Our Aim
One of the key pathological features of Multiple Sclerosis, a human autoimmune disease, is mononuclear cell infiltration within the central nervous system (CNS). Among those cells, autoantigen-specific CD4+ T helper cells play a central role in initiation of CNS inflammation. The aim of our work is to visualize and to understand the cellular and molecular mechanisms of T cell infiltration into the CNS beyond the blood-brain barrier and induction of CNS inflammation.
Our Approach
We use intravital two-photon microscopy to visualize genetically labelled autoreactive T cells in Experimental Autoimmune Encephalomyelitis (EAE), an animal model for Multiple Sclerosis. For T cell labelling, conventional Green Fluorescent Protein (GFP) and Red Fluorescent Protein (RFP) are used. Additionally, we perform functional imaging by using calcium sensing proteins.. Flow cytometry, immune fluorescence confocal imaging and quantitative PCR are used to support findings from intravital imaging.
Some of our current projects
Highly and weakly encephalitogenic T cells
The encephalitogenicity of T cells depends on TCR specificity and host. We have shown previously that the encephalitogenic potential is also related to T cell activation in the CNS. Highly encephalitogenic T cells were activated in the CNS, whereas weakly encephalitogenic T cells were not. However, it is largely unknown what factors contribute to different degrees of T cell activation. We visualized and compared the behaviour of highly and weakly encephalitogenic T cells in the animal. We are especially interested in differences in antigen presentation within the CNS.
Molecular mechanism of encephalitogenic T cell infiltration into the CNS
The encephalitogenic T cells migrate into the CNS through the Blood-Brain barrier, a tight membrane which separates CNS and periphery. Although integrina4, which is the target of Natalizumab, one of the most effective treatments for MS patients, is known to be involved, the detailed molecular mechanism for this step is still largely unknown. We are exploring the molecule which can be used as therapeutic target.
Visualizing T cell activation in vivo
Varying calcium concentrations within migrating T cells were detected using a FRET-based calcium biosensor. Images were acquired by intravital two-photon microscopy. Encephalitogenic T cells are activated in the CNS; however, the percentage of activated T cells goes only up to 40%. By conventional immunological analysis, it is hard to answer where and how T cells are activated within the target organ. In contrast, by using a recently developed calcium sensing protein, we can visualize T cell activation in vivo at single cell level. To this end, we express Fluorescence Resonance Energy Transfer (FRET) based calcium sensing protein in T cells and use them for intravital two-photon imaging.
Intravital imaging in mouse EAE models
Encephalitogenic T cells exist in healthy individuals although they do not induce inflammation in the majority of cases. Recent findings suggest that gut microbiota trigger the first step of inflammation by stimulating T cells to initiate CNS infiltration. By using a transgenic mouse line with high numbers of encephalitogenic T cells in combination with activation sensors and intravital imaging, we try to visualize the stimulation in the gut-associated lymphoid tissues and unveil the mechanism of the process. In addition, we study the phenotypical changes of those encephalitogenic T cells following stimulation.
2021
Behrens G, Edelmann SL, Raj T, Kronbeck N, Monecke T, Davydova E, Wong EH, Kifinger L, Giesert F, Kirmaier ME, Hohn C, de Jonge LS, Pisfil MG, Fu M, Theurich S, Feske S, Kawakami N, Wurst W, Niessing D, Heissmeyer V. Disrupting Roquin-1 interaction with Regnase-1 induces autoimmunity and enhances antitumor responses. Nat Immunol. 2021 Dec;22(12):1563-1576.
Hiltensperger M, Beltrán E, Kant R, Tyystjärvi S, Lepennetier G, Domínguez Moreno H, Bauer IJ, Grassmann S, Jarosch S, Schober K, Buchholz VR, Kenet S, Gasperi C, Öllinger R, Rad R, Muschaweckh A, Sie C, Aly L, Knier B, Garg G, Afzali AM, Gerdes LA, Kümpfel T, Franzenburg S, Kawakami N, Hemmer B, Busch DH, Misgeld T, Dornmair K, Korn T. Skin and gut imprinted helper T cell subsets exhibit distinct functional phenotypes in central nervous system autoimmunity. Nat Immunol. (2021) Jul;22(7):880-892.
2020
Gerhards R, Pfeffer LK, Lorenz J, Starost L, Nowack L, Thaler FS, SchlüterM, Rübsamen H, Macrini C, Winklmeier S, Mader S, Bronge M, Grönlund H, Feederle R, Hsia H, Lichtenthaler SF, Merl-Pham J, Hauck SM, Kuhlmann T, Bauer IJ, Beltran E, Gerdes LA, Mezydlo A, Bar-Or A, Banwell B, Khademi M, Olsson T, Hohlfeld R, Lassmann H, Kümpfel T, Kawakami N*, Meinl E. Oligodendrocyte myelin glycoprotein as a novel target for pathogenic autoimmunity in the CNS. Acta Neuropathologica Communications (2020), accepted. (* co-senior author)
Beltrán E, Paunovic M, Gebert D, Cesur E, Jeitler M, Höftberger R, Malotka J, Mader M, Kawakami N, Meinl E, Bradl M, Dornmair K, Lassmann H. Archeological neuroimmunology: Resurrection of a pathogenic immune response from a historical case sheds light on human autoimmune encephalomyelitis and multiple sclerosis. Acta Neuropathologica (2020), accepted.
