Laboratory of Immunology

Dr Peggy Lymperi | Department of Immunology

Research Director

Επικοινωνία:

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ABOUT

The main research activity of HPI Immunology lab is the study of natural or physiological autoantibodies (NAbs) in both physiological and pathological autoimmunity. NΑbs are the group of antibodies present in healthy organisms without previous external stimulation. They are found in the bloodstream (>90% of circulating Ig of all classes) as well as in other biological fluids and recognize several antigens, of different composition, belonging to the same organism as well as exogenous antigens, a characteristic defined as polyreactivity. This property is closely related to NΑbs participation in multiple phenomena such as the removal of catabolic products, the resistance against infections from pathogenic factors, immunoregulation, homeostasis, transport and functional differentiation of biologically active molecules. Νowadays, it is established that there are autoantibodies able to penetrate in living cells (Cell-Penetrating Antibodies, CPAbs) and locate in the nucleus and/or the cytoplasm. CPAbs are found in patients with autoimmune diseases such as systemic lupus erythematosus and lupus-prone mice, whilst HPI Immunology lab isolated, for the first time worldwide, natural human polyclonal and monoclonal CPAbs from healthy donors. HPI Immunology lab basic research projects are focused on the biological and functional characterization of natural human monoclonal CPAbs. At the applied research level, the lab studies the use of natural human monoclonal CPAbs in the intracellular transport – both in the cytoplasm and in the nucleus-of pharmaceutical agents unable to pass through the cellular membrane, for the targeted therapy of diseases such as cancer.

Recently, by the incorporation of Dr Konstantinos Lazaridis in our team, our activities have included the study of the autoimmune disease myasthenia gravis (MG) and specifically the development of an antigen-specific therapy based on restoring immunological tolerance towards the targeted autoantigens.

The lab is actively involved in students’ education through the supervision  of gradutate, MSc and PhD thesis as well as the organization of, both HPI and national, seminars -for people of all ages – regarding Immunology and its biomedical applications.

CURRENT RESEARCH PROJECTS

1. Cell penetrating polyreactive human autoantibodies (pcPAbs) as innovative therapeutic bio-tools.


The Laboratory of Immunology isolated, from three therapeutic formulations of intravenous immunoglobulin IVIg, natural human polyclonal pcPAbs and effectuated the characterization of their cell-penetrating ability and intracellular action. Based on published data, our Laboratory proceeded with the development of human monoclonal pcPAbs that penetrate into the cytoplasm of living cells and focused on the study of their multiple intracellular role. Those antibodies are indowed with distinct biological properties such as apoptosis induction on cancer cells as well as targeting of molecules with nodal roles in pathologies such as cancer and Alzheimer’s disease. Furthermore, as those antibodies also possess the ability of intracellular transport of diverse biomolecules (e.g. peroxidase, FITC, polystyrene particles) pcPAbs-nanoparticles complexes were created. In this context, the Laboratory is engaged in the making of pcPAbs with biodegradable nanoparticles-carriers of pharmaceutical agents, aiming the realization of targeted therapies against cancer and neurodegenerative diseases.

2. Free range pastured Poultry biochemical Indicators (Novel and Dynamic) to certify Outdoor rearing Systems


This project participants are The Biochemistry Laboratory of the University of Ioannina, the Agricultural Poultry Association of Ioannina (APSI «PINDOS») and the immunology Laboratory of HPI. Those entities carry out together a large industrial research whose goals are:

  1. a) the development of methods, mechanisms and tools of verification of the authenticity of free-range poultry husbandry and consumer protection from fraud and adulteration of high value added as well as b) the establishment of an innovative dietary method of poultry farming, unique in a national level, leading to an increase of the internal value added and a production of high quality end products with exportation potential.

The HPI Immunology Laboratory will undertake the research and development of new immunological and biochemical quality markers in free-range chickens.

The emergence and improvement of the special characteristics of Greek products of primary production by applying innovative production processes is a main step towards fulfilling the Regional development policy in the Agri-food industry, as expressed in the Smart Specialization priorities (RIS3) of the Region of Epirus.

