HCMV Project
Thursday 18 March 2010
Project
The host adaptive immune system is of crucial importance for protection against infections and tumor growth. The immune response itself, however, needs to be tightly regulated to prevent immune-mediated diseases. One important component of the immune regulatory network is the subset of FoxP3pos regulatory T-cells, which can strongly dampen or even prevent unwanted immune responses.
Many viruses co-evolved with their mammalian hosts and have found different ways to adapt to antiviral (immune) responses. We now hypothesize that human cytomegalovirus (HCMV) has developed a novel strategy to target the FoxP3pos regulatory T-cells; in specific, the HCMV protein glycoprotein B (gB) may directly interact with FoxP3pos regulatory T-cells, thereby enhancing their suppressive capacity. Thus, HCMV could have developed a mechanism to dampen the HCMV-specific host response, i.e. by enhancing FoxP3pos regulatory T-cell function.
The aim of this student project is to test this hypothesis and identify a novel HCMV immune evasion strategy. The gene encoding HCMV (soluble) gB will be cloned and expressed in insect cells. Isolated recombinant gB protein will then be added to T-cell suppression assays, in which the suppressive capacity of FoxP3pos regulatory T-cells is determined. Differences in suppressive capacity between FoxP3pos regulatory T-cells incubated in the presence versus absence of recombinant gB will reveal whether gB indeed influences regulatory T-cell function. Subsequently, mechanistic details of this viral interference will be elucidated. Taken together, the experiments of this project may reveal an entirely novel mechanism for viral immune escape.
Techniques
Cellular assays: maintaining, differentiating and propagating mammalian and insect cells in vitro suppression assays
Molecular biology: DNA digestion, ligation, PCR, bacterial transformation, plasmid preps, baculovirus infection and protein expression & purification
Duration
6 or 9 months
Contact
Maaike Ressing, 088 75 506 03, m.ressing@umcutrecht.nl
More info
This project is a collaboration between the department of Medical Microbiology at the UMC Utrecht and the department of Immunology at Veterinary Sciences.
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