Virus Immunology (Prof. Marcus Altfeld)
Prof. Marcus Altfeld, Benjamin Grünhagel, Birgitta Maurer, Dr. Maria Pujantell, Rebecca-jo Thiele, Dr. Kati Tillack
Our immune system defends us from environmental threats, such as bacterial and viral infections, and detects and removes abnormal cells that potentially lead to malignancies. Optimal immunological homeostasis is achieved when the threat (e.g. pathogen) is removed with the highest efficiency whilst avoiding collateral tissue damage for the host. More and more scientific data, however, suggest that this immunological balance differs between women and men. It has been shown that women mount stronger immune responses against pathogens compared to men, leading to more rapid control or clearance of infections. Women also develop stronger immunity in response to vaccinations, and exhibit stronger immune responses against malignancies, including leukemia. Enhanced immune responsiveness in females, however, comes at a cost, including aggravated tissue damage, higher immune activation, persistent inflammation and significantly higher incidences of autoimmune diseases, such as autoimmune hepatitis and multiple sclerosis. However, the precise contribution of sex hormones and sex chromosome-encoded genes to sexspecific differences in immunity, and their relevance for human disease manifestations, remains insufficiently understood. The understanding of these mechanisms will pave the way for novel treatment strategies for infectious and immune-mediated diseases that take the sex of the affected individual into account.
Type I IFN-mediated sex differences in immune response to HIV-1
In acute HIV-1 infection, women control viral replication better than men. In contrast, women develop increased immune activation and faster loss of CD4+ T cells during untreated chronic HIV-1 infection. Increasing data indicate that these sex differences in the manifestation of HIV-1 disease are mediated by sex-specific differences in antiviral immunity. In previous studies we demonstrated that pDCs derived from women produce more IFNα in response to HIV-1 than pDCs from men, potentially contributing to the better control of viremia in acute infection. This increased IFNα production in response to HIV-1 however also led to significantly higher immune activation and faster loss of CD4+ T cells in chronically HIV-1-infected women. Taken together these data strongly suggest that sex differences in Type I IFN production result in differences in HIV-1 disease manifestations between women and men. However, the precise molecular mechanisms underlying these sex differences in HIV-1 and other infectious diseases remain poorly understood. We hypothesize that X chromosome-encoded genes play a critical role in regulating Type I IFN responses, and that gene-dose effects resulting from escape from X chromosome inactivation (XCI) contribute to sex-specific differences in antiviral immunity, using HIV-1 as a model. We will furthermore test the hypothesis that changes in levels of sex hormones can regulate Type I IFN responses of pDCs, using longitudinal samples from a human transgender cohort. Taken together, these studies will identify critical mechanisms underlying sex-specific differences in antiviral immunity, and provide rationale for the design of interventions that take these differences between women and men into account.
DFG: CRU 306 Primary sclerosing cholangitis
DFG: CRC 841 Liver inflammation: Infection, immune regulation und consequences (A7: HCV infection: Mechanisms of NK-cell-mediated control)
EHVA European HIV Alliance
DZIF German Center for Infection Research
Hagen SH, Hennesen J, Altfeld M. (2021) Assessment of escape from X chromosome inactivation and gene expression in single human immune cells. STAR Protoc 2:100641
Sven Hendrik Hagen, Florian Henseling, Jana Hennesen, Hélène Savel, Solenne Delahaye, Laura Richert, Susanne Maria Ziegler, Marcus Altfeld. (2020) Heterogeneous Escape from X Chromosome Inactivation Results in Sex Differences in Type I IFN Responses at the Single Human pDC Level. Cell Rep 33:108485
Adland E, Millar J, Bengu N, Muenchhoff M, Fillis R, Sprenger K, Ntlantsana V, Roider J, Vieira V, Govender K, Adamson J, Nxele N, Ochsenbauer C, Kappes J, Mori L, van Lobenstein J, Graza Y, Chinniah K, Kapongo C, Bhoola R, Krishna M, Matthews PC, Poderos RP, Lluch MC, Puertas MC, Prado JG, McKerrow N, Archary M, Ndung'u T, Groll A, Jooste P, Martinez-Picado J, Altfeld M, Goulder P. (2020) Sex-specific innate immune selection of HIV-1 in utero is associated with increased female susceptibility to infection. Nat Commun 11:1767
Bunders MJ, Altfeld M (2020) Implications of Sex Differences in Immunity for SARS-CoV-2 Pathogenesis and Design of Therapeutic Interventions. Immunity 53:487-495Lotter H, Altfeld M. (2019) Sex differences in immunity. Semin Immunopathol. 41:133-135
Scully EP, Gandhi M, Johnston R, Hoh R, Lockhart A, Dobrowolski C, Pagliuzza A, Milush JM, Baker CA, Girling V, Ellefson A, Gorelick R, Lifson J, Altfeld M, Alter G, Cedars M, Solomon A, Lewin SR, Karn J, Chomont N, Bacchetti P, Deeks SG. (2019) Sex-Based Differences in Human Immunodeficiency Virus Type 1 Reservoir Activity and Residual Immune Activation. J Infect Dis. 219:1084-1094
Rechtien A, Altfeld M. (2019) Sexual dimorphism in HIV-1 infection. Semin Immunopathol 41:195-202Ziegler SM, Beisel C, Sutter K, Griesbeck M, Hildebrandt H, Hagen SH, Dittmer U, Altfeld M. (2017) Human pDCs display sex-specific differences in type I interferon subtypes and interferon α/β receptor expression. Eur J Immunol 47:251-256
Griesbeck M, Ziegler S, Laffont S, Smith N, Chauveau L, Tomezsko P, Sharei A, Kourjian G, Porichis F, Hart M, Palmer CD, Sirignano M, Beisel C, Hildebrandt H, Cénac C, Villani AC, Diefenbach TJ, Le Gall S, Schwartz O, Herbeuval JP, Autran B, Guéry JC, Chang JJ, Altfeld M. (2015) Sex Differences in Plasmacytoid Dendritic Cell Levels of IRF5 Drive Higher IFN-α Production in Women. J Immunol. 195:5327-36
Addo MM, Altfeld M. (2014) Sex-based differences in HIV type 1 pathogenesis. J Infect Dis. 209 Suppl 3:S86-92
Chang JJ, Woods M, Lindsay RJ, Doyle EH, Griesbeck M, Chan ES, Robbins GK, Bosch RJ, Altfeld M. (2013) Higher expression of several interferon-stimulated genes in HIV-1-infected females after adjusting for the level of viral replication. J Infect Dis 208:830-8
Meier A, Chang JJ, Chan ES, Pollard RB, Sidhu HK, Kulkarni S, Wen TF, Lindsay RJ, Orellana L, Mildvan D, Bazner S, Streeck H, Alter G, Lifson JD, Carrington M, Bosch RJ, Robbins GK, Altfeld M. (2009) Sex differences in the Toll-like receptor-mediated response of plasmacytoid dendritic cells to HIV-1. Nat Med 15: 955-9
Clinical Immunology (Prof. Friedrich Haag)
Prof. Friedrich Haag, Gudrun Dubberke, Paul Düngelhoef, Samantha Eiberg, Kilian Keetz, Birgitta Maurer
The focus of our research is on exploring the mechanisms by which extracellular nucleotides, such as ATP, NAD, and their derivatives, regulate the function of immune cells. Topics include - the impact of ATP-gated P2X ion channels on T cell receptor-mediated signaling. - elucidating how and when ATP is released from immune cells, and how it is metabolized to adenosine on the outside. Because of our close connection to the Clinical Immunodiagnostics Laboratory our group is also interested in evaluating and improving immunodiagnostic methods suitable for routine use. Here our focus is currently on the development of flow cytometry protocols for the characterization of immune cells from patients. We also provide assistance and immunodiagnostic services for research projects that need to measure immunological parameters.
