
NATIONAL RESEARCH INFRASTRUCTURE
National Research Infrastructure programs
Currently, 4 National Infrastructure Programs are being implemented in the Research Infrastructures of the HPI. The equipment of the HPI Research Infrastructures is available for use by out of Institute users and the personnel of the Infrastructure Units provides consulting services to all users. The 4 Infrastructure programs are listed and described below.
BIOIMAGING.GR: Greek Research Infrastructure Program for Visualizing and Monitoring Fundamental Biological Processes
BIOIMAGING.GR
BioImaging-GR is a Greek Research Infrastructure Program for Visualizing and Monitoring Fundamental Biological Processes, supported by the Ministry of Education Insfrastructure ESPA 2014-2020. BioImaging-GR aims to create a virtual technological platform of advanced imaging, open to the research community, which will bring together and network the technological infrastructure, research projects and image acquisition and analysis services of the main Imaging facilities in Greece.
The main objective of BioImaging-GR is to continue and strengthen the services already provided by the Light Microscopy Unit of the HPI towards more users a) by the recruitment of three new scientists, b) upgrade of current infrastructure towards live imaging and c) development of new research protocols for live imaging like imaging of fast procedures in cells, middle-throughput imaging and imaging of dynamic processes in live animals. In addition, BioImaging-GR aims to provide image acquisition and analysis service in cutting-edge technologies that are already established or will be developed in HPI LMU through BioImaging-GR, as well as diffusion of research/technological developments, through workshops that will be organized.
INSPIRED: The National Research Infrastructure on Integrated Structural Biology, Drug Screening Efforts and Drug target functional characterization

INSPIRED, will offer a complete set of platforms (techniques) for the integrated Structural Biology research process comprising sample preparation, biophysical characterization, structural analysis, functional analysis & pharmacogenomics.
Joint Research Activities (JRAs) will be carried out, with the aim to deliver solutions to academic and industrial users, driven by the national/regional needs for innovative services and products.
INSPIRED consists of a dynamic infrastructure of complementary core centers of excellence.
HPI Node: devoted to research of Infectious Diseases, Immunology and Neurobiology
HPI is devoted to research of Infectious Diseases, Immunology and Neurobiology. It is also heavily involved in Public Health Services where research efforts focus mainly on the development of novel diagnostic and therapeutic strategies for human applications. In particular, HPI participates in INSPIRED with the following infrastructure:
- Protein sample preparation (Soluble expression and purification)
- Biophysical characterization
- Structural analysis
- In silico analysis and drug design
We have created the basis for synergies to exploit the expertise of the interdepartmental individual scientific groups* establishing a local scientific core capable to provide the know-how for high-level research.
The basic infrastructure is currently available. More specifically, the existing facilities are:
For protein sample preparation
- Heterologous expression of soluble/membrane proteins in bacterial, yeast (Pichia pastoris), protozoan (Leishmania tarentolae), insect & mammalian cell systems
- Phage display technology & chain-shuffling technologiesthat focuses on developing high specific monoclonal antibodies or peptides for research, diagnostic and therapeutic use (unique in Greece)
- Recombinant viruses and viral transduction systems
- Large-scale production of soluble and membrane proteins expressed in yeast and bacteria cells in the HPI fermenter unit Downstream processing of the expressed recombinant proteins, protein purification (affinity, size exclusion or ion exchange chromatography).
For protein characterization
- Size, mass, polydispersity of purified proteins (assessed by DLS) before crystallization trials
- Functional characterization
For structural analysis
- Protein crystallization (crystallization robot) & 3D structure determination
- Modelling and simulation of biological macromolecules.
* Depts. and Labs:
- of Immunology: Lab. of Molecular Biology & Immunobiotechnology (Dr Avgi Mamalaki)
Lab. of Immunology (Dr. K. Lazaridis) - of Neurobiology: Lab. ofMolecular Neurobiology & Immunology (Dr Voula Zisimopoulou)
- of Microbiology: Lab. of Bacteriology (Dr Vivi Miriagou, Dr Stathis Kotsakis)
Lab. of Molecular Virology (Dr Rania Georgopoulou)
Lab. of Molecular Ιntracellular Parasitism (Dr Haralabia Boleti)
Selected Publications that support INSPIRED
- Mamalaki, A., Trakas, N., Tzartos, S. Bacterial expression of a single-chain Fv fragment which efficiently protects the acetylcholine receptor against antigenic modulation caused by myasthenic antibodies. European Journal of Immunology, 23:1831845, 1993.
