中性粒细胞胞外诱捕网抗疣状毛癣菌感染的作用研究

中国真菌学杂志　 2019, Vol. 14 　Issue (6): 326-331.

中性粒细胞胞外诱捕网抗疣状毛癣菌感染的作用研究

张秀军, 郭艳阳, 朱振来, 薛小文, 马翠玲, 王刚, 付萌

第四军医大学西京皮肤医院, 西安 710032

收稿日期:2019-06-01 出版日期:2019-12-28 发布日期:2019-12-28
通讯作者: 付萌,E-mail:fumeng@fmmu.edu.cn E-mail:fumeng@fmmu.edu.cn
作者简介:张秀军,男(汉族),硕士研究生在读.E-mail:lrubus@163.com

Effects of NETs on Trichophyton verrucosum infection

ZHANG Xiu-jun, GUO Yan-yang, ZHU Zhen-lai, XUE Xiao-wen, MA Cui-ling, WANG Gang, FU Meng

Department of Dermatology at Xijing Hospital, the Fourth Military Medical University, Xi'an, Shaanxi 710032, China

Received:2019-06-01 Online:2019-12-28 Published:2019-12-28

摘要/Abstract

摘要：

目的观察到1例疣状毛癣菌（Trichophyton verrucosum，T.verrucosum）致须癣患者的局部皮损中存在大量中性粒细胞。以往文献报道中性粒细胞通过形成胞外诱捕网（Neutrophils extracellular traps，NETs）参与抗真菌免疫，但T.verrucosum能否诱导NETs形成，及NETs是否具有抗T.verrucosum作用尚不清楚，故探究中性粒细胞是否通过形成NETs参与杀灭T.verrucosum。方法收集患者病历资料。从患者组织分离、鉴定T.verrucosum。免疫荧光法检测患者及T.verrucosum感染小鼠组织中的NETs。T.verrucosum刺激正常人外周血中性粒细胞，免疫荧光法检测NETs的形成。NETs与T.verrucosum共孵育，提取T.verrucosum DNA，通过实时荧光定量PCR定量检测真菌数量，观察NETs对T.verrucosum的杀伤作用。结果 T.verrucosum感染患者及小鼠组织可见明显NETs。T.verrucosum与中性粒细胞共孵育后有明显NETs形成。与NETs孵育后T.verrucosum存活率低于对照组（P<0.01）。结论 T.verrucosum可以诱导中性粒细胞产生NETs，NETs对T.verrucosum具有显著杀伤作用。

关键词: 中性粒细胞胞外诱捕网, 疣状毛癣菌, 抗真菌作用

Abstract:

Objective A large number of neutrophils were found in the local lesions of patient with tinea barbae caused by Trichophyton verrucosum (T. verrucosum). Previous literatures reported that neutrophils participate in antifungal immunity through the formation of extracellular traps (NETs), but it was unclear whether T. verrucosum could induce NETs formation and whether NETs were involved in anti-T. verrucosum, so we explored whether neutrophils participate in the killing of T. verrucosum by forming NETs. Methods The patient's clinical data was collected. T. verrucosum was isolated and identified from patient's tissues. NETs in the tissues of patients and T. verrucosum-infected mice were observed by immunofluorescence. T. verrucosum was used to stimulate peripheral neutrophils in normal subjects, immunofluorescence was used to detect the formation of NETs. NETs were incubated with T. verrucosum, and then T. verrucosum DNA was extracted, Real-time quantitative PCR was used to quantitatively detect the killing effect of NETs on T. verrucosum. Results NETs formation could be seen in the tissues of T. verrucosum infected patients and mice. T. verrucosum co-incubation with neutrophils showed NETs formation. The survival rate of T. verrucosum after incubation with NETs was lower than that of the control group (P<0.01). Conclusions T. verrucosum can induce neutrophils to produce NETs, and NETs have a significant killing effect on T. verrucosum.

Key words: NETs, T. verrucosum, antifungal effects

中图分类号:

R379.2

张秀军, 郭艳阳, 朱振来, 薛小文, 马翠玲, 王刚, 付萌. 中性粒细胞胞外诱捕网抗疣状毛癣菌感染的作用研究[J]. 中国真菌学杂志, 2019, 14(6): 326-331.

ZHANG Xiu-jun, GUO Yan-yang, ZHU Zhen-lai, XUE Xiao-wen, MA Cui-ling, WANG Gang, FU Meng. Effects of NETs on Trichophyton verrucosum infection[J]. Chinese Journal of Mycology, 2019, 14(6): 326-331.

参考文献

[1] 崔蕊, 方玲, 谢风. 中性粒细胞胞外诱捕网研究最新进展[J]. 中国实验诊断学, 2018,22(1):178-180.
[2] Delgado-Rizo V, Martínez-Guzmán MA, Iñiguez-Gutierrez L, et al. Neutrophil extracellular traps and its implications in inflammation:an overview[J]. Frontiers in immunology, 2017,8:81.
[3] Brinkmann V, Reichard U, Goosmann C, et al. Neutrophil extracellular traps kill bacteria[J]. Science, 2004,303(5663):1532-1535.
[4] Zhang X, Zhao S, Sun L, et al. Different virulence of Candida albicans is attributed to the ability of escape from neutrophil extracellular traps by secretion of DNase[J]. Am J Transl Res, 2017,9(1):50-62.
[5] Bruns S, Kniemeyer O, Hasenberg M, et al. Production of extracellular traps against Aspergillus fumigatus in vitro and in infected lung tissue is dependent on invading neutrophils and influenced by hydrophobin RodA[J]. PLoS Pathog, 2010,6(4):e1000873.
[6] Burmester A, Shelest E, Glöckner G, et al. Comparative and functional genomics provide insights into the pathogenicity of dermatophytic fungi[J]. Genome biology, 2011,12(1):R7.
[7] Wollina U, Hansel G, Uhrlass S, et al. Deep facial mycosis due to Trichophyton verrucosum-molecular genetic identification of the dermatophyte in paraffin-embedded tissue-case report and review of the literature[J]. Mycoses, 2018,61(3):152-158.
[8] O'Gorman SM, Britton D, Collins P. An uncommon dermatophyte infection:two cases of cutaneous infection with Trichophyton verrucosum[J]. Clin Exp Dermatol, 2015,40(4):395-398.
[9] Shao S, Fang H, Dang E, et al. Neutrophil extracellular traps promote inflammatory responses in psoriasis via activating epidermal TLR4/IL-36R crosstalk[J]. Frontiers in Immunology, 2019,10:746.
[10] 郑艳青, 史娜娜, 曹存巍. 实时荧光定量PCR检测马尔尼菲篮状菌病组织样本中菌载量的研究[J]. 中国真菌学杂志, 2018,13(2):71-74.
[11] Urban CF, Nett JE. Neutrophil extracellular traps in fungal infection[J]. Semin Cell Dev Biol, 2019,89(5):47-57.
[12] Park H J, Wang W, Curlango-Rivera G, et al. A DNase from a fungal phytopathogen is a virulence factor likely deployed as counter defense against host-secreted extracellular DNA[J]. MBio, 2019,10(2):e02805-e02818.
[13] Sørensen OE, Borregaard N. Neutrophil extracellular traps-the dark side of neutrophils[J]. J Clin Invest, 2016,126(5):1612-1620.