Moricin-like peptide C1

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.

Screening for components with antifungal activity in the hemolymph of immune-stimulated Galleria mellonella larvae led to the identification of four novel moricin-like peptides (A, B, C3 and D). Subsequently, eight moricin-like peptide genes (A, B, C1-5 and D) were isolated and shown to code for seven unique peptides (mature C4 and C5 are identical). These genes contained single introns which varied from 180 to 1090bp. The moricin-like peptides were particularly active against filamentous fungi, preventing the growth of Fusarium graminearum at 3 microg/ml, and were also active against yeasts, gram positive bacteria and gram negative bacteria. Searches of the databases identified 30 moricin-like peptide genes which code for 23 unique mature peptides, all belonging to the Lepidoptera (moths and butterflies). The first comprehensive phylogenetic analysis of the moricin-like peptides suggested that they fall into two basic classes which diverged a long time ago. The peptides have since diversified extensively through a high level of gene duplication within species, as seen in G. mellonella and Bombyx mori. The restriction of moricin-like peptides to the Lepidoptera combined with their potent antifungal activity suggests that this diverse peptide family may play a role in the defence response of moths and butterflies.