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ViroGenetics - BSL3 Laboratory of Virology

Group leader
prof. Krzysztof Pyrc

Follow me on ResearchGate   

 

Phone: +48 692 696 863
e-mail: k.a.pyrc@uj.edu.pl
Official website: http://virogenetics.info/
News chanell: https://www.facebook.com/virogenetics/

 

Team members

PhD students

  • Artur Szczepanski
  • Magda Pachota
  • Agnieszka Dabrowska
  • Pawel Botwina
  • Yuliya Chykunova
  • Aleksandra Synowiec
    Agnieszka Suder
    Emilia Barreto Durán
    Kevin Lie

Summary of current research

In our efforts we combine the basic research and utilize it to carry out applicatory projects. Current research of the Virogenetics group focuses on early steps of viral infection, namely (a) identification of viral attachment receptors / co-receptors, (b) identification of internalization pathways to the host cell, and (c) single virus trafficking in the cell. To make an example we mapped early events during infection with a number of coronaviruses.

In order to appropriately study the subject we utilize the most advanced ex vivo culture system HAE (human airway epithelium), which is constructed with fully differentiated, ciliated human epithelium growing on an interphase of air and liquid, to mimic the natural infection microenvironment.

The 3D cultures provide an excellent research model, and in order to excel it and broaden our expertise we are actively collaborating with other research groups, e.g., Amy Sims (University of North Carolina), Katja Wolthers (Universiteit van Amsterdam), Dirk Johmans (Catholic University of Leuven). We plan to further extend our expertise and to start the organoid lung cultures to study the biology of viral infection and to improve translation of ex vivo research to the clinic. As we try to map early events during viral infection, we developed a workbench for such research. This includes methods to visualize single viruses in context of the cell, receptors and markers of certain sub cellular structures / pathways. Even though we have started with members of Coronaviridae family we are now extending the study also to other viral species.

Our research efforts are aimed to understand the virus biology, yet we combine the know how and knowledge gained in such a way to carry out applicatory projects. Consequently, we are working on development of antiviral agents hampering virus entry / internalization. One of the examples of such activity is our fruitful collaboration with chemists from the group of prof. Maria Nowakowska, with whom we formed the interdisciplinary team working on novel polymeric drugs. Combining the know how gained on virus entry and early stages of infection we are able to precisely pinpoint the mechanism of action for such compounds, visualizing their activity in the cell. These efforts are rewarding, as we already developed several effective inhibitors, for which intellectual property is secured (e.g., P416749, P409016, P409015, PCT/PL2013/050032, PCT/PL2013/00064, P401707).

Further, some our innovations are already in the commercialization phase. The StartIt Vet company has been formed by investors to commercialize the HTCC molecule (cationically substituted chitosan), as anticoronaviral drug. Several other compounds were developed, inhibiting entry of influenza virus, human metapneumovirus, human rhinoviruses, human herpesviruses, and flaviviruses. For some of these we are currently negotiating the commercialization route with private investors and venture capital.

The second line of research is related to flaviviruses. This family of viruses encompasses several human and non human species, which cause severe disease in humans. The best known example is hepatitis C, which causes a world-wide pandemic. Yet, there are several other species that are of interest. These include tick-borne encephalitis, dengue virus, West Nile virus, and Zika virus.

In collaboration with excellent teams from AGH Krakow (group of prof. Jerzy Sillbering), Wroclaw (group of prof. Marcin Sienczyk), and Gdansk (group of prof. Adam Lesner), we initiated the work aimed to understand the biology of the virus.

Specific goals:

  1. Characterize flaviviral protease – substrate specificity, structural characterization.
  2. Identify cellular proteins that serve as partners for flaviviruses to understand the pathology of the infection.
  3. Develop novel inhibitors of flaviviral replication and entry.

 

It is also worth to mention that Virogenetics group gathers postdoctoral fellows, PhD students and MSc students. As one of my priorities is to enrich the experience our team we invite researchers to join our team and to work on satellite projects. One of the best examples is dr Anna Kula-Pacurar, who returned to Poland and to start the research within her Polonez grant. She will bring her unique experience in elucidating the interactions between biomolecules. Further, dr Justyna Ciejka, previously working on virus entry inhibitors, currently tries to use the developed biomaterials for fast diagnostics and virus/vaccine purification within her own project. Dr Antonina Naskalska utilizing our experience with coronaviruses aims to develop virus-like particles, which are a perfect tool to safely study early events during coronaviral infection, but also in the future may be used in applicatory projects.