Extracellular nucleotides as regulators of lymphocyte function
Besides their metabolic importance inside cells, adenosine triphosphate (ATP) and other adenine nucleotides fulfill important roles outside of cells as autocrine and paracrine signal mediators. ATP is abundant within cells (3-10 mM), but under steady state conditions it is barely detectable in the extracellular milieu (low nanomolar conentrations). However, cells can actively secrete ATP for signaling purposes, or it can be passively released from cells upon cell damage. Once released into the extracellular space, ATP acts on ionotropic P2X or the metabotropic P2Y receptors present on the cell surface. While P2Y receptors belong to the family of G protein-coupled receptors (GPCRs), P2X receptors are ATP-gated cation channels, which among other actions can mediate the influx of calcium ions (Ca2+) into the cell. Outside the cell, ATP is degraded by the combined actions of ecto-nucleoside triphosphate diphosphohydrolases (ENTPDs) such as CD39 and 5’-nucleotidases like CD73, generating adenosine (ADO), which is a ligand for another family of GPCRs, the P1 receptors. Regulating the balance between extracellular ATP and ADO concentrations is an important mechanism for the regulation of inflammation. The release of ATP is an ancient, evolutionarily conserved danger signal heralding tissue damage, and thus alerting immune cells to initiate or augment an inflammatory reaction. This pro-inflammatory signal is counteracted by the degradation of ATP to ADO. Especially, stimulation of the A2A and A2B P1 receptors acts on many immune cells as a strong inhibitory (and thus anti-inflammatory) signal. In a similar fashion, extracellular nicotinamide adenine dinucleotide (NAD) also initiates signaling cascades by serving as a substrate either for the ADP-ribosyltransferase ARTC2 or for the ecto-NADase CD38. On mouse T lymphocytes, which carry ARTC2, a prominent target for ADP-ribosylation is the P2X7 ion channel, which is activated by this modification. CD38 uses NAD and NADP to generate calcium-mobilizing second messengers such as cyclic ADP-ribose (cADPR) and nicotinic acid adenine dinucleotide phosphate (NAADP) (Fig. 1).
DFG: SFB 1328 Adenine nucleotides in immunity and inflammation (A12 Role of the ectonucleotidases CD39 and CD73 for the modulation of the immune response in chronic human viral infections)
DFG: SFB 1328 Adenine nucleotides in immunity and inflammation (Z02 Antibodies and nanobodies for adenine nucleotide research)
Regulation of immune functions by extracellular nucleotides
Brauneck F, Haag F, Woost R, Wildner N, Tolosa E, Rissiek A, Vohwinkel G, Wellbrock J, Bokemeyer C, Schulze zur Wiesch J, Ackermann C, and Fiedler W. (2021) Increased frequency of TIGIT+CD73-CD8+ T cells with a TOX+ TCF-1low profile in patients with newly diagnosed and relapsed AML. OncoImmunology. 10:193039110.1080/2162402X.2021.1930391.
Libera J, Wittner M, Kantowski M, Woost R, Eberhard JM, de Heer J, Reher D, Huber S, Haag F, and Schulze Zur Wiesch J. (2020) Decreased Frequency of Intestinal CD39(+) γδ(+) T Cells With Tissue-Resident Memory Phenotype in Inflammatory Bowel Disease. Front Immunol. 11:56747210.3389/fimmu.2020.567472.
Haag F, and Buck F. (2015) Identification and analysis of ADP-ribosylated proteins. Curr Top Microbiol Immunol. 384:33-5010.1007/82_2014_424.
Seman M, Adriouch S, Scheuplein F, Krebs C, Freese D, Glowacki G, Deterre P, Haag F, and Koch-Nolte F. (2003) NAD-induced T cell death: ADP-ribosylation of cell surface proteins by ART2 activates the cytolytic P2X7 purinoceptor. Immunity. 19:571-58210.1016/s1074-7613(03)00266-8.
Kaschubowski KE, Kraft AE, Nikolaev VO, and Haag F. (2020) Using FRET-Based Fluorescent Sensors to Monitor Cytosolic and Membrane-Proximal Extracellular ATP Levels. Methods Mol Biol. 2041:223-23110.1007/978-1-4939-9717-6_16.
Johnsen B, Kaschubowski KE, Nader S, Schneider E, Nicola JA, Fliegert R, Wolf IMA, Guse AH, Nikolaev VO, Koch-Nolte F, and Haag F. (2019) P2X7-mediated ATP secretion is accompanied by depletion of cytosolic ATP. Purinergic Signal. 15:155-16610.1007/s11302-019-09654-5.
The immune response to infection and immunization against SARS-CoV-2
Haag F. (2021) Die natürliche und adaptive Immunantwort bei der Covid-19 Erkrankung Hamburger Ärzteblatt. 75:28-30
Rüther DF, Schaub GM, Duengelhoef PM, Haag F, Brehm TT, Fathi A, Wehmeyer M, Jahnke-Triankowski J, Mayer L, Hoffmann A, Fischer L, Addo MM, Luetgehetmann M, Lohse AW, Schulze Zur Wiesch J, and Sterneck M. (2021) SARS-CoV2-specific humoral and T-cell immune response after second vaccination in liver cirrhosis and transplant patients. Clinical Gastroenterology and Hepatology. https://doi.org/10.1016/j.cgh.2021.09.003.
