Z02: SFB 1328

Z02 Project movie

Z02: Summary

Nanobodies - single domain antibodies derived from llama heavy chain antibodies - display a propensity to bind functional epitopes not accessible to conventional antibodies. Such functional nanobodies have emerged as highly specific and potent antagonists and agonists of enzymes, ion channels and receptors. Nanobodies can easily be labeled by fluorochromes at specific sites and can readily be fused to other nanobodies, the Fc domain of a conventional antibody, an ATP-sensor and other proteins. Nanobodies allow high resolution microscopy, and the targeting of specific cells or subcellular structures. Nanobodies are excellent chaperones for the crystallization of membrane proteins. The goal of this Z-project is to provide the PIs of the CRC with 1) monoclonal antibodies and functional nanobodies against the key players of AN signaling, and 2) the nanobody technology platform for targeting AN sensors and fluorescent probes to specific subcelllular structures.

PD Dr. Björn Rissiek

Selected publications

Jaeckstein MY, Fischer AW, Rissiek B, Staehler T, Heine M, Behrens J, Mann O, Pfeifer A, Magnus T, Schlein C, Worthmann A, Scheja L, Koch-Nolte F, Heeren J. Purinergic adipocyte-macrophage crosstalk promotes degeneration of thermogenic brown adipose tissue. EMBO Rep. 2025 Dec;26(24):6460-6493.
Lorenz H, Menzel S, Roshchyna N, Albrecht B, Gebhardt AJ, Schneider E, Haag F, Rissiek B, Oheim R, Koch-Nolte F, Winzer R, Tolosa E. ENPP1/CD203a-targeting heavy-chain antibody reveals cell-specific expression on human immune cells. Cell Mol Life Sci. 2024 Dec 18;82(1):6.
Eden T, Schaffrath AZ, Wesolowski J, Stähler T, Tode N, Richter N, Schäfer W, Hambach J, Hermans-Borgmeyer I, Woens J, Le Gall CM, Wendler S, Linke-Winnebeck C, Stobbe M, Budnicki I, Wanney A, Heitz Y, Schimmelpfennig L, Schweitzer L, Zimmer D, Stahl E, Seyfried F, Gebhardt AJ, Dieckow L, Riecken K, Fehse B, Bannas P, Magnus T, Verdoes M, Figdor CG, Hartlepp KF, Schleer H, Füner J, Tomas NM, Haag F, Rissiek B, Mann AM, Menzel S, Koch-Nolte F. Generation of nanobodies from transgenic 'LamaMice' lacking an endogenous immunoglobulin repertoire. Nat Commun. 2024 Jun 3;15(1):4728.
Sierra-Marquez J, Schaller L, Sassenbach L, Ramírez-Fernández A, Alt P, Rissiek B, Zimmer B, Schredelseker J, Hector J, Stähler T, Koch-Nolte F, Staab- Weijnitz CA, Dietrich A, Kopp R, Nicke A. Different localization of P2X4 and P2X7 receptors in native mouse lung - lack of evidence for a direct P2X4-P2X7 receptor interaction. Front Immunol. 2024 Jun 17;15:1425938.
Menzel S, Duan Y, Hambach J, Albrecht B, Wendt-Cousin D, Winzer R, Tolosa E, Rissiek A, Guse AH, Haag F, Magnus T, Koch-Nolte F, Rissiek B. Generation and characterization of antagonistic anti-human CD39 nanobodies. Front Immunol. 2024 Mar 25;15:1328306.
Wilmes M, Pinto Espinoza C, Ludewig P, Stabernack J, Liesz A, Nicke A, Gelderblom M, Gerloff C, Falzoni S, Tolosa E, Di Virgilio F, Rissiek B, Plesnilla N, Koch-Nolte F, Magnus T. Blocking P2X7 by intracerebroventricular injection of P2X7-specific nanobodies reduces stroke lesions. J Neuroinflammation. 2022 Oct 12;19(1):256.
Brock VJ, Wolf IMA, Er-Lukowiak M, Lory N, Stähler T, Woelk LM, Mittrücker HW, Müller CE, Koch-Nolte F, Rissiek B, Werner R, Guse AH, Diercks BP. P2X4 and P2X7 are essential players in basal T cell activity and Ca²⁺ signaling milliseconds after T cell activation. Sci Adv. 2022 Feb 4;8(5):eabl9770.
Er-Lukowiak M, Duan Y, Rassendren F, Ulmann L, Nicke A, Ufer F, Friese MA, Koch-Nolte F, Magnus T, Rissiek B. A P2rx7 Passenger Mutation Affects the Vitality and Function of T cells in Congenic Mice. iScience. 2020 Nov 27;23(12):101870.
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. Nanobodies that block gating of the P2X7 ion channel ameliorate inflammation. Sci Transl Med. 2016 Nov 23;8(366):366ra162.
Rissiek B, Danquah W, Haag F, Koch-Nolte F. Technical Advance: a new cell preparation strategy that greatly improves the yield of vital and functional Tregs and NKT cells. J Leukoc Biol. 2014 Mar;95(3):543-9.

