Mycobacterial Research Program, Centenary Institute, Sydney, New South Wales, Australia.
PLoS One. 2012;7(5):e34991. doi: 10.1371/journal.pone.0034991. Epub 2012 May 2.
The long-term control of tuberculosis (TB) will require the development of more effective anti-TB vaccines, as the only licensed vaccine, Mycobacterium bovis bacille Calmette-Guérin (BCG), has limited protective efficacy against infectious pulmonary TB. Subunit vaccines have an improved safety profile over live, attenuated vaccines, such as BCG, and may be used in immuno-compromised individuals. MPT83 (Rv2873) is a secreted mycobacterial lipoprotein expressed on the surface of Mycobacterium tuberculosis. In this study, we examined whether recombinant MPT83 is recognized during human and murine M. tuberculosis infection. We assessed the immunogenicity and protective efficacy of MPT83 as a protein vaccine, with monophosphyl lipid A (MPLA) in dimethyl-dioctadecyl ammonium bromide (DDA) as adjuvant, or as a DNA vaccine in C57BL/6 mice and mapped the T cell epitopes with peptide scanning. We demonstrated that rMPT83 was recognised by strong proliferative and Interferon (IFN)-γ-secreting T cell responses in peripheral blood mononuclear cells (PBMC) from patients with active TB, but not from healthy, tuberculin skin test-negative control subjects. MPT83 also stimulated strong IFN-γ T cell responses during experimental murine M. tuberculosis infection. Immunization with either rMPT83 in MPLA/DDA or DNA-MPT83 stimulated antigen-specific T cell responses, and we identified MPT83(127-135) (PTNAAFDKL) as the dominant H-2(b)-restricted CD8(+) T cell epitope within MPT83. Further, immunization of C57BL/6 mice with rMPT83/MPLA/DDA or DNA-MPT83 stimulated significant levels of protection in the lungs and spleens against aerosol challenge with M. tuberculosis. Interestingly, immunization with rMPT83 in MPLA/DDA primed for stronger IFN-γ T cell responses to the whole protein following challenge, while DNA-MPT83 primed for stronger CD8(+) T cell responses to MPT83(127-135). Therefore MPT83 is a protective T cell antigen commonly recognized during human M. tuberculosis infection and should be considered for inclusion in future TB subunit vaccines.
结核分枝杆菌(TB)的长期控制需要开发更有效的抗结核疫苗,因为唯一获得许可的疫苗卡介苗(BCG)对传染性肺结核的保护效果有限。与 BCG 等活减毒疫苗相比,亚单位疫苗具有更好的安全性,可用于免疫功能低下的个体。MPT83(Rv2873)是一种分泌性分枝杆菌脂蛋白,表达于结核分枝杆菌表面。在这项研究中,我们研究了人结核分枝杆菌和鼠结核分枝杆菌感染期间是否会识别重组 MPT83。我们评估了 MPT83 作为蛋白疫苗,用单磷酰脂质 A(MPLA)与二油酰基磷脂酰乙醇胺-聚乙二醇 2000(DOPE-PEG2000)佐剂,或作为 DNA 疫苗在 C57BL/6 小鼠中的免疫原性和保护效力,并通过肽扫描定位 T 细胞表位。我们证明 rMPT83 在活动性结核患者的外周血单核细胞(PBMC)中可被强烈的增殖和干扰素(IFN)-γ分泌性 T 细胞识别,但在健康、结核菌素皮肤试验阴性的对照组中不能被识别。MPT83 也在实验性鼠结核分枝杆菌感染中刺激强烈的 IFN-γ T 细胞反应。用 rMPT83 与 MPLA/DOPE-PEG2000 或 DNA-MPT83 免疫可刺激抗原特异性 T 细胞反应,我们确定 MPT83(127-135)(PTNAAFDKL)是 MPT83 内的主要 H-2(b)-限制性 CD8(+) T 细胞表位。此外,用 rMPT83 与 MPLA/DOPE-PEG2000 或 DNA-MPT83 免疫 C57BL/6 小鼠可刺激肺部和脾脏对结核分枝杆菌气溶胶挑战的显著保护水平。有趣的是,rMPT83 与 MPLA/DOPE-PEG2000 免疫可增强对整个蛋白的 IFN-γ T 细胞反应,而 DNA-MPT83 免疫可增强对 MPT83(127-135)的 CD8(+) T 细胞反应。因此,MPT83 是一种保护性 T 细胞抗原,在人结核分枝杆菌感染期间普遍被识别,应考虑将其纳入未来的结核分枝杆菌亚单位疫苗。