切换至 "中华医学电子期刊资源库"
高级检索
中华肺部疾病杂志(电子版)

中华肺部疾病杂志(电子版) ›› 2019, Vol. 12 ›› Issue (05) : 601 -605. doi: 10.3877/cma.j.issn.1674-6902.2019.05.013

论著

活化的Treg细胞对NKT细胞迁移的调节作用
卢燕鸣1,†(), 李亚琴1, 周文静1, 李小燕1, 于清1   
  1. 1. 201112　上海，上海交通大学医学院附属仁济医院南院儿科
  • 收稿日期:2019-03-10 出版日期:2019-10-20
  • 通信作者: 卢燕鸣
  • 基金资助:
    上海市卫生和计划生育委员会重点课题(201640018)

Regulatory effect of activated Treg cells on migration of NKT cells in mice

Yanming Lu1,†(), Yaqin Li1, Wenjing Zhou1, Xiaoyan Li1, Qin Yu1   

  1. 1. Department of Pediatrics, South Campus, Renji Hospital, School of Medicine, Shanghai Jiaotong University, Shanghai 201112, China
  • Received:2019-03-10 Published:2019-10-20
  • Corresponding author: Yanming Lu
全文 ( ) 下载PDF ( ) 导出引用
引用本文：

卢燕鸣, 李亚琴, 周文静, 李小燕, 于清. 活化的Treg细胞对NKT细胞迁移的调节作用[J]. 中华肺部疾病杂志(电子版), 2019, 12(05): 601-605.

Yanming Lu, Yaqin Li, Wenjing Zhou, Xiaoyan Li, Qin Yu. Regulatory effect of activated Treg cells on migration of NKT cells in mice[J]. Chinese Journal of Lung Diseases(Electronic Edition), 2019, 12(05): 601-605.

目的

探讨Treg细胞活化后对NKT细胞迁移的调控作用。

方法

分离纯化小鼠脾脏Treg细胞及NKT细胞后，通过病毒感染的方式制备Foxp3修饰的Treg细胞；验证Treg细胞的活化情况。体外共培养活化的Treg细胞和NKT细胞，流式细胞仪检测NKT的数量；RT-PCR及ELISA分析NKT细胞胞内因子的分泌水平(IL-4、IFN-γ)。制备卵清白蛋白(OVA)致敏哮喘小鼠模型，体内观察Foxp3修饰的Treg细胞对NKT细胞体内迁移的影响。

结果

经Foxp3修饰后的Treg细胞(Le-Foxp3组)中Foxp3的表达明显增加；CFSE标记法检测显示Le-Foxp3组细胞可显著抑制自体CD4+CD25- T效应T细胞的增殖；同时ELISA检测结果显示相较于空载体转染组(Le-scr组)，Le-Foxp3组细胞上清液中IL-10及TGF-β的水平显著升高。经transwell培养板体外共培养Treg细胞和NKT细胞后，流式细胞仪检测下室中NKT细胞的数目结果显示Le-Foxp3组为(221.15±79.36)，Le-scr组为(649.38±153.97)；同时IL-4及IFN-γ的水平显著下降。在体内实验结果显示，相较于OVA组，OVA+Le-Foxp3组小鼠肺组织中NKT的数量显著降低，而外周血及脾脏中NKT细胞的数量显著升高。

结论

Foxp3修饰可活化体外培养的Treg细胞，此外活化的Treg细胞可抑制NKT细胞的活化及体外迁移并影响OVA哮喘模型小鼠NKT细胞的体内分布。

关键词: Treg细胞, Foxp3, NKT细胞, 迁移, 支气管哮喘
Objective

To investigate the regulatory effect of activated Treg cells on the migration of NKT cells in mice.

Methods

Treg cells and NKT cells were isolated and purified, then Treg cells were modified by the lentivirus carrying Foxp3 and the activation of Treg cells were verified. The activated Treg cells and NKT cells were co-cultured in vitro and flow cytometry was used to detect the number of NKT cells. The secretion of intracellular factors (IL-4 and IFN-γ) in the NKT cells was measured with reverse transcription polymerase chain reaction (RT-PCR) and enzyme-labeled immunosorbent assay (ELISA). Ovalbumin (OVA)-induced asthma mouse models were established and used to observe the effect of Treg cells modified by Foxp3 on the migration of NKT cells in vivo.

