Journal
FRONTIERS IN CELLULAR AND INFECTION MICROBIOLOGY
Volume 11, Issue -, Pages -Publisher
FRONTIERS MEDIA SA
DOI: 10.3389/fcimb.2021.619981
Keywords
Galleria mellonella; Mycobacterium bovis BCG; tuberculosis; innate immunity; in vivo model; proteomics; gene expression
Categories
Funding
- Biotechnology and Biological Sciences Research Centre (BBSRC) [BB/P001262/1]
- National Centre for the Replacement, Refinement and Reduction of Animals in Research (NC3Rs) [NC/R001596/1]
- Maynooth University doctoral studentship
- Science Foundation Ireland (SFI) [12/RI/2346]
- SFI
- European Regional Development Fund [12/RC/2275_P2]
- BBSRC [BB/P001262/1] Funding Source: UKRI
- MRC [MR/P028225/1] Funding Source: UKRI
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The study used the larvae of G. mellonella as a non-mammalian infection model for the Mycobacterium tuberculosis complex, characterizing the immune response through proteomic and transcriptomic approaches. The results showed distinct changes in the proteome at different time points post-infection, with inconsistencies between gene expression and protein abundance. Insights into the innate immune response and potential antimycobacterial agents were gained, validating the insect model as a cost-effective and ethically acceptable tool for studying MTBC members.
The larvae of the insect Galleria mellonella, have recently been established as a non-mammalian infection model for the Mycobacterium tuberculosis complex (MTBC). To gain further insight into the potential of this model, we applied proteomic (label-free quantification) and transcriptomic (gene expression) approaches to characterise the innate immune response of G. mellonella to infection with Mycobacterium bovis BCG lux over a 168 h time course. Proteomic analysis of the haemolymph from infected larvae revealed distinct changes in the proteome at all time points (4, 48, 168 h). Reverse transcriptase quantitative PCR confirmed induction of five genes (gloverin, cecropin, IMPI, hemolin, and Hdd11), which encoded proteins found to be differentially abundant from the proteomic analysis. However, the trend between gene expression and protein abundance were largely inconsistent (20%). Overall, the data are in agreement with previous phenotypic observations such as haemocyte internalization of mycobacterial bacilli (hemolin/beta-actin), formation of granuloma-like structures (Hdd11), and melanization (phenoloxidase activating enzyme 3 and serpins). Furthermore, similarities in immune expression in G. mellonella, mouse, zebrafish and in vitro cell-line models of tuberculosis infection were also identified for the mechanism of phagocytosis (beta-actin). Cecropins (antimicrobial peptides), which share the same alpha-helical motif as a highly potent peptide expressed in humans (h-CAP-18), were induced in G. mellonella in response to infection, giving insight into a potential starting point for novel antimycobacterial agents. We believe that these novel insights into the innate immune response further contribute to the validation of this cost-effective and ethically acceptable insect model to study members of the MTBC.
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