Εργαστήριο Ανοσολογίας

Head of the Laboratory
Lymberi Peggy, Research Director
E-mail: plymberi@pasteur.gr
Tel: +30 210 6478808, 6478807
Fax: +30 210 6478808

About

Autoimmunity, both natural and pathological, is the main research field of the Immunology Laboratory of HPI.

Specifically, as far as natural autoimmunity is concerned we aim to:

  • study the role of natural autoantibodies of human or animal origin in the homeostasis of the immune system, as well as their relationship with pathology
  • investigate the properties and biological role of natural antibodies of IgG class which are able to penetrate into living cells (polyreactive cell-penetrating antibodies; pcPAbs), to be used as therapeutic molecules for intracellular intervention, either per se or as carriers of various active agents

As far as pathological autoimmunity is concerned we aim to:

  • develop animal models for Graves’ disease in order to understand the pathogenic mechanisms underlying this antibody-mediated autoimmune disease of thyroid gland
  • identify the T-cell epitopes of thyroglobulin (the main autoantigen of thyroid gland) and use them for the development of animal models for Hashimoto’s thyroiditis in order to reveal the immunoregulatory mechanisms that impair the development of this T-cell mediated autoimmune thyroid disease
  • identify B-cell epitopes of thyroglobulin in order to use them for the early diagnosis of thyroid autoimmune diseases

The above studies (on thyroid animal models and cell-penetrating antibodies) met the requirement of excellence during HPI’s evaluation by external referees of the General Secretary of Research and Technology in 2005.

Our long-lasting experience in Immunochemistry allowed us to gain significant achievements in applied research, consisting mainly in the development of new immunological products (reagents and immunoassays with a broad spectrum of applications). In this context, we have participated since 1999 in numerous applied research projects (NATO/Science for Stability programme, EU/FAIR, GSRT/EPET ΙΙ) in collaboration with the private sector (Greek pharmaceutical companies Pharmalex and ELPEN). The product range includes: anti-isotypic antibodies and the respective enzyme conjugates, immunodiagnostic assays for viral infections (e.g. hepatitis B, HIV1/2) and autoimmune diseases (e.g. Hashimoto’s thyroiditis), as well as immunoassays for quantitation of various biomolecules (e.g. cytokines) and biochemical quality indicators of meat supplied in the market. In fact, part of these products have been successfully commercialised in the Greek market. More recently, the laboratory produces human thyroglobulin of high quality and supplies it through the HPI website, while it intends to expand its production range with monoclonal antibodies to human thyroglobulin and other proteins.

Great effort is also dedicated to training in Immunology both, undergraduate and postgraduate students as well as scientists at various professional levels. Specifically, 35 (PhD, MSc and BSc) students have successfully completed their dissertation thesis to date and we have successfully organized, in collaboration with the Pasteur Institute of Paris, five 10-day postgraduate seminars on “Modern Diagnostic Methodology used in infectious and autoimmune diseases”, held in HPI for medical doctors and biologists of the Public Health Sector. In parallel, the laboratory organizes and participates in educational programs at other Institutions (Universities / postgraduate courses, Ministry of Health and Social Solidarity and Scientific Societies). Finally, since 2009 the Immunology laboratory has organized the Seminar “The little biologist” for elementary grade students in order to familiarize them with the field of Biology.

Projects

I.  Thyroid Autoimmunity

Background: Autoimmune thyroid diseases (AITD), especially Hashimoto’s thyroiditis (HT) and Graves’ disease (GD) are widely spread throughout the general population* and are characterised by high morbidity, as a large number of patients usually have to be treated pharmaceutically for their entire life-span and require frequent medical attention. This, in turn, greatly increases the health care costs and therefore new methods for prevention and early diagnosis of the diseases are urgently required. HT is caused by the activation of autoreactive T-lymphocytes, which recognize thyroglobulin (Tg) and thyroid peroxidase. These infiltrate and destroy the thyroid gland resulting in reduced hormone production and thus hypothyroidism. On the other hand, GD is caused by autoantibodies which recognize the thyroid stimulating hormone receptor. These antibodies work by mimicking the hormone and therefore forcing the gland to overexpress T3 and T4 hormones, leading to hyperthyroidism. Autoimmune thyroiditis can be induced in mice of appropriate genetic background by Tg or Tg-peptides administration, a model referred to as Experimental Autoimmune Thyroiditis (EAT). EAT is an excellent model of an autoimmune organ-specific disease (because the target-organ is easily accessible and its autoantigens are known) ideal for the study of immunoregulatory mechanisms at T-cell level. In the case of GD, no suitable animal model has been developed so far.

