白念珠菌ALS3、SSA1基因表达在念珠菌性阴道炎免疫机制中的作用

中国真菌学杂志　 2019, Vol. 14 　Issue (2): 65-69.

论著下一篇

白念珠菌ALS3、SSA1基因表达在念珠菌性阴道炎免疫机制中的作用

高盈¹, 王琼¹, 梁官钊¹, 佘晓东¹, 史冬梅^1,2, 沈永年¹, 苏晓红¹, 李冬梅^1,3, 刘维达¹

1. 中国医学科学院北京协和医学院皮肤病研究所, 江苏省皮肤病与性病重点分子生物学实验室, 南京 210042;
2. 济宁市第一人民医院皮肤科, 济宁 272001;
3. Department of Microbiology&Immunology, Georgetown University, Medical Center, Washington DC 20057, USA

收稿日期:2018-11-22 出版日期:2019-04-28 发布日期:2019-04-28
通讯作者: 刘维达,E-mail:liumyco@hotmail.com E-mail:liumyco@hotmail.com
作者简介:高盈,女(汉族),博士,住院医师.E-mail:nanjingpys2013@126.com
基金资助:
国家重点基础研究计划（2013CB531605）；国家自然科学基金（NO.81071332）；江苏省医学科学专项计划（BL2012003）

The role of ALS3 and SSA1 gene expression of Candida albicans in the immune mechanism of vaginal candidiasis

GAO Ying¹, WANG Qiong¹, LIANG Guan-zhao¹, SHE Xiao-dong¹, SHI Dong-mei^1,2, SHEN Yong-nian¹, SU Xiao-hong¹, LI Dong-mei^1,3, LIU Wei-da¹

1. Department of Mycology, Institute of Dermatology, Chinese Academy of Medical Sciences(CAMS) & Peking Union Medical College(PUMC), Jiangsu Key laboratory of Molecular Biology for Skin Disease and STIs, Nanjing 210042, China;
2. Department of Dermatology, Jining No.1 People's Hospital, Jining 272001, China;
3. Georgetown University Medical Center, Department of Microbiology & Immunology, Washington DC 20057, USA

Received:2018-11-22 Online:2019-04-28 Published:2019-04-28

摘要/Abstract

摘要：

目的探讨白念珠菌ALS3、SSA1基因缺失对阴道上皮细胞激发免疫反应的作用。方法培养白念珠菌野生株及ALS3、SSA1基因敲除株（SC5314、Δals3、Δssa1），对其进行形态测定。按不同MOI感染人阴道上皮细胞系VK2/E6E7细胞，通过台盼蓝染色观察和乳酸脱氢酶（LDH）活性检测，评价不同MOI白念珠菌对上皮细胞的损伤作用；使用酶联免疫吸附试验（ELISA）评估感染过程中炎性细胞因子及趋化因子在共培养上清中的差异。结果 ALS3基因的缺失对白念珠菌芽管长度影响差异无统计学意义，而SSA1基因的缺失与其他两个菌株相比芽管长度减少约30%~40%（P<0.001）。台盼蓝染色观察及LDH测定发现，3株菌在感染上皮细胞时，其细胞损伤能力均与菌载量成正比；与野生型相比，Δssa1突变体在相同比率感染上皮细胞时，细胞损伤能力明显降低，且差异有统计学意义（P<0.05），Δals3突变株影响较小，甚至略微升高。检测炎性细胞因子及趋化因子发现，突变株在诱导上皮细胞产生促炎因子及趋化因子（GM-CSF、G-CSF、IL-1α、IL-8）的能力上明显减弱，差异均有统计学意义（P<0.05）。结论 ALS3和SSA1基因表达在阴道上皮细胞抗白念珠菌感染的局部免疫应答过程中可能起到重要作用，且SSA1基因表达意义更大。

关键词: 阴道上皮细胞, 白念珠菌, ALS3, SSA1, 免疫

Abstract:

