Laboratory of Molecular Genetics
Lesley Probert | Immunology Department
Head of the Laboratory
We study the interface between the immune system and the central nervous system (CNS), particularly the participation of the immune system in physiological brain processes such as host defense, and in inflammatory diseases such as multiple sclerosis (MS). The laboratory has a long-standing interest in understanding the functions of a pro-inflammatory cytokine, tumor necrosis factor (TNF), in the CNS. TNF drives inflammatory responses to infection, injury and neurodegeneration, but paradoxically also protects neurons, directly and indirectly by repairing the myelin sheath around demyelinated axons. This diversity of TNF function is now understood to be a direct reflection of its complex biology. “TNF” represents at least a two-ligand (soluble TNF and membrane TNF), two-receptor (TNF receptors 1 and 2/TNFR1 and TNFR2) system with ligands and receptors both differentially expressed and regulated on different cell types. Through the application of sophisticated spatial and temporal gene-targeting techniques in mice, we and others have been able to dissect the individual functions of the two TNFs and their receptors in a number of important brain processes through the study of experimental disease models. In general soluble TNF/TNFR1 interactions dominate inflammatory responses, which often leading to significant secondary tissue damage and, as recently shown by us, also strongly inhibit remyelination.
In contrast, membrane TNF/TNFR2 interactions promote remyelination and neuroprotection, by enhancing oligodendrocyte precursor cells. The ability to separate deleterious and beneficial effects of TNF at a molecular level has direct implications for therapy in human disease. Non-selective TNF inhibitors that block the effects of both soluble and membrane TNF and are blockbuster drugs for peripheral inflammatory diseases such as rheumatoid arthritis, behave badly in the CNS. They exacerbate MS and can even induce de novo demyelinating disease. The experimental data now clearly suggest that selective inhibition of soluble TNF and/or TNFR1, while preserving membrane TNF and/or TNFR2, is a promising future direction for safe immunotherapy in chronic inflammatory diseases of the CNS like MS.
TNF receptors 1 and 2 in neuronal protection
TNF directly protects CNS neurons [1, 2]. Mice deficient in TNF receptor 1 [3, 4]  or receptor 2  develop more severe lesions following acute injuries such as experimental ischemia. Also, pre-conditioning of brain neurons with TNF protects them against a variety of death stimuli that are relevant to neurodegenerative conditions such as stroke and MS. TNF signals its neuroprotective effects through the Akt/protein kinase B pathway and through activation of the transcription factor NF-κB, which in turn increases the production of proteins that inhibit cell death by apoptosis or necrosis. Accordingly, we previously found that mice with a selective depletion of the neuroprotective mediator NF-κB selectively in brain glutamatergic neurons were more seriously affected by an MS-type disease called experimental autoimmune encephalomyelitis (EAE) .
Our current studies focus upon identifying the cellular source of neuroprotective membrane TNF in the CNS during disease, and determining whether anti-inflammatory agents can enhance or inhibit this protective mechanism.
Soluble TNF inhibits and membrane TNF promotes CNS remyelination
Remyelination is a spontaneous regenerative process of the adult CNS that is triggered in response to myelin damage . In MS, remyelination largely fails for unknown reasons, so that demyelinating lesions expand and demyelinated axons are continuously exposed to injury [9, 10]. Our recent studies reveal an unexpected role for soluble TNF as an inhibitor of remyelination. Treatment of mice with XPro1595, a dominant-negative TNF analogue that selectively inactivates soluble TNF , and crosses the blood-brain barrier, did not prevent demyelination in a cuprizone model but permitted early remyelination and neuroprotection due to improved phagocytosis of myelin debris in lesions by CNS macrophages . Genetic studies using knockout mice confirmed that soluble TNF and membrane TNF have opposite effects in remyelination, inhibition and induction respectively . These results suggest that soluble TNF produced by inflammatory cells in MS lesions might be one reason why remyelination fails in this disease. They also suggest that soluble TNF inhibitors that cross the blood-brain barrier might be promising reagents for promoting tissue repair in progressive MS. Our current studies investigate the molecular mechanism by which soluble TNF inhibits myelin phagocytosis by macrophages so that additional therapeutic targets can be identified.
