Project description:The purpose of this project was to identify venom proteins from the venom gland of Leptopilina boulardi (strain Lb17), a parasitoid wasp species that infects fruit flies in the genus Drosophila.
Project description:The purpose of this project was to identify venom proteins from the venom gland of Leptopilina heterotoma (strain Lh14), a parasitoid wasp species that infects fruit flies in the genus Drosophila.
Project description:The purpose of this project was to identify venom proteins from the venom gland of Ganaspis hookeri (strain GhFl, formerly 'G1'), a parasitoid wasp species that infects fruit flies in the genus Drosophila.
Project description:Multinucleated giant hemocytes (MGHs) represent a novel type of blood cell in insects that participate in a highly efficient immune response against parasitoid wasps involving isolation and killing of the parasite. Previously we showed that circulating MGHs have high motility and interaction with the parasitoid rapidly triggers encapsulation, structural and molecular mechanisms behind these processes remained elusive. Here, we use detailed ultrastructural analysis of MGHs and also live cell imaging to study encapsulation in Drosophila ananassae after parasitoid wasp infection. We found dynamic structural changes, mainly driven by the formation of diverse vesicular systems and a large variety of newly developed intracytoplasmic membrane organizations, moreover abundant generation of giant cell exosomes (GCE) in the MGHs. Moreover, we used RNA sequencing to study the transcriptomic profile of MGHs and the activated plasmatocytes 72 hours after infection, as well as the uninduced blood cells. This reveals that differentiation of MGHs is accompanied by broad changes in gene expression. Consistent with the observed structural changes, transcripts mainly related to vesicular function, cytoskeletal organization and adhesion were enriched in MGHs. In addition, transmembrane receptors were upregulated, which may be important for parasitoid recognition. Our results reveal coordinated molecular and structural changes in the course of MGH differentiation and parasitoid encapsulation, providing a mechanistic model for a powerful innate immune response.
Project description:We identified genes regulated by parasitization of the silkworm Bombyx mori by three tachinid parasitoid species, Exorista japonica, Drino inconspicuoides and Pales pavida, using oligonucleotide microarrays. The numbers of genes and their intensity of expression varied with the species of parasitoid, within silkworm hemocytes and fat body.
Project description:Although host-parasitoid interactions are becoming well characterized at the organismal and cellular levels, much remains to be understood of the molecular bases for the host immune response and the parasitoidsâ ability to defeat this immune response. Leptopilina boulardi and L. heterotoma, two closely related, highly infectious natural parasitoids of Drosophila melanogaster, appear to use very different infection strategies at the cellular level. Here, we further characterize cellular level differences in the infection characteristics of these two wasp species using newly derived, virulent inbred strains, and then use whole genome microarrays to compare the transcriptional response of Drosophila to each. While flies attacked by the melanogaster group specialist Leptopilina boulardi (strain Lb17) up-regulate numerous genes encoding proteolytic enzymes, components of the Toll and JAK/STAT pathways, and the melanization cascade as part of a combined cellular and humoral innate immune response, flies attacked by the generalist L. heterotoma (strain Lh14) do not appear to initiate an immune transcriptional response at the time points post-infection we assayed, perhaps due to the rapid venom-mediated lysis of host hemocytes (blood cells). Thus, the specialist parasitoid appears to invoke a full-blown immune response in the host, but suppresses and/or evades downstream components of this response. Given that activation of the host immune response likely depletes the energetic resources of the host, the specialistâs infection strategy seems relatively disadvantageous. However, we uncover the mechanism for one potentially important fitness tradeoff of the generalistâs highly immune suppressive infection strategy. Experiment Overall Design: The parasitoid wasps L. boulardi and L. heterotoma were allowed to attack late second instar D. melanogaster larvae (72 hrs old at 22ËC) in the following manner. Nine petri dishes containing 60 fly larvae were each exposed to six experienced L. boulardi (strain Lb17) female wasps for 2 hrs, another nine plates were exposed to five L. heterotoma (strain Lh14) females, and nine control plates were left uninfected. For each of three time points post-infection (2-5 hrs, 9-12 hrs, 21-24 hrs), 40 larvae from three replicate plates were removed and frozen at -80ËC for RNA extraction and microarray analysis (3 treatments x 3 time points x 3 replicates = 27 samples total).