T −Transfer Experimental Autoimmune Encephalomyelitis: Pillar of and Autoimmunity

This information is current as Reinhard Hohlfeld and Lawrence Steinman of September 24, 2021. J Immunol 2017; 198:3381-3383; ; doi: 10.4049/jimmunol.1700346 http://www.jimmunol.org/content/198/9/3381 Downloaded from

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The Journal of Immunology is published twice each month by The American Association of Immunologists, Inc., 1451 Rockville Pike, Suite 650, Rockville, MD 20852 Copyright © 2017 by The American Association of Immunologists, Inc. All rights reserved. Print ISSN: 0022-1767 Online ISSN: 1550-6606. Th eJournal of Pillars of Immunology Immunology

T Cell–Transfer Experimental Autoimmune Encephalomyelitis: Pillar of Multiple Sclerosis and Autoimmunity Reinhard Hohlfeld*,† and Lawrence Steinman‡

xperimental autoimmune encephalomyelitis (EAE) is autoantigen (MBP) or control Ags (OVA or purified known as the animal model of multiple sclerosis, the derivative of tuberculin). After 3 d of incubation, the cells were E most common disabling neurologic disorder of young collected, and proliferating low-density lymphoblasts were sep- adults (reviewed in Refs. 1–3). Beyond its role as a specific arated from quiescent small cells by centrifugation on discon-

disease model, EAE has served as a key paradigm for inves- tinuous Ficoll density gradients. The lymphoblasts were propagated Downloaded from tigating fundamental problems of autoimmunity and immune and maintained in vitro in culture medium supplemented with tolerance. Classically, EAE is induced by active immuniza- supernatant from ConA-stimulated spleen cells. At regular in- tion with autoantigens, such as myelin basic protein tervals, the cultures were restimulated with the relevant Ag in (MBP), myelin glycoprotein, or proteolipid the presence of irradiated syngeneic thymocytes as feeder cells. protein, in adjuvant. This month’s Pillars of Immunology After Ben-Nun had returned to Israel, the experiment culmi-

article features “T-transfer EAE,” a variant of EAE first de- nated in the first successful transfer of EAE, which was achieved http://www.jimmunol.org/ scribed in 1981 by Ben-Nun et al. (4). In this article (cited by injecting freshly activated MBP-specific T line cells into .800 times), the investigators demonstrated that EAE can be naive recipient rats. About 1 million MBP-specific, but not adoptively transferred by injecting highly purified MBP-specific purified protein derivative of tuberculin–specific, lymphoblasts T line cells into syngeneic naive recipient animals. This effec- 1 were sufficient to induce EAE in the majority of recipients. tively pinpointed myelin-autoreactive CD4 T lymphocytes as One of the authors of this commentary (R.H.), who was a the crucial effector cells of EAE, an observation that had far- postdoctoral fellow in Wekerle’s laboratory at that time, will reaching implications for autoimmunity research, in general, never forget when Irun Cohen’s telegram arrived from Rehovot and for multiple sclerosis, in particular. with just the message “Heureka.” by guest on September 24, 2021 The article resulted from a fruitful Israeli–German collab- The article reporting these findings (4) merits selection as a oration in which each side contributed an essential method- Pillars of Immunology publication for at least four reasons. ological component. Irun R. Cohen and Avraham Ben-Nun First, it illustrates how the technique of culturing lines of the Weizmann Institute in Rehovot, Israel had experience and clones with T cell growth factor opened completely new with classical EAE and were skilled at propagating T cell lines, avenues for autoimmunity research. Earlier studies on adop- which was still a relatively new technique at the time. The tive transfer of lymphoid cells in EAE had taken a different German collaboration partner, Hartmut Wekerle, contributed approach. One experiment showed that repeated transfer of his expertise in the selection and purification of autoreactive myelin-sensitized lymphocytes suppressed actively induced T cells. With a stipend from the Volkswagenstiftung, Ben-Nun paralysis (6). A second experiment showed that T cells sensi- visited Wekerle’s laboratory at the Max-Planck Institute of tized to myelin could induce EAE, but only after the recipi- Immunobiology in Freiburg to learn how to isolate Ag-specific ents were irradiated (7). The featured Pillars of Immunology T cells from primed animals (5). Lymph node cells from im- article used, for the first time, purified autoreactive T cell lines munized Lewis rats were cultured in the presence of myelin for adoptively transferring EAE (4), thereby overcoming the need to irradiate the host. As highlighted in another recent Pillars of Immunology commentary (8), the discovery and ini- *Institute of Clinical Neuroimmunology, Biomedical Center, University Hospital, Campus Martinsried-Grosshadern, Ludwig-Maximilians University, D-81377 Munich, tial characterization of T cell growth factor (9–11), which Germany; †Munich Cluster of Systems Neurology, 81377 Munich, Germany; and ‡De- works across different species (e.g., mouse, rat, and human), partment of Neurology and Neurological Sciences, Stanford University, Stanford, CA 94305 was a major breakthrough in cellular immunology. It allowed ORCID: 0000-0002-2437-2250 (L.S.). the expansion of T cells with specificity for, theoretically, any Address correspondence and reprint requests to Dr. Reinhard Hohlfeld or Dr. Lawrence Ag, including, as first shown by Ben-Nun et al. (4), a defined Steinman, Institute of Clinical Neuroimmunology, Grosshadern Clinic, Ludwig autoantigen of the . Subsequently, similar ap- Maximilians University, Marchioninistrasse 15, D-81377 Munich, Germany (R.H.) or proaches were used for the isolation and propagation of Department of Neurology and Neurological Sciences, Beckman Center for Molecular Medicine, 279 Campus Drive, Stanford, CA 94305-5316 (L.S.). E-mail addresses: autoantigen-specific T cell lines from human patients with [email protected] (R.H.) or [email protected] (L.S.) autoimmune diseases, such as myasthenia gravis (12) and Abbreviations used in this article: EAE, experimental autoimmune encephalomyeli- multiple sclerosis (13–17). tis; MBP, myelin basic protein. Second, the Ben-Nun et al. (4) study touched upon a Copyright Ó 2017 by The American Association of Immunologists, Inc. 0022-1767/17/$30.00 fundamental issue of autoimmunity. According to Burnet’s

