[The effect of Dermatophagoides pteronyssinus group 2 allergen T cell fusion epitope on STAT6 signaling in mice with asthma].

Xiao-dong Zhan, Bin-bin Duan, Ning Tao, Chao-pin Li
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Abstract

Objective: To investigate the alterations of signal transducer and activator of transcription factor 6(STAT6) signaling in a mouse model of asthma receiving treatment with Dermatophagoides pteronyssinus group 2 allergen(Der p 2) T cell fusion epitope and the mechanisms of the specific immunotherapy.

Methods: Thirty mice were randomly divided into three groups by the random number table method: the asthma group, the treatment group receiving immunotherapy with Der p 2 T cell fusion epitope, and the negative control group (PBS group)(n = 10 in each group). Mice in the asthma and the treatment groups received intraperitoneal (i. p.) injection of 100 μl Der p 2 solution [PBS containing 100 μg/ml Der p 2 and 2% Al(OH)3] on days 0,7 and 14, respetively, while mice in the PBS group received same volume of PBS containing 2% Al(OH)3. From day 21, 30-min steam inhalation of 0.5 μg/ml Der p 2 was applied to the asthma and treatment groups (once daily for 7 successive days), and the PBS group inhaled same volume of PBS. From day 25 to day 27, the mice in the treatment group received i. p. injection of 200 μl of Der p 2 T cell fusion epitope (100 μg/ml) while the PBS and the asthma groups received the same volume of PBS. Mice were sacrificed at 24 h after the last inhalation, the bronchoalveolar lavage fluid (BALF) collected, and the total protein was extracted from the lung tissue. The levels of IFN-γ, IL-4 and IL-13 in BALF were determined by ELISA. The expression of STAT6 and phosphorylated STAT6 (p-STAT6) in the lung tissue was detected by Western blotting. Data were analyzed with the one-way variance analysis (ANOVA) method.

Results: The level of IFN-γ in the treatment group[(234.40 ± 24.46) pg/ml] was significantly higher than that in the asthma group[(155.80 ± 20.53) pg/ml](P < 0.01). The levels of IL-4 and IL-13 in the treatment group [(30.00 ± 5.50) pg/ml and (174.50 ± 25.99) pg/ml, respectively] were both significantly lower than those in the asthma group[(53.28 ± 8.26) pg/ml and (308.10 ± 28.32) pg/ml, respectively](P < 0.01). Similarly, the levels of STAT6 and p-STAT6 in the treatment group(0.803 ± 0.221 and 0.966 ± 0.323, respectively) were both significantly lower than those in the asthma group (1.669 ± 0.296 and 1.735 ± 0.298, respectively)(P < 0.01).

Conclusion: The Der p 2 T cell fusion epitope may function through inhibiting STAT6 to treat asthma in mice.

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[鸡翅皮蛾2组变应原T细胞融合表位对哮喘小鼠STAT6信号的影响]。
目的:探讨哮喘小鼠模型接受2组过敏原(Der p2) T细胞融合表位治疗后信号转导和转录因子6(STAT6)信号通路的改变及其特异性免疫治疗的机制。方法:采用随机数字表法将30只小鼠随机分为哮喘组、Der p2t细胞融合表位免疫治疗组和阴性对照组(PBS组),每组10只。哮喘组和治疗组小鼠分别于第0、7、14天腹腔注射100 μl Der p2溶液[含100 μg/ml Der p2和2% Al(OH)3的PBS], PBS组小鼠注射等量含2% Al(OH)3的PBS。自第21天起,哮喘组和治疗组均给予0.5 μg/ml Der p2蒸汽吸入30 min(每日1次,连续7 d), PBS组吸入等量PBS。第25 ~ 27天,治疗组小鼠腹腔注射Der p2 T细胞融合表位200 μl (100 μg/ml), PBS组与哮喘组小鼠腹腔注射等量PBS。末次吸入后24 h处死小鼠,收集支气管肺泡灌洗液(BALF),提取肺组织总蛋白。采用酶联免疫吸附法(ELISA)检测半衰期IFN-γ、IL-4、IL-13水平。Western blotting检测肺组织中STAT6及磷酸化STAT6 (p-STAT6)的表达。数据分析采用单因素方差分析(ANOVA)方法。结果:治疗组血清IFN-γ水平[(234.40±24.46)pg/ml]明显高于哮喘组[(155.80±20.53)pg/ml](P < 0.01)。治疗组IL-4、IL-13水平[分别为(30.00±5.50)pg/ml和(174.50±25.99)pg/ml]均显著低于哮喘组[分别为(53.28±8.26)pg/ml和(308.10±28.32)pg/ml](P < 0.01)。治疗组STAT6、P -STAT6水平(分别为0.803±0.221、0.966±0.323)明显低于哮喘组(分别为1.669±0.296、1.735±0.298)(P < 0.01)。结论:Der p2 T细胞融合表位可能通过抑制STAT6发挥治疗小鼠哮喘的作用。
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