Prof. Dr. Friedrich Nolte (geb. Koch)

Selected publications

Jaeckstein MY, Fischer AW, Rissiek B, Staehler T, Heine M, Behrens J, Mann O, Pfeifer A, Magnus T, Schlein C, Worthmann A, Scheja L, Koch-Nolte F, Heeren J. Purinergic adipocyte-macrophage crosstalk promotes degeneration of thermogenic brown adipose tissue. EMBO Rep. 2025 Dec;26(24):6460-6493.
Lorenz H, Menzel S, Roshchyna N, Albrecht B, Gebhardt AJ, Schneider E, Haag F, Rissiek B, Oheim R, Koch-Nolte F, Winzer R, Tolosa E. ENPP1/CD203a-targeting heavy-chain antibody reveals cell-specific expression on human immune cells. Cell Mol Life Sci. 2024 Dec 18;82(1):6.
Eden T, Schaffrath AZ, Wesolowski J, Stähler T, Tode N, Richter N, Schäfer W, Hambach J, Hermans-Borgmeyer I, Woens J, Le Gall CM, Wendler S, Linke-Winnebeck C, Stobbe M, Budnicki I, Wanney A, Heitz Y, Schimmelpfennig L, Schweitzer L, Zimmer D, Stahl E, Seyfried F, Gebhardt AJ, Dieckow L, Riecken K, Fehse B, Bannas P, Magnus T, Verdoes M, Figdor CG, Hartlepp KF, Schleer H, Füner J, Tomas NM, Haag F, Rissiek B, Mann AM, Menzel S, Koch-Nolte F. Generation of nanobodies from transgenic 'LamaMice' lacking an endogenous immunoglobulin repertoire. Nat Commun. 2024 Jun 3;15(1):4728.
Eggers M, Rühl F, Haag F, Koch-Nolte F (2021) Nanobodies as probes to investigate purinergic signaling. Biochem Pharmacol 87:114394.
Hambach J, Riecken K, Cichutek S, Schütze K, Albrecht B, Petry K, Röckendorf JL, Baum N, Kröger N, Hansen T, Schuch G, Haag F, Adam G, Fehse B, Bannas P, Koch-Nolte F (2020) Targeting CD38-Expressing Multiple Myeloma and Burkitt Lymphoma Cells In Vitro with Nanobody-Based Chimeric Antigen Receptors (Nb-CARs). Cells 9: E321.
Linden J, Koch-Nolte F, Dahl G (2019) Purine Release, Metabolism, and Signaling in the Inflammatory Response. Annu Rev Immunol 37:325-347.
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.
Fumey W, Koenigsdorf J, Kunick V, Menzel S, Schütze K, Unger M, Schriewer L, Haag F, Adam G, Oberle A, Binder M, Fliegert R, Guse A, Zhao YJ, Cheung Lee H, Malavasi F, Goldbaum F, van Hegelsom R, Stortelers C, Bannas P, Koch-Nolte F (2017) Nanobodies effectively modulate the enzymatic activity of CD38 and allow specific imaging of CD38+ tumors in mouse models in vivo. Sci Rep 7:14289.
Danquah W, Meyer-Schwesinger C, Rissiek B, Pinto C, Serracant-Prat A, Amadi M, Iacenda D, Knop JH, Hammel A, Bergmann P, Schwarz N, Assunçao 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. Science Transl Med 8:366ra162
Stark R, Wesselink TH, Behr FM, Kragten NAM, Arens R, Koch-Nolte F, van Gisbergen KPJM, van Lier RAW (2013). TRM maintenance is regulated by tissue damage via P2RX7. Science Immunol 3: eaau1022.

