The interplay of EBV-induced γδ and αβ T cells during chronic EBV reactivation, PTLD, and EBV-directed cellular therapy in transplant recipients

Epstein Barr Virus (EBV) can lead to infectious mononucleosis (“Pfeiffer’s glandular fever”). After a first infection usually in childhood to adolescence, the virus nests in the B cells of the immune system for life and is controlled by the healthy immune system. In case of a weakened immune system, e.g. after transplantation and in patients who need to take immunosupressive drugs, EBV can multiply uncontrollably and can finally lead to the development of malignant tumors (lymphoproliferative disorders).

T cells of the immune system can control EBV and the anti-EBV immune response has been well described for αβ T cells. However, there is increasing evidence that also γδ T cells can control EBV virus control. In this project, we investigate the contribution of γδ T cells to the fight against EBV infection in healthy patients and how this interaction with αβ T cells changes under immunosuppression. One therapeutic option for transplant patients with EBV-associated problems is the transfer of purified EBV-specific T cells from healthy donors. These are usually almost exclusively αβ T cells, in this study we aim at characterizing the role of the patient’s own γδ T cells in the efficacy of the transferred EBV-specific T cells in the patient.

Scientific work programme

For EBV-associated lympho proliferation, we can infect B cells with EBV in the laboratory and then test the immune response in different constellations. In addition to the sheer number of EBV-specific αβ and γδ T cells, their appearance is examined using surface markers and their function in cytokine and cell lysis tests. Sequencing of the T cell receptors will show which specific T cell clones are stimulated and expanded by EBV. Insights gained in this system will be transferred to a cohort of kidney and liver transplant patients who have either successfully overcome EBV infection or chronically detect EBV in their blood among the necessary immunosuppressive drugs.

In a final step, we will follow the progression of αβ and γδ T cells in patients receiving EBV-specific T cell transfer. We will evaluate to what extent γδ T cells, which are not transferred with the preparation, contribute to the success of cell therapy. Through T-cell receptor sequencing, we want to investigate whether there are specific sequences for EBV and compare them to sequences in other viral diseases (e.g., CMV, TP B8, hepatitis) to identify possible common sequences.
In the long term, we expect to gain insights into spurious regulation in immunocompromised patients and to improve the efficacy of T cell preparations by supporting additional cell fractions.

Schematic representation of immune cells contributing to control EBV infection at different stages.

Britta Maecker-Kolhoff is talking about her research at CRC 900

Britta Maecker-Kolhoff joined the CRC in the third funding period in 2018. She is jointly leading the project B11 with Britta Eiz-Vesper and Sarina Ravens, which is concerned with the interactions between certain types of T cells during chronic reactivation of the Epstein-Barr Virus (EBV) and EBV-directed cellular therapy in transplant patients.

Sarina Ravens is talking about her research at CRC 900

Sarina Ravens joined the CRC in the third funding period in 2018. She is jointly leading the project B11 with Britta Eiz-Vesper and Britta Maecker-Kolhoff, which is concerned with the interactions between certain types of T cells during chronic reactivation of the Epstein-Barr Virus (EBV) and EBV-directed cellular therapy in transplant patients.

Britta Eiz-Vesper is talking about her research at CRC 900

Britta Eiz-Vesper joined the CRC in the third funding period in 2018. She is jointly leading the project B11 with Britta Maecker-Kolhoff and Sarina Ravens, which is concerned with the interactions between certain types of T cells during chronic reactivation of the Epstein-Barr Virus (EBV) and EBV-directed cellular therapy in transplant patients.