* Hypothyroidism caused by HT has a prevalence of 4-8% in the general population and up to 20% in the subclinical form. Anti-thyroid autoantibodies are present in 25-30% in women, especially in women older than 70 years. Hyperthyroidism caused by GD has a prevalence of 2.3% among women and 0.3% among men and in its subclinical form the levels are raised up to 40% of the general population.

Aim: The study of immunogenicity of Tg (the main autoantigen of the thyroid gland and the largest known) at B- and T-cell epitope levels has been our main interest in order to develop new diagnostic tools for AITD and establish new animal models for both human diseases, HT and GD. Our ultimate goal is to gain an insight into the pathogenic mechanisms underlying AITD and contribute to their early diagnosis and more efficient therapy.

Our laboratory has accumulated significant knowledge –unique in Greece- during the last decade in developing and studying animal models of AITD.

Projects:

Animal model of Graves’ disease

The research activity of our laboratory was focused on the development of a GD model that could consistently reproduce all the symptoms of the disease in a large number of animals, since up to date no satisfactory model exists. This is mainly due to the low expression of the thyrotropin receptor (TSHR) -main autoantigen of the disease- on thyrocytes which makes it, consequently, difficult to isolate in large quantities, necessary for animal studies. The study consisted in the genetic immunisation, intradermally, of BALB/c mice with the plasmid DNA encoding the human TSHR alone or with cytokine producing plasmids (IL-2, IL-12). Our results indicated that intradermal delivery of hTSHR DNA could break tolerance and elicit GD in inbred mice (Barrett K. et al, 2004). This study was performed in close collaboration with the laboratory of Prof. G. Carayanniotis (St John’s University, Newfoundland, Canada)

B-cell epitopes of Tg and Serology of AITD

In the context of this project, we aim to characterize B-cell epitopes of Tg and use them in the diagnosis of AITD and the differentiation of HT and GD. Up to date, we have identified three 20mer peptides (a.a. 2340-2359, 2472-2491 and 2652-2671) of the C-terminal region of Tg that were associated with GD. It was found that autoantibodies recognizing peptide 2652-2671 were also associated with thyroid ophthalmopathy (Thrasyvoulides A. et al, 2001). Moreover, it was demonstrated that this peptide was immunogenic and induced B-cell epitope spreading (intramolecular and intermolecular) after its administration to rabbits (Thrasyvoulides A. et al, 2003, 2004, 2007). In an ongoing study we perform a B-cell epitope mapping of Tg at the C-terminal as well as at central regions.

Effect of non-enzymic glycation on the immunogenicity of Tg and on the development of EAT

Diabetic patients frequently develop autoimmune thyroiditis and anti-thyroglobulin antibodies (ATGAbs). It is well known that hyperglycemia of diabetic patients results in the non-enzymic glycation of proteins and later on in the formation of advanced glycation end products (AGEs), which are implicated in the development of chronic complications of diabetes.

Nevertheless, there are no data whatsoever correlating the enhanced AGE formation with the production of ATGAbs, or even with the development of Autoimmune thyroiditis. This outcome could be due to the enhancement of Tg’s immunogenicity after its AGE-modification. To initially investigate whether the antigenicity of Tg is affected by accelerated glycation, sera from pregnant women with gestational diabetes, positive for ATGAbs, were tested in parallel against native and in vitro glycated Tg (AGE-Tg). The serum reactivity profiles showed that modification of Tg by AGEs did not alter its antigenicity Further comparison of the antibody titers against AGE-Tg, in the above sera with those in sera from patients with Hashimoto’s thyroiditis (typical thyroid autoimmune disease with high ATGAbs titers) and healthy individuals similarly showed that AGEs cannot affect Tg’s immunogenicity (Hatzioannou A. et al, manuscript in preparation). An ongoing study will shed light on the effect of AGE-rich food on the development of EAT in susceptible animals.