3. Development of an antigen-specific therapy for Myasthenia gravis

Myasthenia gravis (MG) is an antibody-mediated autoimmune disease affecting the neuromuscular junction, causing muscle weakness and fatigability. In the majority of patients (85-90%) it is caused by autoantibodies targeting the muscle acetylcholine receptor (AChR), thus impeding neuromuscular signal transmission. Although the B cell-produced autoantibodies are the pathogenic agents in MG, T cells play a central and indispensable role in disease development, promoting the activity of autoreactive B cells.

The major MG autoantigen, the muscle AChR, is a pentameric transmembrane protein composed of four homologous subunits with the stoichiometry 2α11-ε-δ (or 2α11-γ-δ for embryonic receptors). Interestingly, most of the autoantibodies target the α1 subunit. Furthermore, a number of T cell epitopes have also been identified within the α1 subunit.

Our team (together with the Molecular Neurobiology and Immunology laboratory of the HPI) has developed and characterized in great detail an experimental autoimmune MG (EAMG) model in rats, using immunization with domains of the human AChR subunits, which results in highly reproducible symptom emergence and progression. Due to the use of human proteins for disease induction, this model is ideal for the study of antigen-specific MG immunotherapies.

We are working towards the development of a therapeutic strategy aiming to restore the lost tolerance towards the muscle AChR. The treatment rational is based on chronic administration of the autoantigen or its peptides under specific tolerance-inducing conditions. Recent findings in in vivo studies have shown a significant reduction of symptoms in treated animals compared to untreated or mock-treated ones. Our main efforts currently are focused on elucidating the precise mechanisms and cell populations involved in this response, which will not only increase our understanding and facilitate progression towards clinical application, but also potentially allow the development of even more targeted approaches.

PAST RESEARCH PROJECTS

Αutoimmune thyroid diseases: development of experimental models

The Laboratory of Immunology within three specific research projects, has been engaged on the study of autoimmune thyroid diseases (AITD), especially on Hashimoto’s thyroiditis (HT) and Graves’ disease (GD). The main achievements of the Laboratory are the following:

  • Identification and characterization of epitopes of thyroglobulin epitopes recognized by T lymphocytes (T epitopes) (thyroid gland main autoantigen) and subsequently, through the use of respective synthetic peptides, development of new experimental models for the study of immunoregulatory mechanisms.
  • Development, for the first time worldwide, of a GD model that could consistently reproduce all the symptoms of the disease in a large number of animals. Our research results indicated that intradermal delivery of human thyrotropin receptor (hTSHR) DNA could break tolerance and elicit GD in inbred mice.
  • Characterizatilon of B-cell epitopes of thyroglobulin epitopes recognized by B lymphocytes (B epitopes) and their use on the diagnosis of AITD.

FUNDING

NATIONAL PROGRAMS

2019-2021: National Strategic Reference Framework (NSRF), Human Resources Development, Education and Lifelong Learning, in the context of “Support to researchers with emphasis to young researchers-Round B’” Call, Proposal no: 1091, Research Field: Medical & Biological Sciences, Title: ”Study of human, natural, monoclonal antibodies endowed with cell-penetrating ability as a model of targeted and combinatorial anti-cancer therapy”.

2018-2020: NSRF, IPIROS Prefecture, MIS 5033100, 28/11/2017, Field: AGRI-FOOD. Title: Development of a model unit of industrial poultry rearing/breeding/farming. (IPIROS) 1. University of Ioannina 2. Agricultural, Poultry Farm Cooperative of Ioannina “PINDOS” 3. “AGROZOI” Public Limited Company- Industrial and Commercial Company-Agricultural and Poultry Farming Businesses 4. Immunology Laboratory, Immunology Department, Hellenic Pasteur Institute, Athens, Greece.

2018-2020: Operational Programme Competitiveness, Entrepreneurship and Innovation 2014-2020 (EPAnEK); sectoral operational programme of the Partnership and Cooperation Agreement (the new NSRF) for the period 2014-2020, approved on 18/12/2014 by the European Commission. (Proposal no: Τ1ΕΔΚ-03939, 13-06-2017), Research field: AGRI-FOOD, Title: Free range pastured Poultry biochemical indicators (Novel and Dynamic) to certify Outdoor rearing Systems”. Acronym: P.I.N.D.O.S.