- Tzartos, SJ, Tsantili, P., Papanastasiou, D., Mamalaki, A. Construction of single-chain Fv fragments of anti-MIR monoclonal antibodies. Annals of the New York Academy of Science, 841:475-7, 1998.
- Tsantili, P., Tzartos S. and Mamalaki, A. High affinity scFv antibody fragments protecting the human nicotinic acetylcholine receptor. Journal of Neuroimmunology, 94:15-27, 1999.
- Papanastasiou, D., Mamalaki, A., Eliopoulos, E., Poulas, K., Liolitsas, C. and Tzartos, S. Construction and characterization of a humanized single chain Fv antibody fragment against the main immunogenic region of the acetylcholine receptor. Journal of Neuroimmunology, 94:182-195, 1999. (Corresponding authors: A. Mamalaki and S. Tzartos)
- Tsouloufis, T., Mamalaki, A., Remoundos, M., Tzartos, S.J. Reconstitution of conformationally dependent epitopes of the N-terminal extracellular domain of human muscle AChR α subunit expressed in E.coli: implications for myasthenia gravis therapeutic approaches. International Immunology, 12(9):1255-1265, 2000.
- Kontou, Μ., Leonidas, D.D., Vatzaki, E.H., Acharya, K. R., Mamalaki, A., Oikonomakos, N.G. and Tzartos, S.J. The crystal structure of a Fab fragment of a rat monoclonal antibody against the main immunogenic region of the human muscle acetylcholine receptor. European Journal of Biochemistry, 267(8):2389-2397, 2000.
- Kleinjung, J., Petit, M.C., Orlewski, P., Mamalaki, A., Tzartos, S., Tsikaris, V., Daitsiotis-Sakarellos, M., Sakarellos, C., Marraud, M. and Cung, M.T. The Third-Dimensional Structure of the Complex between an Fv Antibody Fragment and an Analog of the Main Immunogenic Region of the Acetylcholine Receptor: A Combined Two-Dimensional NMR, Homology and Molecular Modeling Approach. Biopolymers, 53(2):113-28, 2000.
- Psaridi, L., Mamalaki, A., Remoundos M., and Tzartos, S. Expression of soluble ligand- and antibody-binding extracellular domain of human muscle acetylcholine receptor alpha subunit in yeast Pichia pastoris. Role of glycosylation in alpha -bungarotoxin binding. J Biol Chem. 277, 30, 26980-26986, 2002.
- Mamalaki A., Gritzapis AD, Kretsovali Α., Belimezi Μ., Papamatheajis J., Perez S., Papamichail M and Baxevanis C. In vitro and in vivo antitumor activity of a mouse CTL hybridoma expressing chimeric receptors bearing the single chain Fv from HER-2/neu- specific antibody and the γ-chain from Fc(ε) RI. Cancer Immunol. Immunoth. 52(8):513-22, 2003.
- Gritzapis A., Mamalaki A, Kretsovali Α., Papamatheajis J., Belimezi Μ, Perez S., Baxevanis C. and Papamichail M. Redirecting mouse T hybridoma against human breast and ovarian carcinomas: in vivo activity against HER-2/neu expressing cancer cells. Br J Cancer, 88(8):1292-300, 2003.
- Psaridi-Linardaki, L., Mamalaki, A., and Tzartos, S. Future therapeutic strategies in autoimmune Myasthenia Gravis. Annals of the New York Academy of Science, 998: 539-548, 2003.
- Avramopoulou, A. Mamalaki and S. Tzartos. Soluble oligomeric and ligand-binding extracellular domain of human neuronal alpha7 acetylcholine receptor expressed in the yeast Pichia pastoris: Replacement of the hydrophobic Cys-loop by the hydrophilic loop of ACh-binding protein enhances protein solubility. J. Biol. Chemistry 279(37):38287-38293, 2004.