T-cell epitopes of Tg and Animal model of Hashimoto’s thyroiditis

One of the main concerns of the laboratory has been the identification of T-cell epitopes on Tg that could induce EAT to genetically different mouse strains. Mouse strains are characterised as susceptible or resistant, based upon their capability to develop EAT at different incidence and extent of infiltration of their gland, depending on their MHC haplotype. Nine years ago we were able to add an extra pathogenic peptide (a.a2340-2359) of Tg to the known ones (Karras E. et al, 2003). This peptide has the unique feature to induce EAT not only to genetically susceptible mouse strains but also to resistant ones (Hatzioannou A. et al, 2007), as well as in transgenic mice that carry the human MHC molecule (DR3) (Karras E. et al, 2005). Minimal T-cell epitope mapping of (2340-2359) revealed a core 9-mer peptide (a.a. 2344-2352, LTWVQTHIR) that is recognized by effector cells in both susceptible and resistant mouse strains. This finding provides support to the notion that, in mouse strains of diverse genetic backgrounds, EAT may be mediated by autoreactive T-cell clones recognizing the same Tg epitope(s) in the context of various MHC class II molecules (Hatzioannou A. et al, 2012). In addition, we have recently identified a second Tg peptide able to induce EAT in both susceptible and resistant mouse strains. This peptide exhibits a unique pathogenic profile; it is more pathogenic in resistant mouse strains than in susceptible ones (Kanistras I. et al, submitted for  publication). Our efforts are now focusing on mapping Tg T-cell epitopes in genetically diverse strains, since they will provide important information when extrapolated to an outbred population, closely resembling the human disease. This finding will support our hypothesis postulated above and if applied to human disease, such “promiscuous” recognition of shared Tg T-cell epitopes may account, in part, for the lack of strong HLA associations with human disease. Moreover such knowledge would expand the study of Tg-peptide-induced EAT to genetic backgrounds that harbour non-MHC alleles useful for immunoregulation studies. In parallel with the above studies, we produced Tg-specific mouse monoclonal antibodies, able 1) to influence Tg processing and, therefore, enhance or suppress the generation of a non-dominant pathogenic T cell epitope (Dai Y. et al, 1999) and 2) to inhibit T cell recognition of a pathogenic Tg epitope and therefore may influence the development of thyroid disease (Dai Y. et al, 2005).

Collaborations:

From Greece: Prof. M. Alevizaki (Endocrine Unit Eugenideion Hospital and Department Medical Therapeutics, University of Athens, School of Medicine), Prof. A. Tsatsoulis (Endocrinology Dept, Medical School, University of Ioannina),  M. Varla-Leftherioti (Laboratory of Immunology and Histocompatibility, “Elena Venizelou” Hospital, Athens)

From USA: Dr E. Vlassara (Division of Experimental Diabetes and Aging, Brookdale Dept of Geriatrics, Mt Sinai School of Medicine, New York),

From Canada: Prof G. Carayanniotis (Endocrinology Dept, Medical School of St John’s University, Newfoundland)