2017-2020: General Secretary of Research & Technology (GSRT), KRIPIS II action: (MIS) 5002486 «Infectious, autoimmune and neurodegenerative diseases: Study of pathogenic mechanisms and development of diagnostic, prognostic and therapeutic strategies» -(P.L.: Responsible for sub-project WP4.4: “Development of human monoclonal IgG autoantibodies able to penetrate living cells, for intracellular targeting of the nucleus or the cytoplasm”)

INDUSTRIAL R&D PROGRAMS

2016-2020: TOLERANZIA ABO R&D Funding “In vivo preclinical studies on immunotherapy approaches for myasthenia gravis” (P.I.: K. Lazaridis)

 

SELECTED PREVIOUS FUNDING

NATIONAL PROGRAMS

2013-2015: General Secretary of Research & Technology (GSRT), KRIPIS action: in the context of InfeNeuTra project of the HPI code no MIS 450598. Action co-funded by Greece & the European Regional Development Fund of the EU, through Operational Program Competitiveness & Entrepreneur-ship of the National Strategic Reference Framework (NSRF) 2007-2013-(P.L.: Responsible for sub-project EE4.8: “Monoclonal polyreactive autoantibodies able to penetrate living cells as new therapeutic tools for intracellular therapy of viral, parasitic & bacterial infections”)

2013-2015: GSRT, Life-Sciences and Biotechnology-BIONIAN Cluster: in the context of the action “Creating Innovative Business Clusters – A Greek Product, One Market, the Planet”. Action co-funded by Greece & the European Regional Development Fund of the EU, through Operational Program Competitiveness & Entrepreneurship of the NSRF 2007-2013- (P.L.: Subcontractor of ICI (Institute for Clinical Investigation) & Responsible for the project “Monoclonal polyreactive cell-penetrating antibodies as new therapeutic tools for infectious diseases”)

2003-2005: GSRT, PRAXE (Phase Α): “Development of immunological tests for the early quality control of bovine meat consisting of the measurement of substances which are related to the quality and the sensory attributes of the final product which is supplied to the market (P.L.: Coordinator)

1999-2001: GSRT, EPET II/Career development program for Greek-speaking researchers working abroad: “Introduction of new methods to study thyroid autoimmunity”-Engagement for three years of Prof. George Carayanniotis (P.L.: Coordinator)

1997-1999: GSRT, EPET II/EKBAN: “Development of infrastructure for the production of recombinant proteins and monoclonal antibodies for diagnosis and therapy”– in Collaboration with ELPEN Pharmaceutical Company (P.L.: Subcontractor)

EUROPEAN-NATO R&D PROGRAMS

1997-1999: Science for Stability program (SfS) Outreach Joint Project/NATO: “Research and Development of Immunological methods: Production of immunodiagnostic assays for the detection of hepatitis and HIV1/2” [P.LYMBERI (P.L): Subcontractor]

1997-1999: FAIR [PL96-1107]: “The prediction of meat quality at the early post-mortem period by detection of novel physical and chemical markers” (P.L: Subcontractor)

1988-1993: SfS/NATO: “Research and Development of Immunodiagnostic Methods” (P.L.: Subcontractor) – Collaboration with Farmalex Pharmaceutical Company

 

 OTHER FUNDING

2017-2019: Donation by I. Kabouris for cancer research: Development of human monoclonal autoantibodies for targeting of cancer cells (P.L.: Coordinator).

2010-2012: ENDOCRINOLOGY SOCIETY: “The role of AGEs (Advanced Glycation End Products) which are present in food, in experimental autoimmune thyroiditis” (P.L.: Responsible for HPI)

2007-2009: ENDOCRINOLOGY SOCIETY: “The role of non-enzymic glycation in the generation of thyroid antibodies in pregnant women with diabetes mellitus” (P.L.:Responsible for HPI).

2006-2008: HPI RESEARCH OF EXCELLENCE: “The study of the role of autoantibodies able to penetrate into living cells” (P.L.:Coordinator).

2000-2002: HPI RESEARCH ACCOUNT 2000: “The role of apoptosis in the pathogenesis of autoimmune thyroididtis: study of Fas/FasL interaction” (P.L.: Coordinator).

