Innate immunity to hantaviruses

HANTA-INNATE

The logo is an schematic representation of the hantavirus, with the three viral RNA genome particles as squares, that as well represent the Croatian chessboard (squares and colours). The circle represents the viral coat, and by being interrupted by the project name on the right side creates the effect of a capital C, from Croatia. 

SUMMARY OF THE PROJECT

Hantaviruses (HTV), belonging to the genus Orthohantavirus, family Hantaviridae, are enveloped RNA viruses, which cause hemorrhagic fever with renal syndrome (HFRS) in Europe and Asia (up to 20% mortality rate), and hantavirus cardiopulmonary syndrome (HPS) in Americas (up to 40% mortality rate). A broad spectrum of HFRS clinical conditions has been recognized, ranging from unapparent or mild to a fulminant hemorrhagic process with severe renal failure and death.
In Europe there are two main HTV which cause HFRS: Puumala (PUUV) and Dobrava (DOBV), while almost whole Croatia is endemic for HTV (except the coastal region), with predomination of PUUV circulation.
The reservoirs for HTV are small rodents with persistent infection and no manifestation of disease. The infection in humans is a consequence to the exposure to aerosols originated in saliva, urine, or faeces of rodents, while one of the most effective preventive methods is the control of animal populations in residential areas.
There is still a significant lack of knowledge about the immune response to HTV. A strong proinflammatory response during acute infection in humans, is probably responsible for viral clearance but also for potentially fatal proinflammatory-mediated disease. In the south-east Europe both DOBV and PUUV virus co-circulate, although these two viruses are more distantly related and cross-protection might not be seen. The lack of an animal model additionally hinders HFRS research. So far, no effective antivirals, vaccines, or immunotherapeutics exist for HFRS.

Goal
The overall objective of this project is to identify some key elements of innate immune response to HTV, their crosstalk, and potential mediation by regulating agents.

Specific Aims
1) To determine the key mechanisms which contribute to the interaction of innate immune players during acute HTV infection.
2) To explore possible regulating pathways of the innate immune response during HTV infection.

Impact and outcomes
As mentioned above, there is a lack of knowledge in the mechanism of immune response to HTV, and no effective antivirals, vaccines, or immunotherapeutics exist for HFRS. Therefore, our study is an unique opportunity in which we plan to detect and compare elements of innate immunity and possible immunosuppressive response in acute HFRS infection, and in experimentally simulated infections. The proposed research will contribute to substantial new knowledge on innate immune response to HTV, and aid to the future potential development of effective treatments for HFRS.

PROJECT DURATION
The project started on July 1st 2014, and was completed on November 30th 2018.


FUNDING

HRZZ - Croatian Science Foundation  
The total budget of the project has been funded by the Croatian Science Foundation for four years with a total amount of 992,495.00 HRK (over 130,000 €). 


PROJECT TEAM MEMBERS
 
University Hospital for Infectious Diseases, Zagreb  
 

Prof. Alemka Markotić, MD, PhD (Principal Investigator)  Curriculum Vitae.pdf
Oktavija Đaković Rode, MD, PhD ( Researcher Curriculum Vitae.pdf
Lidija Cvetko Krajinović, PhD ( Researcher )   Curriculum Vitae.pdf
Ivan-Christian Kurolt, PhD ( Researcher Curriculum Vitae.pdf
Petra Svoboda, BSc (PhD student)  Curriculum Vitae.pdf
Željka Mačak Šafranko, PhD (postodoctoral fellow)  Curriculum Vitae.pdf
Branka Žibrat, BSc (technician)

Hebrew University Faculty of Medicine, Jerusalem, Israel
Professor Ofer Mandelboim, PhD (Researcher) Curriculum Vitae.pdf

Medical Faculty, University of Rijeka
Đurđica Cekinović, MD, PhD (Researcher) Curriculum Vitae.pdf

Medical Faculty, University of Split
Ass. Prof. Dragan Ledina, MD, PhD (Researcher) Curriculum Vitae.pdf

Croatian Institute for Public Health, Zagreb
Ljiljana Žmak, MD, PhD (Researcher) Curriculum Vitae.pdf



COLLABORATIONS



IN THE NEWS


Lidija Cvetko Krajinović and Alemka Markotić visited professor`s Mandelboim Laboratory at the The Lautenberg Center for General and Tumor Immunology, Hadassah Medical School, The Hebrew University, Jerusalem, Israel and proceeded with research on NK cells and hantaviruses.