II. Natural autoantibodies able to penetrate into living cells (polyreactive cell penetrating antibodies: pcPAbs)

Background: During the last six years we initiated a study on natural polyreactive IgG autoantibodies able to penetrate the cytoplasmic and/or nuclear membrane of living cells and to reach their antigen-targets in the respective compartments. PcPAbs are known since the late seventies. Compared to all other antibodies, also termed as penetrating / internalizing / translocating [i.e. structurally modified by chemical or genetic  engineering or coupled to short cationic peptides with membrane translocating properties (e.g. Cell Penetrating Peptides-CPPs), or antibodies monospecific for cell surface receptors] they possess unique properties. More specifically, pcPAbs exhibit polyreactivity, since they recognise antigens of different composition and structure, such as DNA, proteins and haptens and are often encoded by germ line genes, properties common to natural antibodies, a major part of circulating immunoglobulins in healthy individuals and in most animal species. Their penetration mechanism(s) is not yet elucidated, but there is strong evidence of an active process, associated to their polyreactive binding site, which is endocytosis-independent. Once inside the cells, certain pcPAbs do not seem to influence the cell functions, while others induce modifications in cytokines production, cell proliferation or cell apoptosis. Moreover, when conjugated to different molecules of either high or low molecular weight, such as alkaline phosphatase, horseradish peroxidase, avidin, oligonucleic acids, biotin fluorescent dyes, colloidal gold, they retain their penetrating capability, both in vitro and in vivo. Therefore, pcPAbs meet most of the criteria for their use as therapeutic molecules for intracellular intervention, either per se or as carriers of various active agents.

Aim: We aim to investigate the immunochemical and penetrating characteristics, as well as the biological role of pcPAbs present in pathological and healthy conditions, and ultimately develop novel approaches for the intracellular therapy of a wide variety of diseases.

Projects:

Biochemical and immunological properties and cell-penetration characteristics of pcPAbs:

The Immunology Laboratory produced a series of monoclonal pcPAbs derived from mice that spontaneously develop lupus [(NZBxNZW)F1:BW], as well as from healthy mice -that have never been used so far- as a source of such antibodies. These antibodies have been characterized for their ability to recognize nuclear antigens (DNA, SmD, TopoisomeraseI, Histones), haptens (trinitrophenol-TNP) and cytoskeletal antigens (actin, tubulin, myosin). Each of them exhibits a unique polyreactivity profile. Moreover, the optimal conditions (antibody concentration, number of cells, incubation time) that allow a maximum in vitro penetration have been defined using various cancer/non-cancer cell-lines. We have found that these monoclonal pCPAbs are also differentiated in terms of the cellular compartments and cell organelles in which they are located. Moreover, penetration and localisation can be altered in the presence of soluble nuclear antigens. (Zannikou M. et al, submitted for publication)

Mechanism(s) of entry of pcPAbs:

Successful penetration (of cell/nuclear membranes) at both, 37 °C and 4 °C,  indicates that our pcPAbs follow a distinct pathway of entry that differs from classical endocytic pathways, and is, therefore, advantageous to the entrance by phagolysosomes and the subsequent enzymic hydrolysis. Further studies concerning their mechanism of entry are under process.

Biological role of pCPAbs:

Ongoing studies concerning the in vitro mode of action of the above pCPAbs focus on their ability to hydrolyze plasmid and genomic DNA and to induce apoptosis to a series of cell lines. Their role in infections (viral, bacterial, parasitic), immune regulation and angiogenesis is currently also under investigation.

Therapeutic exploitation of pcPAbs:

Preliminary data have shown that pcPAbs retain their penetrating capacity in vitro, after their conjugation to peroxidase or FITC. Imaging analysis by photonic, fluorescence and electron microscopy revealed the localization of the conjugates in the cytoplasm or the nucleus of the cell lines tested. In addition, conjugation of pcPAbs to fluorescent polystyrene beads and gold nanoparticles has been carried out and in vitro penetration of the complexes was achieved. Further studies are focusing on their use as carriers of nanoparticles of different composition loaded with siRNA(s) or doxorubicin, in order to be applied in osteoarthritis or cancer animal models, respectively.

Isolation of pCPAbs from healthy animals and humans:

Monoclonal pCPAbs derived from healthy mice have never been described so far. The Immunology Laboratory produced a series of such antibodies from BALB/c mice; their properties are currently being investigated and compared to the above described disease-associated pcPAbs. As a source of IgG-pcPAbs of human origin we utilize IntraVenous Immunoglobulin (IVIg) commercial preparations, derived from pooled plasma of thousands of donors, and usually applied in the therapy of a wide spectrum of autoimmune/inflammatory diseases. In vitro and in vivo studies concerning the penetrating activity of these commercial preparations, as well as of specific antibody subpopulations isolated from them, have already provided us with interesting results (Sali A. et al, submitted for publication)

 

Confocal microscope images

Monoclonal antibodies localised in the nucleus (a,b), in the cytoplasm (b) and the perinuclear space (c) of HeLa cells. Visualisation via secondary antibody conjugated with fluorescence dye (FITC). The nucleus is marked with blue fluorescent dye.