Microglia in brain physiology and pathophysiology
Microglia are CNS resident immune cells and the main cellular source of TNF and IL-1β in the brain. They have important roles under physiological conditions, such as synaptic pruning and the maintenance of neuronal circuitry during development , and orchestrate adaptive and innate immune responses under conditions of CNS infection or injury . In our laboratory we are interested in understanding the mechanisms of this graded microglial response, which ranges between beneficial roles under physiological and acute pathological conditions, to deleterious roles under chronic pathological conditions such as neurodegeneration, and particularly the involvement of two potent inflammatory cytokines, TNF and IL-1β. We developed mice that are conditionally deficient in IKK-β, the main NF-κB activating kinase triggered by inflammatory stimuli, selectively in myeloid cells including microglia . Under physiological conditions IKK-β-deficient microglia produces less IL-1β, and mice show impaired hippocampal synaptic plasticity and hippocampus-driven behavior during associative learning . These results indicate that microglia, through NF-κB and IL-1β, are critical for normal adult hippocampal function. Our future studies will further develop methodologies for gene targeting specifically in mouse microglia so that we can address the contribution of these cells to host defense against infection and in the pathogenesis of inflammatory neurodegenerative diseases such as MS.
Development of a myelin autoantigen-specific immunotherapy for MS
Our pre-clinical studies show that targeting of myelin antigens to antigen-presenting cells by conjugation to the polysaccharide mannan induces robust peptide-specific peripheral T cell anergy in CD4+ T helper type 1 (IFN-γ-producing; Th1) and Th17 (IL-17-producing) lymphocytes in mice, both of which cell types are implicated in MS. Administration of these mannan-peptides using prophylactic (vaccination) and therapeutic protocols in mice strongly protects them against CNS demyelination in a widely-used experimental model for MS named experimental autoimmune encephalomyelitis (EAE) . More recently we used humanized HLA-DR2b transgenic mice and blood monocytes from MS patients to show that mannan-conjugation reduces the ability of myelin peptides to trigger T cell responses when presented exclusively by human MHC class II molecules, including the MS candidate susceptibility molecule HLA-DR2 (Probert et al., submitted for publication, 2017). We further show that MS patients can be readily screened for their T cell responses to disease-associated myelin peptide epitopes using peripheral blood monocytes in a simple ex vivo lymphocyte response assay. This assay will allow the selection and stratification of MS patients for entry into clinical studies using these peptides. Collectively, our data suggest that mannan-peptides are a promising approach for reducing autoimmune responses in MS, and might prove useful for the treatment of human autoimmune disease. Current studies investigate the precise mechanism of immune tolerance induced by mannan-peptides.
Lab tool box
Our main investigative approach is to develop transgenic or conditionally gene-targeted mice for specific genes and cells of interest and use them in combination with pre-clinical models for MS. The experimental models are mainly experimental autoimmune encephalomyelitis (EAE), which models autoimmune components of MS, and cuprizone-induced demyelination, which is a convenient model for studying the remyelination process as well as key aspects of progressive MS such as oxidative damage and mitochondrial injury in the presence of chronically active microglia. We analyze the function of candidate molecules at clinical, neuropathological, cellular, signaling and molecular levels in vivo and mechanisms are further verified in vitro using isolated primary cells with gene expression and knockdown techniques. Our research is supported by technological platforms of the Institute, mainly the Transgenic Technology Unit, which maintains a mouse repository for immune and disease strains (add LINK to TTU), and the Light Microscope Imaging Unit (add LINK to Imaging).
- Bruce, A.J., et al., Altered neuronal and microglial responses to excitotoxic and ischemic brain injury in mice lacking TNF receptors. Nat Med, 1996. 2(7): p. 788-94.
- Probert, L., TNF and its receptors in the CNS: The essential, the desirable and the deleterious effects. Neuroscience, 2015. 302: p. 2-22.
- Gary, D.S., et al., Ischemic and excitotoxic brain injury is enhanced in mice lacking the p55 tumor necrosis factor receptor. J Cereb Blood Flow Metab, 1998. 18(12): p. 1283-7.
- Taoufik, E., et al., FLIP(L) protects neurons against in vivo ischemia and in vitro glucose deprivation-induced cell death. J Neurosci, 2007. 27(25): p. 6633-46.
- Lambertsen, K.L., et al., Microglia protect neurons against ischemia by synthesis of tumor necrosis factor. J Neurosci, 2009. 29(5): p. 1319-30.
- Fontaine, V., et al., Neurodegenerative and neuroprotective effects of tumor Necrosis factor (TNF) in retinal ischemia: opposite roles of TNF receptor 1 and TNF receptor 2. J Neurosci, 2002. 22(7): p. Rc216.
