Laboratory of Molecular Genetics

Laboratory of Molecular Genetics

Head of the Lab:
Dr Lesley Probert
Tel: +30-210-6478866
Fax: +30-210-6456547
E-mail: lesley@pasteur.gr

The Lab

We are interested in the interactions between the immune system and the central nervous system (CNS) and in how these two complex recognition systems co-operate at the molecular and cellular levels to help maintain brain homeostasis under physiological as well as pathophysiological conditions.

Disturbances of immune-nervous system homeostasis can lead to chronic inflammatory diseases such as multiple sclerosis (MS), a common demyelinating neurodegenerative disease of the CNS that has autoimmune characteristics and is the main cause of neurological disability in young adults. Currently, the majority of treatments for MS induce non-specific immune or T cell suppression but are only partially effective and are often associated with adverse effects either because they suppress important immune functions and therefore the organism’s ability to fight infections, or, as is now becoming more obvious, because they also block critical CNS functions.

In our lab we aim to identify key molecules, mechanisms and cells that are involved in mediating inflammatory neurodegeneration in experimental models for stroke and MS with the aim of finding new, more selective therapeutic targets and neuroprotection strategies for the treatment of inflammatory CNS disorders. To achieve this goal most of our research efforts are directed towards gaining a detailed understanding of the functional interactions between immune cells and cells of the CNS under normal as much as under disease conditions.

Drawing from our previous findings, one example where the refinement of knowledge about functional immune-neuron interactions might aid the development of better therapies for chronic immune pathologies comes from studies of tumour necrosis factor (TNF) in the CNS. TNF is a master pro-inflammatory cytokine and front-line target for the treatment of chronic immune pathologies such as rheumatoid arthritis in humans. However, clinical data that non-selective TNF inhibitors exacerbate MS, trigger demyelination in patients with other diseases and that TNF receptor SF1A represents a MS susceptibility locus, clearly pointed to protective effects of TNF in the CNS and prohibited the non-selective targeting of this cytokine for neuroinflammation. Over the last decade important beneficial neuroactive properties of TNF have been discovered, notably neuroprotection, the facilitation of remyelination and synaptic plasticity. More recently, we and others have shown that the soluble and transmembrane forms of TNF exert opposing deleterious and beneficial effects in the CNS, respectively. Soluble TNF promotes inflammation while transmembrane TNF is neuroprotective and enhances remyelination in a mouse model of MS named experimental autoimmune encephalomyelitis (EAE). These data open the exciting possibility that selective inhibition of soluble TNF, rather than non-selective inhibition of both forms, may provide a new way forward for the treatment of a wide range of chronic immune and neurodegenerative pathologies including MS.

Co-localization of NF-κB p50 subunit (red) with NeuN (green)

Our current and future studies focus on translating basic research findings into new therapeutic leads for the treatment of inflammatory CNS disorders. One approach is to understand the participation of individual brain cell types, especially neurons and macrophage-like cells called microglia, in the defense of the brain during inflammation. By extending earlier studies in our lab which showed that neurons are critical for suppressing autoimmune responses in CNS tissues during EAE we are now looking at whether they regulate brain inflammation in other models of neurodegeneration. Detailed knowledge of the molecular and cellular mechanisms used by neurons to suppress CNS immune responses, or by T cells to trigger neuron damage, should help the design of better neuroprotective therapies. In another approach we are developing more than a decade of work with myelin auto-antigens to produce potential vaccines for MS. Myelin peptides can be targeted to antigen-presenting cells in such a way that they induce long-lasting peptide-specific T cell tolerance and are therapeutic in a preclinical model for MS. This approach is now being developed towards clinical trials in MS patients. Further details of the individual projects and the scientists involved are given below.

Our research is currently supported by an FP7 EU Regional Potential Grant (REGPOT-NeuroSign no: 246083) (www.neurosign.gr) and an ESPA Cooperation Grant from the Greek General Secretariat of Science and Technology, Ministry of Education (Immunotherapy for Multiple Sclerosis no: 09ΣΥΝ-22-609). The laboratory is also actively involved in the running of the European School of Neuroimmunology (ESNI) (www.esni.org).

