A novel biosafety level 2 compliant tuberculosis infection model using a ΔleuDΔpanCD double auxotroph of Mycobacterium tuberculosis H37Rv and Galleria mellonella

Virulence

David Everest¹  William Cooley¹  John Spiropoulos¹  Brian D. Robertson²  Paul R. Langford³  Masanori Asai³  Sandra M. Newton³  Yanwen Li³ 

[1] Department of Pathology, Animal and Plant Health Agency, Addlestone, U;MRC Centre for Molecular Bacteriology and Infection, Department of Infectious Disease, Imperial College London, London, U;Section of Paediatric Infectious Disease, Department of Infectious Disease, Imperial College London, London, U;

关键词: Galleria mellonella;    tuberculosis;    infection model;    Mycobacterium tuberculosis;    drug screening;    antimycobacterial agents;    mycobacteria;    auxotrophic;   

DOI  :  10.1080/21505594.2020.1781486

来源: Taylor & Francis

【 摘 要 】

Mammalian infection models have contributed significantly to our understanding of the host-mycobacterial interaction, revealing potential mechanisms and targets for novel antimycobacterial therapeutics. However, the use of conventional mammalian models such as mice, are typically expensive, high maintenance, require specialized animal housing, and are ethically regulated. Furthermore, research using Mycobacterium tuberculosis (MTB), is inherently difficult as work needs to be carried out at biosafety level 3 (BSL3). The insect larvae of Galleria mellonella (greater wax moth), have become increasingly popular as an infection model, and we previously demonstrated its potential as a mycobacterial infection model using Mycobacterium bovis BCG. Here we present a novel BSL2 complaint MTB infection model using G. mellonella in combination with a bioluminescent ΔleuDΔpanCD double auxotrophic mutant of MTB H37Rv (SAMTB lux) which offers safety and practical advantages over working with wild type MTB. Our results show a SAMTB lux dose dependent survival of G. mellonella larvae and demonstrate proliferation and persistence of SAMTB lux bioluminescence over a 1 week infection time course. Histopathological analysis of G. mellonella, highlight the formation of early granuloma-like structures which matured over time. We additionally demonstrate the drug efficacy of first (isoniazid, rifampicin, and ethambutol) and second line (moxifloxacin) antimycobacterial drugs. Our findings demonstrate the broad potential of this insect model to study MTB infection under BSL2 conditions. We anticipate that the successful adaptation and implementation of this model will remove the inherent limitations of MTB research at BSL3 and increase tuberculosis research output.

【 授权许可】

CC BY