www.jimmunol.org/cgi/doi/10.4049/jimmunol.1700346 3382 PILLARS OF IMMUNOLOGY clonal selection theory (18), the lymphocyte repertoire needs Abs but rather on the elimination of B cells as Ag-presenting to be purged of autoreactive lymphocytes early in development and cytokine-secreting cells (37). to avoid autoimmunity; in other words, self-recognizing clones Like all models, the T cell–transfer model has its limita- are “forbidden.” This would imply that the autoaggressive tions. Despite its tremendous usefulness for studying T cell– T cells isolated by Ben-Nun et al. had arisen by somatic muta- mediated effector mechanisms, the model is obviously not tion. However, using essentially the same method, Schluesener suitable for investigating the initial triggering events of CNS and Wekerle (19) were able to isolate MBP-specific encepha- inflammation. In this regard, genetically engineered sponta- litogenic T cells from lymph nodes of healthy Lewis rats. The neous animal models are more appropriate (38). Finally, it special trick in this case was to eliminate spontaneously pro- should be kept in mind that the demonstration that myelin- 1 liferating T cells via a round of negative selection and then autoreactive CD4 T cells mediate EAE in rodent models to isolate MBP-reactive T cells in a second, positive-selection does not necessarily imply that these T cells are the culprits round (19). Using this approach, the investigators isolated a in human multiple sclerosis, for which a unifying target (auto-) highly encephalitogenic T cell line with which they could antigen has not been identified (reviewed in Refs. 32, 39). transfer disease to naive recipient animals by injecting as few as 1 3 104 T cells. Considering that there is no postthymic somatic hypermutation of TCRs, these observations prove Acknowledgments the presence of potentially autoaggressive T cells in the healthy This article is dedicated to Irun Cohen on the occasion of his 80th birthday. immune repertoire, raising the question of how these cells Downloaded from are normally held in check. Interestingly, the findings (19) were reported at a time when suppressor T cells had fallen Disclosures into complete disregard and only later regained reputation The authors have no financial conflicts of interest. (20). Based on the observation that autoimmune T cells are physiological components of the , Irun Cohen References http://www.jimmunol.org/ proposed that physiological autoimmunity is essential for 1. Steinman, L. 2003. Optic neuritis, a new variant of experimental encephalomyelitis, homeostasis and maintenance, an idea he elaborated with his a durable model for all seasons, now in its seventieth year. J. Exp. Med. 197: 1065– 1071. concept of the “immunological homunculus,” the immune 2. Gold, R., C. Linington, and H. Lassmann. 2006. Understanding pathogenesis system’s “self-image” (21, 22). and therapy of multiple sclerosis via animal models: 70 years of merits and Third, the T cell–transfer model opened a whole new field: culprits in experimental autoimmune encephalomyelitis research. Brain 129: 1953–1971. in vivo imaging of encephalitogenic T cells by multiphoton 3. Ben-Nun, A., N. Kaushansky, N. Kawakami, G. Krishnamoorthy, K. Berer, live microscopy (reviewed in Ref. 23). Using in vitro retroviral R. Liblau, R. Hohlfeld, and H. Wekerle. 2014. From classic to spontaneous and humanized models of multiple sclerosis: impact on understanding pathogenesis and transduction, MBP-specific rat T cells can be labeled with drug development. J. Autoimmun. 54: 33–50.

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