Woo MS, Haag F, Nierhaus A, Jarczak D, Roedl K, Mayer C, Brehm TT, van der Meirschen M, Hennigs A, Christopeit M, Fiedler W, Karagiannis P, Burdelski C, Schultze A, Huber S, Addo MM, Schmiedel S, Friese MA, Kluge S, and Schulze Zur Wiesch J. (2021) Multi-dimensional and longitudinal systems profiling reveals predictive pattern of severe COVID-19. iScience. 24:10275210.1016/j.isci.2021.102752.
Ahmadi P, Hartjen P, Kohsar M, Kummer S, Schmiedel S, Bockmann JH, Fathi A, Huber S, Haag F, and Schulze Zur Wiesch J. (2020) Defining the CD39/CD73 Axis in SARS-CoV-2 Infection: The CD73(-) Phenotype Identifies Polyfunctional Cytotoxic Lymphocytes. Cells. 9:10.3390/cells9081750.
Molecular Immunology (Prof. Friedrich Koch-Nolte)
The Molecular Immunology group studies membrane-bound receptors and enzymes of immune and cancer cells and develops nanobodies and antibodies against these targets.
Prof. Friedrich Koch-Nolte, Dr. rer. nat. Thomas Eden, Marie Eggers, Dr. hum. biol. Carolina Pinto-Espinoza, Josephine Gebhardt, Julia Hambach, Dr. hum. biol. Klaus Kaschubowski, Cerusch Khan, Josephine Krenz, Dr. rer. nat. Anna Marei Mann, Luca Pape, Waldemar Schäfer, Alessa Schaffrath, Fabienne Seyfried, Tobias Stähler, Natalie Tode
DFG: SFB 1192 „Immune-mediated glomerular diseases” (B05: Nanobody-based treatment strategies in glomerulonephritis, Nicola Wanner, Friedrich Nolte)
DFG: SFB 1328 “Adenine Nucleotides in Immunity and Inflammation” (A10: Role of adenine nucleotides and purinergic receptors in adipose tissue, Jörg Heeren, Friedrich Nolte; Z02: Antibodies and nanobodies for adenine nucleotide research Friedrich Haag, Friedrich Nolte)
DFG: FOR 2879 "ImmunoStroke: From Immune Cells to Stroke Recovery" (A1: Role of ATP and NAD as DAMP signals in stroke recovery and as therapy targets, Tim Magnus, Friedrich Nolte)
DFG/ANR: Evaluation of P2X7 as a therapeutic target in autoimmune encephalomyelitis and in tumor immunity (Sahil Adriouch, Univ. Rouen, Friedrich Nolte)
DFG: Engineering the avidity of nanobody-based bispecific diabodies and heavy chain antibodies to specifically target myeloma cells that co-express two membrane proteins (Peter Bannas, Radiologie, Friedrich Nolte)
BMBF COMMUTE Combinatorial and multidisciplinary targeting of effective gene therapy vectors (P5 Targeting glioblastoma and muscle dystrophy with Nanobody-displaying AAV, Friedrich Nolte)
Short CV Prof. Friedrich Koch-Nolte
Deputy Director (since 1997)
Phone: +49 (0) 40 7410 - 53612
Biology, Wesleyan University 1976 (BA)
Medicine, University of Tübingen, 1983 (MD)
Molceular Biology, University of Hamburg, 1986 (Diploma)
Professor associée at the University of Rouen, France (2006-2007)
Visiting scientist at the University of California, San Francisco, CA, USA (1994)
Visiting scientist at the DNAX Research Institute of Molecular Biology, Palo Alto, CA, USA (1997)
Visiting scientist at The Jackson Lab, Bar Harbor, ME, USA (1991, 1999)
Research award of the German Cancer Society (2019)
Mentorship Award of the Simon-Claussen Foundation (2009)
Research Award of the Werner Otto Foundation (1997)
Research Award of the Martini Foundation (1991)
Member of the
Deutsche Gesellschaft für Immunologie (DGfI)
Deutsche Gesellschaft für Biochemie und Molekularbiologie (GBM)
Deutsche Gesellschaft für Zellbiologie (DGZ)
University Medical Center Hamburg-Eppendorf (UKE)
Friedrich Haag, Eva Tolosa, Hans-Willi Mittrücker Institute for Immunology Andreas Guse, Jörg Heeren, Ralf Fliegert Institute for Biochemistry and Molecular Cell Biology Peter Bannas Department of Diagnostic and Interventional Radiology and Nuclear Medicine Tim Magnus, Björn Rissiek Department of Neurology Cathy Meyer-Schwesinger Institute of Cellular and Integrative Physiology Nicola Wanner, Nicola Tomas, Elion Hoxha, Rolf Stahl, Ulf Panzer III. Medical Clinic and Polyclinic Boris Fehse, Kristoffer Rieken Department of Stem Cell Transplantation Irm Hermanns Borgmeyer Center for Molecular Neurobiology Katja Weisel, Walter Fiedler II. Medical Clinic and Polyclinic Thomas Braulke Institute of Osteology and Biomechanics
Sahil Adriouch University of Rouen Normandy Michael Hottiger University of Zurich Pablo Pelegrin University of Murcia Mascha Binder University of Halle Martin Trepel University of Augsburg Dirk Grimm University of Heidelberg Annette Nicke Ludwig Maximilian University of Munich Carsten Watzl Technical University of Dortmund Florian Schmidt University of Bonn Francesco Di Virgilio, Elena Adinolfi University of Ferrara Santina Bruzzone, Paolo Malatesta University of Genoa Jan van Lier University of Amsterdam Jaime Sancho University of Granada Pablo Engel University of Barcelona Ursula Dietrich Georg-Speyer-Haus William Heath University of Melbourne William Petri University of Virginia Steven Mansoor University of Oregon Hidde Ploegh Boston Children's Hospital
The focus of our laboratory is on the molecular characterization of lymphocyte membrane proteins, in particular receptors and enzymes involved in signaling by extracellular nucleotides. We generate monoclonal and single domain antibodies as new research and therapeutic tools. We are interested in ADP-ribosylation as a pathogenic mechanism of bacterial toxins and as a reversible posttranslational modification regulating protein functions. Using genetic immunization and antibody engineering, we strive to develop new tools for combating infections and for treating diseases of the immune system.
1. Lymphocyte membrane proteins: enzymes and receptors involved in signaling by extracellular NAD and ATP
Membrane proteins mediate the communication of cells with their environment. They function as receptors for soluble ligands and counter-receptors on other cells, as ion channels, nutrient transporters, and enzymes. The nucleotides NAD and ATP are key metabolites of energy metabolism found in cells from all kingdoms of life. The cell membrane is impermeable to these nucleotides, but they can exit cells via channels or pores gated by mechanical and/or electrical stimuli. During infection and inflammation injured cells release NAD and ATP through the damaged cell membrane. Extracellular NAD and ATP alert cells of the immune system to sites of tissue damage.