Research plans

  1. Continuation of the current research.
  2. Development of novel antiviral compounds in collaboration with Polish and international research groups.
  3. Improvement of our 3D tissue-culture models and comparatory in vivo – ex vivo studies.
  4. Development of animal 3D tissue cultures to study zoonotic transmission of viruses. 
  5. Development of genetic engineering (lentiviruses, CRISPR-Cas) workbench to modify 3D tissue cultures to understand the biology of the infection.

Grants

 

  1.  2019. Anna Kula-Pacurar. Molecular mechanisms of HIV latency and reactivation from latency
  2. 2019-2022. Horizon2020. ITN networks. Organoids for Virus Research - An innovative training-ETN programme. PI for PL and WP coordinator: K Pyrc.
  3. 2018-2021. NCN Opus. Cellular proteases during coronaviral infection. PI: K Pyrc.
  4. 2018-2021. NCN Opus. Polymer-based systems for inhibition of Zika virus. PI: prof. M Nowakowska (KP: biological research coordinator).
  5. 2018-2020. NCN Preludium. Novel inhibitors of herpes virus protease. PI: M Pachota.6
  6. 2018-2019. NCN Preludium. Analysis of human coronavirus OC43 and Zika virus internalization routes in cells. Inhibition of the early steps of infection by intracellular pH tuning. PI: K Owczarek.
  7. 2017-2020: NCN Opus. Zika virus NS3 protease. PI: K Pyrc.
  8. 2017-2019: NCN Sonata. Hybrid adsorbents for concentrating, purification and detection viruses. PI: Justyna Ciejka.
  9. 2017-2019: NCN Polonez. Molecular dissection of the MATR3/PSF complex in the regulation of HIV-1 posttranscriptional latency: implications for therapy (MATR3PSFHIV) PI: Anna Kula-Pacurar
  10. 2015-2018: NCN Opus. Novel polyelectrolytes as potential inhibitors of HSV-1 virus - synthesis, properties and mechanism of action. PI: M Nowakowska. (KP: biological research coordinator).
  11. 2013-2018: NCN Sonata bis. Coronavirus and the cell: dyplomatic bag. PI: Krzysztof Pyrc.

