细胞因子IL-2、IL-12及IL-18增强结核杆菌ESAT-6 DNA疫苗的免疫保护作用  

Cytokines IL-2, IL-12 and IL-18 enhance the protective immunity of ESAT-6 DNA vaccine against Mycobacterium tuberculosis

作　　者：尹芳[1] 胡月圆[1] 郭琼[1] 李贵南[1] YIN Fang;HU Yueyuan;GUO Qiong;LI Guinan(Department 2 of Neonatology,Hu'nan Children's Hospital,Changsha 410007,China)

机构地区：[1]湖南省儿童医院新生儿二科

出　　处：《免疫学杂志》2019年第11期954-959,共6页Immunological Journal

摘　　要：目的探讨白细胞介素-2(IL-2)、白细胞介素-12(IL-12)以及白细胞介素-18(IL-18)对结核分枝杆菌ESAT-6DNA疫苗免疫保护作用的影响。方法50只雌性Balb/c小鼠随机分为IL-2-pcDNA-ESAT-6、IL-12-pcDNA-ESAT-6、IL-18-pcDNA-ESAT-6、pcDNA-ESAT-6以及PBS对照组,分别于0、2、4周共免疫3次。末次免疫2周后各组处死5只小鼠,ELISA检测小鼠血清IgG、IgA,并分别检测0、2、4、6周小鼠阴道灌洗液中sIgA水平。CCK-8法检测免疫小鼠脾细胞的增殖情况,并测定脾细胞上清中细胞因子IFN-γ、IL-2、TNF-α、IL-6、IL-10的分泌水平。末次免疫2周后,腹腔注射结核杆菌H37Rv感染小鼠,感染后4周取小鼠肺组织进行菌落培养计数;同时通过HE染色以及肺组织炎症因子检测评估其炎症情况。结果细胞因子IL-2、IL-12以及IL-18均能够显著增加小鼠血清IgG、IgA以及阴道灌洗液sIgA的分泌(P<0.05),并促进免疫小鼠脾淋巴细胞的增殖(P<0.05);IL-2、IL-12及IL-18还能够显著增加脾细胞中IFN-γ、IL-2、TNF-α的分泌水平;此外,相较于ESAT-6真核疫苗组以及PBS对照组,IL-2、IL-12及IL-18均能够显著降低小鼠肺组织菌体载量以及炎性侵润,并减少炎性因子分泌。结论细胞因子IL-2、IL-12及IL-18均能不同程度的增强结核杆菌ESAT-6 DNA疫苗的体液以及细胞免疫应答水平,并显著提高其免疫保护效果。To investigate the effect of interleukin-2(IL-2),interleukin-12(IL-12)and interleukin-18(IL-18)on the protective immunity of ESAT-6 DNA vaccine against Mycobacterium tuberculosis,50 female Balb/c mice were randomly divided into IL-2-pcDNA-ESAT-6,IL-12-pcDNA-ESAT-6,IL-18-pcDNA-ESAT-6,pcDNAESAT-6 and PBS control groups,and immunized at weeks 0,2 and 4,respectively.Five mice in each group were sacrificed to tested the serum IgG and IgA by ELISA at week 2 after the last immunization,and the level of vaginal fluid sIgA was detected at weeks 0,2,4 and 6,respectively;the proliferation of spleen cells in immunized mice were detected by CCK-8 method,and the secretion levels of cytokines IFN-γ,IL-2,TNF-α,IL-6 and IL-10 in the supernatant of spleen cells were determined by ELISA.Mice were intraperitoneally infected with Mycobacterium tuberculosis H37 Rv at week 2 after the last immunization,and the lung tissues of mice were collected to determine the bacterial load at week 4 after the infection;the inflammation of mice was evaluated by HE staining and inflammatory factors detection of infected lung tissue.Data showed that cytokines IL-2,IL-12 and IL-18 significantly increased the secretion of serum IgG,IgA and vaginal fluid sIgA,and promoted the proliferation of spleen lymphocytes in immunized mice,as compared to control group(P<0.05).IL-2,IL-12 and IL-18 also significantly increased the secretion levels of IFN-γ,IL-2 and TNF-αin spleen cells of immunized mice;In addition,compared with the ESAT-6 DNA vaccine group and PBS control group,IL-2,IL-12 and IL-18 could significantly reduce the bacterial load and inflammatory invasion in the lung tissue of immunized mice,and reduce the secretion levels of inflammatory factors.In conclusion,cytokines IL-2,IL-12 and IL-18 can effectively enhance the effect of humoral,cellular immunity induced by ESAT-6 DNA vaccine,thus significantly enhancing their protective immunity against Mycobacterium tuberculosis.

关 键 词：细胞因子 结核杆菌 ESAT-6 疫苗 

分 类 号：R392.1[医药卫生—免疫学]