- Emmanouil, M., et al., Neuronal I kappa B kinase beta protects mice from autoimmune encephalomyelitis by mediating neuroprotective and immunosuppressive effects in the central nervous system. J Immunol, 2009. 183(12): p. 7877-89.
- Franklin, R.J. and C. Ffrench-Constant, Remyelination in the CNS: from biology to therapy. Nat Rev Neurosci, 2008. 9(11): p. 839-55.
- Bramow, S., et al., Demyelination versus remyelination in progressive multiple sclerosis. Brain, 2010. 133(10): p. 2983-98.
- Patrikios, P., et al., Remyelination is extensive in a subset of multiple sclerosis patients. Brain, 2006. 129(Pt 12): p. 3165-72.
- Steed, P.M., et al., Inactivation of TNF signaling by rationally designed dominant-negative TNF variants. Science, 2003. 301(5641): p. 1895-8.
- Karamita, M., et al., Therapeutic inhibition of soluble brain TNF promotes remyelination by increasing myelin phagocytosis by microglia. JCI Insight, 2017. 2(8).
- Evangelidou, M., et al., Altered expression of oligodendrocyte and neuronal marker genes predicts the clinical onset of autoimmune encephalomyelitis and indicates the effectiveness of multiple sclerosis-directed therapeutics. J Immunol, 2014. 192(9): p. 4122-33.
- Paolicelli, R.C., et al., Synaptic pruning by microglia is necessary for normal brain development. Science, 2011. 333(6048): p. 1456-8.
- Kyrargyri, V., et al., Differential contributions of microglial and neuronal IKKbeta to synaptic plasticity and associative learning in alert behaving mice. Glia, 2015. 63(4): p. 549-66.
CURRENT GRANTS AND NETWORKS
Multiple Sclerosis Trials Collaboration grant entitled “The role of cellular senescence aging in disability progression in multiple sclerosis”. To Lesley Probert & Dimitris Papadopoulos. 80.400,00 UKP. Duration 36 months (2016-2019).
The laboratory is actively involved in the organization and running of the European School of Neuroimmunology (ESNI) (www.esni.org), and the Hellenic Academy of Neuroimmunology (www.helani.gr).
1. Methods of treating neurological diseases. EPO App. No. 13766804.2-1456 [US/10.09.12/ USP201261699230: US/23.05.13/ USP201361826922]. 10th May 2015.
2. Conjugates comprising mannan and myelin basic protein (MBP). USPO App. No. 14/877.679. 7th October 2015.
3. Therapeutic myelin sheath derived antagonistic peptide conjugates. EPO App. No. 14156495.5-1412 [GB/25.01.08/ GBA0801424: GB/08.02.08/ GBA 0802405: GR/29.02.08/ GRA 20080100151]. 22nd April 2014.
4. Therapeutic vaccines. Australia IP App. No. 2009207345, Ref. 147769. 11th October 2013.
5. Therapeutic myelin sheath derived antagonistic peptide conjugates. USPTO App. No. 12864019 [2011/0243981]. 28th October 2010.
TRANSGENIC MOUSE MODELS DEVELOPED
|Transgenic mouse models developed|
Τg1278 huTNF (regulated human TNF expression)
Keffer et al (1991)
(human TNF in synoviocytes)
|rheumatoid arthritis||Keffer et al (1991)|
(human TNF in T cells)
|normal||Probert et al (1993|
|Tg211 CD2-huTNF-gl (human TNF in T cells)||cachexia/ septic shock||Probert et al (1993)|
|Tg6074 muTNF-gl (murine TNF in oligos)||MS||Probert et al (1995
Akassoglou et al (1998)
(membrane humanTNF in astrocytes)
|MS||Akassoglou et al (1997)
Akassoglou et al (1998)
|TgK742 NFL-huTNF-gl (human TNF in neurons)||encephalitis||Akassoglou et al (1997)|
|TgK3, TgK11, TgK14 NFL-huTNFΔ1-12-gl
(membrane human TNF in neurons)
|normal||Akassoglou et al (1997)|
(human c-FLIP in T cells)
|normal||Tseveleki et al (2004)|
|Tg6988 NFL-muFLIP-IRES-EGFP (murine c-FLIP in neurons)
Tg4617 NFL-dnIκB-IRES-EGFP (dnIκB in neurons)
(Cre recombinase in myeloid cells)
|Taoufik et al (2007)
Ale et al (2016)
Kyrargyri et al (2015)
Karamita M, Barnum C, Mobius W, Tansey MG, Szymkowski DE, Lassmann H, Probert L. (2017). Therapeutic inhibition of soluble brain TNF promotes remyelination by increasing myelin phagocytosis by microglia. J. Clin. Invest. Insight, Apr 20; 2(8). pii: 87455. doi: 10.1172/jci.insight.87455. [Epub ahead of print].2017
Probert L (2015). TNF and its receptors in the CNS: the essential, the desirable and the deleterious effects. Neuroscience Aug 27;302:2-22. doi: 10.1016/ j.neuroscience. 2015.06.038. Epub 2015 Jun 24. Review.