Publications of the project B11

Allogeneic BK Virus-Specific T-Cell Treatment in 2 Patients With Progressive Multifocal Leukoencephalopathy. Hopfner F, Möhn N, Eiz-Vesper B, Maecker-Kolhoff B, Gottlieb J, Blasczyk R, Mahmoudi N, Pars K, Adams O, Stangel M, Wattjes MP, Höglinger G, Skripuletz T. Neurol Neuroimmunol Neuroinflamm 2021 8; 4
Antiviral T-Cell Frequencies in a Healthy Population: Reference Values for Evaluating Antiviral Immune Cell Profiles in Immunocompromised Patients. Schulze Lammers, F.C., Bonifacius, A., Tischer-Zimmermann, S., Goudeva, L., Martens, J., Lepenies, B., von Karpowitz, M., Einecke, G., Beutel, G., Skripuletz, T., Blasczyk, R., Beier, R., Maecker-Kolhoff, B., and Eiz-Vesper, B. J Clin Immunol 2022 42, 546-558. DOI: 10.1007/s10875-021-01205-1
A fetal wave of human type 3 effector gd cells with restricted TCR diversity persists into adulthood. Tan L, Fichtner AS, Bruni E, Odak I, Sandrock I, Bubke A, Borchers A, Schultze-Florey C, Koenecke C, Förster R, Jarek M, von Kaisenberg C, Schulz A, Chu X, Zhang B, Li Y, Panzer U, Krebs CF, Ravens S, Prinz I. Sci. Immunol. 2021 Apr 23; 6, eabf0125 (2021).
COVID-19 immune signatures reveal stable antiviral T cell function despite declining humoral responses. Bonifacius A, Tischer-Zimmermann S, Dragon AC, Gussarow D, Vogel A, Krettek U, Gödecke N, Yilmaz M, Kraft ARM, Hoeper MM, Pink I, Schmidt JJ, Li Y, Welte T, Maecker-Kolhoff B, Martens J, Berger MM, Lobenwein C, Stankov MV, Cornberg M, David S, Behrens GMN, Witzke O, Blasczyk R, Eiz-Vesper B. Immunity 2021 Feb 9 ;54(2):340-354.e6.
CAR-T cells and TRUCKs that recognize an EBNA-3C-derived epitope presented on HLA-B*35 control Epstein-Barr virus-associated lymphoproliferation. Dragon AC, Zimmermann K, Nerreter T, Sandfort D, Lahrberg J, Klöß S, Kloth C, Mangare C, Bonifacius A, Tischer-Zimmermann S, Blasczyk R, Maecker-Kolhoff B, Uchanska-Ziegler B, Abken H, Schambach A, Hudecek M, Eiz-Vesper B. J Immunother Cancer. 2020 Oct ;8(2):e000736
Microbial exposure drives polyclonal expansion of innate γδ T cells immediately after birth. Ravens S, Fichtner AS, Willers M, Torkornoo D, Pirr S, Schöning J, Deseke M, Sandrock I, Bubke A, Wilharm A, Dodoo D, Egyir B, Flanagan KL, Steinbrück L, Dickinson P, Ghazal P, Adu B, Viemann D, Prinz I. Proc Natl Acad Sci U S A. 2020 Aug 4 ;117(31):18649-18660.
Human γδ TCR Repertoires in Health and Disease. Fichtner AS, Ravens S, Prinz I. Cells 2020 Mar 26;9(4):E800.
TCR repertoire analysis reveals phosphoantigen-induced polyclonal proliferation of Vγ9Vδ2 T cells in neonates and adults. Fichtner AS, Bubke A, Rampoldi F, Wilharm A, Tan L, Steinbrück L, Schultze-Florey C, von Kaisenberg C, Prinz I, Herrmann T, Ravens S. J Leukoc Biol. 2020 Feb 17. [Epub ahead of print]
Robust Identification of Suitable T-Cell Subsets for Personalized CMV-Specific T-Cell Immunotherapy Using CD45RA and CD62L Microbeads. Mangare C, Tischer-Zimmermann S,Riese SB, Dragon AC , Prinz I,Blasczyk R, Maecker-Kolhoff B, Eiz-Vesper B. Int. J. Mol. Sci. 2019 March 19; 20,1415
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.