wenting ZHONG Shimin HU Qi ZENG Jing LIAO Wei WANG Jinyu  

Department of Pediatrics, First Affiliated Hospital,  Army Medical University (Third Military Medical University), Chongqing, 400038, China

关键词:
色氨酸代谢物小鼠模型支气管哮喘变应原特异性免疫治疗Th17/Treg
Keywords:
tryptophan metabolites mouse models bronchial asthma allergenspecific immunotherapy Th17/Treg
分类号:
R392-33;R392.32;R562.25
文献标志码:
A
摘要:

目的    对色氨酸代谢物调控Th17/Treg分化在小鼠哮喘变应原特异性免疫治疗中的作用及机制进行研究。方法     将30只BALB/c 鼠随机法分为5组:对照组、哮喘组、OVASIT组(OVA:鸡卵白蛋白;SIT:变应原特异性免疫治疗)、OVASIT+1-MT组(1-MT:IDO抑制剂,1-甲基色氨酸),OVASIT+1MT+KYN组(KYN:色氨酸代谢产物,犬尿氨酸)。哮喘组:第0、7 d予OVA致敏,第6周每天予1% OVA雾化激发,50 d予10% OVA 加强激发;OVASIT组:第4周每天予大剂量OVA皮下注射,余同哮喘组;OVASIT+1MT组:第4周每天在腹腔内注入1-MT,1 h后予大剂量OVA皮下注射h,余同哮喘组;OVASIT+1-MT+KYN组:第3周每天加入1-MT,第4周每天加入 KYN,末次加入KYN后1 h予大剂量OVA行免疫治疗,余同哮喘组。末次激发6 h内检测气道高反应性;对支气管肺泡灌洗液(BALF )进行细胞计数分析;ELISA检测血清IgE及BALF中IL-5、IL-10、IL-17;流式细胞技术检测脾脏CD4+RORγt+T及CD4+Foxp3+T 细胞分化情况。结果     OVA-SIT+1-MT+KYN组较OVA-SIT+1-MT组酸性粒细胞浸润减少,炎症反应明显减轻。前者BALF中Il5为74.8~86.8(83.48±6.02)pg/mL,IL-17为33.8~46.5(38.72±4.61)pg/mL,CD4+RORγt+T 细胞为2.45~2.82(2.60±0.14)%,CD4+Foxp3+T 细胞为7.83~9.09(8.36±0.53)%;后者BALF中Il-5为240.3~285.1(259.65±16.27)pg/mL, IL-17为55.2~65.8(59.97±3.76)pg/mL,CD4+RORγt+T 细胞为4.31~5.34(4.94±0.38)%,CD4+Foxp3+T 细胞为5.93~6.59(6.33±0.28)%,因此OVASIT+1-MT+KYN组中Il-5、IL-17细胞因子以及脾脏中CD4+RORγt+T 细胞均明显低于OVA-SIT+1-MT组,差异均有统计学意义(P<0.01),而前组脾脏中CD4+Foxp3+T 细胞水平明显高于后者,差异有统计学意义(P<0.01)。结论    色氨酸代谢产物有助于特异性免疫治疗减轻气道炎症作用,其机制与通过调控Th17及Treg分化有关。

Abstract:

Objective    To study the role of tryptophan metabolites in regulating Th17/Treg differentiation to alleviate airway inflammation in asthmatic mice receiving allergen-specific immunotherapy (SIT). Methods    Thirty BALB/c mice were randomized equally into control group, asthma group, chicken ovalbumin (OVA)SIT group, OVA-SIT+1-methyltryptophan (1-MT, an indoleamine 2,3 dioxygenase inhibitor) group, and OVASIT+1-MT+kynurenine (KYN, a tryptophan metabolite) group. Except for those in the control group, all the mice were sensitized with OVA on days 0 and 7 followed by daily challenge with 1% OVA in the 6th week and then with 10% OVA on day 50 to induce asthma; In OVA-SIT group, the mice were treated in the 4th week with daily subcutaneous injection of high-dose OVA, and in OVA-SIT+1-MT group, the mice received daily intraperitoneal injection of 1-MT 1 h before subcutaneous high-dose OVA injection; in OVA-SIT+1-MT+KYN group, the mice were treated with daily 1-MT injection in the third week and daily KYN injection in the 4th week, and high-dose OVA immunotherapy was administered at 1 h after the last KYN injection. Airway hyperresponsiveness of the mice was measured within 6 h after the last challenge. Serum levels of IgE and levels of IL-10, IL-5 and IL-17 in bronchoalveolar lavage fluid (BALF) were detected with ELISA, and flow cytometry was performed to analyze the differentiation of CD4+RORγt+and CD4+Foxp3+T cells in the spleen. Results     Compared with those in OVA-SIT+1-MT group, the infiltration of acidic granulocytes and neutrophils decreased and the inflammatory response was significantly reduced in OVA-SIT+1-MT+KYN group. In OVASIT+1-MT group, the mean IL-5 level in BALF was (83.48±6.22) pg/mL, IL-17 level was (38.72±4.61) pg/mL, CD4+Foxp3+T cell percentage was (2.60±0.14)%, and CD4+Foxp3+T cell percentage was (8.36±0.53)%, as compared with the levels of (259.65±16.27) pg/mL, (59.97±3.76) pg/mL, (4.94±0.38)%, and (6.33±0.28)% in OVASIT+1-MT+KYN group, respectively; IL5 and IL-17 levels and spleen CD4+RORγt+T cell percentage was significantly lower (P<0.01) while CD4+Foxp3+T cell percentage was significantly higher  (P<0.01) in OVA-SIT+1-MT+KYN group than in OVA-SIT+1-MT group. Conclusion     Tryptophan metabolites contribute to the efficacy of specific immunotherapy in reducing airway inflammation in asthmatic mice, the mechanism of which may involve the regulation of Th17/Treg cell differentiation.

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更新日期/Last Update: 2018-04-27