Co-targeting EBV lytic as well as latent cycle antigens increases T-cell potency against lymphoma.

The remarkable efficacy of Epstein-Barr virus (EBV) specific T-cells for the treatment of post-transplant lymphomas (PTLD) has not been reproduced for EBV+ malignancies outside the transplant setting. This is due in part to the heterogeneous expression and poor immunogenicity of the viral antigens expressed, namely LMPs 1 and 2, EBNA1, and BARF1 (type-2 (T2) latency). However, EBV lytic cycle proteins are also expressed in certain EBV+ malignancies, and since several EBV lytic cycle proteins are abundantly expressed, have oncogenic activity, and likely contribute to malignancy, we sought and identified viral lytic-cycle transcripts in EBV+ Hodgkin's lymphoma biopsies. This provided the rationale for broadening the target antigen-specific repertoire of EBVSTs for therapy. We stimulated healthy donors and EBV+ lymphoma patients' peripheral blood mononuclear cells (PBMCs) with both lytic and latent cycle proteins to make broad repertoire (BR)-EBVSTs). Compared to T2 Ag-specific (T2-) EBVSTs, BR-EBVSTs more rapidly cleared autologous EBV+ tumors in NSG mice and produced higher levels of pro-inflammatory cytokines that should reactivate the immunosuppressive tumor microenvironment leading to epitope spreading. Our results confirm that lytic cycle antigens are clinically relevant targets for EBV+ lymphoma and underpin the rationale for integrating BR-EBVSTs as a therapeutic approach for relapsed/refractory EBV-positive lymphoma (NCT01555892 and NCT04664179), as well as for other EBV-associated malignancies.