Cells of the immune system are equipped with a variety of sensors for these nucleotides (Fig. 1), including ligand-gated ion channels (P2X purine receptors) and nucleotide-metabolizing ecto-enzymes (CD38, CD296). CD296 (ARTs) functions as NAD-sensors and relay information about the levels of extracellular NAD into ADP-ribosylation of cell surface and secreted proteins. The NAD-hydrolyzing ecto-enzyme CD38 restricts the intensity and duration of NAD-signaling in the extracellular space by hydrolyzing NAD. Opening of the P2X7 ion channel is induced by binding of the soluble ligand ATP, or by NAD-dependent ADP-ribosylation. Passage of ions through P2X7 (calcium into the cell, potassium out of the cell) triggers a cascade of downstream events, including protease activation (caspases and ADAMs), externalization of phosphatidylserine (PS), activation of the inflammasome and cell death. Mice that cannot express the mentioned purine receptors or ecto-enzymes show impaired immune responses. The receptors and enzymes of purinergic signaling, thus, present potential targets for new anti-inflammatory or immune-stimulating drugs. In mouse models, activating the ARTC2 > P2X7 axis can enhance anti-tumor responses, while blocking this axis can reduce inflammation in autoimmune diseases.
2. Antibody engineering: genetic immunization, single domain antibodies
Antibodies are important tools of research, diagnosis and therapy. Antibodies are raised by immunizing experimental animals with proteins, synthetic peptides, or DNA. In the case of DNA immunizations, the protein encoded by the DNA is produced in its native conformation by the cells of the immunized animal and the induced immune response yields antibodies directed against proteins in native conformation (ADAPINCs) (Fig. 2). Such antibodies are useful in many applications were anti-peptide antibodies fail, e.g. affinity purification, flow cytometry, ELISA, FACS, and functional studies. cDNA immunization is offered as a service via the antibody core facility of the UKE.
Llamas produce unusual antibodies composed only of heavy chains. Their antigen-binding domain (VHH) is readily produced as a soluble recombinant protein, also designated nanobody or single domain antibody (sdAb) (Fig. 3). Nanobodies can form finger like extrusions that block clefts on protein surfaces, such as the active site crevice of enzymes, the ligand binding domain of a receptor, and the receptor binding domain of a virus. Nanobodies have great potential as therapeutics and as imaging agents.
We have generated enzyme-blocking nanobodies from llamas immunized with different ARTs: the SpvB Salmonella toxin, the binary clostridium difficile toxin CDTa, and the T cell ecto-enzyme ARTC2 (CD296). These nanobodies protect cells from the cytotoxic effects of SpvB, CDT, and ARTC2. In case of ARTC2, the nanobodies effectively block ARTC2 on the cell surface of regulatory T cells and iNKT cells within 10 minutes after intravenous injection. These nanobodies provide an important tool for protecting these regulatory cells from death induced by NAD released during tissue preparation.
We have also generated nanobodies against CD38, the major NAD-hydrolyzing ecto-enzyme. CD38 is emerging as a therapeutic target in multiple myeloma and other hematopoertic malignancies. Some of our nanobodies outperform the recently licensed CD38-specific monoclonal antibody Daratumumab (Darzalex) in cytotoxicity vs. hematopoetic cancer cell lines.
In cooperation with Ablynx, a Belgian company devoted to developing nanobodies for human therapies, we have generated nanobodies that block or enhance gating of the P2X7 ion channel. The P2X7-blocking nanobodies ameliorated inflammation in mouse models of glomerulonephritis and allergic dermatitis. In endotoxin-treated human blood cells, they effectively blocked LPS/ATP-induced release of the potent pro-inflammatory cytokine IL-1ß.
In order to facilitate the generation of new nanobodies, we have cloned the nanobody-encoding IgH locus from llama and successfully transferred an engineered version of this locus to transgenic mice (Fig. 4). Upon immunization, these mice produce nanobody-based heavy chain antibodies that undergo somatic hypermutation and class switch recombination. This novel platform greatly expands our capacity to generate functional nanobodies against interesting targets.
3. Posttranslational modifications: ADP-ribosylation, glycosylation, lipid anchors
The function of proteins can be regulated via the attachment of chemical moieties. These enzyme-catalyzed modifications, coined posttranslational modifications (PTMs), include phosphorylation, glycosylation, ADP-ribosylation, and the attachment of lipid anchors.
ADP-ribosylation is a reversible PTM, in which ADP-ribosyltransferases (ARTs) transfer the ADP-ribose moiety from NAD onto a specific amino acid side chain in a target protein and ADP-ribosylhydrolases (ARHs) remove the ADP-ribose group (Fig. 5). We have determined the 3D structures of a prototype ART and a prototype ARH. The 3D structure of rat ARTC2 resembles a pacman with a wide-open mouth crunching on the substrate NAD. The 3D structure of human ARH3 resembles a pumpkin in which four alpha helices coordinate two magnesium ions at the bottom of the active site crevice.
ADP-ribosylation is used by pathogenic toxins such as Diphtheria, Pertusssis and Clostridial toxins to modulate host protein functions. Toxin-related ARTs are expressed by cells of the immune system. ADP-ribosylation of membrane proteins can be monitored using labeled analogues of NAD. Using 32P-NAD, ADP-ribosylation results in radiolabeling of the target protein. Using etheno-NAD, ADP-ribosylation of target proteins can be detected with a monoclonal antibody directed against etheno-adenosine. This 1G4 antibody can be used to sort cells on the basis of cell surface ART-activity and to purify etheno-ADP-ribosylated proteins.
ARTC2 itself is posttranslationally modified, e.g. by attachment of a GPI lipid anchor (Fig. 1 above). The GPI-anchor targets ARTC2 to lipid rafts, specialized regions of the cell membrane that play an important role in signal transduction, e.g. during activation of T cells by antigen presenting cells. The association of ARTC2 with lipid rafts focuses ARTC2 onto specific target proteins and thereby may regulate the signaling function of raft-associated proteins. ADP-ribosylation of the P2X7 ion channel on arginine residue 125 activates P2X7 to form a non-selective ion-channel, permitting calcium ions to enter the cell and potassium ions to exit the cell. This induces dramatic alterations of the cell membrane, including the externalization of PS, shedding of L-selectin, and formation of membrane blebs.