Lab Tool Box

Our main investigative approach is to use transgenic or conditionally gene-targeted mice for specific genes and cells of interest in combination with pre-clinical models of CNS inflammation and neurodegeneration, mainly the MS models EAE and cuprizone-induced demyelination. We draw from a plethora of established analytical tools for investigating the functions of candidate molecules. These include analyses of differential gene/protein expression (by DNA microarrays, RealTime RT-PCR, antibody arrays and Western blots), cellularity (by FACS and immunohistochemistry), morphology and cellular interactions (conventional, time-lapse imaging and confocal microscopy) and function (clinical symptoms, immune responses, neuron death/survival, live cell imaging of calcium levels) as well as further validation of molecular mechanisms in primary cells and mice using gene reconstitution or knockdown techniques. Members of the lab run the Institute’s Transgenic Technology Unit, maintain a mouse repository for immune and disease strains and contribute to the Institute’s Experimental CNS Surgery facility for small rodents.

 

Vivi Tseveleki, Ph.D
Development of DC-targeted peptide vaccines for immunotherapy in MS

 

Gene expression profiling of mouse CNS disease models

Ioannis Kanistras, Ph.D and Vivi Tsveleki, Ph.D
Preclinical evaluation of DC-targeted peptide vaccines in a humanized MS model: preparation for clinical trials in MS patients

 

Maria Evangelidou, Ph.D
Live imaging of T cell-neuron interactions and their functional consequences

 

Maria Evangelidou, Ph.D and Sotiris-Spyros Vamvakas, Ph.D
Mini-panel of genes for monitoring the severity of CNS inflammation pathology and the efficacy of novel therapeutics

The aim of this project is to develop a mini-panel of genes, whose expression changes will sensitively monitor the development of neuropathological changes associated with CNS diseases like multiple sclerosis and be useful for the evaluating the efficacy of novel therapeutics in preclinical models. We originally selected and screened a large number of candidate genes based upon their known functions and association with the major pathological processes of multiple sclerosis, specifically inflammation, demyelination, neurodegeneration, adaptive immune responses and CNS plasticity. Using the MOG35-55-EAE model in wild-type C57BL/6 mice we monitored the expression patterns of these genes in the spinal cord over time using RealTime RT-PCR. Gene markers for the mini-panel were selected based upon strict gene expression criteria including sensitivity, reproducibility, clinical score-dependence and statistically-relevant regulation compared to naïve control samples. Our current gene mini-panel comprises 10 genes that sensitively monitor the major pathological processes associated with multiple sclerosis and has accurately predicted the efficacy of the approved multiple sclerosis drug, Copaxone, in a preclinical setting in the MOG35-55-EAE model.

This project received support from the 6th Framework Program of the EU through the integrated project entitled NeuroproMiSe (Grant LSHM-CT-2005-018637).

 

Vasiliki Kyrargyri, B.Sc Biology
Study of the role of the cell-specific inactivation of NF-κB in neuronal function and neurodegenerative diseases

In CNS, activity of the transcription factor NF-κB is modulated by neurotransmitters, neurotrophic factors, electrical activity, oxidative stress and cytokines such as TNF. It is believed to have a pivotal role as a mediator of transcription-dependent enduring changes in the structure and functions of neuronal circuits, such as synaptic plasticity which is inherent to learning and memory, as well as development and response to injury. The pro-inflammatory cytokine TNF is produced in CNS under physiological and pathophysiological conditions and participates in the homeostatic activity-dependent regulation of synaptic connectivity.

The first goal of this project is to understand the role of the TNF-induced NF-κB signaling in neuronal function. The main experimental tools include in vitro studies in primary mouse hippocampal neurons from transgenic mice with neuronal deletion of IKKβ (which is required for NF-κB activation) as well as imaging (confocal microscopy, time-lapse imaging, calcium imaging) and biochemical (western blot, biotinylation) techniques. Electrophysiology recordings are performed in collaboration with the group of Professor Ana Maria Sebastiao and her laboratory at the Faculty of Medicine, University of Lisbon, Portugal. Behavioral tests in mice are being performed in collaboration with Professor Jose Maria Delgado and his laboratory in Pablo de Olavide University in Seville, Spain.

Secondly, this project aims to investigate the role of the NF-κB in neurodegenerative and inflammatory diseases. We use conditional gene-targeted mice in which the IKKβ gene has been selectively deleted from cells of the macrophage lineage and we are investigating its role in the pathogenesis of the experimental model of multiple sclerosis, EAE, as well as in kainic acid induced excitotoxicity. The main experimental tools include neuropathological, immunological, immunohistochemical and gene expression analyses of CNS tissues. This part of this project is a continuing effort that has been started by Mary Emmanouil, PhD.

Mouse hippocampus stained with Cresyl violet

This project received support from the 6th Framework Program of the EU through the integrated project entitled NeuroproMiSe (Grant LSHM-CT-2005-018637), the ESF-COST Action B30, Neural regeneration and plasticity (NEREPLAS) through STSM-B30-3282 mission to the lab of Dr Ana Maria Sebastiao, Universidade de Lisboa, Portugal (2009) and the ESF-COST Action BM0603 entitled Inflammation in Brain Disease (NEURINFNET) through STSM-BM0603-6838 mission to the lab of Prof. David Attwell, University College, London (2010).