- Psaridi-Linardaki, A. Mamalaki, N. Trakas and S. J. Tzartos. Specific immunoadsorption of the autoantibodies from myasthenic patients using the extracellular domain of the human muscle acetylcholine receptor alpha-subunit. Development of an antigen-specific therapeutic strategy. Journal of Neuroimmunology 159(1-2):183-91, 2005.
- Ε. Fostieri, Socrates J. Tzartos, Sonia Berrih-Aknin, David Beeson, and Avgi Mamalaki. Isolation of potent human Fab fragments against a novel highly immunogenic region on human muscle acetylcholine receptor which protect the receptor from myasthenic autoantibodies. Eur J Immunol. 35(2):632-43, 2005.
- Evdokia Protopapadakis, Anna Kokla, Socrates J. Tzartos and Avgi Mamalaki. Isolation and characterization of human anti-acetylcholine receptor monoclonal antibodies from transgenic mice expressing human immunoglobulin loci. Eur J Immunol. 35(6):1960-8, 2005.
- Belimezi, D. Papanastassiou, E. Merkouri, C. Baxevanis and A. Mamalaki Growth inhibition of breast cancer cell lines overexpressing Her2/neu by a novel internalized fully human Fab antibody fragment. Cancer Immunol Immunother. 55: 1091-1099, 2006.
- Tsitoura, U. Georgopoulou and P. Mavromara (2006). HSV-1 based amplicon vectors as an alternative system for the expression of functional HCV proteins Curr Gene Ther 6(3): 393-398 (REVIEW).
- Kostelidou K, Trakas N, Zouridakis M, Bitzopoulou K, Sotiriadis A, Gavra I & Tzartos SJ (2006)Expression and characterisation of soluble forms of the extracellular domains of the β, γ and ε subunits of the human muscle acetylcholine receptor. FEBS Journal 273, 3557-3568
- Zouridakis M, Kostelidou K, Sotiriadis A, Stergiou C, Eliopoulos E, Poulas K & Tzartos SJ (2007)Circular dichroism studies of extracellular domains of human nicotinic acetylcholine receptors provide aninsight into their structure. International Journal of Biological Macromolecules 41(4):423-9
- Zouridakis M, Zisimopoulou P, Eliopoulos E, Jacobson L, Poulas K & Tzartos SJ (2007)Recombinant extracellular domains of human neuronal nicotinic receptors. Preliminary studies on mutant forms for the improvement of solubility. Neurophysiology 39 (4/5) 302-306
- Tsitoura, U. Georgopoulou, S. Petres, A. Varaklioti, A. Karafoulidou, D. Vagena, C. Politis and P. Mavromara (2007). Evidence for cellular uptake of recombinant hepatitis C virus non-enveloped capsid-like particles FEBS Lett 581(21): 4049-4057.
- Kouvatsis, R. Argnani, E. Tsitoura, M. Arsenakis, U. Georgopoulou, P. Mavromara and R.Manservigi (2007). Characterization of herpes simplex virus type 1 recombinants that express and incorporate high levels of HCV E2-gC chimeric proteins Virus Res.123(1):40-49.
- Nianiou, K. Kalantidis, P. Madesis, U Georgopoulou, P.Mavromara and A. Tsaftaris (2008). Expression of an HCV core antigen coding gene in tobacco (N. tabacum L.) Prep Biochem Biotechnol 38(4): 411-421.
- Katsarou, E. Serti, P. Tsitoura ,A. Lavdas ,A. Varaklioti ,A. Pickl-Herk ,D. Blaas ,D. Oz-Arslan, R. Zhu, P.Hinterdoefer, P. Mavromara, U. Georgopoulou (2009). Green-Fluorescent protein-tagged HCV non-enveloped capsid like particles: Development of a new tool for tracking HCV core uptake. Biochimie 91(7):903-915.
- Madesis, P.; Osathanunkul, M.; Georgopoulou, U.; Gisby, MF; Mudd, EA; Nianiou, I; Tsitoura, P; Mavromara, P; Tsaftaris, A; Day, A (2010). A hepatitis C virus core polypeptide expressed in chloroplasts detects anti-core antibodies in infected human sera. Journal of Biotechnology, 145 (4), 377-386.
- Koliaraki, M. Marinou, M. Samiotaki, G. Panayotou, K. Pantopoulos, A. Mamalaki Iron regulatory and bactericidal properties of human recombinant hepcidin expressed in Pichia pastoris. Biochimie 90(5):726-35, 2008.
