IL-18基因修饰的成熟树突状细胞疫苗对哮喘小鼠气道炎症的预防作用

Preventive effect of IL-18 gene modified mature dendritic cells vaccine on airway inflammation in mouse asthma model

目的:观察IL-18基因修饰的成熟树突状细胞(mature dendritic cells,mDC)疫苗对卵白蛋白(OVA)诱导哮喘小鼠气道炎症的预防作用。方法:采用流式细胞术分析IL-18基因修饰的成熟树突状细胞表型变化,通过混合淋巴细胞反应(MLR)检测其诱导同种异体T细胞的应答能力。BALB/c小鼠随机分成6组:健康对照组(Control组)、PBS组、地塞米松治疗组(DXM组)、成熟树突状细胞治疗组(mDC组)、LacZ基因修饰的成熟树突状细胞治疗组(Ad-LacZ-mDC组)、IL-18基因修饰的成熟树突状细胞治疗组(Ad-IL-18-mDC组)。应用卵白蛋白诱导小鼠哮喘模型。HE、AB-PAS染色观察小鼠肺组织病理学变化。光学显微镜下观察小鼠气管肺泡灌洗液中炎性细胞数及嗜酸性粒细胞百分比。酶联免疫吸附试验检测脾细胞培养上清中IFN-γ、IL-4和IL-13的含量。流式细胞细胞术检测脾细胞悬液CD4+CD25+foxP3+调节性T细胞(Treg)百分比。结果:IL-18基因修饰的成熟树突状细胞疫苗免疫哮喘小鼠后,能够有效减少肺组织炎性细胞浸润和杯状细胞增生,降低肺泡灌洗液炎性细胞数和嗜酸性粒细胞比例、脾细胞培养上清中IL-4和IL-13的分泌,提高脾细胞培养上清中IFN-γ的分泌和脾细胞悬液中CD4+CD25+foxP3+Treg的百分比。结论:IL-18基因修饰的成熟树突状细胞疫苗对卵白蛋白诱导的哮喘小鼠气道炎症具有一定的预防作用。

Objective:To investigate the preventive effect of interleukin-18(IL-18) gene modified mature dendritic cells(mDC) vaccine on airway inflammation in mouse asthma model. Methods:The asthma model was induced by injection of ovalbumin(OVA) in BALB/c mice.IL-18 gene modified mouse mature dentritic cells(mDC) were detected by flow cytometry and its capacity of inducing allogeneic T cell responses was examined by mixed lymphocyte reaction(MLR).The OVA-induced asthmatic mice were randomly divided into 6 groups:PBS group,DXM group,mDC group,Ad-LacZ-mDC group,Ad-IL-18-mDC group and control group.The pathological changes in lung tissues were assayed by HE and AB-PAS staining.The numbers of inflammatory cells and percentage of eosinophils(EOS) in bronchoalveolar lavage fluid(BALF) were counted.The levels of IFN-γ,IL-4 and IL-13 in culture supernatant of splenocytes were measured by ELISA method.The percentage of CD4+CD25+Foxp3+ Treg was assessed by flow cytometry analysis. Results:The vaccine was effective in decreasing the infiltration of EOS and accumulation of airway goblet cells in lung tissues,the numbers of inflammatory cells and percentage of EOS in BALF,and the levels of IL-4 and IL-13 in culture supernatant of splenocytes,and in increasing the levels of IFN-γ in culture supernatant of splenocytes and the percentage of CD4+CD25+foxP3+ reg. Conclusion:IL-18 gene modified mDC vaccine has a preventive effect on airway inflammation in OVA-induced asthmatic mice.