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IL-17A抑制Th2细胞分化减轻哮喘小鼠气道嗜酸性粒细胞炎症
IL-17A inhibits Th2 cell differentiation and alleviates eosinophilic airway inflammation in a mouse model of asthma-
- 第三军医大学学报 2017年39卷第10期 页码：954-959
- 作者机构：
400037 重庆，第三军医大学新桥医院全军呼吸内科研究所，全军呼吸病研究重点实验室
- 作者简介：
[王长征，E-mail:czwang@netease.com Wang Changzheng, E-mail: czwang@netease.com
- 基金信息：国家自然科学基金面上项目(81270075)
DOI：10.16016/j.1000-5404.201612143
中图分类号： R392.11;R392.3;R562.25收稿：2016-12-20，
修回：2017-01-30，
纸质出版：2017-05-30
- 稿件说明：
移动端阅览
王熠杰, 赵生涛, 杨旭, 等. IL-17A抑制Th2细胞分化减轻哮喘小鼠气道嗜酸性粒细胞炎症[J]. 第三军医大学学报, 2017,39(10):954-959.
Yijie Wang, Shengtao Zhao, Xu Yang, et al. IL-17A inhibits Th2 cell differentiation and alleviates eosinophilic airway inflammation in a mouse model of asthma-[J]. Journal of Third Military Medical University, 2017, 39(10): 954-959.
王熠杰, 赵生涛, 杨旭, 等. IL-17A抑制Th2细胞分化减轻哮喘小鼠气道嗜酸性粒细胞炎症[J]. 第三军医大学学报, 2017,39(10):954-959. DOI： 10.16016/j.1000-5404.201612143.
Yijie Wang, Shengtao Zhao, Xu Yang, et al. IL-17A inhibits Th2 cell differentiation and alleviates eosinophilic airway inflammation in a mouse model of asthma-[J]. Journal of Third Military Medical University, 2017, 39(10): 954-959. DOI： 10.16016/j.1000-5404.201612143.
摘要
目的
2
研究IL-17A对哮喘小鼠Th2细胞分化及其相关炎症的作用。
方法
2
24只C57BL/6J小鼠按随机数字表法分为对照组、哮喘组和IL-17A处理组（
n
＝ 8）。哮喘组和IL-17A处理组予以卵清蛋白（ovalbumin，OVA）致敏及激发。每次雾化激发前1 h，IL-17A处理组给予重组小鼠IL-17A气道滴入。各步对照均予以生理盐水。末次激发后24 h处死小鼠，收集支气管肺泡灌洗液（bronchoalveolar lavage fluid，BALF）行细胞总数及分类计数。ELISA检测BALF中IL-4、IL-5、IFN-γ、IL-17A的浓度。HE和PAS染色及半定量评分评估小鼠肺部病理变化。流式细胞术检测脾脏和支气管淋巴结Th细胞分化。免疫磁珠分选健康小鼠幼稚CD4
+
T细胞，用Th2极化培养基体外培养，并给予IL-17A或等量PBS干预，检测Th2细胞的增殖
、凋亡和分化。
结果
2
哮喘组较对照组，BALF中细胞总数、嗜酸性粒细胞数及其比例（
P
＜0.05）、IL-4、IL-5、IL-17A浓度均显著增高（
P
＜0.05），IFN-γ浓度显著下降（
P
＜0.05）；支气管、血管周围炎症细胞浸润和杯状细胞化生明显加重（
P
＜0.01）；脾脏和淋巴结Th2细胞分化比例显著增高（
P
＜0.05）。IL-17A处理组较哮喘组，BALF中的细胞总数[（26.00 ± 5.43）× 10
4
/mL
vs
（58.40 ± 26.93）× 10
4
/mL，
P
＜0.05
]
、嗜酸性粒细胞数[（8.04 ± 1.98）× 10
4
/mL
vs
（31.95 ± 12.28）× 10
4
/mL，
P
＜0.05
]
及其比例[（29.93 ± 3.03）%
vs
（53.47 ± 6.62）%，
P
＜0.01
]
显著降低，而中性粒细胞数及其比例无明显变化；BALF中Th2相关因子IL-4浓度[（9.86 ±2.77）pg/mL
vs
（28.13 ± 4.62）pg/mL，
P
＜0.01
]
、IL-5浓度[（7. 30 ± 0. 50）pg/mL
vs
（10.50 ± 1.10）pg/mL，
P
＜0.01
]
均显著降低；支气管、血管周围炎症细胞浸润减轻，HE染色半定量评分降低[（2.00 ± 0.51）
vs
（3.12 ± 0.64），
P
＜0.05
]
，杯状细胞化生减少[（0.80 ± 0.45）
vs
（2.40 ± 0.55），
P
＜0.01
]
；脾脏[（2.24 ± 0.44）%
vs
（4.82 ± 1.83）%，
P
＜0.01
]
和淋巴结[（7.05 ± 0.58）%
vs
（10.57 ± 1.35）%，
P
＜0.05
]
中Th2细胞分化比例显著减少。极化培养的幼稚CD4
+
T细胞，予IL-17A干预后，诱导分化的Th2细胞比例显著减少（
P
＜0.05），而增殖和凋亡无显著变化。
结论
2
IL-17A有抑制Th2细胞分化，减轻哮喘小鼠气道嗜酸性粒细胞炎症的作用。
Abstract
Objective
2
To investigate the effect of IL-17A on the differentiation of Th2 cells and airway inflammation in a mouse model of asthma.
