Foxp3~+调节性T细胞在播散性毛孢子菌病小鼠模型的表达研究
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摘要
目的观察调节性T细胞(regulatory T cells,Tregs)在播散性阿萨希毛孢子菌(Trichosporon asahii,T.asahii)病小鼠模型中的表达,初步探讨Foxp3~+调节性T细胞在播散性毛孢子菌病发病机制中可能发挥的作用。方法 50只雄性BALB/c小鼠随机分为试验组和对照组,每组25只,分别于尾静脉接种阿萨希毛孢子菌悬液和生理盐水。接种后第3,7,14,21及28天,流式细胞仪检测两组小鼠脾脏单个核细胞悬液中CD4~+CD25~+Foxp3~+调节性T细胞比率;实时荧光定量聚合酶链反应(PCR)检测两组小鼠脾脏Foxp3 mRNA水平表达情况。结果试验组小鼠脾脏中CD4~+CD25~+Foxp3~+调节性T细胞比率及Foxp3 mRNA表达随着感染进程先增高后下降至正常水平。接种后第3、28天,试验组小鼠脾脏单个核细胞中CD4~+CD25~+Foxp3~+调节性T细胞的比例与对照组差异无统计学意义(P>0.05);接种后第7、14及21天调节性T细胞的比例较对照组明显上升,差异有统计学意义(P<0.05)。接种后第3、28天,试验组小鼠脾脏Foxp3 mRNA表达水平与正常对照组相比,差异无统计学意义(P>0.05);接种后第7、14、21天与对照组相比,差异有统计学意义(P<0.05)。结论阿萨希毛孢子菌可以诱导小鼠体内CD4~+CD25~+Foxp3~+调节性T细胞在感染中后期表达增高,调节性T细胞可能参与调节播散性毛孢子菌病的免疫应答。
Objective To study the dynamic changes of Foxp3~+regulatory T cells in a mice model of disseminated trichosporonosis,and investigate its possible roles in the pathogenesis of disseminated trichosporonosis.Methods Fifty male BALB/c mice were randomly divided into the experimental group and the control group,then the mice in each group were inoculated with T.asahii suspension and sterile saline respectively through tail vein.On the 3rd,7th,14th,21st and 28th days after inoculation,the ratio of CD4~+CD25~+Foxp3~+regulatory T cells in spleen mononuclear cell suspension was detected by flow cytometry,and the expression of Foxp3 gene in spleen was detected by real-time fluorescence quantitative polymerase chain reaction(PCR).Results The ratio of CD4~+CD25~+Foxp3~+regulatory T cells and the expression of Foxp3 mRNA in spleen of mice in the experimental group increased first and then decreased to normal level with the progress of infection.On the 3rd and 28th day after inoculation,there was no significant difference in the proportion of CD4~+CD25~+Foxp3~+regulatory T cells and Foxp3 gene expression between the experimental group and the control group(P>0.05),but on the 7th,14th and 21st day after inoculation,there was significant difference between the two group(P<0.05).Conclusion T.asahii can induce the expression of CD4~+CD25~+Foxp3~+regulatory T cells in mice at the middle and late stages of infection.Regulatory T cells may be involved in regulating the immune response of disseminated trichosporosis.
Objective To study the dynamic changes of Foxp3~+regulatory T cells in a mice model of disseminated trichosporonosis,and investigate its possible roles in the pathogenesis of disseminated trichosporonosis.Methods Fifty male BALB/c mice were randomly divided into the experimental group and the control group,then the mice in each group were inoculated with T.asahii suspension and sterile saline respectively through tail vein.On the 3rd,7th,14th,21st and 28th days after inoculation,the ratio of CD4~+CD25~+Foxp3~+regulatory T cells in spleen mononuclear cell suspension was detected by flow cytometry,and the expression of Foxp3 gene in spleen was detected by real-time fluorescence quantitative polymerase chain reaction(PCR).Results The ratio of CD4~+CD25~+Foxp3~+regulatory T cells and the expression of Foxp3 mRNA in spleen of mice in the experimental group increased first and then decreased to normal level with the progress of infection.On the 3rd and 28th day after inoculation,there was no significant difference in the proportion of CD4~+CD25~+Foxp3~+regulatory T cells and Foxp3 gene expression between the experimental group and the control group(P>0.05),but on the 7th,14th and 21st day after inoculation,there was significant difference between the two group(P<0.05).Conclusion T.asahii can induce the expression of CD4~+CD25~+Foxp3~+regulatory T cells in mice at the middle and late stages of infection.Regulatory T cells may be involved in regulating the immune response of disseminated trichosporosis.
