Okada M, Kita Y, Nakajima T, Kanamaru N, Hashimoto S, Nishida Y, Nakatani H, Takao K, Kishigami C, Nishimatsu S, Sekine Y, Inoue Y, Nagasawa T, Kaneda Y, Yoshida S, Matsumoto M, Paul Saunderson, Tan E V, Cruz E C Dela, N McMurray D, Sakatani M
Clinical Research Center, National Hospital Organization Kinki-chuo Chest Medical Center, 1180 Nagasone, Kitaku, Sakai, Osaka 591-8555, Japan.
Ikeda Laboratory, GenomIdea Inc.,1-8-31, Midorigaoka, Ikeda, Osaka 530-0043, Japan.
Procedia Vaccinol. 2010;2(1):34-39. doi: 10.1016/j.provac.2010.03.007. Epub 2010 Apr 14.
We have developed a novel tuberculosis (TB) vaccine; a combination of the DNA vaccines expressing mycobacterial heat shock protein 65 (HSP65) and interleukin 12 (IL-12) delivered by the hemagglutinating virus of Japan (HVJ)-envelope and -liposome (HSP65 + IL-12/HVJ). An IL-12 expression vector (IL-12DNA) encoding single-chain IL-12 proteins comprised of p40 and p35 subunits were constructed. This vaccine provided remarkable protective efficacy in mouse and guinea pig models compared to the BCG vaccine on the basis of C.F.U of number of TB, survival, an induction of the CD8 positive CTL activity and improvement of the histopathological tuberculosis lesions. This vaccine also provided therapeutic efficacy against multi-drug resistant TB (MDR-TB) and extremely drug resistant TB (XDR-TB) (prolongation of survival time and the decrease in the number of TB in the lung) in murine models. Furthermore, we extended our studies to a cynomolgus monkey model, which is currently the best animal model of human tuberculosis. This novel vaccine provided a higher level of the protective efficacy than BCG based upon the assessment of mortality, the ESR, body weight, chest X-ray findings and immune responses. All monkeys in the control group (saline) died within 8 months, while 50% of monkeys in the HSP65+hIL-12/HVJ group survived more than 14 months post-infection (the termination period of the experiment). Furthermore, the BCG priming and HSP65 + IL-12/HVJ vaccine (booster) by the priming-booster method showed a synergistic effect in the TB-infected cynomolgus monkey (100% survival). In contrast, 33% of monkeys from BCG Tokyo alone group were alive (33% survival). Furthermore, this vaccine exerted therapeutic efficacy (100% survival) and augmentation of immune responses in the TB-infected monkeys. These data indicate that our novel DNA vaccine might be useful against including XDR-TB and MDR-TB for human therapeutic clinical trials.
我们研发了一种新型结核病（TB）疫苗；它是由日本血凝病毒（HVJ）包膜和脂质体递送的、表达分枝杆菌热休克蛋白65（HSP65）和白细胞介素12（IL-12）的DNA疫苗的组合（HSP65 + IL-12/HVJ）。构建了一种编码由p40和p35亚基组成的单链IL-12蛋白的IL-12表达载体（IL-12DNA）。基于结核杆菌数量的菌落形成单位（C.F.U）、存活率、CD8阳性CTL活性的诱导以及组织病理学结核病变的改善，与卡介苗相比，这种疫苗在小鼠和豚鼠模型中提供了显著的保护效力。这种疫苗在鼠模型中还对耐多药结核病（MDR-TB）和广泛耐药结核病（XDR-TB）具有治疗效果（延长存活时间并减少肺部结核杆菌数量）。此外，我们将研究扩展到了食蟹猴模型，这是目前人类结核病的最佳动物模型。基于死亡率、红细胞沉降率（ESR）、体重、胸部X光检查结果和免疫反应的评估，这种新型疫苗比卡介苗提供了更高水平的保护效力。对照组（生理盐水）的所有猴子在8个月内死亡，而HSP65+hIL-12/HVJ组50%的猴子在感染后存活超过14个月（实验终止期）。此外，通过初免 - 加强方法使用卡介苗初免和HSP65 + IL-12/HVJ疫苗（加强）在感染结核的食蟹猴中显示出协同效应（100%存活）。相比之下，仅接种东京卡介苗组33%的猴子存活（33%存活）。此外，这种疫苗在感染结核的猴子中发挥了治疗效果（100%存活）并增强了免疫反应。这些数据表明，我们的新型DNA疫苗可能对包括XDR-TB和MDR-TB在内的结核病在人类治疗性临床试验中有用。