2018
Spadaro M, Winklmeier S, Beltrán E, Macrini C, Höftberger R, Schuh E, Thaler FS, Gerdes LA, Laurent S, Gerhards R, Brändle S, Dornmair K, Breithaupt C, Krumbholz M, Moser M, Krishnamoorthy G, Kamp F, Jenne D, Hohlfeld R, Kümpfel T, Lassmann H, Kawakami N*, Meinl E*. Pathogenicity of human antibodies against myelin oligodendrocyte glycoprotein. Ann Neurol. 2018 Aug;84(2):315-328. (* co-senior author)
Kawakami N. Intravital Imaging of T Cells Within the Spinal Cord. Methods Mol Biol. 2018;1763:119-127.
2017
Kyratsous NI, Bauer IJ, Zhang G, Pesic M, Bartholomäus I, Mues M, Fang P, Wörner M, Everts S, Ellwart JW, Watt JM, Potter BVL, Hohlfeld R, Wekerle H, Kawakami N. Visualizing context-dependent calcium signaling in encephalitogenic T cells in vivo by two-photon microscopy. Proc Natl Acad Sci U S A. 2017 Aug 1;114(31):E6381-E6389.
Haghayegh Jahromi N, Tardent H, Enzmann G, Deutsch U, Kawakami N, Bittner S, Vestweber D, Zipp F, Stein JV, Engelhardt B. A Novel Cervical Spinal Cord Window Preparation Allows for Two-Photon Imaging of T-Cell Interactions with the Cervical Spinal Cord Microvasculature during Experimental Autoimmune Encephalomyelitis. Front Immunol. 2017 Apr 11;8:406.
Druzd D, Matveeva O, Ince L, Harrison U, He W, Schmal C, Herzel H, Tsang AH, Kawakami N, Leliavski A, Uhl O, Yao L, Sander LE, Chen CS, Kraus K, de Juan A, Hergenhan SM, Ehlers M, Koletzko B, Haas R, Solbach W, Oster H, Scheiermann C. Lymphocyte Circadian Clocks Control Lymph Node Trafficking and Adaptive Immune Responses. Immunity. 2017 Jan 17;46(1):120-132.
2016
Kawakami N. In vivo imaging in autoimmune diseases in the central nervous system. Allergol Int. 2016 Jul;65(3):235-42.
Zilkha-Falb R, Kaushansky N, Kawakami N, Ben-Nun A. Post-CNS-inflammation expression of CXCL12 promotes the endogenous myelin/neuronal repair capacity following spontaneous recovery from multiple sclerosis-like disease. J Neuroinflammation. 2016 Jan 8;13:7.
2014
Koutrolos M, Berer K, Kawakami N, Wekerle H, Krishnamoorthy G. Treg cells mediate recovery from EAE by controlling effector T cell proliferation and motility in the CNS. Acta Neuropathol Commun. 2014 Dec 5;2:163.
Ben-Nun A, Kaushansky N, Kawakami N, Krishnamoorthy G, Berer K, Liblau R, Hohlfeld R, Wekerle H. From classic to spontaneous and humanized models of multiple sclerosis: impact on understanding pathogenesis and drug development. J Autoimmun. 2014 Nov;54:33-50.
2013
Pohl M, Kawakami N, Kitic M, Bauer J, Martins R, Fischer MT, Machado-Santos J, Mader S, Ellwart JW, Misu T, Fujihara K, Wekerle H, Reindl M, Lassmann H, Bradl M. T cell-activation in neuromyelitis optica lesions plays a role in their formation. Acta Neuropathol Commun. 2013 Dec 24;1:85.
Mues M, Bartholomäus I, Thestrup T, Griesbeck O, Wekerle H, Kawakami N*, Krishnamoorthy G. Real-time in vivo analysis of T cell activation in the central nervous system using a genetically encoded calcium indicator. Nat Med. 2013 Jun;19(6):778-83. (* co-senior author)
Pesic M, Bartholomäus I, Kyratsous NI, Heissmeyer V, Wekerle H, Kawakami N. 2-photon imaging of phagocyte-mediated T cell activation in the CNS. J Clin Invest. 2013 Mar;123(3):1192-201.
Naoto Kawakami, Principal Investigator
Read more about the PI on the next tab.
Katrin Lämmle, PhD student
I obtained my bachelor and master degree in Molecular Medicine at the Friedrich-Alexander University in Erlangen. In my master thesis I worked on synovial macrophages in rheumatoid arthritis. I joined the Kawakami lab in 2019 to work on my PhD thesis. My research focuses on the migration of encephalitogenic T cells into the central nervous system at the beginning of experimental autoimmune encephalomyelitis. In my spare time, I enjoy sports, dancing, and baking.
