Sarcotoxin-1C

Insects have the phenomenon of immune priming by which they can have enhanced protection against reinfection with the same pathogen, and this immune protection can be passed on to their offspring, which is defined as "trans-generational immune priming (TGIP)." But whether housefly possesses TGIP is still unclear. Therefore, we used the housefly as the insect model and Candida albicans as the pathogen to explore whether the housefly is capable of eliciting TGIP, and RNA sequencing (RNA-seq) was performed to explore the molecular mechanism of TGIP of the housefly. We found that the housefly possesses TGIP, and adults pre-exposed to heat-killed C. albicans could confer protection to itself and its offspring upon reinfection with a lethal dose of C. albicans. RNA-seq results showed that 30 and 154 genes were differentially expressed after adults were primed with heat-killed C. albicans (CA-A) and after offspring larvae were challenged with a lethal dose of C. albicans (CA-CA-G), respectively. Among the differentially expressed genes (DEGs), there were 23 immune genes, including 6 pattern recognition receptors (PRRs), 7 immune effectors, and 10 immunoregulatory molecules. More importantly, multiple DEGs were involved in the Toll signaling pathway and phagosome signaling pathway, suggesting that the Toll signaling pathway and phagocytosis might play important roles in the process of TGIP of housefly to C. albicans. Our results expanded on previous studies and provided parameters for exploring the mechanism of TGIP.

Some insects display immunological priming as a result of elevated humoral and cellular responses which give enhanced survival against subsequent infection. The humoral immune response of Galleria mellonella larvae following pre-exposure to heat killed Staphylococcus aureus or Candida albicans cells was determined by quantitative mass spectrometry in order to assess the relationship between the humoral immune response and resistance to subsequent bacterial or fungal infection. Larvae pre-exposed to heat killed S. aureus showed increased resistance to subsequent bacterial and fungal infection. Larvae displayed an increased hemocyte density (14.08 ± 2.14 × 106 larva-1 (p < 0.05) compared to the PBS injected control [10.41 ± 1.67 × 106 larva-1]) and increased abundance of antimicrobial proteins (cecropin-D-like peptide (+22.23 fold), hdd11 (+12.61 fold) and prophenol oxidase activating enzyme 3 (+5.96 fold) in response to heat killed S. aureus. Larvae pre-exposed to heat killed C. albicans cells were resistant to subsequent fungal infection but not bacterial infection and showed a reduced hemocyte density (6.01 ± 1.63 × 106 larva-1 (p < 0.01) and increased abundance of hdd11 (+32.73 fold) and moricin-like peptide C1 (+16.76 fold). While immune priming is well recognised in G. mellonella larvae the results presented here indicate distinct differences in the response of larvae following exposure to heat killed bacterial and fungal cells.

PGRP (peptidoglycan recognition protein) is a conserved protein family that recognizes the peptidoglycan in bacterial cell wall and causes the activation of various innate immune responses. Previous studies have reported that PGRP-SCs in Drosophila dampen the activation of Immune Deficiency (Imd) pathway to microbial infection, and participate in the lifespan extension of the insects. To facilitate understanding the function of PGRP-SCs from an evolutionary angle, we identified and functionally characterized the PGRP-SC gene in the housefly Musca domestica, a species that has adapted to a septic environment much harsher than the natural habitat of Drosophila. The gene designated as MdPGRP-SC was found most abundantly expressed in the 3rd instar larvae, and is expressed at this developmental stage predominantly in the gut. MdPGRP-SC was virtually unchanged in whole larvae after a septic injury at the second larval instar, while two antimicrobial peptides (AMPs), diptericin and attacin, were upregulated in the first 24h but not later. Through dsRNA microinjection, MdPGRP-SC was knocked down by RNA interference (RNAi), and caused the significant increased expression of diptericin and attacin. The pupation of MdPGRP-SC-depleted larvae was severely suppressed compared to controls. Opposite to the expression trend of MdPGRP-SC, a spontaneous active expression of diptericin and attacin was found in pre-pupae but not in third instar larvae. Taken together, our study reveals that downregulation of MdPGRP-SC leads to the overexpression of the AMPs, and is involved in the larvae-to-pupa transition of housefly.