Towards the use of antigen-specific γδ T lymphocytes for CMV virus control after allogeneic stem cell transplantation
TCR repertoire analysis
Cytomegalovirus (CMV) remains a significant cause of infectious morbidity and mortality in patients after hematopoietic stem cell transplantation. The goal of this project is to understand how specific immune cells, in particular the so far only incompletely explored γδ T cells, mount an anti-viral response against CMV. On the long run, this should lead the way towards γδ T cell based cellular therapy for control of CMV infection in immunocompromised individuals. To this end, we monitor the kinetics of CMV-reactive γδ T cells after transplantation in a prospective clinical study. To screen the CMV-reactive TCR repertoire of human γδ T cells, we have established a novel next-generation-sequencing approach. After identifiying specific γδ T cell clones we will evaluate their anti-viral function.i. Finally, we will investigate the antigen specificity of CMV- reactive γδ T cells and their potential in cellular therapy.
Scientific work programme
γδ T cells are unconventional lymphocytes, which display both innate and adaptive features. Since γδ T cells recognize their cognate antigen independent of MHC-restriction, they cannot cause graft-versus-host disease and therefore have a high potential for safe cellular therapy. This project addresses the highly diverse repertoire of the γδ T cells’ clonally rearranged T cell receptors (TCRs). We have developed a RNA/cDNA based next generation TCR sequencing technology. Briefly, γδ T-cells are FACS-sorted and the variable (V) and constant (C) regions of the human TRG and TRD genes are targeted by multiplex primers to amplify all CDR3 regions of the gamma and delta chains by PCR. Amplicons will be sequenced and analysed by bioinformatics.
This technology is used to characterize the kinetics of individual γδ TCR repertoires in leukemia patients before and after allogeneic hematopoietic stem cell transplantation (alloHSCT) at high resolution. Our recent findings show that γδ TCR repertoires are quickly reconstituted, highly complex, but are entirely changed after alloHSCT. We anticipate that the results are highly important to understand the role of unconventional T cells for infections after alloHSCT . Most importantly, we aim to further characterize the anti-viral activity and antigen specificity of individual CMV-reactive γδ T cell clones. This might be the basis to design virus-reactive γδ T cells for future immunotherapies in patients at a high risk for CMV reactivation after transplantation.
TCR repertoire visualization
TCR repertoire analysis
Lysing T cells to extract their TCR genes
Immo Prinz and Christian Könecke are talking about their research at CRC 900
Immo Prinz and Christian Könecke have been part of the CRC since the start of the second funding period in 2014. Their project B8 is concerned with the interactions between certain types of T cells and cytomegalovirus in patients after a stem cell transplantation.
Publications of the project B8
- Human γδ TCR Repertoires in Health and Disease. Fichtner AS, Ravens S, Prinz I. Cells 2020 Mar 26;9(4):E800.
- Revisiting the Interaction of γδ T-Cells and B-Cells. Rampoldi, F, Ullrich L, Prinz I. Cells. 2020 Mar; 9(3): 743.
- Focusing of the regulatory T-cell repertoire after allogeneic stem cell transplantation indicates protection from graft-versus-host disease. Odak I, Raha S, Schultze-Florey C, Tavil S, Ravens S, Ganser A, Förster R, Prinz I, Koenecke C. Letters to the editor. Haematologica December 2019 104: e577-e580
- Single-Cell Transcriptomics Identifies the Adaptation of Scart1+ Vγ6+ T Cells to Skin Residency as Activated Effector Cells. Tan L, Sandrock I, Odak I, Aizenbud Y, Wilharm A, Barros-Martins J, Tabib Y, Borchers A, Amado T, Gangoda L, Herold MJ, Schmidt-Supprian M, Kisielow J, Silva-Santos B, Koenecke C, Hovav AH, Krebs C, Prinz I, Ravens S. Cell Rep. 2019 Jun 18; 27(12):3657-3671.e4.
- Mutual interplay between IL-17-producing γδT cells and microbiota orchestrates oral mucosal homeostasis. Wilharm A, Tabib Y, Nassar M, Reinhardt A, Mizraji G, Sandrock I, Heyman O, Barros-Martins J, Aizenbud Y, Khalaileh A, Eli-Berchoer L, Elinav E, Wilensky A, Förster R, Bercovier H, Prinz I, Hovav AH. Proc Natl Acad Sci U S A. 2019 Feb 12;116(7):2652-2661.