Publications

  1. Synowiec A, Gryniuk I, Pachota M, Strzelec L, Roman O, Kłysik-Trzciańska K, Zając M, Drebot I, Gula K, Andruchowicz A, Rajfur Z, Szczubiałka K, Nowakowska K*, Pyrc K*. (2019). Cat flu: broad spectrum polymeric antivirals. Antiviral Research. in press.
  2. Naskalska A*, Dabrowska A, Szczepanski A, Milewska A, Jasik K, Pyrc K*.  (2019). Membrane protein of HCoV-NL63 is responsible for interaction with the adhesion receptor. Journal of Virology. in press.
  3. Pachota M, Kłysik-Trzciańska K, Synowiec A, Yukioka S, Yusa S, Zając, M, Zawilinska B, Dzieciątkowski T, Szczubiałka K, Pyrc K*, Nowakowska M*. (2019). Highly effective and safe polymeric inhibitors of herpes simplex virus in vitro and in vivo. ACS Applied Materials & Interfaces. in press.
  4. Labuz K, Adamowicz P, Kała M, Pyrc K, Reszke E, Mielczarek P, Silberring J, Smoluch M*. (2019). Detection of legal highs in the urine of methadone-treated patient by LC-MS. Basic and clinical pharmacology and toxicology. in press.
  5. Owczarek K, Chykunova Y, Jassoy C, Maksym B, Rajfur Z, Pyrc K. (2019) Zika virus: mapping and reprogramming the entry. (2019). Cell Communication and Signaling. in press.
  6. Szczepanski A, Owczarek K, Bzowska K, Gula K, Drebot I, Ochman M, Maksym B, Rajfur Z, Mitchell JA, Pyrc K. (2019) Canine Respiratory Coronavirus, Bovine Coronavirus and Human Coronavirus OC43: Receptors and attachment factors. Viruses. in press.
  7. Ciejka J, Botwina P, Nowakowska M, Szczubiałka K, Pyrc K. (2019) Synthetic sulfonated derivatives of poly(allylamine hydrochloride) as inhibitors of human metapneumovirus. PLoS One. in press.
  8. Gruba N, Igancio Rodrigez MJ, Wysocka M, Skorenski M, Grzywa R, Dabrowska A, Lecka M, Suder P, Sienczyk M, Pyrc K, Lesner A. (2018). One step beyond: design of substrates spanning primed positions of Zika virus NS2B-NS3 protease. ACS Medicinal Chemistry Letters. in press.
  9. Naskalska A, Dabrowska A, Nowak P, Szczepanski A, Jasik K, Milewska A, Ochman M, Zeglen S, Rajfur Z, Pyrc K. (2018). Novel coronavirus like particles targeting cells lining the respiratory tract. PLoS One. in press.
  10. Skorenski M, Milewska A, Pyrc K, Sienczyk M, Oleksyszyn J. (2018). Phosphonate inhibitors of West Nile virus NS2B/NS3 protease. Journal of Enzyme Inhibition and Medicinal Chemistry. in press.
  11. Szczepanski A, Owczarek K, Milewska A, Baster Z, Rajfur Z, Mitchell J, Pyrc K. (2018) Canine respiratory coronavirus employs caveolin-1-mediated pathway for internalization to HRT-18G cells. Veterinary Research. 49:55
  12. Owczarek K, Szczepanski A, Milewska A, Baster Z, Rajfur Z, Sarna M, Pyrc K. (2018) Early events during human coronavirus OC43 entry to the cell. Scientific Reports. 8(1):7124.
  13. Milewska A, Kindler E, Vkovski P, Zeglen S, Ochman M, Thiel V, Rajfur Z, Pyrc K.(2018) APOBEC3-mediated restriction of RNA virus replication. Scientific Reports. 8(1):5960.
  14. Milewska A, Nowak P, Owczarek K, Szczepanski A, Zarebski M, Hoang-Bujnowicz A, Berniak K, Wojarski J, Zeglen S, Baster Z, Rajfur Z, Pyrc K (2018) Entry of human coronavirus NL63 to the cell. Journal of Virology. 92(3). pii: e01933-17.
  15. Pachota M, Klysik K, Synowiec A, Ciejka J, Szczubialka K, Pyrc K, Nowakowska M. (2017) Inhibition of herpes simplex viruses by cationic dextran derivatives. Journal of Medicinal Chemistry. 60(20):8620-8630.
  16. Burmistrz M, Rodriguez Martinez JI, Krochmal D, Staniec D, Pyrc K. (2017) crRNAs in Porphyromonas gingivalis CRISPR-Cas I-C system. Journal of Bacteriology. 199(23). pii: e00275-17.