Objective To investigate the effect of ALS3 and SSA1 genes in Candida albicans on immune response of vaginal epithelial cells. Methods Wild strains and ALS3, SSA1 gene knockout strains (SC5314, Δals3, Δssa1) of Candida albicans were cultured and their morphology was determined. Human vaginal epithelial cell line VK2/E6E7 was infected in a certain proportion. The damage of different strains to epithelial cells was evaluated by trypan blue staining and lactate dehydrogenase (LDH) activity detection. The co-culture of inflammatory cytokines and chemokines during infection was evaluated by enzyme-linked immunosorbent assay (ELISA).Results There was no significant difference in the effect of Δals3 on the germ tube length of Candida albicans, while the Δssa1 reduced the germ tube length by about 30%~40% compared with the other two strains (P<0.001). Trypan blue staining and LDH assay showed that the cell damage ability of the three strains was proportional to the fungal load when infected with epithelial cells. Compared with wild strain, the cell damage ability of Δssa1 was significantly reduced when infected with epithelial cells at the same MOI, and the difference was statistically significant (P<0.05), while the impact of Δals3 mutant was weak or even slightly higher. Detection of inflammatory cytokines and chemokines showed that the ability of mutants to induce epithelial cells to produce pro-inflammatory factors and chemokines (GM-CSF, G-CSF, IL-1a, IL-8) was significantly weakened (P<0.05).Conclusion ALS3 and SSA1 gene expression played an important role in the local immune response of vaginal epithelial cells to Candida albicans infection, and SSA1 gene expression was more significant.

Key words: vaginal epithelial cells, Candida albicans, ALS3, SSA1, immunity

中图分类号:

R519.3

高盈, 王琼, 梁官钊, 佘晓东, 史冬梅, 沈永年, 苏晓红, 李冬梅, 刘维达. 白念珠菌ALS3、SSA1基因表达在念珠菌性阴道炎免疫机制中的作用[J]. 中国真菌学杂志, 2019, 14(2): 65-69.

GAO Ying, WANG Qiong, LIANG Guan-zhao, SHE Xiao-dong, SHI Dong-mei, SHEN Yong-nian, SU Xiao-hong, LI Dong-mei, LIU Wei-da. The role of ALS3 and SSA1 gene expression of Candida albicans in the immune mechanism of vaginal candidiasis[J]. Chinese Journal of Mycology, 2019, 14(2): 65-69.

参考文献

[1] Sobel JD. Vulvovaginal candidosis[J]. Lancet, 2007,369(9577):1961-1971.
[2] Cenci E, Mencacci A, Spaccapelo R, et al. T helper cell type 1(Th1)-and Th2-like responses are present in mice with gastric candidiasis but protective immunity is associated with Th1 development[J]. J Infect Dis, 1995,171(5):1279-1288.
[3] Fidel PL, Jr., Lynch ME, Redondo-Lopez V, et al. Systemic cell-mediated immune reactivity in women with recurrent vulvovaginal candidiasis[J]. J Infect Dis, 1993,168(6):1458-1465.
[4] Murciano C, Moyes DL, Runglall M, et al. Evaluation of the role of Candida albicans agglutinin-like sequence (Als) proteins in human oral epithelial cell interactions[J]. PLoS One, 2012,7(3):e33362.
[5] Yang W, Yan L, Wu C, et al. Fungal invasion of epithelial cells[J]. Microbiol Res, 2014,169(11):803-810.
[6] Murciano C, Moyes D L, Runglall M, et al. Evaluation of the role of Candida albicans agglutinin-like sequence (Als) proteins in human oral epithelial cell interactions[J]. PLoS One, 2012, 7(3):e33362.
[7] Moyes D L, Runglall M, Murciano C, et al. A biphasic innate immune MAPK response discriminates between the yeast and hyphal forms of Candida albicans in epithelial cells[J]. Cell Host Microbe, 2010, 8(3):225-235.
[8] Zhao X, Oh S H, Cheng G, et al. ALS3 and ALS8 represent a single locus that encodes a Candida albicans adhesin; functional comparisons between Als3p and Als1p[J]. Microbiology, 2004, 150(Pt 7):2415-2428.
[9] Sun J N, Solis N V, Phan Q T, et al. Host cell invasion and virulence mediated by Candida albicans Ssa1[J]. PLoS Pathog, 2010, 6(11):e1001181.
[10] Tutar Y, Arslan D, Tutar L. Heat, pH induced aggregation and surface hydrophobicity of S. cerevesiae Ssa1 protein[J]. Protein J, 2010, 29(7):501-508.
[11] Murciano C, Moyes D L, Runglall M, et al. Evaluation of the role of Candida albicans agglutinin-like sequence (Als) proteins in human oral epithelial cell interactions[J]. PLoS One, 2012, 7(3):e33362.
[12] Dongari-Bagtzoglou A, Fidel PL, Jr. The host cytokine responses and protective immunity in oropharyngeal candidiasis[J]. J Dent Res, 2005,84(11):966-977.
[13] Steele C, Fidel PL, Jr. Cytokine and chemokine production by human oral and vaginal epithelial cells in response to Candida albicans[J]. Infect Immun, 2002,70(2):577-583.
[14] Lee S, Song L, Yang M, et al. Local administration of granulocyte macrophage colony-stimulating factor induces local accumulation of dendritic cells and antigen-specific CD8⁺ T cells and enhances dendritic cell cross-presentation[J]. Vaccine, 2015, 33(13):1549-1555.
[15] Carmi Y, Voronov E, Dotan S, et al. The role of macrophage-derived IL-1 in induction and maintenance of angiogenesis[J]. J Immunol, 2009,183(7):4705-4714.
[16] Homey B, Dieu-Nosjean MC, Wiesenborn A, et al. Up-regulation of macrophage inflammatory protein-3 alpha/CCL20 and CC chemokine receptor 6 in psoriasis[J]. J Immunol, 2000,164(12):6621-6632.