- Genetic models reveal origin, persistence and non-redundant functions of IL-17–producing γδ T cells. Sandrock I, Reinhardt A, Ravens S, Binz C, Wilharm A, Martins J, Oberdörfer L, Tan L, Lienenklaus S, Zhang B, Naumann R, Zhuang Y, Krueger A, Förster R, Prinz I. J Exp Med. 2018 Nov 19. pii: jem.20181439.
- Human γδ T Cell Receptor Repertoires in Peripheral Blood Remain Stable Despite Clearance of Persistent Hepatitis C Virus Infection by Direct-Acting Antiviral Drug Therapy. Ravens S, Hengst J, Schlapphoff V, Deterding K, Dhingra A, Schultze-Florey C, Koenecke C, Cornberg M, Wedemeyer H, Prinz I. Front Immunol. 2018 Mar 16;9:510.
Characterization of high-avidity CMV specific T-cells with differential tetramer binding co-appearing after allogeneic SCT. Ogonek J, Verma K, Schultze-Florey S, Varanasi P, Luther S, Schweier P, Kühnau W, Göhring G, Dammann E, Stadler M, Ganser A, Koehl U, Koenecke C, Weissinger EM, Hambach L. J Immunol. 2017 Jul 15; 199(2):792-805.
Eomes expression reports the progressive differentiation of IFN-γ-producing Th1-like γδ T cells. Lino CNR, Barros-Martins J, Oberdörfer L, Walzer T, Prinz I. Eur J Immunol. 2017 Jun; 47(6):970-981.
Human gd T cells are quickly reconstituted after stem-cell transplantation and show adaptive clonal expansion in response to viral infection. Ravens S, Schultze-Florey C, Raha S, Sandrock I, Drenker M, Oberdörfer L, Reinhardt A, Ravens I, Beck M, Geffers R, von Kaisenberg C, Heuser M, Thol F, Ganser A, Förster R, Koenecke C, Prinz I. Nat Immunol. 2017 Apr; 18(4):393-401.
VH1 family immunoglobulin repertoire sequencing after allogeneic hematopoietic stem cell Transplantation. Sethi MK, Thol F, Stadler M, Heuser M, Ganser A, Koenecke C and Pabst O. PLoS One. 2017 Jan 17;12(1):e0168096.
Disruption of de novo fatty acid synthesis via acetyl-CoA carboxylase 1 inhibition prevents acute graft-versus-host disease. Raha S, Raud B, Oberdörfer L, Castro CN, Schreder A, Freitag J, Longerich T, Lochner M, Sparwasser T, Berod L, Koenecke C, Prinz I. Eur J Immunol. 2016 Sep;46(9):2233-8.
Differential Effects of Gut-Homing Molecules CC Chemokine Receptor 9 and Integrin-β7 during Acute Graft-versus-Host Disease of the Liver. Schreder A, Moschovakis GL, Halle S, Schlue J, Lee CW, Schippers A, David S, Bernhardt G, Ganser A, Pabst O, Förster R, Koenecke C. Biol Blood Marrow Transplant. 2015 Dec; 21(12):2069-78.
MicroRNA-181a/b-1 Is Not Required for Innate γδ NKT Effector Cell Development. Sandrock I, Ziętara N, Łyszkiewicz M, Oberdörfer L, Witzlau K, Krueger A, Prinz I. PLoS One. 2015 Dec 16;10(12):e0145010.
γδ T cells come to stay: Innate skin memory in the Aldara model. Prinz I, Sandrock I. Eur J Immunol. 2015 Nov; 45(11):2994-7.
Impact of the revised International Prognostic Scoring System, cytogenetics and monosomal karyotype on outcome after allogeneic stem cell transplantation for myelodysplastic syndromes and secondary acute myeloid leukemia evolving from myelodysplastic syndromes: a retrospective multicenter study of the European Society of Blood and Marrow Transplantation. Koenecke C, Göhring G, de Wreede LC, van Biezen A, Scheid C, Volin L, Maertens J, Finke J, Schaap N, Robin M, Passweg J, Cornelissen J, Beelen D, Heuser M, de Witte T, Kröger N; MDS subcommittee of the Chronic Malignancies Working Party of the EBMT. Haematologica. 2015 Mar; 100(3):400-8.
A clonotypic Vγ4Jγ1/Vδ5Dδ2Jδ1 innate γδ T-cell population restricted to the CCR6⁺CD27⁻ subset. Kashani E, Föhse L, Raha S, Sandrock I, Oberdörfer L, Koenecke C, Suerbaum S, Weiss S, Prinz I. Nat Commun. 2015 Mar 13;6:6477.