Methods
2
Twenty-four C57BL/6J mice were randomly divided into control group
asthmatic group and IL-17A treatment group. The mice in asthmatic group and IL-17A treatment group were sensitized and challenged with ovalbumin (OVA)
and in the latter group
the mice received intratracheal instillation of IL-17A 1 h before each OVA challenge; normal saline was used for all the control treatments. The mice were sacrificed 2
4 h after the last challenge
and the bronchoalveolar lavage fluid (BALF) were collected for cytological examination and detection of IL-4
IL-5
IL-17A
and interferon-γ(IFN-γ) using ELISA. The lung pathologies were assessed semi-quantitatively using HE and PAS staining. The differentiation of Th cells in the spleen and tracheobronchial lymph nodes were analyzed with flow cytometry. The effects of IL-17A on cell proliferation
apoptosis
and differentiation were examined in naive CD4
+
T cells sorted by immunomagnetic beads from normal mice.
Results
2
Compared with the control mice
the asthmatic mice showed significantly increased total cell and eosinophil counts and eosinophil percentage (
P
<
0.05)
elevated IL-4
IL-5
and IL-17A concentrations (
P
<
0.05)
and lowered IFN-γ concentration in the BALF (
P
<
0.05). The asthmatic mice presented with more obvious inflammatory cell infiltration (
P
<
0.01) and goblet cell hyperplasia (
P
<
0.05) and significantly increased proportion of Th2 cells in the spleen and lymph nodes (
P
<
0.05). IL-17A treatment of the asthmatic mice significantly reduced the total cell number [(26.00 ± 5.43) × 10
4
/mL
vs
(58.40 ± 26.93) × 10
4
/mL
P
<
0.05
]
eosinophil count [(8.04 ± 1.98) × 10
4
/mL
vs
(31.95 ± 12.28) × 10
4
/mL
P
<
0.05
]
and eosinophil percentage [(29. 93 ± 3. 03)%
vs
(53.47 ± 6.62)%
P
<
0.01
]
in the BALF without affecting neutrophil count or percentage. IL-17A treatment also resulted in lowered levels of IL-4 (9.86 ± 2.77 pg/mL
vs
28.13 ± 4.62 pg/mL
P
<
0.01) and IL-5 (7.30 ± 0.50 pg/mL
vs
10.50 ± 1.10 pg/mL
P
<
0.01) in the BALF
alleviated inflammatory cell infiltration (2. 00 ± 0.51
vs
3.12 ± 0.64
P
<
0.05) and goblet cell hyperplasia (0.80 ± 0.45
vs
2. 40 ± 0.55
P
<
0.01)
and reduced the percentages of Th2 cells in the spleen [(2.24 ± 0.44)%
vs
(4.82 ± 1.83)%
P
<
0.01
]
and lymph nodes [(7.05 ± 0.58)%
vs
(10.57 ± 1.35)%
P
<
0.05
]
. In naive CD4
+
T cells from normal mice
IL-17A treatment obviously inhibited cell differentiation into Th2 cells in vitro [(4.15 ± 0.55)%
vs
(2.53 ± 0.47)%
P
<
0.05
]
without affecting the cell proliferation or apoptosis.
Conclusion
2
IL-17A can inhibit the differentiation of Th2 cells and alleviate eosinophilic airway inflammation in mice.
关键词
Keywords
references
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