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Honeybee (Apis Mellifera) and Bumblebee (Bombus Terrestris) Venom: Analysis and Immunological Importance of the Proteome

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Honeybee (Apis Mellifera) and Bumblebee (Bombus Terrestris) Venom: Analysis and Immunological Importance of the Proteome Department of Physiology (WE15) Laboratory of Zoophysiology Honeybee (Apis mellifera) and bumblebee (Bombus terrestris) venom: analysis and immunological importance of the proteome Het gif van de honingbij (Apis mellifera) en de aardhommel (Bombus terrestris): analyse en immunologisch belang van het proteoom Matthias Van Vaerenbergh Ghent University, 2013 Thesis submitted to obtain the academic degree of Doctor in Science: Biochemistry and Biotechnology Proefschrift voorgelegd tot het behalen van de graad van Doctor in de Wetenschappen, Biochemie en Biotechnologie Supervisors: Promotor: Prof. Dr. Dirk C. de Graaf Laboratory of Zoophysiology Department of Physiology Faculty of Sciences Ghent University Co-promotor: Prof. Dr. Bart Devreese Laboratory for Protein Biochemistry and Biomolecular Engineering Department of Biochemistry and Microbiology Faculty of Sciences Ghent University Reading Committee: Prof. Dr. Geert Baggerman (University of Antwerp) Dr. Simon Blank (University of Hamburg) Prof. Dr. Bart Braeckman (Ghent University) Prof. Dr. Didier Ebo (University of Antwerp) Examination Committee: Prof. Dr. Johan Grooten (Ghent University, chairman) Prof. Dr. Dirk C. de Graaf (Ghent University, promotor) Prof. Dr. Bart Devreese (Ghent University, co-promotor) Prof. Dr. Geert Baggerman (University of Antwerp) Dr. Simon Blank (University of Hamburg) Prof. Dr. Bart Braeckman (Ghent University) Prof. Dr. Didier Ebo (University of Antwerp) Dr. Maarten Aerts (Ghent University) Prof. Dr. Guy Smagghe (Ghent University) Dean: Prof. Dr. Herwig Dejonghe Rector: Prof. Dr. Anne De Paepe The author and the promotor give the permission to use this thesis for consultation and to copy parts of it for personal use. Every other use is subject to the copyright laws, more specifically the source must be extensively specified when using results from this thesis. Please refer to this work as: Van Vaerenbergh, M. (2013) Honeybee (Apis mellifera) and bumblebee (Bombus terrestris) venom: analysis and immunological importance of the proteome. PhD thesis, Ghent University. Funding: This study was financially supported by a PhD grant for Strategic Basic Research from the agency for Innovation by Science and Technology (IWT). Author’s email address: [email protected] Front cover Photographs © http://www.abeezhoney.net/wp-content/uploads/2011/12/HoneyStock1.jpg © http://commons.wikimedia.org/wiki/File:Bumblebee_2007-04-19.jpg Design: Quinten De Corte Curriculum vitae Curriculum vitae PERSONAL DETAILS Name Matthias Van Vaerenbergh Date of birth 19/04/1986 Nationality Belgian Telephone 0498/679827 E-mail [email protected] EDUCATION 2009-2013 Ph.D. candidate, Biochemistry and Biotechnology, Ghent University (Grant: IWT-Flanders) PhD thesis: Honeybee (Apis mellifera) and bumblebee (Bombus terrestris) venom: analysis and immunological importance of the proteome. 2007-2009 Master in Science (MSc), Biology, Ghent University (magna cum laude) Master thesis: Venom of honeybees (Apis mellifera): immunological importance of new venom components. 2004-2007 Bachelor in Science (BSc), Biology, Ghent University (cum laude) Bachelor thesis: Study of zebrafish (Danio rerio) mutants with an aberrant tooth phenotype. 1998-2004 Secondary school: Science-mathematics, KTA “De Rijdtmeersen”, Brakel SCHOLARSHIPS AND AWARDS Ph.D. scholarship of the Institute for the Promotion of Innovation by Science and Technology in Flanders (IWT-Flanders), Belgium, 01/01/2010 - 31/12/2013 INTERNSHIPS 28/01/2012 - 10/03/2012: Institute of Biochemistry and Molecular Biology, Hamburg University, Germany Curriculum vitae SCIENTIFIC OUTPUT A1 peer reviewed publications 1. Cardoen D., Ernst U. R., Van Vaerenbergh M., Boerjan B., de Graaf D. C., Wenseleers T., Schoofs, L. and Verleyen, P. (2011) Differential proteomics in dequeened honeybee colonies reveals lower viral load in hemolymph of fertile worker bees. PLoS ONE 6(6): e20043. 2. Van Vaerenbergh M., Cardoen D., Formesyn E. M., Brunain M., Van Driessche G., Blank S., Spillner E., Verleyen P., Wenseleers T., Schoofs L., Devreese B., de Graaf D. C. (2013) Extending the honey bee venome with the antimicrobial peptide apidaecin and a protein resembling wasp antigen 5. Insect Molecular Biology 22(2): 199–210. 3. Van Vaerenbergh M., Debyser G., Devreese B., de Graaf D. C. Exploring the hidden honeybee (Apis mellifera) venom proteome by integrating a combinatorial peptide ligand library approach with FTMS. Journal of Proteomics: in press. 