 

Maria Karamita, B.Sc Biology
Study of the role of the cell-specific inactivation of NF-κB during inflammation and demyelination  of neurodegenerative diseases  using transgenic mice

Conditionalgene targeting of vital NF-κB components, such as the activating kinase IKKβ,in selected cell lineages of mice has allowed the study of cell-specificNF-κB functions in adult mice under physiological and pathologicalconditions. In CNS pathology the role of NF-κB is controversial. It becomes activatedin most cell types, particularly neurons, astrocytes, and microglia,and both neurotoxic and neuroprotective effectshave been described. The mechanisms that underlie such diverseeffects of NF-κB and one of its main activating ligands TNF inthe CNS are not yet fully understood. The research goal of this project is to investigate the cellular basis of NF-κB function in the CNS in two experimental models for multiple sclerosis, EAE and cuprizone-induced demyelination. We use conditional gene-targeted mice in which the gene ΙΚΚβ gene has been selectively inactivated in neurons to investigate the contribution of NF-κB in the progression of inflammation, demyelination and the mobilization of oligodendrocyte precursor cells (OPCs). The main experimental tools are neuropathological, immunohistochemical and gene expression analyses of CNS tissues.

This project received support from the 6th Framework Program of the EU through the integrated project entitled NeuroproMiSe (Grant LSHM-CT-2005-018637) and the ESF-COST Action BM0603 “Inflammation in Brain Disease” (NEURINFNET) through attendance to the Braga Summer School “Neuroimmune Interactions”, Universidade do Minho, Braga, Portugal (2009) organized by Dr Joana Palha and through COST-STSM-BM0603-7919 mission to the lab of Prof Hans Lassmann, Medical University of Vienna, Austria (2011).

Eirini Papazian, MSc and Maria Evangelidou, Ph.D
Study of phenotypic changes in cortical neuron cultures induced by mesenchymal stem cells CM during neuroprotection against glutamate-induced cell death

Several studies indicate that mesenchymal stem cells (MSC) have potent immune regulatory and neuroprotective properties. Specifically, they can limit CNS inflammation, reduce neurological defects and promote endogenous CNS repair mechanisms. Their therapeutic efficacy has been assessed in experimental disease models for stroke, multiple sclerosis and CNS injury. Recent work from our lab shows that mouse MSC protect neurons against glutamate-induced cell death, by reducing the expression of glutamate receptors and glutamate-induced calcium mobilization. Also, a short 24 h exposure of neurons to conditioned medium from MSC (MSC CM) was enough to shift gene expression of the neuronal cultures towards a ‘less mature” pattern suggesting that MSC might induce changes consistent with regeneration plasticity (Voulgari-Kokota et al 2012). To further investigate the neuroprotective effects of MSC we are performing longer-term cultures of primary mouse cortical neurons with MSC CM and measuring the effects on neuron morphology and function, as measured by live cell imaging of calcium influxes. We are also monitoring the representation of different cell lineages, including astrocytes, OPC and neurons, over time within these cultures by immunocytochemistry and correlating changes observed with changes in gene expression, as measured by quantitative RT-PCR. Through these studies our aim is to gain further insight into the mechanisms by which MSC CM exerts its beneficial effects in the CNS in various models of neurodegeneration.

Nadia Kavrohorianou, B.Sc and Sylva Haralambous, Ph.D

TNF and type I interferons (IFN-a and IFN-β) are amongst the primary cytokines produced during an infection which mediate innate immune responses and provide host defense. In macrophages the signaling pathways induced by these different cytokines are integrated into a newly-recognized IFN-β-mediated autocrine loop that sustains and amplifies TNF-induced expression of inflammatory mediators and IFN response genes thereby enhancing immune responses to pathogens. The group of Dr Sylva Haralambous is investigating the role of IFN-a receptor 1 in T lymphocytes during immune responses, in various infections (viral, bacterial, parasitic), in neurodegenerative diseases like multiple sclerosis, in cancer and as an adjuvant for influenza vaccines in collaboration with other national and international research teams .

