摘要
目的探讨多房棘球蚴感染小鼠脾脏细胞中辅助性T细胞17比例,转录因子RORγτ和IL-17因子的表达及其意义。方法选用6~8周龄BALB/c小鼠共30只,随机分为多房棘球蚴感染组(Em)、多房棘球蚴感染阿苯达唑治疗组(Em+Albendazole,Em+ABZ)和健康对照组(healthy controls,HC),每组10只。通过腹腔注射法建立泡型包虫病小鼠模型。Em+ABZ治疗组给予灌胃阿苯达唑治疗(100μl/d,35d)通过流式细胞术检测小鼠脾脏细胞中辅助性T17细胞(CD4+IL-17+Th17细胞/CD4+T细胞)比例,采用实时荧光定量PCR检测脾脏细胞中维甲酸孤独受体(retinoid acid orphan receptor-γτ,RORγτ)mRNA的表达;采用酶联免疫吸附试验(ELISA)检测IL-17水平。结果 CD4+IL-17+Th17细胞/CD4+T细胞比例,Em组为(2.10±1.00)%,与HC组(0.85±0.71)%比较,差异有统计学意义(F=3.07,t=2.66,P〈0.01)。Em+ABZ组为(1.54±1.19)%,与HC组和Em组比较,差异无统计学意义(F=3.07,P〉0.05);脾脏细胞RORγτmRNA在Em组中的表达(6.1×10-4±5.8×10-4)高于在Em+ABZ组(6.0×10-4±3.5×10-4)和HC组(3.4×10-4±2.4×10-4)中的表达,但差异无统计学意义(F=0.66,P〉0.05)。Em+ABZ组与HC组比较,差异亦无统计学意义(F=0.66,P〉0.05)。脾脏细胞培养上清液中IL-17A,Em组为(10.71±1.85)ng/ml,明显高于Em+ABZ组[(5.58±3.56)ng/ml]和HC组[(7.31±1.22)ng/ml],差异有统计学意义(F=9.82,t1=3.12,t2=5.57,P〈0.01)。Em+ABZ组与HC组比较差异无统计学意义(F=9.82,t=0.84,P〉0.05)。结论 Th17细胞参与BALB/c小鼠多房棘球蚴感染免疫和炎性应答过程,阿苯达唑治疗能够减轻免疫损伤和炎症反应。Th17细胞及IL-17A可能是潜在的多房棘球蚴病新的分子免疫干预靶点。
Objective To explore the alteration and significance of T helper cell 17,transcription factor RORγτ,and IL-17 in BALB/c mice infected with Echinococcus multilocularis. Methods Thirty 6-8week-old BALB/c mice were randomly divided into three groups:mice infected with E.multilocularis(n=10),mice infected with E.multilocularis+treated with albendazole(ABZ)(n=10),and healthy controls(HCs,n=10).A model of alveolar echinococcosis was created by intraperitoneally infecting mice with E.multilocularis.Mice infected with E.multilocularis+treated with ABZ were administered ABZ(100ul/d for 35days)via a gastric tube.The proportion of Th17 cells in the spleen was determined using flow cytometry,expression of RORγτmRNA was determined using real-time PCR,and levels of IL-17 in splenic cell supernatant were detected using ELISA. Results The ratio of CD4+IL-17A+Th17cells/CD4+T cells in the spleen was significantly higher in mice infected with E.multilocularis(2.10±1.00%)than in the HCs(0.85±0.71%,F=3.07,t=2.66,P〈0.01).The ratio of CD4+IL-17A+Th17cells/CD4+T was higher in mice infected withE.multilocularis+treated with ABZ(1.54±1.19%)than in HCs but lower than that in mice infected with E.multilocularis,although the difference was not statistically significant(F=3.07,P〉0.05).The level of expression of the transcription factor RORγτin splenic cells was higher in mice infected with E.multilocularis(6.1×10-4±5.8×10-4)than in HCs(3.4×10-4±2.4×10-4)and in mice infected with E.multilocularis+treated with ABZ(6.0×10-4±3.5×10-4),but the difference was not statistically significant.The level of RORγτexpression was higher in mice infected with E.multilocularis+treated with ABZ than in HCs,but the difference was not statistically significant(F=0.66,P〉0.05).The level of IL-17 Aexpression in splenic cell supernatant was significantly higher in mice infected with E.multilocularis(10.71±1.85ng/ml)than in mice infected with E.multilocularis+treated with ABZ(5.58±3.56ng/ml)and in HCs(7.31±1.22ng/ml),and the difference was statistically significant(F=9.82,t1=3.12,t2=5.57,P〈0.01).However,the level of IL-17 Aexpression did not differ significantly in mice infected with E.multilocularis+treated with ABZ and HCs(F=9.82,t=0.84,P〉0.05). Conclusion Th17 cells are actively involved in immunity from infection and inflammatory response in a murine model of alveolar echinococcosis.ABZ reduced the number of Th17 cells and related cytokine levels,possibly alleviating immunopathological injury and inflammation.Th17 cells and related cytokines may be a potential therapeutic molecular target for alveolar echinococcosis.
出处
《中国病原生物学杂志》
CSCD
北大核心
2014年第8期694-698,共5页
Journal of Pathogen Biology
基金
国家自然科学基金项目(No.U1303222
81160201)
长江学者和创新团队发展计划(No.IRT1181)
新疆包虫病基础医学重点实验室开放课题资助项目(No.XJDX0202-2011-03)
关键词
多房棘球蚴
感染免疫
辅助性T细胞
免疫损伤
Echinococcus multilocularis
immunity from infection
T helper cell
Immunological injury