- Koliaraki V, Marinou M, Vassilakopoulos TP, Vavourakis E,Tsochatzis E, Pangalis GA, Papatheodoridis G, Stamoulakatou A, Swinkels D, PapanikolaouG, Mamalaki A. A novel immunological assay for hepcidin quantification in human serum. PLoS One. 2009;4(2):e4581.
- Alexios Dimitriadis, Chrysanthi Gontinou, Constantin N. Baxevanis and Avgi Mamalaki The mannosylated extracellular domain of Her2/neu produced in P. pastoris induces protective antitumor immunity..BMC cancer, 2009 Oct 30;9:386
- Zouridakis M, Zisimopoulou P, Eliopoulos E, Poulas K & Tzartos SJ (2009) Design and expression of human α7 nicotinic acetylcholine receptor extracellular domain mutants with enhanced solubility and ligand-binding properties Biochim Biophys Acta1794(2):355-66 doi: 10.1016/j.bbapap.2008.11.002
- Kotsakis SD, Papagiannitsis CC, Tzelepi E, Tzouvelekis LS, Miriagou V. Extended-spectrum properties of CMY-30, a Val211Gly mutant of CMY-2 cephalosporinase. Antimicrobial Agents and Chemotherapy 2009, 53:3520-3.
- Kotsakis SD, Miriagou V, Tzelepi E, Tzouvelekis LS. Comparative biochemical and computational study of the role of naturally occurring mutations at Ambler’s positions 104 and 170 in GES β-lactamases. Antimicrobial Agents and Chemotherapy 2010, 54:4864-71
- Stefan Gattenlohner, Hannah Jörißen, Michael Huhn, Angela C. Vincent, David Beeson, S. Tzartos, Avgi Mamalaki, Benjamin Etschmann, Hans-Konrad Müller-Hermelink, Eva Koscielniak, Stefan Barth and Alexander Marx. A Human Recombinant Autoantibody-based Immunotoxin Specific for the Fetal Acetylcholine Receptor Inhibits Rhabdomyosarcoma Growth in vitro and in a Murine Transplantation Model. Journal of Biomedicine and Biotechnology, 2010:187621
- Sidera K, El Hamidieh A, Mamalaki A, Patsavoudi E. The 4C5 cell-impermeable anti-HSP90 antibody with anti-cancer activity, is composed of a single light chain dimer. PLoS One. 2011;6(9):e23906.
- Stergiou, Ch., Zisimopoulou, P., and Tzartos, S.J. (2011).Expression of water-soluble, ligand-binding concatameric extracellular domains of the human neuronal nicotinic receptor alpha4 and beta2 subunits in the yeast pichia pastoris. J. Biol. Chem. 286: 8884-92.
- Papagiannitsis CC, Kotsakis SD, Petinaki E, Vatopoulos AC, Tzelepi E, Miriagou V, Tzouvelekis LS. Characterization of metallo-β-lactamase VIM-27, an A57S mutant of VIM-1 associated with Klebsiella pneumoniae ST147. Antimicrobial Agents and Chemotherapy 2011,55:3570-2.
- Kotsakis SD, Tzouvelekis LS, Petinaki E, Tzelepi E, Miriagou V. Effects of the Val211Gly substitution on molecular dynamics of CMY-2 cephalosporinase: implications on hydrolysis of expanded-spectrum cephalosporins. Proteins 2011, 79:3180-92.
- Kotsakis SD, Tzouvelekis LS, Zerva L, Liakopoulos A, Petinaki E. Staphylococcus lugdunensis strain with a modified PBP1A/1B expressing resistance to β-lactams. Eur J Clin Microbiol Infect Dis 2012 31(2):169-172.
- Niarchos A, Zouridakis M, Douris V, Georgostathi A, Kalamida D, Sotiriadis A, Poulas K, Iatrou K, Tzartos SJ(2013) Expression of a highly antigenic and native-like folded extracellular domain of the human α1 subunit of muscle nicotinic acetylcholine receptor, suitable for use in antigen specific therapies for Myasthenia Gravis. PLoS One 2013 Dec 20;8(12) doi: 10.1371/journal.pone.0084791
- Kotsakis SD, Caselli E, Tzouvelekis LS, Petinaki E, Prati F, Miriagou V. (2013) Interactions of oximino-substituted boronic acids and β-lactams with the CMY-2-derived extended-spectrum cephalosporinases CMY-30 and CMY-42. Antimicrobial Agents &Chemotherapy; 57(2): 968-76.