 

Ioannis Grivas, M.Sc and Sylva Haralambous, Ph.D

 

We currently have active collaborations with the laboratories of:

José M Delgado- García- Division of Neurosciences, Pablo de Olavide University, Seville, Spain. http://uponeurociencias.pacifico-meetings.com

Hans Lassmann- Centre for Brain Research, Division of Neuroimmunology, Medical University of Vienna, Austria. www.meduniwien.ac.at/typo3/?id=5265

Vijay Kuchroo- Brigham and Women’s Hospital Ctr for Neurological Diseases, Boston, MA, USA. www.hms.harvard.edu/dms/immunology/fac/Kuchroo.php

Xavier Navarro – Institute of Neurosciences, Universitat Autonoma de Barcelona, Bellaterra, Spain. http://inc.uab.cat/index.php?option=com_content&view=article&id=122:xavier-navarro&catid=46:grupos&Itemid=112&lang=en

Ana M Sebastiao – Institute of Pharmacology & Neurosciences, University of Lisbon, Portugal. www.imm.fm.ul.pt/web/imm/neurosciencesunit

Theodore Tselios and John Matsoukas – Department of Chemistry, University of Patras, Rio, Greece. http://www.chem.upatras.gr/index.php?lang=en

Hartmut Wekerle- Department of Neuroimmunology, Max Planck Institute of Neurobiology, Martinsried, Germany. www.neuro.mpg.de/english/people/wekerle_h.shtml

Members

Head of the Lab: Lesley Probert, Ph.D, Researcher A’
Tel: +30-210-6478866, Fax: +30-210-6456547
Email: lesley@pasteur.gr, lesley.probert@gmail.com

Researchers

Sylva Haralambous, Ph.D, Researcher B’ (Head of Transgenic Technology Unit) sharalambous@pasteur.gr

 

Post-Doctoral Research Fellows

Vivian Tseveleki, Ph.D vtseveleki@hotmail.com
Maria Evangelidou, Ph.D  meuagelidou@gmail.com
Ioannis Kanistras, PhD ikanistras@pasteur.gr

Research Assistant

Ioanna Stergiatou, B.Sc Biology (Transgenic Technology Unit)

 

Technologist

Fotis Bantounas (Transgenic Technology Unit)

 

Ph.D Students

Vasiliki Kyrargyri, B.Sc Biology,  kyrargyriv@pasteur.gr
Maria Karamita, B.Sc Biology, mariakaramita@gmail.com
Nadia Kavrohorianou, B.Sc Biology, nkavrohorianou@pasteur.gr
Ioannis Grivas, M.Sc Biology,  grgns@yahoo.gr

 

Undergraduate & Masters Students

Eirini Papazian, B.Sc Biology (M.Sc project)
Melina Makroyiannaki (undergraduate project)
Marina Papoulia (undergraduate project)

 

Past members of the lab (1998-2012)

Research Associates

Anda Voulgari-Kokota, Ph.D, Era Taoufik, Ph.D, Sotiris-Spyros Vamvakas, Ph.D, Mary Emmanouil, Ph.D, Stavroula Alexopoulou, Voula Stavropoulou, Afroditi Kartouna, David Plows, Ph.D, Chengmai Ruan, Ph.D, Antonis Vylliotis, Ioanna Dagliani, Ph.D, George Kassiotis, Ph.D, Baosheng Ge, Ph.D, Katerina Akassoglou, Ph.D.

Undergraduate and Masters Students

Panayiotis-Orpheas Protopsaltis (practical training 2012), Michael Makryllos (undergraduate project 2009-2011), Eleni Malaktari (undergraduate project 2011-2012), Konstantina Ioannidou (practical training 2011), Amalia Stantzou (practical training 2008), Areti Manda (undergraduate project 2007-2008), Sophia Beina (undergraduate project 2007-2008), Christos Georgiadis (practical training 2007), Michael Koutrolos (undergraduate project 2006-2007), Nikos Simos, B.Sc Bioinformatics (M.Sc project 2006), Athanassia Stathopoulou (undergraduate project 2005), Thomas Alexandris (undergraduate project 2004-2005), Eirini Papadaki (practical training 2004), Orestis Argyros (undergraduate project 2002-2003), Marianna Georgakopoulou (practical training 2002-2003), Elena Odiatou (undergraduate project 2001-2002), Sandrine Billet (practical training 2000).

Funding

Current grants and networks

FP7 Development of a Center of Excellence in Neurosignalling(NeuroSign). 2010-2013

FP6 Neuroprotective Strategies for Multiple Sclerosis (NeuroproMiSe). 2005-2010

ESF-COST Action BM0603, Inflammation in brain disease (NEURINFNET). 2007-2011.

