Immune Tolerance

The goal of this unit is the development of cell- and antibody-based therapeutic strategies to treat complications following hematopoietic stem cell transplantation. Novel concepts of immunological tolerance oriented towards immunologic and therapy associated complications (e.g. GVHD) are being tested in new, in-house developed models.

Analysis of new stem cell sources regarding their hematopoietic potential

Analysis of non-adherent bone marrow cells (NA-BMC) by electron microscopy.

Analysis of non-adherent bone marrow cells (NA-BMC) by electron microscopy.

In this project the emphasis lies on the investigation of new stem cell sources as therapy option for hematopoietic stem cell transplantations (HSCT). These are to be examined concerning their potential for the regeneration of hematopoiesis. The background is that fast hematopoiesis could decrease the risk of complications (e.g. infections, tumor relapse) in immunosuppressed and transplanted patients.

The research unit examined non-adherent bone marrow-derived stem cells (NA-BMC) and developed special competencies in cell culture models and immunological methods (e.g. CBA, ELISpot). By in vitro and in vivo experiments it could be shown that the mixed cell fraction of NA-BMCs possesses characteristics of hematopoietic and mesenchymal cells and supports hematopoiesis. Using humanized mice with a human CD4 and HLA-DR and a knock-out of the murine CD4 it can be exactly differentiated between donor and receptor cells. Thus, information on the therapeutic effectiveness of new cell stem cell sources can be determined.

The influence of tolerance-inducing anti-human CD4 antibodies on the graft-versus-tumor effect (GvT) following allogeneic hematopoietic stem cell trans­plantation

Reconstituted bone marrow of recipient mice treated with antibodies following allogenic stem cell transplantation.
© Photo Fraunhofer IZI

Reconstituted bone marrow of recipient mice treated with antibodies following allogenic stem cell transplantation.

The main complication following allogeneic hematopoietic stem cell transplantation is acute graft-versus-host-disease (aGvHD). Besides conventional immunosuppresive agents, OKT3-antibodies, interleukin-2 receptor antibodies or anti-thymocyte globulin to name just a few are also being used to treat this disease, though they are often associated with low-level long-term success and toxic side effects. In addition, these types of therapy suppress the entire immune system and with this also the immunologically significant anti-tumor effect of the immune cells (graft-versus-leukemia effect). This GvL effect must however be maintained in order to prevent the risk of the underlying disease (leukemia) returning in the patient.

Due to the need for innovative and less stressful forms of therapy, murine GvHD and leukemia transplantation models were developed within the unit. These models were able to be used, among other things, to establish human immune systems in mice. In due consideration of the GvL effect, these models enable the testing of active agents for the prevention and treatment of GvHD, which would be immediately conceivable for clinical application on human subjects. The focus is on the investigation of the influence of various anti-human CD4 antibodies on the GvHD with due regard to the GvL effect. So far, it could be demonstrated, for example, that the emergence of aGvHD following allogeneic hematopoietic stem cell transplantation could be prevented in the long term through the ex vivo incubation of a transplant with the anti-human CD4 antibody MAX.16H5. This involved the use of a mouse model which also displays features of the human immune system.

A GvHD-NOD/SCID mouse model and a leukemia mouse model will be further developed at a later stage in the project. Furthermore, anti-CD4 antibodies will be introduced in the treatment and prevention of GvHD as therapeutic agents until clinical application is possible. New findings from antibody therapy for the transplantation of solid organs and for other immunological disease patterns (e.g. autoimmune diseases) will also be deduced. Besides a new type of antibody therapy, insights into immunological processes of GvHD and the GvL effect are to be expected. These insights may be extremely valuable not just for hematopoietic stem cell transplantation, but also for the transplantation of solid organs and for application in other indications (e.g. autoimmune diseases).

Induction of immune tolerance after transplantation by anti-human CD4 antibody therapy

Immunohistology for human CD4 in the gut of humanized mice.
© Photo Fraunhofer IZI

Immunohistology for human CD4 in the gut of humanized mice.

Hematopoietic stem cell transplantation (HSCT) is often the only curative regenerative treatment for diseases of the hematopoietic system (e.g. leukemia). HSCT is often accompanied by life-threatening complications (bacterial, viral and fungal infections). One of the main complications is the graft-versus-host disease (GVHD) which affects up to 80 percent of the patients. Donor T cells, which are located in the graft, recognize the recipient tissues as foreign and destroy them.