TEACHING ACTIVITIES / DISTRIBUTION OF KNOWLEDGE

2000 – to date: Participation every year in two MSc Interdepartmental Postgraduate Courses “Clinical Biochemistry & Molecular Diagnosis”, and “Applications of Biology in Medicine “of National and Kapodistrian University of Athens, teaching “Natural and Pathological autoimmunity” (3h theory) and “Diagnostic methodology used in thyroid autoimmune diseases: as an example of organ-specific autoimmune diseases” (3h practice) in the context of the Chapter of “Immunology”

2018: Participation in MSc Postgraduate Course of Medical School University of Thessaly “Molecular Medicine Clinical Applications”, talk «The know-thyself of the Immune System: Its significance and applications in Clinical Practice”

2012- to date: Supervision of undergraduate, MSc and PhD students.

2012 – to date: Three-month or six-month training programs of undergraduate students.

2012-2017: The Laboratory of Immunology hosted the visit of high school baccalauréat-candidate students with a scientific orientation from the Lycée Franco-Hellénique Eugene Delacroix (LFH-ED). The main purpose of the visit was to assist these students in their professional career as biologists/researchers at French universities. They were introduced to the main concepts of Immunology and to the research interests of the Laboratory, and were also trained on immunological techniques. Overall, their 5 h- visit and their interaction with the researchers, along with the familiarization on the techniques, was very successful and LFH wishes to expand its visitation to our Laboratory, on an annual basis, given the impact of such meetings on the students’ professional career.

2013 – to date: Participation in the “Forum des Metiers” organized by the LFH-ED -inviting parents with different professions- to assist high school students in their professional career as biologists at French universities and future researchers.

2012 and 2013: Participation of our laboratory to the event “Night with the Researcher” that is open to the public and held at Demokritos Research Institution, with interesting videos, mini experiments and an interactive snake board game for elementary school students based on knowledge about vaccines and their importance. It was highly appreciated by the people that visited HPI exhibition stand and all the children received an illustrated copy of the game in order to test in their classroom the knowledge acquired.

2015 and 2017: Participation of our laboratory to the event “Athens science festival” that is open to the public and held at Technopolis City of Athens with easy and funny experiments for elementary and secondary school students, with videos and posters based on knowledge about the immune system key players and the vaccines and their importance.

ACHIEVEMENTS

  1. Basic research

2010   Hellenic Society of Immunology for the best study presented in poster during the 8th Hellenic Congress in Immunology

2009   European Society for Reproductive Immunology for the best study presented orally during the 7th European Congress on Reproductive Immunology

2005   General Secretary of Research and Technology: Project of excellence “Autoantibodies able to penetrate living cells”

  1. Αpplied research achievements

2004-2007 Supply of human thyroglobulin in the market

1999         Transfer of the technology developed in the context of FAIR PL96-1107 EU program

to the National Food Centre of Dublin –Ireland

PUBLICATIONS

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ZANNIKOU M, BELLOU S, ELIADES P, HATZIOANNOU A, MANTZARIS M, CARAYANNIOTIS G, AVRAMEAS S, LYMBERI P. DNA-histone complexes as ligands amplify cell penetration and nuclear targeting of anti-DNA antibodies via energy-independent mechanisms. Immunology 2016, 147: 73-81

2016
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SALI AD, KARAKASILIOTIS I, EVANGELIDOU M, AVRAMEAS S, LYMBERI P. Immunological evidence and regulatory potential for cell-penetrating antibodies in intravenous immunoglobulin. ClinTransl Immunology 2015; Volume:4 Issue:10 Pages: e42; doi: 10.1038/cti.2015.18

HATZIOANNOU A, KANISTRAS I, MANTZOU E, ANASTASIOU E, PEPPA M, SARANTOPOULOU V, LYMBERI P, ALEVISAKI M. A Effect of Advanced Glycation End Products on Human Thyroglobulin's Antigenicity as Identified by the Use of Sera from Patients with Hashimoto's Thyroiditis and Gestational Diabetes Mellitus. Int J Endocrinol 2015; 2015:849615. doi: 10.1155/2015/849615. Epub 2015 Jul 2.142(2):300-6

2015
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KANISTRAS J, HATZIOANNOU A, LYMBERI P. A novel pathogenic peptide of thyroglobulin (aa 2208-2207) induces autoreactive T-cell and B-cell responses in both high and low responder mouse strains Immunology 2014; 142(2):300-6

