- Molecular virology and cell biology of RNA virus infections
- Antiviral innate immunity
- Viral innate immune evasion
- Development of antiviral strategies
Department of Medical Microbiology
PI Innate immunity and innate immune evasion
PI Ultrastructure and function of viral replication complexes
PI Vaccine development
The importance of innate immunity for the combat against viruses has become very clear in the past years. These responses commence very soon after the beginning of infection, and ultimately lead to an “antiviral state” which is very unfavourable for the replication of viruses. Among the gene products induced by this system are factors that suppress translation, or which recognize and degrade viral RNA species. Also induction of a proper adaptive immune response largely depends on a full-blown innate immune reaction.
Virtually all viruses possess mechanisms to evade or suppress innate immune responses, in order to create a “window-of-opportunity” to enable efficient replication of the viral genome and ensure a successful start of the infection. We study (innate) immune evasion strategies for viruses with a +RNA genome, particularly the Nidoviruses, among which are the severe acquired respiratory syndrome coronavirus (SARS-CoV), Middle East Respiratory coronavirus (MERS-CoV), equine arteritis virus (EAV) and porcine reproductive and respiratory syndrome virus (PRRSV). We focus especially on viral inhibition of ubiquitin-regulated innate immune signalling.
One of the first processes in the viral life-cycle after entry onto the cell is the formation of a replication-enzyme complex. For viruses with a +RNA genome this complex is associated with elaborate intracellular membrane modifications, which involve morphological change of specific cellular organelles into virus replication “organelles”. These structures are essential to the viral replication, negatively influence cellular processes, and they ultimately contribute to cytopathic effect (CPE) and cell death. We study the morphogenesis and architecture of the replication organelles induced by +RNA viruses, as well as their interactions with cellular pathways and processes such as the innate immune response.
Potential targets for antiviral strategies, which yield from our research results, are occasionally developed further, in most cases together with a commercial or academic partner.
Marjolein Kikkert (1969) studied Molecular Sciences, and graduated from Wageningen University and Research Center (WUR) with a specialisation in Molecular Biology, in 1993. She subsequently started a PhD research project at the Department of Virology of this university, under supervision of Dr. Richard Kormelink and late Prof. Rob Goldbach. In 1999, this resulted in a PhD degree and a thesis entitled “Role of the envelope glycoproteins in the infection of tomato spotted wilt virus”. Dr. Kikkert became a postdoc in the laboratory of Prof. Emmanuel Wiertz at the National Institute of Health and the Environment (RIVM) in Bilthoven, and later at Leiden University Medical Center (LUMC). She switched to the laboratory of Prof. Eric Snijder at the LUMC in 2006, where she started as a postdoc, and developed into a senior researcher. In 2011 she became an Assistant Professor in the department of Medical Microbiology of the LUMC, and her research group currently investigates replication-associated membrane modifications of the Nidoviruses, as well as antiviral innate immunity and immune evasion strategies of these viruses. In part, this research is supported by grants from the Netherlands Organisation for Scientific Research (NWO), and by a research contract with a large company. Dr. Kikkert supervised more than 20 undergraduate students during lab rotations (3-9 months each) in the course of her carreer. She is the daily supervisor and anticipated co-promotor of two or three PhD students and she also supervises research technicians. She reviews manuscripts for a number of scientific journals on an ad hoc basis and participates in selection committees for NWO grants. She participates in several teaching activities, mostly for bachelor and master Biomedical Sciences students at the LUMC. Together with colleagues she for example organises and teaches the bachelor course “Infectious agents and Immunity”, and she is co-coordinator of the frontiers-of-science (FOS) course "Molecular Virology of RNA viruses" in the master of Biomedical Sciences.
Oudshoorn D, Rijs K, Limpens RWAL, Groen K, Koster AJ, Snijder EJ, Kikkert M, Bárcena M. (2017) Expression and Cleavage of Middle East Respiratory Syndrome Coronavirus nsp3-4 Polyprotein Induce the Formation of Double-Membrane Vesicles That Mimic Those Associated with Coronaviral RNA Replication. MBio. Nov 21;8(6). pii: e01658-17. doi: 10.1128/mBio.01658-17.
Scutigliani EM, Kikkert M. (2017) Interaction of the innate immune system with positive-strand RNA virus replication organelles. Cytokine Growth Factor Rev. Oct;37:17-27. doi:10.1016/j.cytogfr.2017.05.007.
