A novel biosafety level 2 compliant tuberculosis infection model using a ΔleuDΔpanCD double auxotroph of Mycobacterium tuberculosis H37Rv and Galleria mellonella
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 mai...
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Taylor & Francis Group
2020
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oai:doaj.org-article:572ae8bfa51e40ebb1855c127c6572d02021-11-17T14:21:58ZA novel biosafety level 2 compliant tuberculosis infection model using a ΔleuDΔpanCD double auxotroph of Mycobacterium tuberculosis H37Rv and Galleria mellonella2150-55942150-560810.1080/21505594.2020.1781486https://doaj.org/article/572ae8bfa51e40ebb1855c127c6572d02020-12-01T00:00:00Zhttp://dx.doi.org/10.1080/21505594.2020.1781486https://doaj.org/toc/2150-5594https://doaj.org/toc/2150-5608Mammalian 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.Masanori AsaiYanwen LiJohn SpiropoulosWilliam CooleyDavid EverestBrian D. RobertsonPaul R. LangfordSandra M. NewtonTaylor & Francis Grouparticlegalleria mellonellatuberculosisinfection modelmycobacterium tuberculosis complexdrug screeningantimycobacterial agentsmycobacteriaauxotrophicInfectious and parasitic diseasesRC109-216ENVirulence, Vol 11, Iss 1, Pp 811-824 (2020) |
| institution |
DOAJ |
| collection |
DOAJ |
| language |
EN |
| topic |
galleria mellonella tuberculosis infection model mycobacterium tuberculosis complex drug screening antimycobacterial agents mycobacteria auxotrophic Infectious and parasitic diseases RC109-216 |
| spellingShingle |
galleria mellonella tuberculosis infection model mycobacterium tuberculosis complex drug screening antimycobacterial agents mycobacteria auxotrophic Infectious and parasitic diseases RC109-216 Masanori Asai Yanwen Li John Spiropoulos William Cooley David Everest Brian D. Robertson Paul R. Langford Sandra M. Newton A novel biosafety level 2 compliant tuberculosis infection model using a ΔleuDΔpanCD double auxotroph of Mycobacterium tuberculosis H37Rv and Galleria mellonella |
| description |
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. |
| format |
article |
| author |
Masanori Asai Yanwen Li John Spiropoulos William Cooley David Everest Brian D. Robertson Paul R. Langford Sandra M. Newton |
| author_facet |
Masanori Asai Yanwen Li John Spiropoulos William Cooley David Everest Brian D. Robertson Paul R. Langford Sandra M. Newton |
| author_sort |
Masanori Asai |
| title |
A novel biosafety level 2 compliant tuberculosis infection model using a ΔleuDΔpanCD double auxotroph of Mycobacterium tuberculosis H37Rv and Galleria mellonella |
| title_short |
A novel biosafety level 2 compliant tuberculosis infection model using a ΔleuDΔpanCD double auxotroph of Mycobacterium tuberculosis H37Rv and Galleria mellonella |
| title_full |
A novel biosafety level 2 compliant tuberculosis infection model using a ΔleuDΔpanCD double auxotroph of Mycobacterium tuberculosis H37Rv and Galleria mellonella |
| title_fullStr |
A novel biosafety level 2 compliant tuberculosis infection model using a ΔleuDΔpanCD double auxotroph of Mycobacterium tuberculosis H37Rv and Galleria mellonella |
| title_full_unstemmed |
A novel biosafety level 2 compliant tuberculosis infection model using a ΔleuDΔpanCD double auxotroph of Mycobacterium tuberculosis H37Rv and Galleria mellonella |
| title_sort |
novel biosafety level 2 compliant tuberculosis infection model using a δleudδpancd double auxotroph of mycobacterium tuberculosis h37rv and galleria mellonella |
| publisher |
Taylor & Francis Group |
| publishDate |
2020 |
| url |
https://doaj.org/article/572ae8bfa51e40ebb1855c127c6572d0 |
| work_keys_str_mv |
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