(The Lautenberg Center for General and Tumor Immunology, Hadassah Medical School, The Hebrew University, Jerusalem, Israel)


- A new Postdoctoral Fellow (Juan Jose Gonzalez Plaza) joined our team in June 2015.
 
- In 2015 the Hanta-Innate research team became a part of the Center of Excellence for Virus Immunology and Vaccines under the coordination of Prof Stipan Jonjić.


- Postodoctoral Fellow (Juan Jose Gonzalez Plaza) left our team at the end of March 2016.


- Finally, after three long years we're off for our long trip to the Xth International Conference on HFRS, HPS & Hantaviruses organized in Fort Collins, Colorado, USA to present some of our data. This is the biggest scientific meeting for researches in the field of Hantaviruses and the diseases they cause which is held every three years. 


- Two of our team members, Lidija Cvetko Krajinović, PhD and Ivan-Christian Kurolt, PhD, have been awarded a travel grant of $1000 each by the International Society for Hantaviruses (HFRS and HPS) to present their results at the Xth International Conference on HFRS, HPS & Hantaviruses. The picture below shows them with the other awardees in the beautiful Organ Recital Hall (ORC) at the University Center for the Arts at Colorado State University.



- We welcome our new Postdoctoral Fellow (Željka Mačak Šafranko) who joined our team in July 2016.


PROJECT ACTIVITIES


Research stay
• The Lautenberg Center for General and Tumor Immunology, Hadassah Medical School, The Hebrew University, Jerusalem, Israel (26.05.-05.06.2015.)
- establishment of scientific cooperation with Prof. Offer Mandelboim and short research stay of Alemka Markotić and Lidija Cvetko Krajinović related to research on NK cells and hantaviruses 

Dissemination
• 2014 Annual Meeting of Croatian Immunological Society, Krk, Croatia (17.–18.10.2014.)
- oral presentation: Cvetko Krajinović L. Early immunoreactions in peripheral blood mononuclear cells induced by Puumala virus.

• 1st Croatian Virus Workshop, Rijeka, Croatia (14.11.2014.)
- oral presentation: Cvetko Krajinović L. Early immunoreactions in peripheral blood mononuclear cells induced by Puumala virus.

• 82nd Scientific and Professional Symposium - Zoonoses, Slavonski Brod, Croatia (28.-30.05.2015.)
- poster: Cvetko Krajinović L, et al. Unspecific and specific immunity to hantaviruses.

• 1st Inter-Academy Scientific Symposium Emerging and re-emerging zoonoses of the 21st century, Zagreb, Croatia (15.10.2015.)
- invited lecture: Markotić A. Emerging and re-emerging zoonoses in Croatia.

• X International Conference on HFRS, HPS &Hantaviruses, Fort Collins, Colorado, USA (31.05.-03.06.2016.)
- oral presentation: Cvetko Krajinović L. Early slowdown of the peripheral immune response triggered by Puumala virus infection.
- oral presentation: Kurolt IC. Micro RNAs in urine as potential biomarkers for severity of hemorrhagic fever with renal syndrome.
- poster: Svoboda P, et al. Cytokine and chemokine kinetics and their potential as biomarkers in patients with Puumala virus infection.

• Hantavirus Reunion Symposium, U.S. Army Medical Research Institute of Infectious Diseases (USAMRIID), Frederick, Maryland, USA (06.06.2016.)
- invited lecture: Markotić A. Clinical studies of HFRS.

• 2016 Annual Meeting of Croatian Immunological Society, Ogulin, Croatia (14.-15.10.2016.)
- oral presentation: Kurolt IC. Urinary microRNAs as new early indicators for disease severity in hemorrhagic fever with renal syndrome.
- poster: Svoboda P, et al. Kinetics of selected cytokines and chemokines in patients with Puumala virus infection. 

• 2nd Inter-Academy Scientific Symposium Emerging zoonoses, Slovenian Academy of Science and Arts, Ljubljana, Slovenia (27.10.2016.)
- invited lecture: Markotić A. Multiple zoonotic infections – clinical, diagnostic and epidemiological impact.
- invited lecture: Cvetko Krajinović L. Parameters of innate immune response in peripheral circulation of patients infected with Puumala virus. 
- invited lecture: Kurolt IC. MicroRNAs in urine - biomarkers for disease severity in patients infected with Puumala virus.