  17. Skalniak L, Zak K, Guzik K, Magiera K, Musielak B, Pachota M, Szelazek B, Kocik J, Grudnik P, Tomala M, Krzanik S, Pyrc K, Dömling A, Dubin G, Holak T. (2017) Small-molecule inhibitors of PD-1/PD-L1 immune checkpoint alleviate the PD-L1-induced exhaustion of T-cells. Oncotarget. 8(42):72167-72181.
  18. Sheahan TP, Sims AC, Graham RL, Menachery VD, Gralinski LE, Case JB, Leist SR, Pyrc K, Feng JY, Trantcheva I, Bannister R, Park Y, Babusis D, Clarke MO, Mackman RL, Spahn JE, Palmiotti CA, Siegel D, Ray AS, Cihlar T, Jordan R, Denison MR, Baric RS (2017) Broad-spectrum antiviral GS-5734 inhibits both epidemic and zoonotic coronaviruses. Science Translational Medicine, Vol. 9, Issue 396, eaal3653.
  19. Skorenski M, Pachota M, Pyrc K, Sienczyk M, Oleksyszyn J (2017) Novel Peptidyl α-Aminoalkylphosphonates as Inhibitors of Hepatitis C Virus NS3/4A Protease. Antiviral Research. 144:286-298.
  20. Ciejka J, Wolski K, Nowakowska M, Pyrc K¥, Szczubiałka K¥. (2017) Biopolymeric nano/microspheres for selective and reversible adsorption of coronaviruses. Materials Science & Engineering C. 76, 735–742.
  21. Szelazek B, Kabala W, Kus K, Zdzalik M, Twarda-Clapa A, Golik P, Burmistrz M, Florek D, Wladyka B, Pyrc K, Dubin G¥. (2017) Structural characterization of HCoV‑NL63 N-protein. Journal of Virology. 91(11). pii: e02503-16
  22. Gruba N*, Rodriguez Martinez JI*, Grzywa R, Wysocka M, Skoreński M, Burmistrz M, Łęcka M, Lesner A¥, Sieńczyk M¥, Pyrc K¥. (2016) Substrate profiling of Zika virus NS2B-NS3 protease. FEBS Letters. 590(20):3459-3468.
  23. Szychowska M, Siwek W, Pawolski D, Karani AA, Pyrc K, Bochtler M¥ (2016) Type III CRISPR complexes from Thermus thermophilus. Acta Biochimica Polonica. 63(2):377-86.
  24. Milewska A, Kaminski K, Ciejka J, Kosowicz K, Zeglen S, Wojarski J, Nowakowska M, Szczubialka K¥, Pyrc K¥ (2016) HTCC: broad range inhibitor of coronavirus entry. PLoS One. 11(6):e0156552.
  25. Ciejka J, Milewska A, Wytrwal M, Wojarski J, Golda A, Ochman M, Nowakowska M, Szczubialka K¥, Pyrc K¥. (2016) Novel polyanions inhibiting replication of influenza viruses. Antimicrobial Agents and Chemotherapy (AAC). 60(4):1955-66.
  26. Szczubialka K., Pyrc K., Nowakowska M¥. (2016) In search for effective and definitive treatment of Herpes Simplex Virus Type 1 (HSV-1) infections. RSC Advances. 6, 1058-1075.
  27. Benedyk M., Mydel P., Płaza K., Gawron K., Milewska A., Maresz K., Koziel J., Jotwani R., Pyrc K., Potempa J¥. (2016) Gingipains: critical factors in the development of aspiration pneumonia caused by Porphyromonas gingivalis. Journal of Innate Immunity. 8(2):185-98..
  28. Burmistrz M, Dudek B, Staniec D, Rodriguez Martinez JI, Bochtler M, Potempa J, Pyrc K¥. (2015). Functional analysis of Porphyromonas gingivalis W83 CRISPR-Cas systems. Journal of Bacteriology. 197(16):2631-41.
  29. Burmistrz M, Pyrc K¥. (2015). CRISPR/Cas systems in prokaryotes. Polish Journal of Microbiology. 64(3):193-202.
  30. Huang X, Dong W, Milewska A, Golda A, Qi Y, Zhu Q, Marasco W, Baric R, Sims A¥, Pyrc K¥, Li W¥, Sui J¥. (2015). HCoV-HKU1 Spike protein uses O-acetylated sialic acid as an attachment receptor determinant and employs HE protein as a receptor-destroying enzyme. Journal of Virology. 89(14):7202-13.