Tseveleki V, Tselios T, Kanistras I, Koutsoni O, Karamita M, Vamvakas SS, Apostolopoulos V, Dotsika E, Matsoukas J, Lassmann H, Probert L (2015). Mannan-conjugated myelin peptides prime non-pathogenic Th1 and Th17 cells and ameliorate experimental autoimmune encephalomyelitis. Exp. Neurol. 267:254-67; doi: 10.1016/j.expneurol.2014.10.019. Epub 2014 Oct 30.
Kyrargyri V, Vega-Flores G, Gruart A, Delgado-Garcia JM, Probert L (2015). Differential contributions of microglia and neuronal IKKβ to synaptic plasticity and associative learning in alert behaving mice. Glia Apr; 63 (4): 549-566; doi: 10.1002/glia.22756. Epub 2014 Oct 9.
Evangelidou M, Karamita M, Vamvakas SS, Szymkowski DE, Probert L (2015). Altered expression of oligodendrocyte and neuronal marker genes predicts the clinical onset of autoimmune encephalomyelitis and indicates the effectiveness of multiple sclerosis-directed therapeutics. J Immunol. May ;192(9):4122-33. doi: 10.4049/jimmunol.1300633. Epub 2014 Mar 28.
Schuh C, Wimmer I, Hametner S, Haider L, Van Dam A-M, Liblau RS, Smith KJ, Probert L, Binder CJ, Bauer J, Bradl M, Mahad D, Lassmann H (2014). Oxidative tissue injury in multiple sclerosis is only partly reflected in experimental disease models. Acta Neuropathol. Aug;128(2):247-66. doi: 10.1007/s00401-014-1263-5. Epub 2014 Mar 13.2014
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. Jul;236(1):161-70. doi: 10.1016/j.expneurol.2012.04.011. Epub 2012 Apr 25.2012
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 Sep;134(Pt 9):2722-35. doi: 10.1093/brain/awr203.
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.691:575-81; doi: 10.1007/978-1-4419-6612-4_60.
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. Jun 19;375(9732):2142.
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 changes. Genomics Aug;96(2):82-91. Epub 2010 May 7.
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.
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):7877-89.2009
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.
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: 11387-11396.2006
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): 709-714.2003
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): 275-283.2002
Probert L, Akassoglou K (2001). Glial expression of cytokines in transgenic animals- how do these models reflect the “normal situation”. Invited Review. Glia , 36, 212-219.2001
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: 397-403.1994
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
Past researchers of the lab
Katerina Akassoglou, Ph.D (1994-1998)
George Kassiotis, Ph.D (1995-2000)
Baosheng Ge, Ph.D (1998-2000)
Era Taoufik, PhD (2000-2012)
Vivian Tseveleki, PhD (2000-2014)
Chengmai Ruan, Ph.D (2001-2002)
David Plows, Ph.D (2002-2004)
Mary Emmanouil, Ph.D (2004-2009)
Anda Voulgari-Kokota (2005-2010)
Sotiris-Spyros Vamvakas, Ph.D (2007-2009)
Vasiliki Kyrargyri, Ph.D (2008-2015)
Ioannis Kanistras, Ph.D (2012-2014)
We maintain long-term collaborations with:
Prof. David Atwell – Neuroscience, Physiology & Pharmacology, Faculty of Life Sciences, University College London, UK.
Prof. Dr. Hans Lassmann– Center for Brain Research, Division of Neuroimmunology, Medical University of Vienna, Austria.
Dr David Szymkowski – Xencor, Monrovia, CA, USA.
Prof. Malu Tansey – Department of Physiology, Emory University School of Medicine,
Atlanta, GA, USA.
Prof. Theodore Tselios – Department of Chemistry, University of Patras, Rio, Greece.