The focus of the project is the development of new therapeutic strategies using human CD4 antibodies. These antibodies should induce immune tolerance after transplantation, in particular hematopoietic stem cell transplantation, in patients and ensure their survival. In the model system the application of human CD4 antibodies leads to a block of the human CD4 receptors on the T helper cells and a significant reduction of GVHD.

  • ACOMED Statistik
  • Affimed Therapeutics AG
  • Alcyomics Ltd.
  • Charité – Universitätsmedizin Berlin, Campus Benjamin Franklin (CBF)
  • Charité – Universitätsmedizin Berlin, Campus Charité Mitte (CCM)
  • Consiglio Nazionale delle Ricerche (CNR), Institute for Physical and Chemical Processes (IPCF)
  • Dr. med. Steffi Fricke, Specialist in Neurology / Psychiatry / Psychotherapeutic Medicine
  • Euroderm AG
  • GuangzhoGuangzhou Gendustry Inc.
  • Herzzentrum Leipzig, Klinik für Kardiologie
  • Julius-Maximilians University Würzburg, Institute for Virology and Immunobiology
  • Praxis PD Dr. Hoheisel Leipzig
  • Southern Medical University China, South Genomics Research Center
  • University of Leipzig, Faculty of Medicine, Institute for Pathology
  • University of Leipzig, Institute of Anatomy
  • University of Leipzig, Institute for Clinical Immunology and Transfusion Medicine (IKIT)
  • University of Leipzig, Institute for Microbiology and Infection Epidemiology
  • University of Leipzig, Institute for Virology
  • University of Leipzig, Medical Experimental Center of the Faculty of Medicine
  • University of Leipzig, Translational Center for Regenerative Medicine (TRM) Leipzig
  • University Hospital Leipzig, Clinic and Polyclinic for Gastroenterology and Rheumatology
  • University Hospital Leipzig, Clinic for Radiation Therapy and Radio Oncology
  • University Hospital Leipzig, Independant Department for Hematology, Medical Oncology and Hemostaseology
  • University Hospital Rostock, Clinic for Radiation Therapy
  • University Hospital Schleswig-Holstein, Clinic for Applied Cell Therapy