Publications

2012

Voulgari-Kokota A, Fairless R, Karamita M, Kyrargyri V, Tseveleki V, Evangelidou M, Delorme B, Charbord P, Diem R, Probert L (2012). Mesenchymal stem cells protect CNS neurons against glutamate excitotoxicity by inhibiting glutamate receptor expression and function. Exp. Neurol. 236: 161-170.

2011

Emmanouil M, Taoufik E, Tseveleki V, Vamvakas S-S, Probert L (2011). A Role for Neuronal NF-κB in Suppressing Neuroinflammation and Promoting Neuroprotection in the CNS. Adv Exp Med Biol. 2011;691:575-81.

Ghezzi P, Bernaudin M, Bianchi R, Blomgren K, Brines M, Campana W, Cavaletti G, Cerami A, Chopp M, Coleman T, Digicaylioglu M, Ehrenreich H, Erbayraktar S, Erbayraktar Z, Gassmann M, Genc S, Gokmen N, Grasso G, Juul S, Lipton SA, Hand CC, Latini R, Lauria G, Leist M, Newton SS, Petit E, Probert L, Sfacteria A, Siren AL, Talan M, Thiemermann C, Westenbrink D, Yaqoob M, Zhu C (2011). Erythropoietin: not just about erythropoiesis. Lancet. 2010 Jun 19;375(9732):2142.

Taoufik E, Tseveleki V, Chu SY, Tselios T, Karin M, Szymkowski DE, Lassmann H, Probert L (2011). Transmembrane TNF is neuroprotective and regulates experimental autoimmune encephalomyelitis via neuronal NF-κB. Brain, 134: 2722-2735.

2010

Evangelidou M, Tseveleki V, Vamvakas S-S, Probert L (2010). TNFRI is a positive T-cell costimulatory molecule important for the timing of cytokine responses. Immunol. Cell Biol. Jul; 88(5): 586-95. Epub 2010 Mar 9.

Tseveleki V, Rubio R, Vamvakas S-S, White J, Taoufik E, Petit E, Quackenbush J & Probert L (2010). Comparative gene expression analysis in mouse models for multiple sclerosis, Alzheimer’s disease and stroke for identifying commonly regulated and disease-specific gene changes. Genomics, Aug;96(2):82-91. Epub 2010 May 7.

2009

Emmanouil M, Taoufik E, Tseveleki V, Vamvakas S-S, Tselios T, Karin M, Lassmann H, Probert L (2009). Neuronal IkappaB kinase beta protects mice from autoimmune encephalomyelitis by mediating neuroprotective and immunosuppressive effects in the central nervous system. J Immunol. Dec 15;183(12):7877-89.

Moumtzi SS, Roberts ML, Joyce T, Evangelidou M, Probert L, Frillingos S, Fotsis T, Pintzas A (2009). Gene Expression Profile Associated with Oncogenic Ras-induced Senescence, Cell Death, and Transforming Properties in Human Cells. Cancer Invest. Nov 2. [Epub ahead of print].

Oikonomou E, Makrodouli E, Evagelidou M, Joyce T, Probert L, Pintzas A (2009). BRAF(V600E) efficient transformation and induction of microsatellite instability versus KRAS(G12V) induction of senescence markers in human colon cancer cells. Neoplasia Nov;11(11):1116-31.

1990-2008

Quinones MP, Kalkonde Y, Estrada CA, Jimenez F, Ramirez R, Mahimainathan L, Mummidi S, Choudhury GG, Martinez H, Adams L, Mack M, Reddick RL, Maffi S, Haralambous S, Probert L, Ahuja SK, Ahuja SS (2008).Role of astrocytes and chemokine systems in acute TNFalpha induced demyelinating syndrome: CCR2-dependent signals promote astrocyte activation and survival via NF-kappaB and Akt. Mol.Cell.Neurosci. 37(1):96-109.

Taoufik E, Petit E, Divoux D, Tseveleki V, Mengozzi M, Roberts ML, Valable S, Ghezzi P, Quackenbush J, Brines M, Cerami A & Probert L (2008). TNF receptor I sensitizes neurons to erythropoietin- and VEGF-mediated neuroprotection after ischemic and excitotoxic injury. PNAS USA 105: 6185-6190.

Taoufik E & Probert L (2008). Ischemic neuronal damage. Curr.Pharm.Des. 14: 3565-3573.

Taoufik E, Tseveleki V, Euagelidou M, Emmanouil M, Voulgari-Kokota A, Haralambous S, Probert L (2008). Positive and negative implications of tumor necrosis factor neutralization for the pathogenesis of multiple sclerosis. Neurodegener.Dis. 5(1):32-7.

