Novartis Institute for Tropical Diseases, Singapore, Singapore.
Antimicrob Agents Chemother. 2011 Aug;55(8):3677-83. doi: 10.1128/AAC.00136-11. Epub 2011 May 31.
Mycobacterium tuberculosis is the causative agent of a pulmonary epidemic that is estimated to infect one-third of the world's population and that has an increased incidence of multidrug resistance. The evaluation of new chemical entities against M. tuberculosis is hampered by the lack of biological tools to help predict efficacy, from early drug development to clinical trials. As the rat is the animal species of choice in the pharmaceutical industry, we have developed a rat model of acute and chronic phases of M. tuberculosis infection for drug efficacy testing. In this model, we have evaluated the impact of tuberculosis drugs on T cell response using the enzyme-linked immunospot assay methodology. Infected rats treated with isoniazid (INH) or rifampin (RIF) responded to therapy, the potency of which was comparable to that seen in the mouse. Peripheral blood mononuclear cells from infected rats produced gamma interferon (IFN-γ) in response to RD-1 antigens, such as the 6-kDa early secretory antigen target (ESAT-6) and the 10-kDa culture filtrate protein (CFP-10). A decrease in IFN-γ spot-forming cells (SFCs) was consistently observed in response to drug treatment. In both the acute- and chronic-phase models, the T cell response was more sensitive to ESAT-6 than to CFP-10. The SFC count in response to ESAT-6 appears to be an indicator of bacterial killing in the rat. Collectively, our data suggest that the ESAT-6 response could be used as a potential surrogate of drug efficacy in the rat and that such a readout could help shorten drug testing during preclinical development.
结核分枝杆菌是一种肺部传染病的病原体,据估计,这种传染病感染了世界人口的三分之一,而且其多药耐药性的发病率正在上升。由于缺乏有助于预测疗效的生物工具,从早期药物开发到临床试验,对结核分枝杆菌的新化学实体的评估受到了阻碍。由于大鼠是制药行业中首选的动物物种,我们已经开发了一种大鼠急性和慢性结核分枝杆菌感染模型,用于药物疗效测试。在该模型中,我们使用酶联免疫斑点测定法评估了结核药物对 T 细胞反应的影响。用异烟肼(INH)或利福平(RIF)治疗的感染大鼠对治疗有反应,其效力与在小鼠中观察到的相当。感染大鼠的外周血单核细胞对 RD-1 抗原(如 6-kDa 早期分泌抗原靶标(ESAT-6)和 10-kDa 培养滤液蛋白(CFP-10))产生伽马干扰素(IFN-γ)。在用药物治疗时,IFN-γ 斑点形成细胞(SFC)的数量持续减少。在急性和慢性模型中,T 细胞反应对 ESAT-6 的敏感性均高于 CFP-10。对 ESAT-6 的 SFC 计数似乎是大鼠中细菌杀伤的指标。总的来说,我们的数据表明 ESAT-6 反应可作为大鼠药物疗效的潜在替代指标,并且这种检测结果可以帮助缩短临床前开发期间的药物测试。