Eggers M, Rühl F, Haag F, Koch-Nolte F. (2021) Nanobodies as probes to investigate purinergic signaling. Biochem Pharmacol. 87:114394. review PMID: 33388283.
Linden J, Koch-Nolte F, Dahl G. (2019) Purine Release, Metabolism, and Signaling in the Inflammatory Response. Annu Rev Immunol. 37:325-347. review. PMID: 30676821
Stortelers C, Pinto-Espinoza C, Van Hoorick D, Koch-Nolte F. (2018) Modulating ion channel function with antibodies and nanobodies. Curr Opin Immunol. 52:18-26. Review PMID: 29579624
Bannas P, Hambach J, Koch-Nolte F. (2017) Nanobodies and Nanobody-Based Human Heavy Chain Antibodies As Antitumor Therapeutics. Front Immunol. 8:1603. Review PMID: 29213270
Wesolowski J, Alzogaray V, Reyelt J, Unger M, Juarez K, Urrutia M, Cauerhff A, Danquah W, Rissiek B, Scheuplein F, Schwarz N, Adriouch S, Boyer O, Seman M, Licea A, Serreze DV, Goldbaum FA, Haag F, Koch-Nolte F. (2009) Single domain antibodies: promising experimental and therapeutic tools in infection and immunity. Med Microbiol Immunol. 198:157-74. Review PMID: 19529959
Koch-Nolte F, Kernstock S, Mueller-Dieckmann C, Weiss MS, Haag F. (2008) Mammalian ADP-ribosyltransferases and ADP-ribosylhydrolases. Front Biosci. 13:6716-29. PMID: 18508690
Hambach J, Stähler T, Eden T, Wendt D, Tode N, Haag F, Tolosa E, Altfeld M, Fathi A, Dahlke C, Addo MM, Menzel S, Koch-Nolte F. (2021) A simple, sensitive, and low-cost FACS assay for detecting antibodies against the native SARS-CoV-2 spike protein. Immun Inflamm Dis. 9:905-917. PMID: 33979020.
Schriewer L, Schütze K, Petry K, Hambach J, Fumey W, Koenigsdorf J, Baum N, Menzel S, Rissiek B, Riecken K, Fehse B, Röckendorf JL, Schmid J, Albrecht B, Pinnschmidt H, Ayuk F, Kröger N, Binder M, Schuch G, Hansen T, Haag F, Adam G, Koch-Nolte F, Bannas P. (2020) Nanobody-based CD38-specific heavy chain antibodies induce killing of multiple myeloma and other hematological malignancies. Theranostics. 10:2645-2658. PMID: 32194826
Eichhoff AM, Börner K, Albrecht B, Schäfer W, Baum N, Haag F, Körbelin J, Trepel M, Braren I, Grimm D, Adriouch A, Koch-Nolte F. (2019) Nanobody-enhanced targeting of AAV gene therapy vectors Mol Ther Methods Clin Dev. 15:211-220. PMID: 31687421
Eden T, Menzel S, Wesolowski J, Bergmann P, Nissen M, Dubberke G, Seyfried F, Albrecht B, Haag F, Koch-Nolte F. (2018) A cDNA Immunization Strategy to Generate Nanobodies against Membrane Proteins in Native Conformation. Front Immunol. 8:1989. PMID: 29410663
Danquah W, Meyer-Schwesinger C, Rissiek B, Pinto C, Serracant-Prat A, Amadi M, Iacenda D, Knop JH, Hammel A, Bergmann P, Schwarz N, Assunção J, Rotthier W, Haag F, Tolosa E, Bannas P, Boué-Grabot E, Magnus T, Laeremans T, Stortelers C, Koch-Nolte F. (2016) Nanobodies that block gating of the P2X7 ion channel ameliorate inflammation. Sci Transl Med. 8:366ra162. PMID: 27881823
Adriouch S, Bannas P, Schwarz N, Fliegert R, Guse AH, Seman M, Haag F, Koch-Nolte F. (2008) ADP-ribosylation at R125 gates the P2X7 ion channel by presenting a covalent ligand to its nucleotide binding site. FASEB J. 22:861-9. PMID: 17928361
Koch-Nolte F, Reyelt R, Schössow B, Schwarz N, Scheuplein F, Rothenburg S, Haag F, Alzogaray V, Cauerhff A, Goldbaum F. (2007) Single domain antibodies from llama effectively and specifically block T cell ecto-ADP-ribosyltransferase ART2.2 in vivo. FASEB J. 21:3490-8. PMID: 17575259
Mueller-Dieckmann C, Kernstock S, Lisurek M, von Kries JP, Haag F, Weiss MS, Koch-Nolte F. (2006) The structure of human ADP-ribosylhydrolase 3 (ARH3) provides insights into the reversibility of protein ADP-ribosylation. Proc Natl Acad Sci U S A. 103:15026-31. PMID: 17015823
Otto H, Reche PA, Bazan F, Dittmar K, Haag F, Koch-Nolte F. (2005) In silico characterization of the family of PARP-like poly(ADP-ribosyl)transferases (pARTs). BMC Genomics. 6:139. PMID: 16202152
Bannas P, Adriouch S, Kahl S, Braasch F, Haag F, Koch-Nolte F. (2005) Activity and specifity of toxin-related mouse T cell ecto-ADP-ribosyltransferase ART2.2 depends on it association with lipid rafts. Blood. 105:3663-70. PMID: 15657180
Seman M, Adriouch S, Scheuplein F, Krebs C, Freese D, Glowacki G, Deterre P, Haag F, Koch-Nolte F. (2003) NAD-induced T cell death: ADP-ribosylation of cell surface proteins by ART2 activates the cytolytic P2X7 purinoceptor. Immunity. 19:571-82. PMID: 14563321
Otto H, Tezcan-Merdol D, Girisch R, Haag F, Rhen M, Koch-Nolte F. (2000) The spvB gene-product of the Salmonella enterica virulence plasmid is a mono(ADP-ribosyl)transferase. Mol Microbiol. 37:1106-15. PMID: 10972829
Koch-Nolte F, Petersen D, Balasubramanian S, Haag F, Kahlke D, Willer T, Kastelein R, Bazan F, Thiele HG. (1996) Mouse T cell membrane proteins Rt6-1 and Rt6-2 are arginine/protein mono(ADPribosyl)transferases and share secondary structure motifs with ADP-ribosylating bacterial toxins. J Biol Chem. 271:7686-93. PMID: 8631807
Koch F, Haag F, Kashan A, Thiele HG. (1990) Primary structure of rat RT6.2, a nonglycosylated phosphatidylinositol-linked surface marker of postthymic T cells. Proc Natl Acad Sci U S A. 87:964-7. PMID: 2300588
Koch F, Thiele HG, Low MG. (1986) Release of the rat T cell alloantigen RT-6.2 from cell membranes by phosphatidylinositol-specific phospholipase C. J Exp Med. 164:1338-43. PMID: 3489808
Infection Immunology (Prof. Hans-Willi Mittrücker)
Prof. Dr. Hans-Willi Mittrücker, Dr. Friederike Raczkowski, Peter Bradtke, Stefanie Fertig, Katrin Heider, Niels Lory, Jonas Rösch, Joanna Schmid, Leonie Voss, Dennis Yüzen
T cells play a central role in immune responses against pathogens, yet these cells are also crucial for the formation of autoimmune diseases. We use different mouse models for bacterial infection and for autoimmune kidney diseases to characterise the function and regulation of T cells. Main focuses of research are:
● The function and regulation of CD4 and CD8 T cells during infection with intracellular bacteria. For these studies, we use infection models for Listeria monocytogenes, Staphylococcus aureus and Salmonella typhimurium.