Oikonomou E, Kothonidis K, Taoufik E, Probert E, Zografos G, Nasioulas G, Andera L, Pintzas A (2007). Newly established tumourigenic primary human colon cancer cell lines are sensitive to TRAIL-induced apoptosis in vitro and in vivo. Br.J.Cancer 97(1): 73-84.

Taoufik E, Valable S, Muller G, Roberts ML, Divoux D, Tinel A, Voulgari-Kokota A, Tseveleki V, Altruda F, Lassmann H, Petit E & Probert L (2007). FLIPL protects neurons against in vivo ischemia and in vitro glucose-deprivation-induced cell death. J.Neurosci. 27 (25): 6633-6646.

Tseveleki V, Tsagosis P, Koutsoni O, Dotsika E & Probert L (2007). Cellular FLIP long isoform transgenic mice overcome inherent Th2-biased immune responses to efficiently resolve Leishmania major infection. Int.Immunol. 19: 1183-1189.

Matronardi FG, Wood DD, Mei J, Raijmakers R, Tseveleki V, Dosch H-M, Probert L, Casaccia-Bonnefil P & Moscarello MA (2006). Increased citrullination of histone H3 in multiple sclerosis brain and animal models of demyelination: a role for tumor necrosis factor-induced peptidylarginine deiminase 4 translocation. J.Neurosci. 26: 11387-11396.

Akassoglou K, Adams R, Bauer J, Mercado P, Tseveleki V, Probert L, Lassmann H & Strickland S (2004). Fibrin depletion decreases inflammation and delays the onset of demyelination in a tumor factor transgenic mouse model for multiple sclerosis. PNAS USA, 101: 6698-6703.

Tsagozis P, Tseveleki V, Probert L, Dotsika E, Karagouni E (2004). Vaccination with plasmids encoding the Leishmania major gp63 glycoprotein and CD40L results in a partial suppression of the inflammatory reaction after experimental infection. Eur.J.Inflamm. 2 (2): 1-6.

Tseveleki V, Bauer J, Taoufik E, Ruan C, Leondiadis L, Haralambous S, Lassmann H & Probert L (2004). c-FLIPL overexpression in T cells is sufficient to drive Th2 effector responses and immunoregulation of experimental autoimmune encephalomyelitis. J.Immunol. 173: 6619-6626.

Akassoglou K, Douni E, Bauer J, Lassmann H, Kollias G & Probert L (2003). Exclusive tumor necrosis factor (TNF) signaling by the p75TNF receptor triggers inflammatory ischaemia in the CNS of transgenic mice. PNAS USA, 100(2): 709-714.

Tselios T, Apostolopoulou V, Daliani I, Deraos S, Grdadolnik S, Mavromoustakos T, Melachrinou M, Thymianou S, Probert L, Mouzaki A, Matsoukas J (2002). Antagonistic effectes of human cyclic MBP (87-99) altered peptide ligands in experimental allergic encephalomyelitis and human T-cell proliferation. J.Med.Chem. 45(2): 275-283.

Probert L, Akassoglou K (2001). Glial expression of cytokines in transgenic animals- how do these models reflect the “normal situation”. Invited Review. Glia , 36, 212-219.

Fiore M, Angelucci F, Alleva E, Branchi I, Probert L, Aloe L (2000). Learning performances, brain NGF distribution and NPY levels in transgenic mice expressing TNF-alpha. Beh.Brain Res. 112: 165-175.

Probert L, Eugster H-P, Akassoglou K, Bauer J, Frei K, Lassmann H & Fontana A (2000). TNFR1signalling is critical for demyelination and the limitation of T-cell responses during immune-mediated CNS disease. Review, Brain 123, 2005-2019.

Tselios T, Daliani I, Deraos S, Thymianou S, Matsoukas E, Troganis A, Gerothanassis I, Mouzaki A, Mavromoustakos T, Probert L, & Matsoukas J (2000). Treatment of experimental allergic encephalomyelitis (EAE) by  a rationally designed cyclic analogue of myelin basic protein (MBP) epitope 72-85. Bioorgan.Med.Chem.Lett. 10: 2713-2717.

Tselios T, Daliani I, Probert L, Deraos S, Matsoukas E, Roy S, Pires J, Moore G & Matsoukas J (2000). Treatment of experimental allergic encephalomyelitis (EAE) induced by guinea pig myelin basic protein epitope 72-85 with a human MBP 87-99 analogue and effects of cyclic peptides. Bioorgan.Med.Chem. 8: 1903-1909.

Akassoglou K, Bauer J, Kassiotis G, Lassmann H, Kollias G & Probert L (1999). Transgenic models of TNF induced demyelination. Adv.Exp.Med.Biol. 468: 245-259.