2014
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HATZIOANNOU A, ALEVIZAKI M, CARAYANNIOTIS G, LYMBERI P Fine epitope mapping within the pathogenic thyroglobulin peptide 2340-2359: minimal epitopes retaining antigenicity across various MHC haplotypes are not necessarily immunogenic Immunology 2012; 135: 245-253

VASILEIOUV, SARANDOPOULOU V, PHILIPPOU G, LYMBERI P, ET AL. Does the presence of thyroid antibodies affect the course and outcome of pregnancy in type 1 diabetic women? Diabetologia 2012; 55: S436-7

2012
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MATHIOUDAKI K, SCORILAS A, ARDAVANIS A, LYMBERI P, TSIAMBAS E, DEVETZI M, APOSTOLAKI A, TALIERI M, Clinical evaluation of PRMT1 gene expression in breast cancer. Tumor Biology 2011; 32:575-582

LYMBERI P, TERPOS E, HATZIOANNOU A, ET AL. High frequency of monoclonal immunoglobulins exhibiting natural autoantibody-like activity in patients with multiple myeloma and Waldenstrommacroglobulinemia. Blood 2011; 118(21): 1248-1249

2011
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HATZIOANNOU A, BOBOU G, LYMBERI P, BARNEA E. Suppressive effect of an embryo-derived factor to pathogenic T-cells causing thyroid autoimmunity. J ReprodImmunol 2009; 81(2): 141-141

2009
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AVRAMEAS S, TERNYNCK T, TSONIS IA, LYMBERI P. Naturally occurring B-cell autoreactivity: a critical overview. J Autoimmun 2007; 29:213-218

LIVADITI O, GIAMARELLOS-BOURBOULIS EJ, KAKKAS I, KAPSIMALI V, LYMBERI P, PAPASTARIADES C, DOUZINAS EE. Grouping of patients with common variable immunodeficiency based on immunoglobulin biosynthesis: comparison with a classification system on CD4-naïve cells. Immunol.Lett 2007; 114:103-9

HATZIOANNOU A, LIAKATA E, KARRAS E, THRASYVOULIDES A, ALEVIZAKI M, LYMBERI P. Pathogenicity of a human thyroglobulin peptide (2340-2359) in mice with high or low genetic susceptibility to thyroiditis. Immunology. 2007; 122: 343-349

THRASYVOULIDES A, LIAKATA E, LYMBERI P. Spreading of antibody reactivity to non-thyroid antigens during experimental immunization with human thyroglobulin. ClinExpImmunol 2007; 147:120-127

2007
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PHILIPPOU G, SARANDOPOULOU V, ALEVIZAKI M, LYMPERI P ET AL. Type 1 diabetic pregnant women may develop postpartum thyroiditis independently of the presence of anti-thyroid antibodies. Diabetologia 2006; 49: 571-572

2006
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KARRAS E, YANG H, LYMBERI P, CHRISTADOSS P. Human thyroglobulin peptide p2340 induces autoimmune thyroiditis in HLA-DR3 transgenic mice. J Autoimmun 2005; 24: 291-6

DAI Y, ELIADES P, CARAYANNIOTIS K, MCCORMICK DJ, KONG Y-CM, MAGAFFA V, CORDOPATIS P, LYMBERI P, CARAYANNIOTIS G. Thyroxine-binding antibodies inhibit T cell recognition of a pathogenic thyroglobulin epitope. J Immunol 2005; 174: 3105-10

2005
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THRASYVOULIDES A, LYMBERI P. Antibodies cross-reacting with thyroglobulin and thyroid peroxidase are induced by immunization of rabbits with an immunogenic thyroglobulin 20mer peptide. ClinExpImmunol 2004; 138: 423-9

BARRETT K, LIAKATA E, RAO PV, WATSON P, WEETMAN A, LYMBERI P, BANGA JP, CARAYANNIOTIS G. Induction of hyperthyroidism in mice by intradermal immunization with DNA encoding the thyrotropin receptor. ClinExpImmunol 2004; 136: 413-422

TSITSILONIS OE, THRASYVOULIDES A, BALAFAS A, VOUTSAS JF, PAPAMICHAIL M, LYMBERI P. Serological detection of hepatitis B viral infection by a panel of solid-phase enzyme-linked imunosorbent assays (ELISA). J Pharmaceut Biomed Analysis 2004; 34: 811-822