白念珠菌ALS3、SSA1基因表达在念珠菌性阴道炎免疫机制中的作用

The role of ALS3 and SSA1 gene expression of Candida albicans in the immune mechanism of vaginal candidiasis

PDF (PC)

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

编辑推荐

Metrics

本文评价

[1]	孙青刚, 张海燕, 刘楠, 王明辉, 时洪川, 王海兵. 非造血干细胞移植血液病患者并发肺孢子菌肺炎7例临床分析[J]. 中国真菌学杂志, 2020, 15(6): 344-348.
[2]	刘丹, 邢丽枝, 杨心茹, 罗伟, 何荣霞. 唑类药物治疗外阴阴道假丝酵母菌病研究进展[J]. 中国真菌学杂志, 2020, 15(5): 318-320.
[3]	黄春阳, 潘开素, 李秀楹, 曹存巍, 郑艳青. Calcineurin A基因对马尔尼菲篮状菌致病力的影响[J]. 中国真菌学杂志, 2020, 15(3): 138-144.
[4]	马玉帛, 张逸, 黄晨, 王晓雯, 李若瑜. 自然杀伤细胞在真菌感染免疫的研究进展[J]. 中国真菌学杂志, 2020, 15(3): 175-178.
[5]	郑晓丽, 陈官芝, 刘维达. 抗真菌药物的免疫调节作用[J]. 中国真菌学杂志, 2020, 15(3): 179-182.
[6]	邱熙然, 陈思敏, 侯炜彤, 张玉, 郭诗雨, 姜远英, 安毛毛. 白念珠菌侵袭宿主毒力因子研究进展[J]. 中国真菌学杂志, 2020, 15(3): 183-188.
[7]	张静云, 彭靖雯, 王琼, 高盈, 陈万鑫, 沈永年, Li Dong-mei, 佘晓东, 刘维达. 表皮生长因子受体在阴道念珠菌病发病过程中的作用初探[J]. 中国真菌学杂志, 2020, 15(2): 65-71.
[8]	刘光荣, 翟春涛, 金敏蓉, 李成亮. 天然抗菌剂对5种致病菌的抑制作用研究[J]. 中国真菌学杂志, 2020, 15(2): 88-92.
[9]	谭静文, 徐红, 宋金凤, 高志琴, 杨虹, 杨连娟, 温海. 聚六亚甲基双胍体外抗真菌作用研究[J]. 中国真菌学杂志, 2020, 15(2): 93-96.
[10]	张静云, 佘晓东, 刘维达. 白念珠菌糖代谢与毒力[J]. 中国真菌学杂志, 2020, 15(1): 61-64.
[11]	王晓东, 迪丽努尔·塔西买买提, 迪丽达尔·地里夏提, 哈地利亚·哈斯木, 帕丽达·阿布利孜. 球形孢子丝菌刺激小鼠树突状细胞表达前炎症细胞因子的研究[J]. 中国真菌学杂志, 2019, 14(6): 321-325.
[12]	赵晓琴, 冯文莉. 转录因子在白念珠菌生物膜形成过程中的调控机制[J]. 中国真菌学杂志, 2019, 14(6): 381-384.
[13]	黄进宝, 李红艳, 兰长青, 吕骁, 林志来, 王新航, 张宏英, 邹盛华, 翁恒. 两种血清隐球菌荚膜多糖抗原检测方法在肺隐球菌病中的应用研究[J]. 中国真菌学杂志, 2019, 14(5): 264-269.
[14]	王钰婷, 王玉柱, 刘锦燕, 孟玲宁, 李卫华, 项明洁. 酵母相和菌丝相白念珠菌感染阴道上皮细胞免疫反应的比较[J]. 中国真菌学杂志, 2019, 14(4): 193-199.
[15]	蔡晴, 杨连娟, 高志琴, 任弘瑾, 杨虹, 谭静文. 罗米地辛对氟康唑体外抗白念珠菌影响的研究[J]. 中国真菌学杂志, 2019, 14(4): 200-203,216.