Aloe L, Fiore M, Probert L, Turrini P & Tirassa P (1999). Overexpression of tumour necrosis factor alpha in the brain of transgenic mice differentially alters nerve growth factor levels and choline acetyltransferase activity. Cytokine 11: 45-54.

Aloe L, Properzi F, Probert L, Akassoglou K, Kassiotis G, Micera A & Fiore M (1999). Learning abilities, NGF and BDNF brain levels in two lines of TNF-α transgenic mice, one characterized by neurological disorders, the other phenotypically normal. Brain Res. 840: 125-137.

Glosli H, Veiby OP, Fjerdingstad H, Mehlum A, Probert L, Kollias G, Gjernes E, Prydz H. (1999). Effects of hTNF alpha expression in T cells on haematopoiesis in transgenic mice. Eur. J. Haematol. 63: 50-60.

Kassiotis G, Bauer J, Akassoglou K, Lassmann H, Kollias G & Probert L (1999). A tumor necrosis factor-induced model of human primary demyelinating diseases develops in immunodeficient mice. Eur.J.Immunol. 29: 912-917.

Kassiotis G, Pasparakis M, Kollias G & Probert L (1999).TNF accelerates the onset but does not alter the incidence and severity of myelin basic protein-induced experimental autoimmune encephalomyelitis. Eur.J.Immunol. 29: 774-780.

Tselios T, Probert L, Kollias G, Daliani I, Matsoukas E, Troganis A, Gerothanassis IP, Mavromoustakos T, Moore GJ & Matsoukas JM (1999). Design and synthesis of a potent cyclic analogue of the myelin basic protein epitope MBP72-85: Importance of the Ala81 carboxyl group and of a cyclic conformation for induction of experimental allergic encephalomyelitis. J.Med.Chem. 42: 1170-1177.

Akassoglou K, Bauer J, Kassiotis G, Pasparakis M, Lassmann H, Kollias G & Probert L (1998). Oligodendrocyte apoptosis and primary demyelination induced by local  TNF/p55TNF receptor signaling in the CNS of transgenic mice: Models for Multiple Sclerosis with primary oligodendropathy. Am.J.Pathol. 153:  801-813.

Alonzi T, Fattori E, Lazzaro D, Costa P, Probert L, Kollias G, De Benedetti F, Poli V & Ciliberto G (1998) Interleukin 6 is required for the development of collagen-induced arthritis. J.Exp.Med. 187: 461-468.

Fiore M, Alleva E, Probert L, Kollias G, Angelucci F & Aloe L (1998) Exploratory and displacement behaviour in transgenic mice expressing high levels of brain TNF alpha. Physiol.Behav. 63: 571-576.

Tselios T, Probert L, Kollias G, Matsoukas E, Roumelioti P, Alexopoulos K, Moore GJ & Matsoukas J (1998). Design and synthesis of small semi-mimetic peptides with immunomodulatory activity based on myelin basic protein (MBP). Amino Acids 14: 333-341.

Akassoglou K, Probert L, Kontogeorgos G & Kollias G (1997) Astrocyte- but not Neuron-Specific Transmembrane TNF Triggers Inflammation, Demyelination and Neuronal Degeneration in the CNS of Transgenic Mice. J.Immunol. 158: 438-445.

Cope AP, Liblau RS, Yang X-D, Congia M, Laudanna C, Schreiber RD, Probert L, Kollias G & McDevitt HO (1997) Chronic tumor necrosis factor (TNF) alters T cell responses by attenuating T cell receptor signalling. J.Exp.Med. 185: 1573-1584.

Probert L & Selmaj K (1997) TNF and related molecules: Trends in neuroscience and clinical applications. J.Neuroimmunol. 72: 113-117. Review.

Probert L, Akassoglou K, Kassiotis G, Pasparakis M, Alexopoulou L & Kollias G (1997) TNF alpha transgenic and knockout models of CNS inflammation and degeneration. J.Neuroimmunol. 72 137-141. Review.

Tselios T, Deraos S, Matsoukas E, Panagiotopoulos D, Matsoukas J, Moore G J, Probert L, Kollias G, Hilliard B, Rostami A & Monos D (1997) Myelin basic protein peptides: induction and inhibition of experimental allergic encephalomyelitis. Rev.Clin.Pharmacol.Pharmokin. 11: (2&3) 60-64.

Douni E, Akassoglou K, Alexopoulou L, Georgopoulos S, Haralambous S, Hill S, Kassiotis G, Kontoyiannis D, Pasparakis M, Plows D, Probert L & Kollias G (1996) Transgenic and knockout analyses of the role of TNF in immune regulation and disease pathogenesis. J.Inflamm. 47: 27-38. Review.

