T cells specific for different latent and lytic viral proteins efficiently control Epstein-Barr virus-transformed B cells.
OBJECTIVE
Epstein-Barr virus (EBV)-associated post-transplant lymphoproliferative disorders (PTLD) belong to the most dreaded complications of immunosuppression. The efficacy of EBV-specific T-cell transfer for PTLD has been previously shown, yet the optimal choice of EBV-derived antigens inducing polyclonal CD4(+) and CD8(+) T cells that cover a wide range of human leukocyte antigen types and efficiently control PTLD remains unclear.


METHODS
A pool of 125 T-cell epitopes from seven latent and nine lytic EBV-derived proteins (EBVmix) and peptide pools of EBNA1, EBNA3c, LMP2a and BZLF1 were used to determine T-cell frequencies and to isolate T cells through the use of the interferon (IFN)-γ cytokine capture system. We further evaluated the phenotype and functionality of the generated T-cell lines in vitro.


RESULTS
EBVmix induced significantly higher T-cell frequencies and allowed selecting more CD4(+)IFN-γ(+) and CD8(+)IFN-γ(+) cells than single peptide pools. T cells of all specificities expanded similarly in vitro, recognized cognate antigen, and, to a lower extent, EBV-infected cells, exerted moderate cytotoxicity and showed reduced alloreactivity. However, EBVmix-specific cells most efficiently controlled EBV-infected lymphoblastoid cell lines (LCLs). This control was mainly mediated by EBV-specific CD8(+) cells with an oligoclonal epitope signature covering both latent and lytic viral proteins. Notably, EBV-specific CD4(+) cells unable to control LCLs produced significantly less perforin and granzyme B, probably because of limited LCL epitope presentation.


CONCLUSIONS
EBVmix induces a broader T-cell response, probably because of its coverage of latent and lytic EBV-derived proteins that may be important to control EBV-transformed B cells and might offer an improvement of T-cell therapies.
Infection Biology Laboratory, Department of Biomedicine, University and University Hospital of Basel, Switzerland.
Justyna
J
Nowakowska
Infection Biology Laboratory, Department of Biomedicine, University and University Hospital of Basel, Switzerland.
Claudia
C
Stuehler
Infection Biology Laboratory, Department of Biomedicine, University and University Hospital of Basel, Switzerland; Clinical Microbiology, University Hospital of Basel, Switzerland.
Adrian
A
Egli
Division of Infectious Diseases and Hospital Epidemiology, Department of Biomedicine and Clinical Research University Hospital of Basel, Switzerland.
Manuel
M
Battegay
Research and Development, Miltenyi Biotec, Bergisch-Gladbach, Germany.
Georg
G
Rauser
Immunobiology, Department of Biomedicine, University and University Hospital of Basel, Switzerland.
Glenn Robert
GR
Bantug
AIDS Research Institute-IrsiCaixa and AIDS Unit, Hospital Germans Trias i Pujol, Autonomous University of Barcelona, Badalona, Spain, Institució Catalana de Recerca i Estudis Avançats, Barcelona, Spain; University of Vic and Central Catalonia, Vic, Spain.
Christian
C
Brander
Immunobiology, Department of Biomedicine, University and University Hospital of Basel, Switzerland.
Christoph
C
Hess
Infection Biology Laboratory, Department of Biomedicine, University and University Hospital of Basel, Switzerland; Division of Infectious Diseases and Hospital Epidemiology, Department of Biomedicine and Clinical Research University Hospital of Basel, Switzerland. Electronic address: nina.khanna@usb.ch.
Nina
N
Khanna
D001402:B-Lymphocytes / Q000276:immunology / Q000821:virology*; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology* / Q000637:transplantation; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology* / Q000637:transplantation; D002472:Cell Transformation, Viral / Q000276:immunology*; D002478:Cells, Cultured; D018984:Epitopes, T-Lymphocyte / Q000276:immunology; D020031:Epstein-Barr Virus Infections / Q000628:therapy*; D019309:Epstein-Barr Virus Nuclear Antigens / Q000276:immunology; D053804:Granzymes / Q000378:metabolism; D004854:Herpesvirus 4, Human / Q000276:immunology*; D006801:Humans; D007371:Interferon-gamma / Q000276:immunology; D054353:Perforin / Q000096:biosynthesis; D015534:Trans-Activators / Q000276:immunology; D014763:Viral Matrix Proteins / Q000276:immunology
C063676:BZLF1 protein, Herpesvirus 4, Human; C498774:EBNA-3C, epstein-barr virus; C037662:EBV-associated membrane antigen, Epstein-Barr virus; D018984:Epitopes, T-Lymphocyte; D019309:Epstein-Barr Virus Nuclear Antigens; D015534:Trans-Activators; D014763:Viral Matrix Proteins; D054353:Perforin; D007371:Interferon-gamma; D053804:Granzymes; C417029:EBV-encoded nuclear antigen 1
Epstein-Barr virus; IFN-γ selection; PTLD; T-cell transfer; immunotherapy