● The role of the transcription factor Interferon Regulatory Factor 4 (IRF4) in activation and differentiation of T cells.
● The regulation of immune responses by Interleukin-6, in particular, the impact of different Interleukun-6 signalling pathways on activation and differentiation of immune cells.
● Autoimmune kidney diseases and the role of T cells in formation and regulation of immune response against components of the glomerulus. Studies are conducted in mouse models for anti-glomerular basement nephritis.
● The role of adenine nucleotides, ectoenzymes and purinegic receptors for the regulation of T cells.
DFG:SFB 1328 “Adenine Nucleotides in Immunity and Inflammation” (A03: “Adenine nucleotide-modulated T cell differentiation and effector Functions”, Prof. H.-W. Mittrücker, Prof. Samuel Huber)
DFG: SFB 1192 „Immune-mediated glomerular diseases” (A4: “Autoreactive T cell responses in glomerulonephritis”)
DFG: KFO 296 „Feto-maternal immune cross talk: Consequences for maternal and offspring’s health” (P2 "Identification of viral determinants and maternal immune responses underlying influenza A virus disease severity during pregnancy“, Prof. G. Gabriel, Prof. H.-W. Mittrücker, Prof. P. Arck)
Harberts A, Schmidt C, Schmid J, Reimers D, Koch-Nolte F, Mittrücker H-W*, Raczkowski F*. Interferon Regulatory Factor 4 controls effector functions of CD8+ memory T cells. (2021) Proc Natl Acad Sci USA. 118:e2014553118. (*shared last authorship)
Krebs CF, Reimers D, Zhao Y, Paust HJ, Bartsch P, Nuñez S, Rosemblatt MV, Hellmig M, Kilian C, Borchers A, Enk LUB, Zinke M, Becker M, Schmid J, Klinge S, Wong MN, Puelles VG, Schmidt C, Bertram T, Stumpf N, Hoxha E, Meyer-Schwesinger C, Lindenmeyer MT, Cohen CD, Rink M, Kurts C, Franzenburg S, Koch-Nolte F, Turner JE, Riedel JH, Huber S, Gagliani N, Huber TB, Wiech T, Rohde H, Bono MR, Bonn S, Panzer U*, Mittrücker H-W*. (2020) Pathogen-induced tissue-resident memory TH17 (TRM17) cells amplify autoimmune kidney disease. Sci Immunol. 5:eaba4163. (*shared last authorship)
Klinge S, Yan K, Reimers D, Brede KM, Schmid J, Paust HJ, Krebs CF, Panzer U, Hopfer H, Mittrücker H-W. (2019) Role of regulatory T cells in experimental autoimmune glomerulonephritis. Am J Physiol Renal Physiol. 316:F572-F581.
Mahnke J, Schumacher V, Ahrens S, Käding N, Feldhoff LM, Huber M, Rupp J, Raczkowski F, Mittrücker H-W. (2016) Interferon Regulatory Factor 4 controls TH1 cell effector function and metabolism. Sci Rep. 6:35521.
Hünemörder S, Treder J, Ahrens S, Schumacher V, Paust H-J, Menter T, Matthys P, Kamradt T, Meyer-Schwesinger C, Panzer U, Hopfer H, Mittrücker H-W. (2015) TH1 and TH17 cells promote crescent formation in experimental autoimmune glomerulonephritis. J Pathol. 237:62-71.
Raczkowski F, Ritter J, Heesch K, Schumacher V, Höcker L, Raifer H, Klein M, Bopp T, Harb H, Kesper DA, Pfefferle PI, Grusdat M, Lang PA, Mittrücker H-W*, Huber M*. (2013) The transcription factor Interferon Regulatory Factor 4 is required for the generation of protective effector CD8+ T cells. Proc Natl Acad Sci USA. 110:15019-15024. (*shared last authorship)
Mittrücker H-W, Steinhoff U, Köhler A, Krause M, Lazar D, Mex P, Miekley D, Kaufmann SHE. (2007) Poor correlation between BCG vaccination-induced T cell responses and protection against tuberculosis. Proc Natl Acad Sci USA. 104:12434-12439.
Kursar M, Höpken UE, Koch M, Köhler A, Lipp M, Kaufmann SHE, Mittrücker H-W. (2005) Differential requirements for the chemokine receptor CCR7 in T-cell activation during Listeria monocytogenes infection. J Exp Med. 201:1447-1457.
Kursar M, Bonhagen K, Fensterle J, Köhler A, Hurwitz R, Kamradt T, Kaufmann SHE, Mittrücker H-W. (2002) Regulatory CD4+ CD25+ T cells restrict memory CD8+ T cell responses. J Exp Med. 196:1585-1592.
Mittrücker H-W, Matsuyama T, Grossman A, Kündig TM, Potter J, Shahinian A, Wakeham A, Patterson B, Ohashi PS, Mak TW. (1997) Requirement for the transcription factor LSIRF/IRF4 for mature B and T lymphocyte function. Science. 275:540-543.