  • Oelkrug C, Sack U, Boldt A, Nascimento IC, Ulrich H, Fricke S. Antibody- and aptamer-strategies for GvHD prevention. Journal of Cellular and Molecular Medicine. 2015 Jan;19(1):11-20. DOI: dx.doi.org/10.1111/jcmm.12416.
  • Boldt A, Borte S, Fricke S, Kentouche K, Emmrich F, Borte M, Kahlenberg F, Sack U. Eight-color immunophenotyping of T-, B-, and NK-cell subpopulations for characterization of chronic immunodeficiencies. Cytometry B Clinical Cytometry. 2014 May;86(3):191-206.DOI: dx.doi.org/10.1002/cyto.b.21162.
  • Boldt A, Kahlenberg F, Fricke S, Rasche FM, Sack U. Flow cytometric phenotyping of lymphocytes in patients with systemic lupus erythematosus. Cytometry A. 2014 Jul;85(7):567-9.DOI: dx.doi.org/10.1002/cyto.a.22477.
  • Boldt A, Kentouche K, Fricke S, Borte S, Kahlenberg F, Sack U. Differences in FOXP3 and CD127 expression in Treg-like cells in patients with IPEX syndrome. Clinical Immunoloy. 2014 Jul;153(1):109-11. DOI: dx.doi.org/10.1016/j.clim.2014.04.001.
  • Fricke S, Hilger N, Fricke C, Schönfelder U, Behre G, Ruschpler P, Boldt A, Oelkrug C, Sack U, Emmrich F. Prevention of graft-versus-host-disease with preserved graft-versus-leukemia-effect by ex vivo and in vivo modulation of CD4(+) T-cells. Cellular and Molecular Life Science. 2014 Jun;71(11):2135-48.DOI: dx.doi.org/10.1007/s00018-013-1476-0.
  • Fricke S, Pfefferkorn C, Wolf D, Riemschneider S, Kohlschmidt J, Hilger N, Fueldner C, Knauer J, Sack U, Emmrich F, Lehmann J. Characterization of the murine myeloid precursor cell line MuMac-E8. PLoS One. 2014 Dec 29;9(12):e113743. DOI: dx.doi.org/10.1371/journal.pone.0113743.
  • Oelkrug C, Hilger N, Schönfelder U, Boltze J, Sack U, Fricke C, Hildebrandt G, Keller T, Emmrich F, Fricke S. Modelling hematological parameters after total body irradiation. International Journal of Radiation Biology. 2014 Jul;90(7):538-46. DOI: dx.doi.org/10.3109/09553002.2014.899443.
  • Oelkrug C, Lange CM, Wenzel E, Fricke S, Hartke M, Simasi J, Schubert A. Analysis of the tumoricidal and anti-cachectic potential of curcumin. Anticancer Research. 2014 Sep;34(9):4781-8.
  • Weis S, Schlaich TC, Dehghani F, Carvalho T, Sommerer I, Fricke S, Kahlenberg F, Mössner J, Hoffmeister A. p8 Deficiency causes siderosis in spleens and lymphocyte apoptosis in acute pancreatitis. Pancreas. 2014 Nov;43(8):1277-85. DOI: dx.doi.org/10.1097/MPA.0000000000000172.
  • Fricke S, Rothe K, Hilger N, Ackermann M, Oelkrug C, Fricke C, Schönfelder U, Niederwieser D, Emmrich F, Sack U. Allogeneic bone marrow grafts with high levels of CD4(+) CD25(+) FoxP3(+) T cells can lead to engraftment failure. Cytometry A. 81 (2012), 6, S. 476-88. doi: 10.1002/cyto.a.22061.
  • Fricke S, Fricke C, Oelkrug C, Blatz R, Schönfelder U, Niederwieser D, Hilger N, Ruhnke M, Rodloff AC. A real-time PCR for the detection and characterisation of Aspergillus species. Mycoses. 55 (2012), 5, S. 416-25. doi: 10.1111/j.1439-0507.2011.02161.x.
  • Grey D, Sack U, Scholz M, Knaack H, Fricke S, Oppel C, Luderer D, Fangmann J, Emmrich F, Kamprad M. Increased CD64 expression on polymorphonuclear neutrophils indicates infectious complications following solid organ transplantation. Cytometry A. 79 (2011), 6, S. 446 - 60. doi: 10.1002/cyto.a.21049.
  • Fricke S, Fricke C, Oelkrug C, Hilger N, Schönfelder U, Kamprad M, Lehmann J, Boltze J, Emmrich F, Sack U. Characterization of murine non-adherent bone marrow cells leading to recovery of endogenous hematopoiesis. Cell Mol Life Sci. 67 (2010), 23, S. 4095-4106.
  • Fricke S, Fricke C, Schimmelpfennig C, Oelkrug C, Schönfelder U, Blatz R, Zilch C, Faber S, Hilger N, Ruhnke M, Rodloff AC. A real-time PCR assay for the differentiation of Candida species. J Appl Microbiol. 109 (2010), 4, S 1150-1158.
  • Fricke S, Ackermann M, Stolzing A, Schimmelpfennig C, Hilger N, Jahns J, Hildebrandt G, Emmrich F, Ruschpler P, Pösel C, Kamprad M, Sack U. Allogeneic non-adherent bone marrow cells facilitate hematopoietic recovery but do not lead to allogeneic engraftment. PLoS One (2009), Jul 7, 4(7), e6157.
  • Fricke S, Fricke C, Schimmelpfennig C, Oelkrug C, Schönfelder U, Blatz R, Faber S, Zilch C, Hilger N, Ruhnke M, Rodloff A. New Real-time PCR assay for the differentiation of Candida species. Appl Microbiol (2009) (accepted JAM-2009-2135).
  • Ueberham E, Lindner R, Kamprad M, Hiemann R, Hilger N, Woithe B, Mahn D, Cross M, Sack U, Gebhardt R, Arendt T, Ueberham U. Oval cell proliferation in p16INK4a expressing mouse liver is triggered by chronic growth stimuli. J Cell Mol Med 12 (2008), 2, S. 622-638.
  • Hiemann R, Hilger N, Michel J, Nitschke J, Bohm A, Anderer U, Weigert M, Sack U. Automatic analysis of immunofluorescence patterns of HEp-2 cells. Ann NY Acad Sci 1109 (2007), 1, S. 358-371.
  • Hiemann R, Hilger N, Sack U, Weigert M. Objective quality evaluation of fluorescence images to optimize automatic image acquisition. Cytom Part A, 69 (2006), 3, S. 182-184.
  • Pohl S, Fricke S, Hildebrandt G, Uharek L, Braun JM, Emmrich F. Permanent induction of specific immunotolerance to allograft and prevention of graft-versushost-disease. Zeitschrift für Regenerative Medizin (2006), 1, S. 60.
  • Schneider A, Sack U, Rothe K, Bennek J. Peritoneal taurolidine lavage in children with localised peritonitis due to appendicitis. Pediatr Surg Int 21 (2005), 6, S. 445-448.