 

Collaborations:

From Greece: Prof. T. Fotsis (Dept of Biochemistry, Medical School, University of Ioannina), Prof. C. Stournaras (Dept of Biochemistry, Medical School, University of Crete), Assoc. Prof. G. Thyphronitis (Dept of Biological applications and technologies, University of Ioannina), Assoc. Prof. C. Avgoustakis (Dept of Pharmacy, University of Patras), Αssoc. Prof. Α.Tsezou (Dept of Biology and Genetics, Medical School, University of Thessaly), Assist. Prof O. Tsitsilonis (Dept of Biology, University of Athens), Dr. A. Tsirogianni (Laboratory of Immunology and Histocompatibility, “Evangelismos” Hospital, Athens)

From France: Dr L. Mouthon (Service de Medecine Interne, Hopital Cochin, Paris)

From Japan: Dr K. Terashima (Pathology Dept, Medical School, University of Tokyo)

Education

Education

  • Supervision of PhD Students in collaboration with Greek Universities
  • Laboratory workshops – Education of under- / post-graduate students
  • Organisation and participation in postgraduate seminars on “Modern Diagnostical Methodology in infectious and autoimmune diseases”. They are frequently organised at IPH and are directed to doctors and biologists within the Public Health Sector
  • Organisation and participation in Educational programmes of other Institutions (Universities / postgraduate programmes, Ministry of Health and Social Solidarity, Scientific Societies)
  • Compilation of notes and laboratory methodology in the context of seminars and chapters or translations in educational books
  • Organisation of the seminar entitled “The Young Biologist” for elementary grade students, in the context of which the children are familiarised with the researcher’s study, learn about the microbes and the vaccination and participate in practical exercises, by observing various samples under the microscope
Members

Head of the Laboratory
Lymberi Peggy, Research Director
E-mail: plymberi@pasteur.gr
Tel: +30 210 6478808, 6478807
Fax: +30 210 6478808

Researchers
Liakata Elisavet, Researcher D΄, liakata@pasteur.gr

Technicians
Balafas Apostolos, balafas@pasteur.gr

Postgraduate students
Kanistras Ioannis, MSc,  ikanistras@pasteur.gr
Sali Aggeliki, BSc, asali@pasteur.gr
Zannikou Markella, BSc, mzannikou@pasteur.gr

Visiting Professor
Avrameas Stratis, Honorary Professor of Medicine, avrameasstratis@pasteur.gr

Publications

PUBLICATIONS IN INTERNATIONAL JOURNALS

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(3):245-53.

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. Tumour Biol. 2011; 32(3):575-82.

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; 114:103-109.

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. Clin Exp Immunol 2007; 147:120-127.

Karras E, Yang H, Lymberi P, Christadoss P. Human thyroglobulin peptide p2340 induces autoimmune thyroiditis in HLA-DR3 transgenic mice. J Autoimmun 2005; 24: 291-296

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: 3105-3110

Thrasyvoulides A, Lymberi P. Antibodies cross-reacting with thyroglobulin and thyroid peroxidase are induced by immunization of rabbits with an immunogenic thyroglobulin 20mer peptide. Clin Exp Immunol 2004; 138: 423-429

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. Clin Exp Immunol 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-immunosorbent assays (ELISA) J Pharmaceut Biomed Analysis 2004; 34: 811-822

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: 631-636

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 experimental immunization with an immunogenic thyroglobulin peptide. Clin Exp Immunol 2003; 132:401-407

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

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; 268:141-148

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

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 Prakt Chem 2000; 342: 179-191

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

Haralambous S, Blackwell C, Mappouras DG, Weir D, Lymberi P. Increased natural autoantibody activity to cytoskeleton proteins in sera from patients with necrobiosis lipoidica, with or without insulin-dependent diabetes mellitus. Autoimmunity 1995; 20: 267-275