La Zhuo, PhD student
My bachelor's and master's degrees are in clinical medicine and neurology at Jilin University, where I gained a systematic understanding of neurological diseases and witnessed many clinical cases that showed how severe the effects of the immune system can be. Since I was very interested in the interaction between immune cells and the central nervous system, I joined the lab of Naoto Kawakami in 2021 to do my PhD thesis. My research mainly focused on the interaction of T cells and microglia under EAE disease model. In my free time, I like to go swimming, hiking, and having fun with friends
Alumni
Isabel Bauer
Cuilien Du
Clara Seizer
Mary Claire Tuohy
Ping Fang, post doctoral fellow, UCLA, USA
Max Frank
Mariam Wörner
Nikolaos Kyratsous
Marija Pesic
Elham Saghaei
Ingo Barholomäus
PD. Dr. rer. nat. Naoto Kawakami
I have been working on the function of encephalitogenic T cells during CNS inflammation for almost 20 years. I would like to understand how those encephalitogenic T cells migrate into the CNS, become activated and cause local inflammation. To answer these questions, I have establihsed and still use fluorescent protein based activation sensors and intravital imaging. Key research achievements include the first imaging of encephalitogenic T cells in a living animal (Nature 2009) and the detection of encephalitogenic T cell activation by using NFAT-GFP fusion proteins (JCI, 2013) and calcium sensing proteins (PNAS 2017).
Training
1992 – 1996 Bachelor Pharmaceutical Sciences, Osaka University, Japan
1996 – 1998 Master Pharmaceutical Sciences, Osaka University, Japan
1998 – 2001 PhD Pharmaceutical Sciences, Osaka University, Japan
2001 – 2002 Postdoctoral fellow at Dept. of Neuroimmunology, Max Planck Institute of Neurobiology, Martinsried, Germany
Academic positions & appointments
2003 – 2011 Group leader at Dept. of Neuroimmunology, Max Planck Institute of Neurobiology, Martinsried, Germany
Since 2011 Group leader at Institute of Clinical Neuroimmunology, LMU Munich, Germany
2013 Habilitation in Neuroimmunology, LMU Munich, Germany
2015 – 2020 Heisenberg fellowship
since 2015 Member and research group leader at the Biomedical Center, LMU Munich, Germany
Collaborative Research
2008 – 2011 Member and PI of the Collaborative Research Center 571 “Autoimmune reactions: From manifestations and mechanisms to therapy”
since 2016 Member and PI of the Collaborative Research Center 128 “Initiating/effector versus regulatory mechanisms in Multiple Sclerosis – progress towards tackling the disease”
Awards & honors
2001 – 2002 Research fellowship from Alexander von Humboldt Foundation
2006 Helmut Bauer prize for a Young Scientist in Multiple Sclerosis Research
2015 – 2020 Heisenberg fellowship
5 key papers
Spadaro M, Winklmeier S, Beltrán E, Macrini C, Höftberger R, Schuh E, Thaler FS, Gerdes LA, Laurent S, Gerhards R, Brändle S, Dornmair K, Breithaupt C, Krumbholz M, Moser M, Krishnamoorthy G, Kamp F, Jenne D, Hohlfeld R, Kümpfel T, Lassmann H, Kawakami N*, Meinl E. Pathogenicity of human antibodies against myelin oligodendrocyte glycoprotein. Ann Neurol. 2018 Aug;84(2):315-328. (* co-senior author)
Kyratsous NI, Bauer IJ, Zhang G, Pesic M, Bartholomäus I, Mues M, Fang P, Wörner M, Everts S, Ellwart JW, Watt JM, Potter BVL, Hohlfeld R, Wekerle H, Kawakami N. Visualizing context-dependent calcium signaling in encephalitogenic T cells in vivo by two-photon microscopy. Proc Natl Acad Sci U S A. 2017 Aug 1;114(31):E6381-E6389.
Mues M, Bartholomäus I, Thestrup T, Griesbeck O, Wekerle H, Kawakami N*, Krishnamoorthy G. Real-time in vivo analysis of T cell activation in the central nervous system using a genetically encoded calcium indicator. Nat Med. 2013 Jun;19(6):778-83. (* co-senior author)
Pesic M, Bartholomäus I, Kyratsous NI, Heissmeyer V, Wekerle H, Kawakami N. 2-photon imaging of phagocyte-mediated T cell activation in the CNS. J Clin Invest. 2013 Mar;123(3):1192-201.
Bartholomäus I, Kawakami N, Odoardi F, Schläger C, Miljkovic D, Ellwart JW, Klinkert WE, Flügel-Koch C, Issekutz TB, Wekerle H, Flügel A. Effector T cell interactions with meningeal vascular structures in nascent autoimmune CNS lesions. Nature. 2009 Nov 5;462(7269):94-8.
Christmas 2019
Octoberfest 2019
Conference in Cuba 2019
Group picture 2018
Thanksgiving 2017
Autumn 2017
