LC3-LpqH增强Ag85B抗结核分枝杆菌DNA疫苗的免疫保护作用  

LC3-LpqH enhances the immunoprotection of Ag85B DNA vaccine against Mycobacterium tuberculosis

作　　者：胡东[1] 杨小康[1] 陈功[1] 吴静[1] 王婉[1] 戴京京 陈天义[2] 张荣波[1] 

机构地区：[1]安徽理工大学医学院免疫与检验教研室免疫与感染研究所,安徽淮南232001 [2]安徽理工大学医学院组织病理学教研室,安徽淮南232001

出　　处：《细胞与分子免疫学杂志》2014年第12期1243-1246,1250,共5页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家自然科学基金(81302524;81202294;81172778);安徽省自然科学基金(1208085QH162;KJ2013A105);国家级大学生创新创业训练计划(201210361121;201310361096)

摘　　要：目的探讨微管相关蛋白轻链3-脂蛋白前体-LpqH(LC3-LpqH)DNA佐剂对Ag85B抗结核分枝杆菌(MTB)DNA疫苗免疫保护作用的影响。方法分别用纯化的pCMV-Ag85B、pCMV-LpqH/pCMV-Ag85B、pCMV-LC3-LpqH/pCMV-Ag85B、pCMV质粒DNA对BALB/c小鼠进行3次肌肉注射免疫。末次免疫2周后,ELISA检测血清中抗Ag85B IgG亚型及滴度。Ag85B刺激下培养各组小鼠脾脏淋巴细胞,ELISA检测血清白细胞介素2(IL-2)、干扰素-γ(IFN-γ)、IL-4、IL-10分泌水平。末次免疫3个月后,用MTB标准毒株H37Rv尾静脉注射感染小鼠,计数肺和脾脏MTB载量并观察肺组织病理变化。结果与Ag85B免疫组相比,LC3-LpqH/Ag85B免疫组抗Ag85B的IgG2a水平显著提高,而IgG1水平显著降低,并伴随IL-2、IFN-γ分泌增加及IL-4、IL-10减少,肺和脾脏内MTB载量降低,肺组织病变程度减轻。结论 LC3-LpqH可显著增强Ag85B抗MTB疫苗的Th1型免疫应答,并显示较好的免疫保护效应。Objective To explore the effect of microtubule-associated protein light chain 3-lipoprotein antigen precursor（pCMV-LC3-LpqH） as an DNA adjuvant on the immunoprotection of the anti-tuberculosis DNA vaccine encoding Ag85 B antigen from Mycobacterium tuberculosis（MTB）. Methods BALB /c mice were randomly divided into four groups that were immunized three times with pCMV-Ag85 B, pCMV-LpqH /pCMV-Ag85 B, pCMV-LC3-LpqH /pCMV-Ag85 B and pCMV,respectively. Two weeks after the final vaccination,the titers and subtypes of anti-Ag85 B IgG of BALB /c mice were detected by ELISA. In vitro,the levels of interleukin 2（ IL-2）,interferon γ（ IFN-γ）,IL-4 and IL-10 that were secreted from Ag85B-stimulated splenocytes were also measured by ELISA. Three months after the final vaccination,mice were challenged with virulent MTB H37 Rv strain through the tail veins. The mice spleens and lungs were dissected out for counting colony-forming unit（ CFU） and observing pulmonary histopathological changes after MTB infection. Results LC3-LpqH /Ag85B-immunized mice showed the significant increase of IgG2 a,IL-2 and IFN-γ,along with the decrease of IgG1,IL-4 and IL-10,lower MTB CFU in the spleens and lungs and a marked relief in pulmonary injury. Conclusion LC3-LpqH could enhance Ag85B-specific Th1 immune response that exerts a superior immune protection,which might be a new practical strategy for developing MTB vaccines.

关 键 词：结核分枝杆菌 DNA疫苗 自噬 免疫保护 

分 类 号：R378.911[医药卫生—病原生物学] R392.33[医药卫生—基础医学]