Mappouras DG, Philippou G , Haralambous S , Tzartos S, Balafas A , Souvatzoglou A, Lymberi P. Antibodies to acetylcholinesterase cross-reacting with thyroglobulin in myasthenia gravis and Graves’ disease. Clin Exp Jmmunol 1995; 100: 336-343

Kourtis AP, Boussiotis VA, Lymberi P, Pangalis GA. Increased natural antibody activity in sera of patients with malignant non-Hodgkin’s lymphomas containing paraproteins. Am J Hematol 1994; 46: 283-288

Kourtis A, Pangalis G, Borche L, Boussiotou V, Dighiero G, Lymberi P. Monoclonal and/or oligoclonal immunoglobulins in sera of patients with non-Hodkin’s Lymphomas determined by isoelectric focusing. Leukemia and Lymphoma 1991; 5: 255-262

Anagnostides S, Alevras A, Lymberi P, Tsiganos C. Isolation and characterization of two glycoproteins from hyaline cartilage. Eur J Biochem (FEBS) 1990; 193: 905-912

Lymberi P, Aessopos A, Karageorga M, Hadziyianni D, Loukopoulos D, Kaklamanis Ph. Increased IgA natural autoantibody activity in sera of patients with β‑thalassemia. Autoimmunity 1990; 8: 81-82

Toft AD, Blackwell CC, Saadi AT, Wu P, Lymberi P, Soudjidelli M, Weir DM. Secretor status and infection in patients with Graves’ disease. Autoimmunity 1990; 7: 279-289

Dighiero G, Lymberi P, Monot C, Abuaf N. Sera with high levels of anti-smooth muscle and anti-mitochondrial antibodies frequently bind to cytoskeleton proteins. Clin Exp Immunol 1990; 82(1): 52-56

Lymberi P, Barbouche R. Assays for autoantibodies. Curr Opin Immunol 1990; 2: 917-922

Blancher A, Oksman F, Lymberi P, Calvas P, Cambon-Tomsen A, Clanet M, Ducos J. Human monoclonal autoantibodies produced by hybridomas derived from Multiple Sclerosis patients lymphocytes. Res Immunol (Inst Pasteur) 1989; 140: 711-715

Baxevanis C, Reclos G, Arsenis P, Anastasopoulos E, Katsiyiannis A, Lymberi P, Matikas N, Papamichail M. Decreased expression of HLA-DR antigens on monocytes in patients with multiple sclerosis. J Neuroimmunol 1989; 22: 177-184

Lymberi P, Blancher A, Calvas P, Avrameas S. Natural autoantibodies in nude and normal outbred (SWISS) and inbred (BALB/c) mice. J Autoimmunity 1989; 2: 283-295

Guery JC, Druet E, Glotz D, Hirsch F, Kuhn J, Lymbery P, Druet P. Idiotypes of autoantibodies in mercury-induced autoimmunity in the rat. Monogr Allergy (Karger, Basel) 1987; 22: 99-108

Dighiero G, Lymberi P, Guilbert B, Ternynck T, Avrameas S. Natural autoantibodies constitute a substantial part of normal circulating immunoglobulins. Ann NY Acad Sci 1986; 475: 135-145

Lymberi P, Hirsch F, Kuhn J, Ternynck T, Druet P, Avrameas S. Autoimmunity induced by HgCl2 in Brown-Norway rats. II. Monoclonal antibodies sharing specificities and idiotypes with mouse natural monoclonal antibodies. J Immunol 1986; 136: 3277-3281

Karray S, Lymberi P, Avrameas S, Coutinho A. Quantitative evidence against inactivation of self-reactive B-cell clones. Scand J Immunol 1986; 23: 475-480

Dighiero G, Lymberi P, Guilbert B. Les autoanticorps naturels produits par des clones autoréactifs constituent une partie substantielle des immunoglobulines circulantes, et les protéines monoclonales correspondent fréquemment à l’expansion incontrôlée de ces clones. Nouv Rev Franç Hematol 1985; 27: 59

Lymberi P, Dighiero G, Ternynck T, Avrameas S. A high incidence of cross-reactive idiotypes among murine natural autoantibodies. Eur J Immunol 1985; 15: 702-707

Dighiero G, Lymberi P, Holmberg D, Lundquist I, Coutinho A, Avrameas S. High frequency of natural autoantibodies in normal newborn mice. J Immunol 1985; 134: 765-771.

