Real-Time Bioluminescence Imaging of Mixed Mycobacterial Infections

被引:18
作者
Chang, MiHee [1 ]
Anttonen, Katri P. [1 ]
Cirillo, Suat L. G. [1 ]
Francis, Kevin P. [2 ]
Cirillo, Jeffrey D. [1 ]
机构
[1] Texas A&M Hlth Sci Ctr, Dept Microbial Pathogenesis & Immunol, Bryan, TX 77807 USA
[2] PerkinElmer, Alameda, CA USA
来源
PLOS ONE | 2014年 / 9卷 / 09期
基金
美国国家卫生研究院;
关键词
IN-VIVO; BACTERIAL PATHOGENS; BOVIS BCG; TUBERCULOSIS; IDENTIFICATION; EXPRESSION; REPORTER; MACROPHAGE; CONSTRUCTION; REPLICATION;
D O I
10.1371/journal.pone.0108341
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
Molecular analysis of infectious processes in bacteria normally involves construction of isogenic mutants that can then be compared to wild type in an animal model. Pathogenesis and antimicrobial studies are complicated by variability between animals and the need to sacrifice individual animals at specific time points. Live animal imaging allows real-time analysis of infections without the need to sacrifice animals, allowing quantitative data to be collected at multiple time points in all organs simultaneously. However, imaging has not previously allowed simultaneous imaging of both mutant and wild type strains of mycobacteria in the same animal. We address this problem by using both firefly (Photinus pyralis) and click beetle (Pyrophorus plagiophthalamus) red luciferases, which emit distinct bioluminescent spectra, allowing simultaneous imaging of two different mycobacterial strains during infection. We also demonstrate that these same bioluminescence reporters can be used to evaluate therapeutic efficacy in real-time, greatly facilitating our ability to screen novel antibiotics as they are developed. Due to the slow growth rate of mycobacteria, novel imaging technologies are a pressing need, since they can they can impact the rate of development of new therapeutics as well as improving our understanding of virulence mechanisms and the evaluation of novel vaccine candidates.
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页数:13
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