- Papadimitropoulou A, Mamalaki A. The glycosylated IgII extracellular domain of EMMPRIN is implicated in the induction of MMP-2. Mol Cell Biochem. 2013 Jul;379 (1-2):107-13.
- Lazaridis, K., Zisimopoulou, P., Giastas, P., Bitzopoulou, K., Evangelakou, P., Sideri, A., Tzartos, S.J. Expression of human AChR extracellular domain mutants with improved characteristics (2014) International Journal of Biological Macromolecules, 63, pp. 210-217. DOI: 10.1016/j.ijbiomac.2013.11.003
- Zouridakis M, Giastas P, Zarkadas E, Chroni-Tzartou D, Bregestovski P & Tzartos SJ (2014) Crystal structures of free and antagonist-bound states of human α9 nicotinic receptor extracellular domain. Nature Struct Mol Biol. doi: 10.1038/nsmb.2900
- Kouvatsos N, Niarchos A, Zisimopoulou P, Eliopoulos E, Poulas K, and Tzartos(2014) Purification and functional characterization of a truncated human α4β2
- Kotsakis SD, Miriagou V, Vetouli EE, Bozavoutoglou E, Lebessi E, Tzelepi E, Tzouvelekis LS. (2015) Increased hydrolysis 1 of oximino-β-lactams by CMY-107, aTyr199Cys mutant of CMY-2 produced by Escherichia coli. Antimicrobial Agents & Chemotherapy; 59(12): 7894-8.
- Azam L, Papakyriakou A, Zouridakis M, Giastas P, Tzartos SJ & McIntosh JM (2015) Molecular interaction of α-conotoxin RgIA with the rat α9 nAChR. Mol Pharm doi: 10.1124/mol.114.096511
- Arnaouteli, S., Giastas, P., Andreou, A., Tzanodaskalaki, M., Aldridge, C., Tzartos, S.J., Vollmer, W., Eliopoulos, E., Bouriotis, V. Two putative polysaccharide deacetylases are required for osmotic stability and cell shape maintenance in Bacillus anthracis (2015) Journal of Biological Chemistry, 290 (21), pp. 13465-13478. DOI: 10.1074/jbc.M115.640029
- Lykhmus O, Koval L, Pastuhova D, Zouridakis M,Tzartos S, Komisarenko S, Skok M (2016) The role of carbohydrate component of recombinant α7 nicotinic acetylcholine receptor extracellular domain in its immunogenicity and functional effects of resulting antibodies. Immunobiology doi: 10.1016/j.imbio.2016.07.012
- Kouvatsos, N., Giastas, P., Chroni-Tzartou, D., Poulopoulou, C., Tzartos, S.J. Crystal structure of a human neuronal nAChR extracellular domain in pentameric assembly: Ligand-bound α2 homopentamer (2016) Proc. Natl. Acad. Sci., USA, 113 (34), pp. 9635-9640. DOI: 10.1073/pnas.1602619113
- Giastas P, Zouridakis M& Tzartos SJ¶ (2017) Understanding structure-function relationships of the human neuronal acetylcholine receptor: insights from the first crystal structures of neuronal subunits. Br J Pharmacol. doi: 10.1111/bph.13838
- Ivanova II, Mihaylova NM, Manoylov IK, Makatsori D, Lolov S, Nikolova MH, Mamalaki A, Prechl J, Tchorbanov AI. Targeting of Influenza Viral Epitopes to Antigen-Presenting Cells by Genetically Engineered Chimeric Molecules in a Humanized NOD SCID Gamma Transfer Model. Hum Gene Ther. 2018 Sep;29(9):1056-1070.
