Tom
van der Most

Project

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Description PhD project

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Research questions / objectives

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Timeline

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Tags matching with the contents of track 14/24

Methods

• Labwork

  • PCR: Presence of USUV & WNV in field-caught Dutch Culex pipiens will be determined with PCR. PCR also used for regular laboratory work.
  • RNA/DNA extraction: RNA (and DNA) will be isolated from mosquito samples for RNA-seq (virome analysis) and PCR. Similar work for mosquito cell-lines.
  • In vivo: The immune response of Culex pipiens to arboviruses will be assayed in laboratory-held mosquito colony in Wageningen. Vector competence studies may also be performed with isolated insect-specific viruses.
  • In vitro: Culex cell-lines will be used for determining the transcriptome of the Toll, IMD and/or JAK-STAT pathway. Cell-lines also for culturing of viruses from field-caught mosquitoes.
  • Next generation sequencing (NGS): RNA-seq will be used for detection of insect-specific viruses in Dutch mosquito samples and to determine the transcriptome in response to Toll, IMD or JAK-STAT pathway activation.
  • Molecular Cloning: Cloning work will be performed to create Ago2-KO cell-lines and in vitro reporter assays for immune pathway activation.
  • Plaque Assay: For in vivo studies and in vitro virus culture, plaque assays will be used (or equivalent techniques) to determine virus presence and amount.

  Fieldwork

  • Mosquito trapping: Mosquitoes caught by trapping will be analysed for arboviral presence and virome analysis.

  Literature

  • Literature review: Review will be written to provide an overview of anti-viral immunity genes in mosquito species and Drosophila.

Topics

• Vector:

  • Vector immune response: Vector immune response will be tested on laboratory-held Culex pipiens and interventions in the immune signalling pathways will be performed to investigate their role in the immune response.
  • Vector susceptibility: Vector susceptibility/Vector competence studies will be performed in the acquirement of an insect-specific virus from Dutch Culex pipiens to see potential for intervention.
  • Vector competence: Vector susceptibility/Vector competence studies will be performed in the acquirement of an insect-specific virus from Dutch Culex pipiens to see potential for intervention.

  Virus:

  • Virus-mosquito interaction: The impact of viruses (both arboviruses and insect-specific viruses) on the mosquito immune response will be analysed in laboratory-held Culex pipiens colony.

  Interventions:

  • Other interventions: In the mapping of the virome for Dutch Culex pipiens, we hope to acquire an insect-specific virus that can be used to restrict vector competence of arboviruses.

Species

• Species:

  • Mosquito: The project looks at arboviral presence and naturally-infecting viruses in Dutch Culex pipiens and the Culex pipiens anti-viral immune response.

Virus

• Virus:

  • Virus (general): This project will analyse the Dutch Culex pipiens samples for the presence of all viruses deemed relevant within the One Health pact.
  • West Nile virus: Presence of WNV in Dutch Culex pipiens will be determined in this project. WNV will also be used as a model arbovirus when looking at the anti-viral immune response and the effect of insect-specific viruses on vector competence.
  • Usutu Virus: Presence of USUV in Dutch Culex pipiens will be determined in this project. USUV will also be used as a model arbovirus when looking at the anti-viral immune response and the effect of insect-specific viruses on vector competence.