Results

The expression of Foxp3 was significantly up-regulated in the Treg cells modified by Foxp3 (Le-Foxp3 group). CFSE assay showed that the Treg cells carrying Foxp3 significantly inhibited the proliferation of autologous CD4+ CD25- T cells (Teff cells), and ELISA showed that the levels of IL-10 and TGF-β were increased markedly in the Le-Foxp3 group compared with the control group (the cells were infected with the lentivirus carrying scrambled sequence, Le-scr group). After co-culture of the Treg cells and NKT cells in a transwell culture plate, the flow cytometer showed that the NKT cells in the lower chamber were (221.15±79.36) in the Le-Foxp3 group and (649.38±153.97) in the Le-scr group. Meanwhile, the levels of IL-4 and IFN-γdecreased remarkably. The results from OVA-induced asthma model showed that the forced expression of Foxp3 remarkably increased the NKT cells in the spleen and the peripheral blood, while decreased the NKT cells in the lungs.

Conclusion

Infection of lentivirus carrying Foxp3 can activate Treg cells. The activated Treg cells can inhibit the activation and migration of NKT cells in vitro and affect the distribution of NKT cells in OVA-induced asthma mouse model.

Key words: Treg cells, Foxp3, NKT cells, Migration, Bronchial asthma
图1 Le-Foxp3转染效率检测；注：A：RT-PCR检测Treg细胞中Foxp3的表达；B：Western blot检测Treg细胞中Foxp3的蛋白表达
图2 表达Foxp3对Treg细胞功能的影响；注：A：CFSE标记法检测Treg细胞对Teff细胞的增殖抑制作用；B：ELISA检测细胞因子IL-10和TGF- β的水平
图3 RT-PCR(A)和ELISA(B)检测表达Foxp3的Treg细胞对NKT细胞胞内因子IL-4和IFN-γ分泌的影响
图4 OVA哮喘模型中Treg细胞对NKT细胞体内分布的影响
1
Khan DA. Allergic rhinitis and asthma: epidemiology and common pathophysiology[J]. Allergy Asthma Proc, 2014, 35(5): 357-361.
2
Divekar R, Lal D. Recent advances in biologic therapy of asthma and the role in therapy of chronic rhinosinusitis[J]. F1000Res, 2018, 7: 412.
3
Gandhi GR, Neta MTSL, Sathiyabama RG, et al. Flavonoids as Th1/Th2 cytokines immunomodulators: A systematic review of studies on animal models[J]. Phytomedicine, 2018, 44: 74-84.
4
Kanagaratham C, Radzioch D. Allergic asthma: A summary from genetic basis, mouse studies, to diagnosis and treatment[J]. Curr Pharm Des, 2016, 22(41): 6261-6272.
5
Wang J, Jin RG, Xiao L, et al. Anti-asthma effects of synthetic salidroside through regulation of Th1/Th2 balance[J]. Chin J Nat Med, 2014, 12(7): 500-504.
6
Liang P, Peng S, Zhang M, et al. Huai Qi Huang corrects the balance of Th1/Th2 and Treg/Th17 in an ovalbumin-induced asthma mouse model[J]. Biosci Rep, 2017, 37(6). pii: BSR20171071.
7
Huang X, Tang L, Wang F, et al. Astragaloside Ⅳ attenuates allergic inflammation by regulation Th1/Th2 cytokine and enhancement CD4(+)CD25(+)Foxp3 T cells in ovalbumin-induced asthma[J]. Immunobiology, 2014, 219(7): 565-571.
8
卢燕鸣，曹兰芳，薛海燕，等. 支气管哮喘患儿外周血自然杀伤T淋巴细胞的变化及临床意义[J]. 中华实用儿科临床杂志，2012, 28(10): 775-777.
9
卢燕鸣，曹兰芳，李　琛，等. 特异性免疫治疗对屋尘螨致敏哮喘小鼠NKT细胞的影响[J]. 现代生物医学进展，2012, 12(7): 1201-1208.
10