- Andreou, A., Giastas, P., Christoforides, E., Eliopoulos, E.E. Structural and evolutionary insights within the polysaccharide deacetylase gene family of bacillus anthracis and bacillus cereus (2018) Genes, 9 (8), art. no. 386, DOI: 10.3390/genes9080386
- Giastas, P., Andreou, A., Papakyriakou, A., Koutsioulis, D., Balomenou, S., Tzartos, S.J., Bouriotis, V., Eliopoulos, E.E. Structures of the Peptidoglycan N-Acetylglucosamine Deacetylase Bc1974 and Its Complexes with Zinc Metalloenzyme Inhibitors (2018) Biochemistry, 57 (5), pp. 753-763 DOI:10.1021/acs.biochem.7b00919
- Kryukova, E.V., Ivanov, I.A., Lebedev, D.S., Spirova, E.N., Egorova, N.S., Zouridakis, M.,et al. (2018). Orthosteric and/or Allosteric Binding of alpha-Conotoxins to Nicotinic Acetylcholine Receptors and Their Models. Mar Drugs 16(12). doi: 10.3390/md16120460.
- Doukas A., Karena E, Papakostas K, Papadaki A., Tziouvara O. Xingi E., Frillingos S., Boleti H.“Heterologous expression of the mammalian sodium-nucleobase transporter rSNBT1 in Leishmania tarentolae (under revision).
ELI - LASERLAB Europe Synergy, HiPER & IPERION-CH.gr
The research infrastructures HiPER, ELI and LASERLAB Europe and IPERION-CH.gr, now united as (HELLAS-CH), are an interdisciplinary National Research Infrastructure (NRI) that will allow access to advanced technological experimental infrastructures supporting research in the area of Laser and its applications. The Research Infrastructure HELLAS-CH is hosted at FORTH, at the Institute of Electronic Structure and Laser, with Dr Dimitrios Charalambides as the head of this scientific project. The consortium is of particular interest for biomedical applications of the use of this radiation.
The participation of the Hellenic Pasteur Institute (HPI) concerns (a) the exploration and development of applications of the above NRI, in the field of biomedical research. The Inflammation Research Laboratory and the Department of Animal Biomedical Research of HPI will use the infrastructure of the School of Applied Sciences of the Electronics Department of the Technological Educational Institute of Crete, a member of the consortium to study biomarkers of inflammation under the influence of laser radiation used for therapeutic purposes in biological material, and (b) contributing to the development of the Operation Regulation of the NRI, its organizational structure and user access planning, which are the deliverables of the project.
Co-financed by Greece and the European Union
ΚRIPIS II
2014-2020 Action “Research-Create-Innovate”
Hepcidin, the central regulator of iron homeostasis, as diagnostic biomarker and personalized therapeutic agent
The main aims of this project are briefly the following: a. the development of the 2nd generation immunodiagnostic test for hepcidin determination b. Validation of the new hepcidin immnunoassay kit and analysis of a large number of control samples and determination of reference values for the Greek population. c. Evaluation of the diagnostic and prognostic use of measuring hepcidin in patients with several disorders, in order to develop/appraise personalized treatment options. d. Cloning and structural characterization of anti-hepcidin monoclonal antibodies. e. Evaluation of hepcidin agonist or antagonist administration as means to treat serious infections in an experimental animal model.
Development of α PCR microarray for assessment of differential gene expression related to resistance of Sea Bass to Viral Nervous Necrosis caused by b-nodavirus, with potential for commercial exploitation NGS/ PCR Array (MIS 5010925)

The objective of the Project is the development of PCR microarrays for assessment of gene expression profiles related to sea bass resistance against viral nervous necrosis. The European sea bass (Dicentrarchus labrax) is one of the most important farmed species of aquaculture in the Mediterranean, with 153.182 tons of annual production. One of the main problems in sea bass farming is the development of diseases, including viral nervous necrosis (VNN). VNN is caused by beta-nodavirus strains and there is no effective treatment or widely available vaccine for the disease. Selective reproduction of resistant individuals is an alternative approach towards disease elimination, with the potential for increase of survival at least 12.5% per generation. In the context of the present Project, PCR microarrays will be developed as a molecular tool to assess the resilience of farmed sea bass populations to VNN for genetic improvement, with commercial exploitation potential. The microarrays will be based on the determination of genes and – by extension- the signaling pathways and cellular functions, which are related to the natural resistance of sea bass against the virus. Results will be obtained by application of Next Generation Sequencing (NGS) to analyze the whole cell transcriptome of the sea bass immune system. Aim of the present action is to identify differentially expressed genes that are responsible or act cooperatively in order to protect sea bass against the disease.