2004
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LIAKATA E, PHILIPPOU G, LYMBERI P,CARAYANNIOTIS G.Assessment of the frequency of mutant (hprt-) T lymphocytes from peripheral blood of patients with Hashimoto’s thyroiditis. Thyroid 2003;13: 631-6

ZAMANOU A, SAMIOTAKI M, PANAYOTOU G, MARGARITIS L, LYMBERI P. Fine specificity and subclass of IgG anti-actin autoantibodies differ in health and disease. J Autoimmun 2003; 20: 333-344

THRASYVOULIDES A, LYMBERI P. Evidence for intramolecular B-cell epitope spreading during experi-mental immunization with animmunogenic thyroglobulin peptide.ClinExpImmunol 2003;132: 401-7

KARRAS E, CARAYANNIOTIS G, LYMBERI P. Induction of murine thyroiditis by a non-dominant Ek –restricted peptide of human thyroglobulin. Immunology 2003; 108: 556-561

2003
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ZAMANOU A, TSIROGIANNI A, TERZOGLOU C, BALAFAS A, ECONOMIDOU I, LYMBERI P. Anti-smooth muscle antibodies (ASMA) and anti-cytoskeleton antibodies (ACTA) in liver diseases: A comparison of classical indirect immunofluorescence with ELISA. J Clin Lab Analysis 2002; 16(4):194-200

TSITSILONIS OE, STOEVA S, ECHNER H, MARGOMENOU L, BALAFAS A, TROY DJ, VOELTER W, PAPAMICHAIL M, LYMBERI P. A skeletal muscle troponin T ELISA based on the use of an antibody against the soluble troponin T (16-31) fragment. J Immunol Methods 2002; 268:141-8

2002
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THRASYVOULIDES A, SAKARELLOS-DAITSIOTIS M, PHILIPPOU G, SOUVATZOGLOU A, SAKARELLOS C, LYMBERI P. B-cell autoepitopes on the acetylcholinesterase-homologous region of human thyroglobulin: association with Graves’disease and thyroid eye disease.Eur J Endocrinology 2001; 145(2): 119-127

2001
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VOELTER W, STOEVA S, SCHICK M, ECHNER H, BECK A, LEHMAN R, MULLEN A.M, CASSERLY U, TROY D.J, TSITSILONIS O.E, LYMBERI P, BAXEVANIS C.N, PAPAMICHAIL M. Αnalytical tools for rapid, sensitive, quantitative identification of potential meat quality markers: application of immunoassays, HPLC and capillary electrophoresis. J PraktChem 2000; 342: 179-191

DAI Y, CARAYANNIOTIS KA, ELIADES P, LYMBERI P, ET AL. Blockade of T-cell recognition of a pathogenic thyroglobulin epitope by thyroxine-specific antibodies. Faseb J 2000; 14(6): A999-A999

2000
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DAI Y, CARAYANNIOTIS K, ELIADES P, LYMBERI P, SEPHERD P, KONG Y-CM, CARAYANNIOTIS G. Enhancing or suppressive effects of antibodies on processing of a pathogenic T-cell epitope in thyroglobulin. J Immunol 1999; 162: 6987-6992

1999
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PUBLICATIONS IN GREEK SCIENTIFIC JOURNALS

• AVRAMEAS S, LYMBERI P. The immune system from the perspective of natural autoimmunity. Immunity (Edition of the Hellenic Society of Immunology) 2014, 10, 2: 99-103.

• LYMBERI P, PHILIPPOU G. Thyroid autoimmnity: autoantigens, autoantibodies, autoreactive T lymphocytes, pathogenicity. Αrchives of Hellenic Medicine 1999, 16: 337-351.

MEMBERS

Peggy Lymperi

Research Director
Head of Laboratory

Konstantinos Lazaridis

Principal Investigator
Τηλ.: 210-6478807

Apostolos Balafas

Technician
Τηλ.: 210-6478807
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Theodora Stivarou

Post-Doctoral Fellow
Τηλ.: 210-6478807

Ioannis Sarrigeorgou

PhD Student

Vasiliki Baltatzidou

PhD Student
Τηλ.: 210-6478807
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Eleni Doukaki

BSc Student

PAST MEMBERS

 

ALUMNI

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