Nguyen KD, Vanichsarn C, Nadeau KC. Increased cytotoxicity of CD4+ invariant NKT cells against CD4+CD25hiCD127lo/- regulatory T cells inallergic asthma[J]. Eur J Immunol, 2008, 38(7): 2034-2045.
11
Thorburn AN, Foster PS, Gibson PG, et al. Components of Streptococcus pneumoniae suppress allergic airways disease and NKT cells by inducing regulatory T cells[J]. J Immunol, 2012, 188(9): 4611-4120.
12
Lu Y, Guo Y, Xu L, et al. Foxp3 regulates ratio of Treg and NKT cells in a mouse model of asthma[J]. Mol Cell Biochem, 2015, 403(1-2): 25-31.
13
Zhang M, Qian YY, Chai SJ, et al. Enhanced local Foxp3 expression in lung tissue attenuates airway inflammation in a mouse model of asthma[J]. J Asthma, 2014, 51(5): 451-458.
14
Marques CR, Costa RS, Costa GNO, et al. Genetic and epigenetic studies of FOXP3 in asthma and allergy[J]. Asthma Res Pract, 2015, 1: 10.
15
Lu Y, Li Y, Xu L, et al. Bacterial lysate increases the percentage of natural killer T cells in peripheral blood and alleviates asthma in children[J]. Pharmacology, 2015, 95(3-4): 139-144.
16
Mortuaire G, Gengler I, Vandenhende-Szymanski C, et al. Immune profile modulation of blood and mucosal eosinophils in nasal polyposis with concomitant asthma[J]. Ann Allergy Asthma Immunol, 2015, 114(4): 299-307.e2.
17
何　青，聂汉祥，杨巧玉，等. NKT细胞与调节性T细胞在免疫调节中相互作用的研究进展[J]. 免疫学杂志，2016, 32(6): 535-539.
18
惠　郁，王晓明，周韵娇. 支气管哮喘儿童急性发作期诱导痰液T细胞亚群及自然杀伤T细胞的变化[J/CD]. 中华哮喘杂志(电子版), 2010, 4(1): 30-33.
[1] 陈金业, 凌潜龙, 朱冰, 骆杰. 补体B因子在结直肠癌中的表达及临床意义[J]. 中华普通外科学文献(电子版), 2024, 18(03): 192-198.
[2] 张生军, 赵阿静, 李守博, 郝祥宏, 刘敏丽. 高糖通过HGF/c-met通路促进结直肠癌侵袭和迁移的实验研究[J]. 中华普外科手术学杂志(电子版), 2024, 18(01): 21-24.
[3] 白若靖, 郭军. 维生素D对肺部疾病临床意义的研究进展[J]. 中华肺部疾病杂志(电子版), 2024, 17(04): 659-662.
[4] 赵蒙蒙, 黄洁, 余荣环, 王葆青. 过表达小GTP酶Rab32抑制非小细胞肺癌细胞侵袭性生长[J]. 中华肺部疾病杂志(电子版), 2024, 17(04): 512-518.
[5] 陈华萍, 陈晓龙, 胡明冬. 难治性哮喘的发病机制及诊治进展[J]. 中华肺部疾病杂志(电子版), 2024, 17(01): 144-147.
[6] 朱斯悦, 张晓莹, 严玉茹, 陈绯. 介入支气管镜在肺部疾病诊断和治疗中的应用[J]. 中华肺部疾病杂志(电子版), 2024, 17(01): 148-151.
[7] 薛春娥, 赵娟娟, 刘韦成, 张义亮, 马毓晗, 李迎巧. CD4+CD25+foxp3+调节性T细胞通过NKG2D增强急性髓性白血病中NK细胞毒性[J]. 中华细胞与干细胞杂志(电子版), 2024, 14(04): 212-219.
[8] 曾聿理, 雷发容, 肖慧, 邱德亮, 谢静, 吴寻. 氯普鲁卡因通过调控circRNA-ZKSCAN1表达抑制肝癌Huh-7细胞体外生长和转移的研究[J]. 中华细胞与干细胞杂志(电子版), 2024, 14(04): 220-228.
[9] 李晶, 潘侠, 周芳, 汪晶, 洪佳. 普鲁卡因通过上调lncRNA DGCR5抑制胃癌细胞增殖、迁移和侵袭[J]. 中华细胞与干细胞杂志(电子版), 2024, 14(03): 151-158.
[10] 刘杜先, 张杰东, 付鲁渝, 熊志强, 龚程, 张小雅, 高明悦, 孟俊宏, 刘兰侠. 沉默circXPO1抑制肝癌细胞恶性生物学行为[J]. 中华细胞与干细胞杂志(电子版), 2024, 14(03): 159-166.
[11] 李博, 马秀岩, 孙杰. lncRNA TINCR对滋养层HTR-8/SVneo细胞生物学行为的影响及其机制[J]. 中华细胞与干细胞杂志(电子版), 2024, 14(03): 167-172.
[12] 夏辉, 戴斌, 冉君, 王威, 龚昭, 周程. DEP结构域蛋白1B在肝细胞癌中的表达及功能[J]. 中华肝脏外科手术学电子杂志, 2024, 13(02): 205-213.
[13] 崔精, 鲍一帆, 沈晓明, 杨增辉, 高森, 鲍传庆. 结直肠癌中circMFSD12对肿瘤细胞功能及5-FU敏感性的调控[J]. 中华结直肠疾病电子杂志, 2024, 13(04): 294-302.
[14] 邵文哲, 孙倩男, 王道荣. Galectin-9在结直肠癌中的表达及其临床意义[J]. 中华结直肠疾病电子杂志, 2024, 13(02): 112-120.
[15] 周泽恺, 刘宝胤, 聂云韬, 孟化. 肥胖对肺功能影响的研究进展[J]. 中华肥胖与代谢病电子杂志, 2024, 10(01): 58-65.
阅读次数
全文


摘要

版权所有 中华医学会 中华医学电子音像出版社有限责任公司  京ICP备14006079号-1  网络出版服务许可证：(署)网出证(京)字第075号