The implementation of the Project will be achieved under the supervision of University of the Aegean, through the collaboration of three partners which are active in complementary fields such as fish pathology, immunology, molecular and systems biology, and sciences involved in fish-breeding programs, the development of new products that could be commercially exploited and promoted via domestic and export activities. The Project’s deliverables are innovative technological achievements of high interest to the international scientific community and manufacturers, since purpose of this project is the production of a molecular tool for resistant sea bass immune profile determination, and also for improved control rearing farmed species, breeders’ selection or juveniles sales with certification for resistance to VNN. In fact, the successful completion of the Project could enrich the company’s product pipeline with a new product. Moreover, four new jobs are created, two positions for highly qualified scientists (HPI) and two positions that offer the possibility for new scientists training (University of the Aegean), enhancing their academic and professional profile.
Development of peptide-based nanovaccines against leishmaniasis NanoVacLeish (MIS 5031816)


The objective of the current Project is the development of innovative peptide-based nanovaccines for preventing and controlling leishmaniasis caused by protozoan parasites of the genus Leishmania which are endemic in tropical and subtropical areas, as well as in the Mediterranean basin, and are transmitted through the bite of infected phlebotomine sandflies. Current treatment based on chemotherapy is expensive and highly toxic, and often ineffective due to parasite’s drug resistance. The development of a safe vaccine is thought to be, worldwide, the most effective and economic way to eliminate and control the disease, and thereby it attracts social and financial interest. In the context of this Project, candidate vaccines are been constructed using biocompatible self-nanoemulsifying drug delivery systems with adjuvant properties for the encapsulation of multi-epitope peptides derived from immunogenic Leishmaniaproteins upon performing a combined approach based on reverse vaccinology and proteomics.
The implementation of the Project is been achieved under the supervision of Hellenic Pasteur Institute through the collaboration of three partners with activity in complementary scientific fields, such as nanomaterial science, parasitology, molecular biology and immunology, as well as Public Health-related services serving the concept of the “One Health Initiative”, the development of new products that could be commercially exploited and promoted via domestic and export activities. The Project’s deliverables are innovative technological achievements of high interest to the international scientific community and manufacturers. Beyond their contribution to the efforts made towards the development of a vaccine against leishmaniasis, these products will provide novel patterns in vaccinology which can be expanded in other infectious diseases caused by intracellular pathogens or in cancer. The successful completion of the Project can enrich company’s product pipeline with a new candidate product. Moreover, 5 new jobs (5 HPI, 2 CERTH) are also being created, thus offering highly specialized training to new scientists and enhancement of their academic and professional profile.
Our research unveils that administration of a chimeric protein with inherent immunostimulatory capacity, alone or incorporated in cationic liposomes, provides significant long-term protection against experimental VL, mediated by the differentiation of Ag-specific multi-functional CD4+ and CD8+ T memory cells1,2,3. On the contrary, the use of multi-epitope oligopeptides incorporated into PLGA or SNEDDS in combination with MPLA or squalene/tocopherol as adjuvants, respectively, fails to induce long lasting immunity4,5. Aiming to discover further candidate protein molecules and / or epitopes to be used for the rational design of effective peptide vaccines, we conducted analysis of the protein load of exosomes isolated from the blood of dogs with leishmaniasis, from promastigote or amastigote origin or from infected macrophages cultures by LC-MS/MS6. First step of analysis revealed a set of common proteins in exosomes from promastigote and amastigote origin, highly enriched in HTL and CTL epitopes. L. infantum challenge of previously vaccinated BALB/c mice with promastigote exosomes, in combination with Addavax, resulted to significant reduction of parasitic load, confirming their prophylactic potential as vaccine candidates7. Exosomes derived from the infected macrophages analysis unveiled their immunomodulatory role by polarizing macrophages towards M2 phenotype and promoting infection establishment via IL-17 and IFNα-regulation signaling pathway proteins8,9.
Publications – Conferences
- Agallou, M., M. Margaroni, S.D. Kotsakis, E. Karagouni (2020). A canine-directed chimeric multi-epitope vaccine induced protective immune responses in BALB/c mice infected with Leishmania infantum. Vaccines; 8(3), 350. doi: 10.3390/vaccines8030350.