University Medical Center Hamburg-Eppendorf (UKE)
Marcus Altfeld, Friedrich Koch-Nolte, Friedrich Haag, Eva Tolosa Institute for Immunology Ulf Panzer, Christian Krebs, Oliver Steinmetz, Nicola Tomas, Elion Hoxha, Rolf Stahl III. Medical Clinic and Polyclinic Samuel Huber, Nicola Gagliani, Peter Hübener I. Medical Clinic and Polyclinic Petra Arck Experimental feto-maternal medicine Gisa Tiegs Institute of Experimental Immunology and Hepatology Andreas Guse, Björn-Philipp Dierks Institute for Biochemistry and Molecular Cell Biology Tim Magnus, Björn Rissiek Department of Neurology Cathy Meyer-Schwesinger Institute of Cellular and Integrative Physiology Manuel Friese Institute of Neuroimmunology and Multiple Sclerosis
Helmut Hopfer Universitätsspital Basel Minka Breloer, Hanna Lotter Bernhard Nocht Institute for Tropical Medicine, Hamburg Magdalena Huber, Ulrich Steinhoff, Michael Lohoff Philipps University of Marburg Dirk Schmidt-Arras, Stefan Rose-John Christian-Albrechts University of Kiel Santina Bruzzone University of Genova, Italy Gülsah Gabriel Leibniz Institute for Experimental Virology, Hamburg
Immune Regulation (Prof. Eva Tolosa)
Prof. Eva Tolosa, Elena Billeb, Annika Boxnick, Julian Drese, Dr. Anna Gieras, Laura Glau, Romy Hackbusch, Zoe-Isabella Junginger, Manuela Kolster, Jonathan May, Du-Hanh Nguyen, Friederike Thiele, Dr. Kati Tillack, Dr. Riekje Winzer
Adenine nucleotides and immune regulation
Expression, function and regulation of purinergic receptors and adenine nucleotide-metabolizing ectoenzymes.
Adenine nucleotides play a key role in the modulation of the immune response. In our research group, we investigate the release of adenine nucleotides during inflammatory responses, the activation of purinergic receptors, and the degradation of adenine nucleotides by ectoenzymes.
The concentration of extracellular adenosine triphosphate (ATP) is low under physiological conditions, but increases significantly during inflammatory responses or cellular stress. Higher concentrations of ATP are also present in some tissues, such as the intestine. Extracellular ATP serves as a ligand for P2 receptors, and stimulation of P2X7 receptors on immune cells promotes the release of pro-inflammatory cytokines. The coordinated action of the ectoenzymes CD39 and CD73 converts ATP into adenosine (ADO), providing a dual function, which is the elimination of the danger signal ATP and the generation of the anti-inflammatory mediator ADO. This illustrates a mechanism to control the outcome of an immune response.
In some diseases, such as tumor diseases, it is therapeutically beneficial to enhance the immune response, whereas other disease conditions, such as autoimmune diseases, require immunosuppressive therapy. Modulating the ATP-ADO-axis by regulation of ectoenzymes and purinergic receptors thus represents an interesting research target for the first step in developing immunomodulatory therapies. Using inhibitors and knock-out systems, we want to modulate the cell-specific influence of these ectoenzymes and receptors and thus characterize potential targets for the regulation of immune responses.
Current projects in the lab related to adenine nucleotide signaling are:
Dissecting the role of ATP and adenosine receptor signaling forimmune adaptation of intestinal Th17 cells, PI: Prof. Eva Tolosa, Prof. Nicola Gagliani; CRC 1328 subproject A14.
In inflamed tissue, Th1 and Th17 cells mediate tissue-damaging processes by secreting pro-inflammatory cytokines. Some of these effector cells possess a certain degree of plasticity and can adopt an anti-inflammatory phenotype (a process referred to as immune adaptation), contributing to the resolution of inflammation. Our hypothesis is that the hydrolysis of extracellular ATP to ADO by Th1 and Th17 cells, and ADO receptor-mediated signaling pathways promote their conversion into regulatory cells. To decipher the role of ectonucleotidases CD39 and CD73 in controlling the availability of ADO, and the role of ADO receptor signaling in immune adaptation, we will perform experiments in vitro using human Th1 and Th17 cells, and, in collaboration with Prof. Nicola Gagliani, use cytokine reporter mice in the context of intestinal inflammation.
Here you can find more information about the CRC 1328
Generation of adenosine and its role in post-stroke immune suppression, PIs: PI: Prof. Eva Tolosa (Immunology, UKE Hamburg), Prof. Götz Thomalla (Neurology, UKE Hamburg), and Profs. Luisa Klotz and Heinz Wiendl (Neurology, Uni. Muenster); RU 2879 subproject C1
Ischemic stroke induces a rapid immunologic response characterized by the invasion of leukocytes from the periphery into the ischemically damaged region. At the same time, clinically relevant immunosuppression can be observed in stroke patients. The aim of this project is to elucidate the dynamics of stroke-related changes in the immune signature and functional properties of peripheral blood immune cells, with a focus on purinergic signaling as potential mechanism underlying stroke-induced immune suppression. Changes in the human immunome will be evaluated in relation to stroke subtype, characteristics of ischemic stroke lesions and disease outcome, with the aim of an early identification and specific treatment of risk-patients.
Here you can find more information about the RU 2879
Thymus function in health and disease
The thymus, a primary lymphoid organ, plays a crucial role in T cell development. Tightly controlled selection processes ensure the establishment of a functional but self-tolerant immune response against pathogens or tumors, without leading to autoimmunity. Around birth, thymic function is at its peak and therefore, events affecting the thymus during this sensitive period of time may have serious consequences for the child’s immunity.
In our lab, we explore a range of early life factors that lead to a decline or even loss of thymic function. Causes can be various and range from medical interventions (e.g. prenatal steroids, early life thymectomy) to inborn disorders (e.g. Down syndrome). The consequences may persist well beyond the first months of life and lead to an impaired immunity (e.g. development of chronic or autoimmune diseases).
Mechanisms of prenatal glucocorticoid-induced changes on the offspring's immune system PI: Prof. Eva Tolosa, CRU 296 subproject 7
Over the past 30 years, prenatal corticosteroid treatment (betamethasone) has been administered to thousands of women at risk of premature birth to accelerate fetal lung maturation in the preterm neonates. A remarkably higher survival-rate of premature and infants with very low birth-weight highlights the efficiency of this treatment. However, long term effects have not been analyzed in detail, especially possible effects on the immune system, considering that steroids are known to induce cell death of developing T cells.
Consequences of congenital heart disease and early thymectomy on immune development PI: Dr. Anna Gieras, supported by the German Heart Foundation
Early life medical interventions can be detrimental to thymic development and need to be cautiously evaluated to prevent unwanted adverse effects. We are investigating the impact of early life thymectomy on thymic function and development of the adaptive immune system.
Mechanistic insights into thymic function and immune dysfunction in Down Syndrome PI: Dr. Anna Gieras, supported by the Werner-Otto Stiftung
We study the impact of congenital diseases on poor thymic function with a major focus on patients with Down syndrome. These patients exhibit an altered peripheral immune system but the nature of the defect causing changes in the peripheral T cell pool are not yet known and a detailed analysis of thymic function is missing. Our preliminary data show a defective development of immature thymocytes (ex vivo and in vitro) and we are currently investigating underlying molecular mechanisms.
