T cell re-targeting to EBV antigens following TCR gene transfer: CD28-containing receptors mediate enhanced antigen-specific IFNgamma production.
EBV is associated with a broad range of malignancies. Adoptive immunotherapy of these tumors with EBV-specific CTL proved useful. We generated a panel of primary human T cells specific to various EBV antigens (i.e. Epstein-Barr nuclear antigen 3A, 3B and BamHI-M leftward reading frame) via transfer of modified TCR genes that are either coupled to CD3zeta or Fc(epsilon)RIgamma. TCR-transduced T cells from 20-60% of donors (total number of 25) demonstrated specific lysis of EBV peptide-loaded target cells, whereas lymphoblastoid cell lines expressing native EBV antigens were not killed by any of the EBV-specific T cell populations. This non-responsiveness, confirmed at the level of nuclear factor of activated T cells activation, is not due to receptor configuration since identical receptor formats specific for melanoma antigens successfully re-targeted T cells to native melanoma cells. In an effort to generate a more potent receptor, we introduced a CD28 domain into one of the EBV-specific TCR. This TCR did not affect the cytotoxic response of re-targeted T cells, but dramatically enhanced antigen-specific IFNgamma production. We therefore conclude that these novel CD28-containing EBV-specific TCRs provide a basis for further development of TCR gene transfer to treat EBV-induced diseases.
Schaft
Niels
N
Laboratory of Tumor Immunology, Department of Medical Oncology, Erasmus MC-Daniel den Hoed Cancer Center, Groene Hilledijk 301, 3075 EA Rotterdam, the Netherlands.
Lankiewicz
Birgit
B
Drexhage
Joost
J
Berrevoets
Cor
C
Moss
Denis J
DJ
Levitsky
Victor
V
Bonneville
Marc
M
Lee
Steven P
SP
McMichael
Andrew J
AJ
Gratama
Jan-Willem
JW
Bolhuis
Reinier L H
RL
Willemsen
Ralph
R
Debets
Reno
R
D018106:CD28 Antigens / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay; D019309:Epstein-Barr Virus Nuclear Antigens / Q000276:immunology*; D005434:Flow Cytometry; D018014:Gene Transfer Techniques; D019672:Genes, T-Cell Receptor; D015316:Genetic Therapy / Q000379:methods; D006801:Humans; D007371:Interferon-gamma / Q000096:biosynthesis*; D019169:Jurkat Cells; D011948:Receptors, Antigen, T-Cell / Q000235:genetics / Q000276:immunology; D013601:T-Lymphocytes / Q000276:immunology*; D014161:Transduction, Genetic
D018106:CD28 Antigens; D019309:Epstein-Barr Virus Nuclear Antigens; D011948:Receptors, Antigen, T-Cell; D007371:Interferon-gamma