- Agallou M., M. Margaroni, E. Tsanaktsidou, O. Kammona, E. Karagouni (2021). A novel cationic liposome for validation of a multi‐epitope chimeric protein for vaccine development against visceral leishmaniasis. European Journal of Immunology; 51, 1-448, Suppl. 1. doi: 10.1002/eji.202170200.
- Agallou, M., M. Margaroni, E. Tsanaktsidou, O. Kammona, C. Kiparissides, E. Karagouni (2022). A liposomal vaccine promotes immune responses via dendritic cell activation in draining lymph nodes, submitted.
- Tsanaktsidou, Ε., O. Kammona, C. Kiparissides. SNEDDS Incorporating two Adjuvants as Potential Nanocarriers for Vaccines.17th International Conference on Nanosciences & Nanotechnologies (NN20), 7-10 July 2020, Thessaloniki, Greece.
- Margaroni M., M. Agallou, E. Tsanaktsidou, O. Kammona, C. Kiparissides, E. Karagouni. Immunoinformatics-aided design and immunogenicity evaluation of a multi-epitope vaccine against visceral leishmaniasis (Poster). 71th Congress of Hellenic Society for Biotechnology and Molecular Biology, Athens, Greece, 26-28 November 2021.
- Efstathiou A., Toubanaki D.K., Karagouni E. Evaluation of exosome isolation methods from canine biological fluids. 7th YSF, 28.11.19, 70th National Conference of HSBMB, 29.11.19-01.12.19, Eugenides Foundation, Athens.
- Efstathiou A., M. Agallou, D.K. Toubanaki, E. Karagouni. An exosome-based vaccination protocol with protective immune responses in Leishmania infantum experimental model of BALB/c mice. Virtual Congress of Hellenic Society for Biotechnology and Molecular Biology, January-June 2021.
- Efstathiou A., D.K. Toubanaki, M. Samiotaki, G. Stamatakis, E. Karagouni (2021). Proteomic analysis of exosomes highlights their role in specific‐signaling pathways’ modulation during Leishmania infection in macrophages. European Journal of Immunology; 51, 1-448, Suppl. 1. doi: 10.1002/eji.202170200
- Efstathiou A., D.K. Toubanaki, M. Samiotaki, E. Karagouni. IL-17 and IFNa regulation signalling pathways’ enrichment highlights a potent immunomodulatory role of exosomes in Leishmania-infected macrophages (Oral presentation). 71th Congress of Hellenic Society for Biotechnology and Molecular Biology, Athens, Greece, 26-28 November 2021.
Total budget: 839.856,90 € / HPI budget: 415.308,90 €
Systematic development and commercial exploitation of novel aggregation inhibitors of the protein α-synuclein, AlphaSyn, MIS 5131418
Title: Systematic development and commercial exploitation of novel aggregation
inhibitors of the protein α-synuclein
Parkinson’s disease (PD) is the second most frequent neurodegenerative disease after Alzheimer’s disease. It causes kinetic dysfunctions, such as limb tremors, but also non-motor symptoms. Approved treatments against PD are symptomatic and only partially relieve the kinetic issues. To date, there is no medication available that treats or delays the progressive neurodegeneration of the disease. Therefore, there is a great need for the development of therapies and treatments that will directly target the pathogenesis of the disease, will actually be neuroprotective and will delay the progression of the disease. PD is characterized by the accumulation of aggregated forms of the protein α-synuclein (αSyn) in the brain, which exhibit increased neurotoxicity. It is believed that inhibiting the formation of these aggregates would restrict the disease progression in the early stages, where the pathology is relatively localized and therefore more susceptible to therapeutic regimens. In the framework of the AlphaSyn project, we will develop new and effective small-molecule aSyn aggregation inhibitors. These will result from screening (1) libraries of cyclic oligopeptides comprising hundreds of millions of different molecules, and (2) natural product libraries from Greek marine organisms. The most effective inhibitors of αSyn aggregation and neurotoxicity will be preclinically developed with the ultimate goal of developing and exploiting commercially (1) a neuroprotective nutritional supplement that promotes the kinetic and cognitive function of elderly people, as well as (2) lead compounds with potentially therapeutic properties against PD.