In vitro studies of thymic function in humans are challenging, mainly due to limitations in the availability of thymic tissue. In order to mimic the thymic environment in vitro we have established 3D human thymic organoids to assess and manipulate the development of T lymphocytes from patient-derived progenitor cells.
High resolution multiparameter flow cytometry for profiling the immune system in health and disease
Profiling the immune system is to get a snapshot of an individual’s immune health, and allows the identification of immune traits associated to a disease condition, genetic defect, ageing, etc. While immune cells in diseased organs can show substantial differences compared to their physiological state, changes in peripheral blood are often very subtle. Therefore, deep immune phenotyping including assessment of function and project-specific bioinformatics analysis are necessary to detect disease- or group-associated traits ( RU 2879, C1 ). In our lab we have developed panels that are specific to address the immune status in relation to age, with specific panels developed for cord blood analysis ( CRU 296, project S1 ); response to vaccination; for comparison of peripheral blood to organ- and tumor-infiltrating cells, for assessment of disease-related immune traits, etc.. We use a flow bioinformatics pipeline for quick evaluation of changes in the immune status of individuals, and for unbiased analysis of raw data for the quantification of differentially abundant clusters in pre-defined groups of patients or cellular compartments.
DFG: CRC 1328 Adenine nucleotides in immunity and inflammation
DFG: RU 2879 ImmunoStroke
DFG: CRU 296 Feto-maternal immune cross talk: Consequences for maternal and offspring's
Winzer R, Serracant-Prat A, Brock V, Pinto-Espinoza C, Rissiek B, Amadi M, Eich N, Rissiek A, Schneider E, Magnus T, Guse A, Diercks B, Koch-Nolte F, Tolosa E (2022) P2X7 is expressed on human innate-like T lymphocytes and mediates susceptibility to ATP-induced cell death EEUR J IMMUNOL. 2022;52(11):1805-1818.
Schneider E#, Winzer R#, Rissiek A, .., Magnus T, Fliegert R, Müller CE, Gagliani N, Tolosa E (2021) CD73-mediated adenosine production by CD8 T cell-derived extracellular vesicles constitutes an intrinsic mechanism of immune suppression in humans. Nat Commun. 8;12:5911.
Poch T, Krause J, Casar C, Liwinski T, Glau L, …Huber S, Tolosa E#, Gagliani N#, Schramm C# (2021) Single-cell atlas of hepatic T cells reveals expansion of liver-resident naive-like CD4+ T cells in primary sclerosing cholangitis. J Hepatol. S0168-8278(21)00219-1 3.
Bremer SJ, Glau L, Gehbauer C, Boxnick A, Biermann D, Sachweh JS, Tolosa E, Gieras A. (2021) OMIP 073: Analysis of human thymocyte development with a 14-color flow cytometry panel. Cytometry A. 99(9):875-879.
Lessel D, Gehbauer C, Bramswig NC, Schluth-Bolard C, Venkataramanappa S, van Gassen KLI,… Agrawal PB, Britsch S, Tolosa E#, Kubisch C# (2018) BCL11B mutations in patients affected by a neurodevelopmental disorder with reduced type 2 innate lymphoid cells. Brain; 141(8):2299-311
Mohme M, Schliffke S, Maire CL, Runger A, Glau L, … Westphal M, Binder M#, Tolosa E#, Lamszus K# (2018) Immunophenotyping of Newly Diagnosed and Recurrent Glioblastoma Defines Distinct Immune Exhaustion Profiles in Peripheral and Tumor-infiltrating Lymphocytes. Clin Cancer Res; 24(17):4187-200
Gieras A, Gehbauer C, Perna-Barrull D, Engler JB, Diepenbruck I, Glau L, Joosse SA, Kersten N, Klinge S, Mittrücker HW, Friese MA, Vives-Pi M, Tolosa E. (2017) Prenatal administration of betamethasone causes changes in the T cell receptor repertoire influencing development of autoimmunity. Frontiers in Immunology. 8:1505.
Thom V, Schmid S, Gelderblom M, Hackbusch R, Kolster M, .., Pless O, Bernreuther C, Glatzel M, Wegscheider K, Gerloff C, Magnus T#, Tolosa E# (2016) IL-17 production by CSF lymphocytes as a biomarker for cerebral vasculitis. Neurol Neuroimmunol Neuroinflamm; 3(2):e214.
Rissiek A, Baumann I, Cuapio A, Mautner A, Kolster M, Arck PC, Dodge-Khatami A, Mittrücker HW, Koch-Nolte F, Haag F, Tolosa E (2015) The expression of CD39 on regulatory T cells is genetically driven and further upregulated at sites of inflammation. J Autoimmun; 58:12-20.
Adamopoulou E, Tenzer S, Hillen N, Klug P, Rota IA, Tietz S, Gebhardt M, Stevanovic S, Schild H, Tolosa E, Melms A, Stoeckle C (2013) Exploring the MHC-peptide matrix of central tolerance in the human thymus. Nat Commun. 4:2039. doi:10.1038/ncomms3039
Stoeckle C, Quecke P, Ruckrich T, Burster T, Reich M, Weber E, Kalbacher H, Driessen C, Melms A, Tolosa E (2012) Cathepsin S dominates autoantigen processing in human thymic dendritic cells. J Autoimmun. 38 (4):332-343.
Brucklacher-Waldert V, Stuerner K, Kolster M, Wolthausen J, Tolosa E (2009) Phenotypical and functional characterization of T helper 17 cells in multiple sclerosis. Brain. 132:3329-3341.
Feger U*, Tolosa E#, Huang Y-H, Waschbisch A, Biedermann T, Melms A, Wiendl H (2007) HLA-G expression defines a novel regulatory T-cell subset present in human peripheral blood and sites of inflammation. Blood. 110 (2):568-577.
Tolosa E, Li W, Yasuda Y, Wienhold W, Denzin LK, Lautwein A, Driessen C, Schnorrer P, Weber E, Stevanovic S, Kurek R, Melms A, Bromme D (2003) Cathepsin V is involved in the degradation of invariant chain in human thymus and is overexpressed in myasthenia gravis. Journal of Clinical Investigation. 112 (4):517-526.
Tolosa E, King LB, Ashwell JD (1998) Thymocyte Glucocorticoid Resistance Alters Positive Selection and Inhibits Autoimmunity and Lymphoproliferative Disease in MRL-lpr/lprMice. Immunity. 8 (1):67-76. 15.