Prof. Dr. Friedrich Haag ✝

Selected publications

Wildner NH, Ahmadi P, Schulte S, Brauneck F, Kohsar M, Lütgehetmann M, Beisel C, Addo MM, Haag F, Schulze Zur Wiesch J. B cell analysis in SARS-CoV-2 versus malaria: Increased frequencies of plasmablasts and atypical memory B cells in COVID-19. J Leukoc Biolc 2021; 109: 77-90.
Libera J, Wittner M, Kantowski M, Woost R, Eberhard JM, de Heer J, Reher D, Huber S, Haag F, Schulze Zur Wiesch J. Decreased Frequency of Intestinal CD39(+) γδ(+) T Cells With Tissue-Resident Memory Phenotype in Inflammatory Bowel Disease. Front Immunol 2020; 11: 567472.
Ahmadi P, Hartjen P, Kohsar M, Kummer S, Schmiedel S, Bockmann JH, Fathi A, Huber S, Haag F, Schulze Zur Wiesch J. Defining the CD39/CD73 Axis in SARS-CoV-2 Infection: The CD73(-) Phenotype Identifies Polyfunctional Cytotoxic Lymphocytes. Cells 2020; 9.
Johnsen B, Kaschubowski KE, Nader S, Schneider E, Nicola JA, Fliegert R, Wolf IMA, Guse AH, Nikolaev VO, Koch-Nolte F, Haag F. P2X7-mediated ATP secretion is accompanied by depletion of cytosolic ATP. Purinergic Signal 2019; 15: 155-166.
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. Nanobodies that block gating of the P2X7 ion channel ameliorate inflammation. Sci Transl Med 2016; 8: 366ra162.
Haag F, Buck F. Identification and analysis of ADP-ribosylated proteins. Curr Top Microbiol Immunol 2015; 384: 33-50.
Hubert S, Rissiek B, Klages K, Huehn J, Sparwasser T, Haag F, Koch-Nolte F, Boyer O, Seman M, Adriouch S. Extracellular NAD+ shapes the Foxp3+ regulatory T cell compartment through the ART2-P2X7 pathway. J Exp Med 2010; 207: 2561-2568.
Adriouch S, Dox C, Welge V, Seman M, Koch-Nolte F, Haag F. Cutting edge: a natural P451L mutation in the cytoplasmic domain impairs the function of the mouse P2X7 receptor. J Immunol 2002; 169: 4108-4112.
Seman M, Adriouch S, Scheuplein F, Krebs C, Freese D, Glowacki G, Deterre P, Haag F, and Koch-Nolte F. NAD-induced T cell death: ADP-ribosylation of cell surface proteins by ART2 activates the cytolytic P2X7 purinoceptor. Immunity 2003; 19: 571-582.
Adriouch S, Dox C, Welge V, Seman M, Koch-Nolte F, Haag F. Cutting edge: a natural P451L mutation in the cytoplasmic domain impairs the function of the mouse P2X7 receptor. J Immunol 2002; 169: 4108-4112.