In vivo Bioluminescence Imaging of Burkholderia mallei Respiratory Infection and Treatment in the Mouse Model.
In vivo Bioluminescence Imaging of Burkholderia mallei Respiratory Infection and Treatment in the Mouse Model.
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DOI：
10.3389/fmicb.2011.00174
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发表时间：
2011
影响因子：
5.2
通讯作者：
Torres AG
中科院分区：
文献类型：
--
作者：
Massey S;Johnston K;Mott TM;Judy BM;Kvitko BH;Schweizer HP;Estes DM;Torres AG
生物发光成像(BLI)技术是监测传染病进展和治疗方法的有力工具。BLI对于追踪可能很难从某些器官恢复的挑剔的细胞内病原体特别有用。马来伯克霍尔德氏菌(Burkholderia Mallei)是一种兼性的细胞内病原菌，由于其高度传染性和潜在的生物武器用途，已被美国疾控中心列为B类选择性病原体。对扁桃体的发病机制或治疗方法知之甚少。我们研究了使用生物发光报告结构来实时监测感染的动态以及治疗方法的疗效。利用pUTmini-Tn5：：LuxKm2构建了一株稳定的发光报告菌株B.Mallei，并用于监测BALB/c小鼠模型中的腺体。通过鼻腔途径感染5 × 10 3细菌，在24、48和72 h进行BLI监测。我们验证了我们的报告构建物与野生型Mallei保持了相似的毒力和生长动力学，并证实了它在有或没有抗生素选择的情况下都保持了发光稳定性。这种发光信号最初出现在肺部，在感染过程中逐渐传播到肝脏和脾。我们证明，感染后24 小时的抗生素治疗导致了生物发光的减少，这可以归因于靶器官中细菌负荷的减少。这些发现表明，BLI可用于监测腺体感染期间的疾病进展和治疗效果。最后，我们报告了一种替代mini-TN5：：LuxKm2转座子的方法，该方法使用mini-TN7-lux元件，该元件在已知的基因组连接位置特异性地插入，也可用于标记细菌。
Bioluminescent imaging (BLI) technology is a powerful tool for monitoring infectious disease progression and treatment approaches. BLI is particularly useful for tracking fastidious intracellular pathogens that might be difficult to recover from certain organs. Burkholderia mallei, the causative agent of glanders, is a facultative intracellular pathogen and has been classified by the CDC as a Category B select agent due to its highly infectious nature and potential use as a biological weapon. Very little is known regarding pathogenesis or treatment of glanders. We investigated the use of bioluminescent reporter constructs to monitor the dynamics of infection as well as the efficacy of therapeutics for B. mallei in real-time. A stable luminescent reporter B. mallei strain was created using the pUTmini-Tn5::luxKm2 plasmid and used to monitor glanders in the BALB/c murine model. Mice were infected via the intranasal route with 5 × 103 bacteria and monitored by BLI at 24, 48, and 72 h. We verified that our reporter construct maintained similar virulence and growth kinetics compared to wild-type B. mallei and confirmed that it maintains luminescent stability in the presence or absence of antibiotic selection. The luminescent signal was initially seen in the lungs, and progressed to the liver and spleen over the course of infection. We demonstrated that antibiotic treatment 24 h post-infection resulted in reduction of bioluminescence that can be attributed to decreased bacterial burden in target organs. These findings suggest that BLI can be used to monitor disease progression and efficacy of therapeutics during glanders infections. Finally, we report an alternative method to mini-Tn5::luxKm2 transposon using mini-Tn7-lux elements that insert site-specifically at known genomic attachment sites and that can also be used to tag bacteria.
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