• Symposium Hemorrhagic fever with renal syndrome - introduction to the new epidemic in 2017?, Zagreb, Croatia (07.04.2017.)
- oral presentation: Markotić A. Clinical presentation of the hemorrhagic fever with renal syndrome.
- oral presentation: Cvetko Krajinović L. Molecular diagnostics of hemorrhagic fever with renal syndrome. 
- oral presentation: Mačak Šafranko Ž. Hemorrhagic fever with renal syndrome in Croatia in 2017. 
- oral presentation: Đaković Rode O. „Point-of-care“ tests for quick diagnostics of hemorrhagic fever with renal syndrome.

• 1st Croatian Congress on Biosecurity and Biosafety, Zagreb, Croatia (07.-09.10.2017.)
- oral presentation: Markotić A. Microbial Forensics, Sciences and Technology and Biological and Toxin Weapons Convention.
- oral presentation: Kurolt IC. Biosafety infrastructure and capabilities at the University Hospital for Infectious Diseases „Dr. Fran Mihaljević”. 
- oral presentation: Đaković Rode O. Point‐of‐care tests for emerging and reemerging pathogens.
- poster: Cvetko Krajinović L. Biosecurity - Organization, Documentation, Inspection, Certification and Recertification of BSL3 and BSL4 Laboratories.
- poster: Mačak Šafranko Ž, et al. Outbreak of hemorrhagic fever with renal syndrome in Croatia in 2017.
- poster: Svoboda P. Personal protection and regulatory aspects of infectious samplesm packing and transport.

• 2017 Annual Meeting of Croatian Immunological Society, Zagreb, Croatia (20.-21.10.2017.)
- oral presentation: Markotić A.

• 3rd Inter-Academy Scientific Symposium Emerging and re-emerging zoonoses of the 21st century, The Academy of Sciences and Arts of Bosnia and Herzegovina, Sarajevo, Bosnia and Herzegovina (28.10.2017.)
- invited lecture: Markotić A. Zoonoses, bioterrorism and microbial forensics.
- invited lecture: Cvetko Krajinović L. Deregulation of monocyte related cytokine/chemokine profile in serum milieu during Puumala virus infection. 
- invited lecture: Mačak Šafranko Ž. HFRS outbreak in Croatia in 2017.

• 16th Croatian Conference on Laboratory Diagnosis, Zagreb, Croatia (11.–13.05.2018.)
- invited lecture: Cvetko Krajinović L. In search of laboratory markers for estimating hemorrhagic fever with renal syndrome clinical presentation severity.
- invited lecture: Mačak Šafranko Ž. Possibilities of using NK cells receptor expression analysis for diagnostic and therapeutic purposes.
- invited lecture: Kurolt IC, et al. MicroRNA in urine in urogenital tract diseases.

• ECI 2018: 5th European Congress of Immunology, Amsterdam, Netherlands (02.-05.09.2018.)
- poster: Cvetko Krajinovic L, et al. Deregulation of monocyte related cytokine/chemokine profile in serum milieu during Puumala virus infection.

• 2018 Annual Meeting of Croatian Immunological Society, Zadar, Croatia (19.-20.10.2018.) 
- poster: Svoboda P, et al. The expression level changes of lineage-specific cell surface molecules on innate immune cells in response to orthohantavirus infection.

• 4th Inter-Academy Scientific Symposium Zoonoses and vector-borne diseases - principles of biosafety and biosecurity, HAZU, Zagreb, Croatia (06.12.2018.)
- invited lecture: Cvetko Krajinović L. Exploring host viral diversity and discovery of novel viruses.

