PuSH - Publikationsserver des Helmholtz Zentrums München: Generation of Epstein-Barr virus antigen-specific T cell receptors recognizing immunodominant epitopes of LMP1, LMP2A, and EBNA3C for immunotherapy.

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Dudaniec, K.* ; Westendorf, K.* ; Nößner, E. ; Uckert, W.*

Generation of Epstein-Barr virus antigen-specific T cell receptors recognizing immunodominant epitopes of LMP1, LMP2A, and EBNA3C for immunotherapy.

Hum. Gene Ther. 32, 919-935 (2021)

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Epstein-Barr virus (EBV) infections in healthy individuals are usually cleared by immune cells, wherein CD8+ T cells play the most important role. However, in some immunocompromised individuals, EBV infections can lead to the development of cancer in B, T, NK cells and epithelial cells. Most EBV-associated cancers express a limited number of virus-specific antigens such as latent membrane proteins (LMP1, LMP2) and nuclear proteins (EBNA1, -2, EBNA3A, -B, -C, EBNA-LP). These antigens represent true tumor-specific antigens and can be considered useful targets for TCR gene therapy to treat EBV-associated diseases. We used a TCR isolation platform based on a single major histocompatibility class I complexe (MHC I) K562 cell library for the detection, isolation, and re-expression of TCRs targeting immunodominant peptide-MHC (pMHC) complexes. Mature dendritic cells (mDCs) were pulsed with in vitro-transcribed (ivt) RNA encoding for the selected antigen to stimulate autologous T cells. The procedure allowed the mDCs to select an immunogenic epitope of the antigen for processing and presentation on the cell surface in combination with the most suitable MHC I molecule. We isolated eight EBV-specific TCRs. They recognize various pMHC complexes of EBV antigens LMP1, LMP2A, and EBNA3C, some of them described previously and some newly identified in this study. The TCR genes were molecularly cloned into retroviral vectors and the resultant TCR-engineered T cells secreted interferon-γ after antigen contact and were able to lyse tumor cells. The EBV-specific TCRs can be used as a basis for the generation of a TCR library, which provides a valuble source of TCRs for the production of EBV-specific T cells to treat EBV-associated diseases in patients with different MHC I types.

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