Avrameas S, Dighiero G, Lymberi P, Guilbert B. Studies on natural antibodies and autoantibodies. Ann Immunol (Inst. Pasteur) 1983; 134D: 103-113

Dighiero G, Lymberi P, Mazie JC, Rouyre S, Butler-Browne GS, Whalen RG, Avrameas S. Murine hybridomas secreting natural monoclonal antibodies reacting with self antigens. J Immunol 1983; 131: 2267-2272

Dighiero G, Guilbert B, Fermand J P, Lymberi P, Danon F, Avrameas S. Thirty-six human monoclonal immunoglobulins with antibody activity against cytoskeleton proteins, thyroglobulin and native DNA: Immunological studies and clinical correlations. Blood 1983; 62: 264-270

 

PUBLICATIONS IN GREEK JOURNALS

Lymberi P, Philippou G. Thyroid autoimmunity: autoantigens, autoantibodies, autoreactive T lymphocytes, pathogenicity. Αrchives of Hellenic Medicine 1999, 16: 337-351

 

CHAPTERS IN SCIENTIFIC BOOKS 

  • INTERNATIONAL

Thrasyvoulides A, Lymberi P. A human thyroglobulin peptide able to drive intramolecular B cell epitope spreading when administered in rabbits. In: Sadia Rashid (ed) Essays on Science. Felicitation Volume in honour of Prof. Wolfgang Voelter, Hamdard Foundation Pakistan, 2006, Volume 8, pp.1-17.

Amital-Teplizki H, Lymberi P, Tomer Y, Shoenfeld Y. The idiotypic network, autoimmunity, and natural autoantibodies. In: Shoenfeld Y and Isenberg DA (eds) Natural autoantibodies: their physiological role and regulatory significance, CRC Press, 1993; ch. 10, pp. 175-204.

  • GREEK

Lymberi P, Philippou G, LIAKATA Ε, Hatzioanou A,  Souvatzoglou Α. Thyroglobulin and thyroid peroxidase as autoantigens. Felicitation Volume in honour of Prof. Dimitris Koutras, Scientific Editions Parissianou, Αthens 2006, chapt. 15:pp. 101-108

Germenis ΑΕ, Lymberi p. Autoimmunity – Immunological laboratory methods– English-Greek dictionary of immunological terms. In: Germenis ΑΕ. Medical Immunology, Edit. Papazisis, Αthens 2000, chapt. 14:pp. 237-262, chapt. 20: pp. 383-412, appendix. D: pp. 469-480.

Lymberi P, Philippou G . Thyroid Antibodies. In: Intensive Education in Endocrinology – 7th Section: Thyroid, Hellenic Endocrinology Society Edition, Αthens 2004, pp. 131-144.

TRANSLATIONS

Translation from french to greek the laboratory manual “Techniques immunoenzymatiques” by Τ.Ternynck and  S.Avrameas”.  HPI Edition –1989

Funding

EUROPEANNATO 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”   (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” (Subcontractor

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

 

NATIONAL PROGRAMS

2003-2005
General Secretary of Research & Technology (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 (Coordinator)

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

1999-2000
KESY (National Health Council), Ministry of Health:  Serological investigation of postpartum thyroiditis in women with insulin-dependent diabetes mellitus

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

1993-1995
KESY:Isolation and immunological characterization of thyroid antigens – Development of diagnostic immunoassays for thyroid diseases (Responsible for HPI)

1993-1995 (part 1) and 1996-1998 (part 2)
KESY: Studies on natural autoantibodies in patients with renal failure, before and after transplantation”.

1990-1992 (Responsible for HPI)
KESY: Studies on the immunological disturbances at the molecular and cellular in patients with autoimmune hepatopathies (Responsible for HPI)

 

OTHER PROGRAMS

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

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

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

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

 

OWN RESOURCES

2004-up to date
Commercialisation of immunological products (human Thyroglobulin)