Education
• Flow Cytometry Course Principles and Applications, Zagreb, Croatia (31.08.-04.09.2015.) - Lidija Cvetko Krajinović

• Workshop: APHL/SECID/CDC Biosafety Workshop, Klinika za infektivne bolesti “Dr. Fran Mihaljević”, Zagreb, Croatia (10.-13.05.2016.) - Lidija Cvetko Krajinović, Petra Svoboda, Branka Žibrat

• Advanced EFIS-EJI Course: Metchnikoff’s Legacy: Tissue Phagocytes and Functions, EFIS-EJI Ruggero Ceppellini Advanced School of Immunology, Napoli, Italy (12.–14.10.2016.) - Lidija Cvetko Krajinović 

• Annual Flow Cytometry Course 2016: From First Principles to Polychromatic Applications, Zagreb, Croatia (17.-21.10.2016.) - Petra Svoboda, Željka Mačak Šafranko

• Innate Immunity and Infectious Diseases Course, Pasteur Institute, Paris, France (10.-25.04.2017.) - Lidija Cvetko Krajinović 

• International Workshop "Governance of Dual Use Research in the Life Sciences: Advancing Global Consensus on Research Oversight", HAZU, Zagreb, Croatia (11.-13.06.2018.) - Lidija Cvetko Krajinović 

• 2nd EVD-LabNet Course: Laboratory diagnostics and surveillance of emerging rodent-borne viral infections, Tvärminne Biological Station, Hanko, Finland (19.-21.09.2018) - Ivan-Christian Kurolt 

• International Workshop on Biosafety Laboratory Management and Techniques, Wuhan Institute of Virology, Wuhan, China (15.-25.10.2018.) - Petra Svoboda


Organization of workshops, courses, symposia

• 1st Inter-Academy Scientific Symposium: Emerging and reemerging zoonoses in the 21st century, HAZU, Zagreb, Croatia (15.10.2015.) 
- Organizing Committee (members): Alemka Markotić, Ivan-Christian Kurolt, Oktavija Đaković Rode

• APHL/SECID/CDC Biosafety Workshop, University Hospital for Infectious Diseases “Dr. Fran Mihaljević”, Zagreb, Croatia (10.-13.05.2016.)
- Organizing Committee (members): Alemka Markotić, Ivan-Christian Kurolt

• WHO symposium Neglected tropical diseases, University Hospital for Infectious Diseases “Dr. Fran Mihaljević”, Zagreb, Croatia (13.05.2016.)
- Organizing Committee (president): Alemka Markotić
- Scientific Committee (members): Ivan-Christian Kurolt, Oktavija Đaković Rode

• Symposium and round table: Applied Genomics in the Republic of Croatia, HAZU, Zagreb, Croatia (17.05.2016.) 
- Organizing Committee (president): Alemka Markotić

• 2nd Inter-Academy Scientific Symposium: Emerging zoonoses, SAZU, Ljubljana, Slovenia (27.10.2016.) 
- Scientific Committee (members): Alemka Markotić

• Symposium: Hemorrhagic fever with renal syndrome – An introduction  to a new epidemic in 2017?, Zagreb, Croatia (07.04.2017.)
- Organizing Committee (president): Alemka Markotić
- Organizing Committee (members): Ivan-Christian Kurolt, Petra Svoboda

• 1st Croatian congress on Biosecurity and Biosafety with international participation, Zagreb, Croatia (7.-9.10.2017.)
- Organizing Committee (president): Alemka Markotić
- Organizing Committee (members): Ivan-Christian Kurolt, Oktavija Đaković Rode

• 3rd Inter-Academy Scientific Symposium: Emerging and re-emerging zoonoses of the 21st century, ANUBiH, Sarajevo, Bosnia and Hercegovina (28.10.2017.) 
- Scientific Committee (member): Alemka Markotić 

• Measles epidemic – global biosecurity threat, symposium with international participation, Zagreb, Croatia (05.05.2018.)
- Organizing Committee (president): Alemka Markotić
- Organizing Committee (member): Oktavija Đaković Rode

• Governance of Dual Use Research in the Life Sciences: Advancing Global Consensus on Research Oversight, Zagreb, Croatia (05.05.2018.)
- Local Organizing Committee (member): Alemka Markotić

• 16th Conference on Laboratory Diagnosis (with international participation), Zagreb, Croatia (11.-13.05.2018.)
- Organizing Committee (vice-president): Alemka Markotić

• 4th Inter-Academy Scientific Symposium: Zoonoses and Vector-borne diseases – Biosafety and Biosecurity approch, HAZU, Zagreb, Croatia (06.12.2018.) 
- Organizing Committee (president): Alemka Markotić
- Organizing Committee (members): Ivan-Christian Kurolt 