Fiore M, Probert L, Kollias G, Akassoglou K, Alleva E & Aloe L (1996) Neuro-behavioural alterations in developing transgenic mice expressing human TNF alpha in the brain. Brain Behav.Immun. 10: 126-138.

Probert L, Akassoglou K, Alexopoulou L, Douni E, Haralambous S, Hill S, Kassiotis G, Kontoyiannis D, Pasparakis M, Plows D & Kollias G (1996) Dissection of the pathologies induced by transmembrane and wild-type tumor necrosis factor in transgenic mice. J.Leuk.Biol. 59: 518-525. Review.

Aloe L, Probert L, Kollias L, Micera A & Tirassa P (1995). Effect of NGF antibodies on mast cell distribution, histamine and substance P levels in the knee joint of TNF-arthritic transgenic mice. Rheumat.Int. 14: 249-252.

Plows D, Probert L, Georgopoulos S, Alexopoulou L & Kollias G. The role of tumour necrosis factor (TNF) in arthritis: studies in transgenic mice. Proceedings of the XIIIth Eur.Congr.Rheumatol. (ERASS) 20-23rd June 1995. Vol 24 suppl. 2: 51-54.

Probert L, Plows D, Kontogeorgos G & Kollias G. (1995) The type I IL-1 receptor acts in series with TNF to induce arthritis in TNF transgenic mice. Eur.J.Immunol. 25: 1794-1797.

Probert L, Akassoglou K, Pasparakis M, Kontogiorgos G & Kollias G (1995) Spontaneous inflammatory demyelinating disease in transgenic mice showing CNS-specific expression of tumor necrosis factor. PNAS USA 92: 11294-11298.

Siegel SA, Shealy DJ, Nakada MT, Le J, Wolfe DS, Probert L, Kollias G, Ghrayeb J, Vilcek J & Daddona PE (1995) The mouse/human chimeric monoclonal antibody Ca2 neutralizes TNF in vitro and protects transgenic mice from cachexia and TNF lethality in vivo. Cytokine 7: 15-25. *Pre-clinical trial of Ca2 anti-TNF antibody now used in RA patients.

Taverne J, Sheikh N, de Souza JB, Playfair JHL, Probert L & Kollias G (1994) Anaemia and resistance to malaria in transgenic mice expressing human TNF. Immunol. 82: 397-403.

Aloe L, Probert L, Kollias G, Bracci-Laudiero L, Micera A, Mollinari C & Levi-Montalcini R (1993). Level of nerve growth factor and distribution of mast cells in the synovium of tumour necrosis factor transgenic arthritic mice. Int.J.Tissue Reactions 15: 139-143.

Aloe L, Probert L, Kollias G, Bracci-Laudiero L, Spillantini MG & Levi-Montalcini R (1993). The synovium of transgenic arthritic mice expressing tumor necrosis factor contains a high level of nerve growth factor. Growth Factors 9: 149-155.

Probert L, Keffer J, Corbella P, Cazlaris H, Patsavoudi E, Stephens S, Kaslaris E, Kioussis D & Kollias G (1993). Wasting, ischaemia and lymphoid abnormalities in mice expressing T cell-targeted human tumour necrosis factor transgenes. J. Immunol. 151: 1894-1906.

Keffer J, Probert L, Cazlaris H, Georgopoulos S, Kaslaris E, Kioussis D & Kollias G (1991). Transgenic mice expressing human tumour necrosis factor: a predictive genetic model of arthritis. EMBO J. 10: 4025-4031. *Selected as a classic paper in Rheumatoid Arthritis, PPS Europe Ltd, 1994.

 

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We are interested in the interactions between the immune system and the central nervous system (CNS), particularly in how the immune system contributes to inflammatory neurodegeneration, and conversely how neurons control immune responses to maintain brain homeostasis. Recent work has centered on identifying key molecules, mechanisms and cells that are involved in mediating inflammatory neurodegeneration in experimental models for stroke and multiple sclerosis and to find new therapeutic targets and neuroprotection strategies for the treatment of inflammatory CNS disorders.

Research is supported by an FP6 EU Grant (NeuroproMiSe)(www.neuropromise.eu), an FP7 EU Regional Potential Grant (NeuroSign) and the European Cooperation in Science & Technology (COST) Action, Inflammation in Brain Disease (NEURINFNET)(www1.sdu.dk/multi/neurinfnet).The laboratory is also actively involved in the running of the European School of Neuroimmunology (ESNI) (www.esni.org).