Bailey-Elkin BA, Knaap RCM, Kikkert M, Mark BL (2017) Structure and Function of Viral Deubiquitinating Enzymes. J Mol Biol. Nov 10;429(22):3441-3470. doi: 10.1016/j.jmb.2017.06.010.
Zhang W, Bailey-Elkin BA, Knaap RCM, Khare B, Dalebout TJ, Johnson GG, van Kasteren PB, McLeish NJ, Gu J, He W, Kikkert M, Mark BL, Sidhu SS. (2017) Potent and selective inhibition of pathogenic viruses by engineered ubiquitin variants. PLoS Pathog. May 18;13(5):e1006372. doi: 10.1371/journal.ppat.1006372.
Oudshoorn D, van der Hoeven B, Limpens RW, Beugeling C, Snijder EJ, Bárcena M, Kikkert M. (2016) Antiviral Innate Immune Response Interferes with the Formation of Replication-Associated Membrane Structures Induced by a Positive-Strand RNA Virus. MBio. Dec 6;7(6). pii: e01991-16. doi: 10.1128/mBio.01991-16.
Rabouw HH, Langereis MA, Knaap RC, Dalebout TJ, Canton J, Sola I, Enjuanes L, Bredenbeek PJ, Kikkert M, de Groot RJ, van Kuppeveld FJ. (2016) Middle East Respiratory Coronavirus Accessory Protein 4a Inhibits PKR-Mediated Antiviral Stress Responses. PLoS Pathog. Oct 26;12(10):e1005982. doi: 10.1371/journal.ppat.1005982.
van der Hoeven B, Oudshoorn D, Koster AJ, Snijder EJ, Kikkert M, Bárcena M. (2016) Biogenesis and architecture of arterivirus replication organelles. Virus Res. Jul 15;220:70-90. doi: 10.1016/j.virusres.2016.04.001.
de Wilde AH, Wannee KF, Scholte FE, Goeman JJ, Ten Dijke P, Snijder EJ, Kikkert M, van Hemert MJ. (2015) A Kinome-Wide Small Interfering RNA Screen Identifies Proviral and Antiviral Host Factors in Severe Acute Respiratory Syndrome Coronavirus Replication, Including Double-Stranded RNA-Activated Protein Kinase and Early Secretory Pathway Proteins. J Virol. Aug;89(16):8318-33. doi: 10.1128/JVI.01029-15
Bailey-Elkin BA, Knaap RC, Johnson GG, Dalebout TJ, Ninaber DK, van Kasteren PB, Bredenbeek PJ, Snijder EJ, Kikkert M, Mark BL. (2014) Crystal structure of the Middle East respiratory syndrome coronavirus (MERS-CoV) papain-like protease bound to ubiquitin facilitates targeted disruption of deubiquitinating activity to demonstrate its role in innate immune suppression. J Biol Chem. Dec 12;289(50):34667-82. doi: 10.1074/jbc.M114.609644.
Snijder EJ, Kikkert M, Fang Y. (2013) Arterivirus molecular biology and pathogenesis. J Gen Virol. 2013; 94:2141-63.
van Kasteren PB, Bailey-Elkin BA, James TW, Ninaber DK, Beugeling C, Khajehpour M, Snijder EJ, Mark BL, Kikkert M. (2013) Deubiquitinase function of arterivirus papain-like protease 2 suppresses the innate immune response in infected host cells. Proc Natl Acad Sci U S A. 2013; 110:E838-47.
Knoops K, Kikkert M, Worm SH, Zevenhoven-Dobbe JC, van der Meer Y, Koster AJ, Mommaas AM, Snijder EJ. (2008) SARS-coronavirus replication is supported by a reticulovesicular network of modified endoplasmic reticulum. PLoS Biol. 2008; 6:e226.
Frias-Staheli N, Giannakopoulos NV, Kikkert M, Taylor SL, Bridgen A, Paragas J, Richt JA, Rowland RR, Schmaljohn CS, Lenschow DJ, Snijder EJ, García-Sastre A, Virgin HW 4th. (2007) Ovarian tumor domain-containing viral proteases evade ubiquitin- and ISG15-dependent innate immune responses. Cell Host Microbe. 2007; 2:404-16.
Leids Universitair Medisch Centrum
2333 ZA Leiden
2300 RC Leiden