引文
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[13]Netea MG1,Sutmuller R,Hermann C,et al.Toll-like receptor 2suppresses immunity against Candida albicans through induction of IL-10 and regulatory T cells[J].Immunol,2004,172(6):3712-3718.
[14]Silva AA1,Sotto MN,Duarte MI,et al.Regulatory T cells in cutaneous lesions of patients with Paracoccidioidomycosis[J].Microb Pathog,2013,65:36-40.
[15]杨冬倩,敖俊红,廖勇,等.白细胞介素10和转化生长因子β在播散性毛孢子菌病小鼠模型中的动态研究[J].实用皮肤病学杂志,2015,8(3):161-166.
[2]Chagas-Neto TC,Chaves GM,Melo AS,et al.Bloodstream infections due to Trichosporon:species distribution,T.asahii genotypes based on rDNA IGS1 sequencing and antifungal susceptibility testing[J].JClin Microbiol,2009,47(4):1074-1081.
[3]Tamayo Lomas L,Domínguez-Gil González M,Martín Luengo AI,et al.Nosocomial infection due to Trichosporon asahii in a critical burned patient[J].Rev Iberoam Micol,2014,S1130-1406(14):83-87.
[4]Magee K,Hata DJ,Meza D.Trichosporon asahii infection in a patient with metastatic prostate cancer as an example of an emerging fungal pathogen[J].Lab Med,2015,46(3):e74-78.
[5]Kumar A,Udayakumaran S,Babu R,et al.Trichosporon asahii infection presenting as chronic meningo-ventriculitis and intra ventricular fungal ball:a case report and literature review[J].Mycoses,2015,58(2):99-103.
[6]Colombo AL,Padovan AC,Chaves GM.Current knowledge of Trichosporon spp.and Trichosporonosis[J].Clin Microbiol Rev,2011,24(4):682-700.
[7]Gavin MA,Rasmussen JP,Fontenot JD,et al.Foxp3-dependent programme of regulatory T-cell differentiation[J].Nature,2007,445(7129):771-775.
[8]Sakaguchi S.Naturally arising CD4+regulatory T cells for immunologic self-tolerance and negative control of immune responses[J].Annu Rev Immunol,2004,22:531-562.
[9]Rowe JH,Ertelt JM,Way SS.Foxp3(+)regulatory T cells,immune stimulation and host defence against infection[J].Immunology,2012,136(1):1-10.
[10]Campbell DJ,Koch MA.Phenotypic and functional specialization of FOXP3+regulatory T cells[J].Nat Rev Immunol,2011,11(2):119-130.
[11]Vignali DA,Collison LW,Workman CJ.How regulatory T cells work[J].Nat Rev Immunol,2008,8(7):523-532.
[12]Wang R,Wan Z,Li R.Th and Treg response induced by Aspergillus fumigatus pulsed dendritic cells in vitro[J].Chin Med J(Engl),2014,127(20):3616-3622.
[13]Netea MG1,Sutmuller R,Hermann C,et al.Toll-like receptor 2suppresses immunity against Candida albicans through induction of IL-10 and regulatory T cells[J].Immunol,2004,172(6):3712-3718.
[14]Silva AA1,Sotto MN,Duarte MI,et al.Regulatory T cells in cutaneous lesions of patients with Paracoccidioidomycosis[J].Microb Pathog,2013,65:36-40.
[15]杨冬倩,敖俊红,廖勇,等.白细胞介素10和转化生长因子β在播散性毛孢子菌病小鼠模型中的动态研究[J].实用皮肤病学杂志,2015,8(3):161-166.
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