  31. Zabieglo K, Majewski P, Majchrzak-Gorecka M, Wlodarczyk A, Grygier B, Zegar A, Kapinska-Mrowiecka M, Naskalska A, Pyrc K, Dubin A, Wahl S, Cichy J¥. (2015). The inhibitory effect of secretory leukocyte protease inhibitor (SLPI) on formation of neutrophil extracellular traps. Journal of Leukocyte Biology. 98(1):99-106.
  32. Pyrc K¥. (2015). Ludzkie koronawirusy. Postępy Nauk Medycznych. t. XXVIII, nr 4B, 2015.
  33. Naskalska A, Pyrc K¥. (2015) Virus Like Particles as Immunogens and Universal Nanocarriers. Polish Journal of Microbiology. 64(1):3-13.
  34. Zuwala K, Golda A, Kabala W, Burmistrz M, Zdzalik M, Nowak P, Kedracka-Krok S, Zarebski M, Dobrucki J, Florek D, Zeglen S, Wojarski J, Potempa J, Dubin G, Pyrc K¥. (2015) The nucleocapsid protein of human coronavirus NL63. PLoS One. 20;10(2):e0117833.
  35. Milewska A, Zarebski M, Nowak P, Stozek K, Potempa J, Pyrc K¥. (2014) Human coronavirus NL63 utilize heparan sulfate proteoglycans for attachment to target cells. Journal of Virology. 88(22):13221-30.
  36. Gawron K, Łazarz–Bartyzel K, Łazarz M, Stęplewska K, Pyrc K, Potempa J, Chomyszyn–Gajewska M¥. (2014) In vitro testing the potential of a novel chimeric IgG variant for inhibiting collagen fibrils formation in recurrent hereditary gingival fibromatosis. Journal of Physiology and Pharmacology. 65(4):585-91.
  37. Koziel J¥, Bryzek D, Sroka A, Maresz K, Glowczyk I, Bielecka E, Kantyka T, Pyrc K, Svoboda P, Pohl J, Potempa J. (2014) Citrullination Alters Immunomodulatory Function of LL-37 Essential for Prevention of Endotoxin-Induced Sepsis. Journal of Immunology. 2014 Apr 25.
  38. Florek D, Burmistrz M, Potempa J, Pyrc K¥. Stability of infectious human coronavirus NL63 (2014) Journal of Virological Methods. S0166-0934(14)00136-0.
  39. Maresz K, Hellvard A, Sroka A, Adamowicz K, Bielecka E, Koziel J, Gawron K, Mizgalska D, Marcinska K, Benedyk M, Pyrc K, Quirke A, Jonsson R, Alzabin S, Venables P, Nguyen K, Mydel P¥, Potempa J¥. Porphyromonas gingivalis Facilitates the Development and Progression of Destructive Arthritis through Its Unique Bacterial Peptidylarginine Deiminase (PAD). (2013) PloS Pathogens. 9(9): e1003627.
  40. Pyrc K¥, Stozek K, Galan W, Potempa J. (2013) HexaPrime: a novel method for detection of coronaviruses. Journal of Virological Methods. 188 (1–2), 29–36.
  41. Milewska A, Ciejka J, Kaminski K, Karewicz A, Bielska D, Zeglen S, Karolak W, Nowakowska M, Potempa J, Bosch BJ, Pyrc K¥, Szczubialka K¥ (2013) Novel polymeric inhibitors of HCoV-NL63. Antiviral Research. 97(2):112-21.
  42. Kantyka T*, Pyrc K*¥, Gruca M, Smagur J, Plaza K, Guzik K, Zeglen S, Ochman M, Potempa J. (2013) Staphylococcus aureus proteases degrade lung surfactant protein A potentially impairing innate immunity of the lung. Journal of Innate Immunity. 5(3):251-60.
  43. Pyrc K¥, Milewska A, Kantyka T, Sroka A, Maresz K, Kozieł J, Nguyen KA, Enghild JJ., Knudsen AD, Potempa J. (2013) Inactivation of epidermal growth factor by Porphyromonas gingivalis as a potential mechanism for periodontal tissue damage. Infection and Immunity, 81(1):55.
  44. Milewska A, Pyrc K¥, Human RIG-1/Mda5 and TLR3 signaling pathways and their role in antiviral response during mucosal infections, Recent Research Developments in Virology, (2012) ISBN: 978-81-7895-563-6, Transworld Research Network.
  45. Dubin G¥, Koziel J., Pyrc K., Wladyka B., Potempa J. (2013) Bacterial proteases in disease – role in intracellular survival, evasion of coagulation/fibrinolysis innate defenses, toxicoses and viral infections. Current Pharmaceutical Design. 19(6):1090-113.