Doctoral thesis
• Petra Svoboda: The influence of pathogenic Puumala and apathogenic Tula orthohantavirus on the innate immune response of primary monocytes and macrophages
- Defence of thesis: Faculty of Science, University of Zagreb (to be defended in 2019.)
- Supervisor: Lidija Cvetko Krajinović, PhD

Graduation thesis
• Petra Čikeš: Detection of interleukin 15 in peripheral circulation of patients with hemorrhagic fever with renal syndrome
- Defence of thesis: Faculty of Science, University of Zagreb (26.02.2016.)
- Supervisor: Prof Alemka Markotić, PhD

• Filip Šijanski: Detection of interferon γ and chemokine CXCL10 in the primary culture of monocytes infected with ortohantaviruses
- Defence of thesis: Faculty of Science, University of Zagreb (to be defended in 2019.)
- Supervisor: Lidija Cvetko Krajinović, PhD

Awards
• Lidija Cvetko Krajinović: Fellowship from the European Federation of Immunological Societies (EFIS) and European Journal of Immunology (EJI) for the participation at the Advanced EFIS-EJI Course: Metchnikoff’s Legacy: Tissue Phagocytes and Functions, EFIS-EJI Ruggero Ceppellini Advanced School of Immunology, Napoli, Italy (2016.)

• Lidija Cvetko Krajinović and Ivan-Christian Kurolt: Travel Award from the Organizing Committee for the participation at the 10th International Conference on HFRS, HPS and Hantaviruses, Fort Collins, Colorado, SAD (2016.)

Establishment of scientific collaborations 
• Croatian Institute for Transfusion Medicine, Zagreb, Croatia
• Fidelta Ltd, Zagreb, Croatia
• Srebrnjak Children's Hospital, Zagreb, Croatia

Procurement of equipment
• StepOnePlus™ Real-Time PCR System

New scientific projects
• Znanstveni centar izvrsnosti „Centre of Exellence for Research in Viral Immunology and the Development of New Vaccines (CerVirVac)
(Ministarstvo znanosti i obrazovanja, PI: Stipan Jonjić) 
– project collaborator

• Šumski ekosustavi kao prirodna staništa uzročnika zoonoza 
(Ministarstvo poljoprivrede, PI: Josip Margaletić) 
– projekt prihvaćen za financiranje 2016.; Klinika za infektivne bolesti „Dr. Fran Mihaljević“, Zagreb suradnička ustanova na projektu (suradnici na projektu: Alemka Markotić, Lidija Cvetko Krajinović, Petra Svoboda)

• Centar kompetencije molekularne dijagnostike 
(Ministarstvo gospodarstva, PI: Gordan Lauc) 
– projektna prijava u tijeku; Klinika za infektivne bolesti „Dr. Fran Mihaljević“, Zagreb član zajednice prijavitelja

• Jačanje kapaciteta CerVirVac-a za istraživanja u virusnoj imunologiji i vakcinologiji
(EU/Europski fond za regionalni razvoj: KK.01.1.01.0006, PI: Stipan Jonjić) 
- (project collaborators: Alemka Markotić, Lidija Cvetko Krajinović, Petra Svoboda, Željka Mačak Šafranko, Ivan-Christian Kurolt)

• Utjecaj nepatogenoga Tula hantavirusa na urođeni imunosni odgovor primarnih monocita
(Hrvatsko katoličko sveučilište: HKS-2018-1, PI: Alemka Markotić)
– projekt prihvaćen za financiranje 2018.; Klinika za infektivne bolesti „Dr. Fran Mihaljević“, Zagreb suradnička ustanova na projektu (suradnici na projektu: Lidija Cvetko Krajinović, Petra Svoboda)

• New Cytomegaloviral vaccine vector concepts (MCMVaccine)
(Helmholtz Association: PIE-0008, PI: Luka Čičin-Šain) 
- project collaborator


CONTACT


 

Prof. Alemka Markotić, MD, PhD
Head of the Research Department, University Hospital for Infectious Diseases "Dr. Fran Mihaljević"
Mirogojska 8, 10 000 
Zagreb, Croatia
Tel/fax: 385-1-2826-148
Mobile phone: 385-(0)91-4012 648
E-mail: res.dept.bfm@gmail.com