Derrick Steven C, Yang Amy Li, Morris Sheldon L
Laboratory of Mycobacterial Diseases and Cellular Immunology, Building 29/Room 511, CBER/FDA, 29 Lincoln Dr., Bethesda, MD 20892, USA.
Infect Immun. 2005 Nov;73(11):7727-35. doi: 10.1128/IAI.73.11.7727-7735.2005.
To improve DNA vaccination against Mycobacterium tuberculosis, we evaluated the effectiveness of a Sindbis virus-based DNA construct expressing the tuberculosis antigen 85B (Sin85B). The protective efficacy of Sin85B was initially assessed by aerogenically challenging immunized C57BL/6 mice with virulent Mycobacterium tuberculosis. At 1 and 7 months postinfection, the lung bacterial burdens were considerably reduced and the lung pathology was improved in vaccinated mice compared to naive controls. Furthermore, the mean survival period for Sin85B-immunized mice (305 +/- 9 days) after the tuberculous challenge was extended 102 days relative to the naive mice (203 +/- 13 days) and was essentially equivalent to the survival time of Mycobacterium bovis BCG-vaccinated mice (294 +/- 15 days). The essential role of gamma interferon (IFN-gamma) in Sin85B-mediated protection was established by showing that significantly increased levels of IFN-gamma mRNA were present postinfection in lung cells from vaccinated mice relative to control mice and by demonstrating that IFN-gamma depletion prior to challenge abolished the vaccine-induced protection. The substantial antituberculosis protective responses induced by Sin85B immunization of CD4-/- mice strongly suggested that CD8 cells partially mediate Sin85B-induced protective immunity. Interestingly, Sin85B vaccination did not protect RNase L-/- (a key enzyme in the innate antiviral response) mice while significant protection was detected in RNase L-/- mice immunized with either BCG or a conventional DNA plasmid expressing antigen 85B. These data show that immunization with Sin85B offers protection similar to BCG in a murine model of pulmonary tuberculosis and suggest that Sin85B-induced protection is dependent upon both innate and acquired immune mechanisms.
为了改进针对结核分枝杆菌的DNA疫苗接种,我们评估了一种表达结核抗原85B的基于辛德毕斯病毒的DNA构建体(Sin85B)的有效性。最初通过用强毒结核分枝杆菌对免疫的C57BL/6小鼠进行气溶胶攻击来评估Sin85B的保护效力。在感染后1个月和7个月时,与未免疫的对照小鼠相比,接种疫苗的小鼠肺部细菌载量显著降低,肺部病理状况得到改善。此外,结核攻击后,Sin85B免疫小鼠的平均存活期(305±9天)相对于未免疫小鼠(203±13天)延长了102天,并且基本上等同于牛分枝杆菌卡介苗接种小鼠的存活时间(294±15天)。通过显示接种疫苗小鼠的肺细胞在感染后相对于对照小鼠存在显著升高水平的γ干扰素(IFN-γ)mRNA,并通过证明攻击前IFN-γ的耗竭消除了疫苗诱导的保护作用,确定了IFN-γ在Sin85B介导的保护中的重要作用。Sin85B免疫CD4-/-小鼠诱导的大量抗结核保护反应强烈表明CD8细胞部分介导Sin85B诱导的保护性免疫。有趣的是,Sin85B疫苗接种不能保护RNase L-/-(先天抗病毒反应中的关键酶)小鼠,而在用卡介苗或表达抗原85B的传统DNA质粒免疫的RNase L-/-小鼠中检测到显著的保护作用。这些数据表明,在肺结核小鼠模型中,用Sin85B免疫提供的保护与卡介苗相似,并表明Sin85B诱导的保护依赖于先天和后天免疫机制。