Modifying the anti-influenza A virus activity of collectins using recombinant protein technology
Friday 3 July 2009
Prof.dr Henk Haagsman and dr Edwin Veldhuizen
Project
Influenza is a highly contagious respiratory disease of humans and animals and recent developments with (almost) pandemic Mexican H1N1 illustrate the major threat of this fast spreading disease to public health all over the globe. An important class of mammalian innate immune proteins known as “the collectins”, is involved in the early pulmonary response to limit infection and spread of influenza A virus (IAV) in the airways. These proteins help to prevent viral entry into lung cells via different mechanisms (e.g. aggregation, opsonization) to fight the virus infection.
The repertoire of collectins and effectivity of collectin-mediated neutralization of IAV can seriously differ between animal species. Recent studies in our lab on porcine innate immune proteins indicate that glycan-modifications of lung collectins can result in substantially enhanced IAV neutralization activity in vitro. In addition, it was found for porcine surfactant protein D (pSP-D), by far the most effective collectin molecule in terms of IAV-neutralization described so far, that several porcinespecific structural elements in the lectin domain are likely to be involved in generating the distinct inhibitory activity of pSP-D against IAV. This project will focus on the identification and combination of these structural features by designing and expressing modified recombinant human collectins. We will produce our recombinant proteins in collaboration with U-Protein Express BV (also based at Utrecht University premises) by use of their HEK293 mammalian cell expression system. During the course of this “student’s project”, we will design novel collectins, prepare constructs by cDNA cloning and various DNA manipulation techniques using various vector systems, protein purification strategies, biochemical characterization and protein analysis of the final product (purity, size, content).
The anti-IAV activity and the spectrum of IAV strains that can be inhibited by our modified collectins will be assessed in vitro and evaluated to gain insight into the structural features that are important in providing strong and acute protection against a broad-range of IAV strains. These experiments will be done in collaboration with the Virology Department of Erasmus Medical Center Rotterdam (Dr. Guus Rimmelzwaan) and will reflect the final stage of this project.
Contact
Dr. M. van Eijk, 030 - 253 5361, m.vaneijk@uu.nl
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