Using Bioluminescence To Monitor Treatment Response in Real Time in Mice with Mycobacterium ulcerans Infection
Using Bioluminescence To Monitor Treatment Response in Real Time in Mice with Mycobacterium ulcerans Infection
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DOI：
10.1128/aac.01260-10
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发表时间：
2011-01-01
影响因子：
4.9
通讯作者：
Nuermberger, Eric L.
中科院分区：
文献类型：
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作者：
Zhang, Tianyu;Li, Si-Yang;Nuermberger, Eric L.
溃疡分枝杆菌引起布鲁里溃疡，这是一种可能致残的溃疡性皮肤病。直到最近，抗菌疗法才被证明是有效的。在小鼠足垫模型实验证明具有杀菌活性后，首次提出用利福平加链霉素治疗 2 个月。尽管较大的溃疡可能需要辅助手术，但这种治疗现在被认为是首选治疗方法。较短的口服方案是理想的，但溃疡分枝杆菌生长非常缓慢，需要 3 个月才能产生可计数的菌落，从而阻碍了对小鼠药物活性的评估。我们创建了表达哈维弧菌 luxAB 的重组生物发光溃疡分枝杆菌菌株，用于实时评估体内抗菌效果。小鼠足垫注射野生型溃疡分枝杆菌 1059 (WtMu) 或重组生物发光菌株 (rMu)。两周后，小鼠接受利福平加链霉素、单独卡那霉素（rMu 具有耐药性）或单独链霉素治疗 4 周，并观察足垫肿胀（预防模型）。未经治疗的对照组和卡那霉素治疗的 rMu 感染小鼠在出现足垫肿胀后接受利福平加链霉素 4 周（治疗模型）。与WtMu相比，rMu表现出相似的体内生长和毒力以及相似的药物敏感性。脚垫匀浆中的发光（以相对光单位测量）和活细菌数量（以 CFU 测量）之间观察到良好的相关性。还展示了对活体小鼠药物活性的连续实时评估的概念验证。这些结果表明生物发光作为小鼠足垫中活细菌的实时替代标记的潜力，可以加速布鲁里溃疡新疗法的鉴定。
Mycobacterium ulcerans causes Buruli ulcer, a potentially disabling ulcerative skin disease. Only recently was antimicrobial therapy proven effective. Treatment for 2 months with rifampin plus streptomycin was first proposed after experiments in the mouse footpad model demonstrated bactericidal activity. This treatment is now considered the treatment of choice, although larger ulcers may require adjunctive surgery. Shorter, oral regimens are desired, but evaluating drug activity in mice is hampered by the very slow growth of M. ulcerans, which takes 3 months to produce countable colonies. We created a recombinant bioluminescent M. ulcerans strain expressing luxAB from Vibrio harveyi for real-time evaluation of antimicrobial effects in vivo. Mouse footpads were injected with wild-type M. ulcerans 1059 (WtMu) or the recombinant bioluminescent strain (rMu). Two weeks later, mice received rifampin plus streptomycin, kanamycin alone (to which rMu is resistant), or streptomycin alone for 4 weeks and were observed for footpad swelling (preventive model). Untreated controls and kanamycin-treated rMu-infected mice received rifampin plus streptomycin for 4 weeks after developing footpad swelling (curative model). Compared to WtMu, rMu exhibited similar growth and virulence in vivo and similar drug susceptibility. A good correlation was observed between luminescence (measured as relative light units) and number of viable bacteria (measured by CFU) in footpad homogenates. Proof of concept was also shown for serial real-time evaluation of drug activity in live mice. These results indicate the potential of bioluminescence as a real-time surrogate marker for viable bacteria in mouse footpads to accelerate the identification of new treatments for Buruli ulcer.