  46. Dijkman R, Jebbink MF, Deijs M, Milewska A, Pyrc K, Buelow E, van der Bijl A, van der Hoek L¥. (2012) Replication dependent downregulation of cellular ACE2 protein expression by Human Coronavirus NL63. Journal of General Virology. 93(Pt 9):1924-9.
  47. Pyrc K¥, Stozek K, Wojcik K, Gawron K, Zeglen S, Karolak W, Wojarski J, Ochman M, Hubalewska-Mazgaj M, Bochenek G, Sanak M, Zembala M, Szczeklik A, Potempa J. (2012) Use of Sensitive, Broad-Spectrum Molecular Assays and Human Airway Epithelium Cultures for Detection of Respiratory Pathogens. PLoS ONE 7(3): e32582. 
  48. Pyrc K¥, Strzyz P, Milewska A, Golda A, Schildgen O, Potempa J. (2011) Porphyromonas gingivalis enzymes enhance infection with human metapneumovirus in vitro. Journal of General Virology. 92(10):2324-32.
  49. Pyrc K¥, Milewska A, Potempa J. (2011) Development of loop-mediated isothermal amplification assay for detection of human coronavirus-NL63. Journal of Virological Methods. 175(1): 133-136.
  50. Golda A, Malek N, Dudek B, Zeglen S, Wojarski J, Ochman M, Kucewicz E, Zembala M, Potempa J, Pyrc K¥ (2011) Infection with human coronavirus NL63 enhances streptococcal adherence to epithelial cells. Journal of General Virology. 92(6):1358-68.
  51. Pyrc K, Sims AC, Dijkman R, Jebbink M, Long C, Deming D, Donaldson E, Vabret A, Baric R, van der Hoek L, Pickles R¥ (2010) Culturing the Unculturable: Human Coronavirus HKU1 Infects, Replicates, and Produces Progeny Virions in Human Ciliated Airway Epithelial Cell Cultures. Journal of Virology. 84(21):11255-63.
  52. Pyrc K¥ (2008) Ludzki koronawirus NL63 - Obraz kliniczny i charakterystyka molekularna. Postepy Mikrobiologii, 47(3) 257-262.
  53. Dijkman R, Jebbink MF, El Idrissi NB, Pyrc K, Muller MA, Kuijpers TW, Zaaijer HL, van der Hoek L¥. (2008) Human coronavirus NL63 and 229E seroconversion in children. Journal of Clinical Microbiology. 46(7):2368-73.
  54. Golda A, Pyrc K¥ (2008) Recent antiviral strategies against human coronavirus – related respiratory illness. Current Opinions in Pulmonary Medicine. 14(3):248-53.
  55. de Vries M, Pyrc K, Berkhout R, Vermeulen-Oost W, Dijkman R, Jebbink MF, Bruisten S, Berkhout B, van der Hoek L¥ (2008) Human parechovirus type 1, 3, 4, 5 and 6 detection in picornavirus cultures. Journal of Clinical Microbiology. 46(2):759-62.
  56. Pyrc K¥, Jebbink MF, Berkhout B, van der Hoek L. (2008) Detection of new viruses by VIDISCA: virus discovery based on cDNA-amplified fragment length polymorphism. in: SARS-and Other Coronaviruses, Laboratory Protocols. Methods in Molecular Biology, Vol. 454, Ed. Cavanagh, Dave, ISBN: 978-1-58829-867-6.
  57. Pyrc K, van der Hoek L¥ (2007) Human Coronavirus NL63, a Long Lost Brother. Coronaviruses: Molecular and Cellular Biology, Chapter 15, Ed. Volker Thiel, ISBN: 978-1-904455-16-5
  58. Pyrc K¥, Berkhout B, van der Hoek L.; Identification of new human coronaviruses; Expert Review of Anti-infective Therapy. 5(2):245-253.
  59. Pyrc K¥, Berkhout B, van der Hoek L (2007) Antiviral Strategies Against Human Coronaviruses; Infectious Disorders - Drug Targets. 7(1): 59-66.
  60. Dijkman R¥, Jebbink MF, Wilbrink B, Pyrc K, Zaaijer HL, Minor PD, Franklin S, Berkhout B, Thiel V, van der Hoek L (2006) Human coronavirus 229E encodes a single ORF4 protein between the spike and the envelope genes. Virology Journal. 28;3(1):106. 
  61. Pyrc K, Berkhout B., and van der Hoek L¥. The novel human coronaviruses NL63 and HKU1; Journal of Virology. 81(7):3051-7.
