Effect of nutrient deprivation on the susceptibility ofGalleria mellonellalarvae to infection

Banville, Nessa; Browne, Niall; Kavanagh, Kevin

doi:10.4161/viru.21972

Cited by 63 publications

(62 citation statements)

References 32 publications

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“…Hemocyte depletion has previously been correlated with insect mortality in infection 3,52 , although recent results suggest that this picture is more complex than first belived 37 . Recently, it has been shown that starvation of larvae leads to increased susceptibility to infection through a suppression of immune responses 53 . In the assays described here, larvae were not fed for the duration of the study and it is not known how well fed larvae would respond to L. pneumophila infection.…”

Section: Discussionmentioning

confidence: 99%

Use of Galleria mellonella as a Model Organism to Study Legionella pneumophila Infection

Harding

Schroeder

Collins

et al. 2013

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Citation: Harding, C.R., Schroeder, G.N., Collins, J.W., Frankel, G. Use of Galleria mellonella as a Model Organism to Study Legionella pneumophila Infection. J. Vis. Exp. (81), e50964, doi:10.3791/50964 (2013). AbstractLegionella pneumophila, the causative agent of a severe pneumonia named Legionnaires' disease, is an important human pathogen that infects and replicates within alveolar macrophages. Its virulence depends on the Dot/Icm type IV secretion system (T4SS), which is essential to establish a replication permissive vacuole known as the Legionella containing vacuole (LCV). L. pneumophila infection can be modeled in mice however most mouse strains are not permissive, leading to the search for novel infection models. We have recently shown that the larvae of the wax moth Galleria mellonella are suitable for investigation of L. pneumophila infection. G. mellonella is increasingly used as an infection model for human pathogens and a good correlation exists between virulence of several bacterial species in the insect and in mammalian models. A key component of the larvae's immune defenses are hemocytes, professional phagocytes, which take up and destroy invaders. L. pneumophila is able to infect, form a LCV and replicate within these cells. Here we demonstrate protocols for analyzing L. pneumophila virulence in the G. mellonella model, including how to grow infectious L. pneumophila, pretreat the larvae with inhibitors, infect the larvae and how to extract infected cells for quantification and immunofluorescence microscopy. We also describe how to quantify bacterial replication and fitness in competition assays. These approaches allow for the rapid screening of mutants to determine factors important in L. pneumophila virulence, describing a new tool to aid our understanding of this complex pathogen.

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Section: Discussionmentioning

confidence: 99%

Use of Galleria mellonella as a Model Organism to Study Legionella pneumophila Infection

Harding

Schroeder

Collins

et al. 2013

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“…G. mellonella larvae were acquired from Alcotán Valencia SLU and kept at 21°C in darkness with a nonrestricted diet. It has been reported that food deprivation of G. mellonella larvae leads to reductions in cellular and immune responses (38). Larvae were used within 12 Infection of G. mellonella larvae.…”

Section: Methodsmentioning

confidence: 99%

Modeling Klebsiella pneumoniae Pathogenesis by Infection of the Wax Moth Galleria mellonella

et al. 2013

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The implementation of infection models that approximate human disease is essential for understanding pathogenesis at the molecular level and for testing new therapies before they are entered into clinical stages. Insects are increasingly being used as surrogate hosts because they share, with mammals, essential aspects of the innate immune response to infections. We examined whether the larva of the wax moth Galleria mellonella could be used as a host model to conceptually approximate Klebsiella pneumoniae-triggered pneumonia. We report that the G. mellonella model is capable of distinguishing between pathogenic and nonpathogenic Klebsiella strains. Moreover, K. pneumoniae infection of G. mellonella models some of the known features of Klebsiella-induced pneumonia, i.e., cell death associated with bacterial replication, avoidance of phagocytosis by phagocytes, and the attenuation of host defense responses, chiefly the production of antimicrobial factors. Similar to the case for the mouse pneumonia model, activation of innate responses improved G. mellonella survival against subsequent Klebsiella challenge. Virulence factors necessary in the mouse pneumonia model were also implicated in the Galleria model. We found that mutants lacking capsule polysaccharide, lipid A decorations, or the outer membrane proteins OmpA and OmpK36 were attenuated in Galleria. All mutants activated G. mellonella defensive responses. The Galleria model also allowed us to monitor Klebsiella gene expression. The expression levels of cps and the loci implicated in lipid A remodeling peaked during the first hours postinfection, in a PhoPQ-and PmrAB-governed process. Taken together, these results support the utility of G. mellonella as a surrogate host for assessing infections with K. pneumoniae. In 1890, Robert Koch formulated Koch's postulates as general guidelines for identifying disease-causing organisms. One century later, Stanley Falkow established the molecular version of Koch's postulates, this time to guide the identification of microbial genes encoding virulence factors. One of the key points of the molecular postulates is to test the virulence of a microorganism with an inactivated candidate virulence gene in an appropriate animal model. Therefore, the use of animal models to identify the virulence factors of human pathogens is indispensable. Currently, identification and characterization of novel virulence factors rely largely on assessing mutant bacteria for growth in the organs of infected mice. The dependence on mouse infection models limits large-scale analysis of virulence due to the large number of animals needed to obtain statistically significant results.To circumvent these issues, the search for alternative host models is ongoing. Ideally, these alternative models should be easy to maintain and infect, should be amenable to genetic manipulation, and should model aspects of vertebrate defenses upon infection, chiefly the immune response. The immune defense consists of two main parts, an innate and an adaptive response, wit...

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“…G. mellonella is an established and powerful model for the investigation of gut microbe homeostasis and foodborne diseases. [14][15][16] This species is also being investigated as a source for novel anti-infectives 17 and the availability of a comprehensive transcriptome allows the detailed molecular analysis of its immune response by transcriptional profiling and mass spectrometry. 18 We added bacteria to the G. mellonella larval diet to experimentally mimic the most common routes of natural exposure to microbes.…”

Section: Introductionmentioning

confidence: 99%

The maternal transfer of bacteria can mediate trans-generational immune priming in insects

et al. 2014

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Effect of nutrient deprivation on the susceptibility ofGalleria mellonellalarvae to infection

Cited by 63 publications

References 32 publications

Use of <em>Galleria mellonella</em> as a Model Organism to Study <em>Legionella pneumophila</em> Infection

Use of <em>Galleria mellonella</em> as a Model Organism to Study <em>Legionella pneumophila</em> Infection

Modeling Klebsiella pneumoniae Pathogenesis by Infection of the Wax Moth Galleria mellonella

The maternal transfer of bacteria can mediate trans-generational immune priming in insects

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