4. Van Vaerenbergh M., Debyser G., Smagghe G., Devreese B., de Graaf D. C. Unraveling the venom proteome of the bumblebee (Bombus terrestris) by integrating a combinatorial peptide ligand library approach with FT-ICR MS. Toxicon: in press. Participation at international conferences 1. BEEDOC Workshop: Diagnostics in honeybees: from sampling to data analyses, Ghent, Belgium, August 2010. Van Vaerenbergh M., Jacobs F. J., Devreese B., de Graaf D. C. Molecular cloning reveals the existence of multiple alternative splice variants of hyaluronidase and icarapin in honeybee venom glands. p. 22. (poster) 2. 4th International Symposium on Molecular Allergology, Munich, Germany, October 2010, p. 26. Van Vaerenbergh M., Devreese B., de Graaf D. C. Exploring the honeybee (Apis mellifera) venom transcriptome using a PCR-based cloning approach. (poster) 3. 17th Benelux Congress of Zoology: Classic Biology in Modern Times, Ghent, Belgium, October 2010, p. 156. Van Vaerenbergh M., Devreese B., de Graaf D. C. Identification of novel isoforms of PVF-1 and icarapin in honeybee venom glands using an RT-PCR-based approach. (poster) 4. Bee–together: conference on pollinators with emphasis to stimulate interactions within the field, Ghent, Belgium, December 2010, p. 17. Van Vaerenbergh M., Devreese B., de Graaf D. C. Mass spectrometry as a powerful tool for Hymenoptera venom analysis. (oral presentation) 5. 17th Congress of the European Section of the International Society on Toxinology. Valencia, Spain, September 2011, p. 235. Van Vaerenbergh M. An in-depth analysis of the honeybee (Apis mellifera) and bumblebee (Bombus terrestris) venom proteome. (poster) 6. EAACI-WAO World Allergy & Asthma Congress. Milan, Italy, 22-26 June 2013, p. 44. Van Vaerenbergh M., De Smet L., Blank S., Spillner E., Ebo D., Jakob T., Devreese B., de Graaf D.C. IgE recognition of multiple novel Api m 10 isoforms evaluated by protein array technology. (oral presentation) Curriculum vitae GUIDANCE OF STUDENTS Educational support (practicals, exercises, seminars) First Bachelor of Medicine and Dentistry: Aspects of peripheral blood research: 2009-2010, 2010- 2011 Second Bachelor of Biochemistry and Biotechnology: Practical exercises of biomedical physiology: 2009-2010, 2010-2011, 2012-2013 Thesis support Master thesis: The venom of honeybees (Apis mellifera): immunological importance of the venom compounds icarapin and PVF1. (Kim De Crem, 2009-2010) Master project: Recombinant expression of honeybee venom compounds in an insect cell expression system. (Nele Van Der Steen, 2010-2011) Bachelor thesis: Determination of the serum IgG4-titers against honeybee venom and Api m 10 variants. (Sarah Delbaere, Stef Deyaert, Lieven Lancksweerdt, 2012-2013) Dankwoord Dankwoord Ongelooflijk, de tijd vliegt! Meer dan vier jaar geleden stond ik vol zenuwen dit doctoraatsproject te verdedigen voor de IWT-jury. Met succes, zo bleek achteraf. Nu, met een afgewerkt proefschrift voor mij, kan ik met grote voldoening terugkijken op de fantastische ervaring die dit doctoraatsproject is geweest. Natuurlijk zou dit niet gerealiseerd zijn zonder de hulp van anderen. Bij deze wil ik dus beginnen om iedereen te bedanken die een bijdrage heeft geleverd aan dit proefschrift. Eerst en vooral bedank ik mijn beide promotoren, Prof. Dirk de Graaf en Prof. Bart Devreese, voor hun excellente begeleiding, hun vertrouwen in mij, hun niet-aflatende steun en het ter beschikking stellen van hun labo. Mijn dank gaat ook uit naar de overige leden van mijn examencommissie en begeleidingscommissie: Prof. Grooten, Prof. Baggerman, Prof. Braeckman, Prof. Ebo, Prof. Smagghe, Dr. Aerts en Dr. Blank. Dankzij jullie terechte opmerkingen is de kwaliteit van dit werk aanzienlijk verbeterd. Hiernaast dank ik IWT-Vlaanderen voor de financiële steun die dit project mogelijk maakte. Ook dank ik alle co-auteurs van de verschillende hoofdstukken van dit proefschrift nog eens voor hun bijdrage aan dit onderzoek. In het bijzonder wil ik Griet Debyser bedanken om mij wegwijs te maken in de wereld van massaspectrometrie en Dr. Lina De Smet voor de praktische begeleiding doorheen het project. Of course I also want to thank Dr. Simon Blank and Prof. Edzard Spillner to allow me to do a 6-week internship in the Institute of Biochemistry and Molecular Biology at Hamburg University. Special thanks to Simon, Frankie and Yvonne who guided me through the lab work. Thanks Laura, Julia, Thorsten, Yvonne, Andrea, Helene, Simon, Edzard, Frankie, Melanie, Sara, Tim, Dirk and Thomas for the pleasant stay in the lab, for teaching me very interesting German words and for organizing social activities such as walking on the frozen Dankwoord Alster lake, movie night, for having a spicy lunch in the Mensa and going out for dinner. It was a great experience for me to live and work in Hamburg! Verder bedank ik de collega’s van het Laboratorium voor Zoöfysiologie voor alle hulp tijdens het doctoraat, maar ook voor het aangenaam gezelschap, de goede werksfeer en toffe babbels. Marleen, bedankt voor de administratieve
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