  62. van der Hoek L, Sure K, Ihorst G, Stang A, Pyrc K, Jebbink MF, Petersen G, Forster J, Berkhout B, Uberla K¥ (2006) Human coronavirus NL63 infection is associated with croup. Advances in experimental medicine and biology. 581:485-91.
  63. Pohlmann S, Gramberg T, Wegele A, Pyrc K, van der Hoek L, Berkhout B, Hofmann H¥ (2006) Interaction between the spike protein of human coronavirus NL63 and its cellular receptor ACE2. Advances in experimental medicine and biology. 581:281-4. 
  64. Hofmann H, Marzi A, Gramberg T, Geier M, Pyrc K, van der Hoek L, Berkhout B, Pohlmann S¥ (2006) Attachment factor and receptor engagement of SARS coronavirus and human coronavirus NL63. Advances in experimental medicine and biology. 581:219-27.
  65. Piotrowski Y, van der Hoek L, Pyrc K, Berkhout B, Moll R, Hilgenfeld R¥ (2006) Nonstructural proteins of human coronavirus NL63. Advances in experimental medicine and biology. 581:97-100.
  66. Pyrc K., Dijkman R., Deng L., Jebbink MF., Ross HA., Berkhout B. van der Hoek L¥ (2006) Mosaic structure of human coronavirus NL63, one thousand years of evolution. Journal of Molecular Biology. 364(5):964-73.
  67. Schildgen O, Jebbink MF, de Vries M, Pyrc K, Dijkman R, Simon A, Muller A, Kupfer B, van der Hoek L¥ (2006) Identification of cell lines permissive for human coronavirus NL63. Journal of Virological Methods. 138(1-2):207-10.
  68. van der Hoek L., Pyrc K., Berkhout B¥ (2006) Human coronavirus NL63, a new respiratory virus. FEMS Microbiology Reviews. 30(5): 760-773.
  69. van der Hoek L, Pyrc K., Berkhout B¥ (2006) Coronavirus NL63, the new Dutch. Tijdschrift voor Infectieziekten. 1(3):108-114.
  70. Pyrc K., Bosch BJ., Berkhout B., Jebbink MF., Dijkman R., Rottier P., and van der Hoek L¥ (2006) Inhibition of Human Coronavirus NL63 Infection at Early Stages of the Replication Cycle. Antimicrobial Agents and Chemotherapy. 50(6): 2000-2008.
  71. Pyrc K., Berkhout B, van der Hoek L¥ (2005) Molecular characterization of human coronavirus NL63. Recent Research in Infection and Immunity. 3(2005): 25-48 ISBN: 81-7895-182-7, Transworld Research Network.
  72. van der Hoek L, Sure K, Ihorst G, Stang A, Pyrc K., Jebbink MF, Petersen G, Forster J, Berkhout B, Uberla K¥ (2005) Croup Is Associated with the Novel Coronavirus NL63., PLoS Medicine. 23;2(8):e240.
  73. Hofmann H, Pyrc K., van der Hoek L, Geier M, Berkhout B, Pohlmann S¥ (2005) Human coronavirus NL63 employs the severe acute respiratory syndrome coronavirus receptor for cellular entry. PNAS Proceedings of the National Academy of Sciences, USA. 102(22):7988-93.
  74. Moes E, Vijgen L, Keyaerts E, Zlateva K, Li S, Maes P, Pyrc K., Berkhout B, van der Hoek L, Van Ranst M¥ (2005) A novel pancoronavirus RT-PCR assay: frequent detection of human coronavirus NL63 in children hospitalized with respiratory tract infections in Belgium. BMC infectious diseases. 1;5(1):6.
  75. Pyrc K., Jebbink MF, Berkhout B, van der Hoek L¥. (2004) Genome structure and transcriptional regulation of human coronavirus NL63. Virology Journal. 1(1):7.
  76. van der Hoek L, Pyrc K., Jebbink MF, Vermeulen-Oost W, Berkhout RJ, Wolthers KC, Wertheim-van Dillen PM, Kaandorp J, Spaargaren J, Berkhout B¥ (2004) Identification of a new human coronavirus. Nature Medicine. 10(4):368-73.
  77. Kapiszewska M., Pyrc K. (2003) Anticancer and antiartherioslosis action of flavonoids. Vegetables and fruits in the nutrition. Medycyna i Spoleczenstwo, Acta Academiae Modrevianae, pp. 101-108.