795. The Use of Autologous LMP2-Specific Cytotoxic T Lymphocytes for the Treatment of Relapsed EBV +ve Hodgkin Disease and Non-Hodgkin Lymphoma
Authors: Catherine M. Bollard, Stephen Gottschalk, Elizabeth Buza, Helen Huls, Vicky Gresik, George Carrum, Malcolm K. Brenner, Cliona M. Rooney, Helen E. Heslop
Journal: Molecular Therapy (2005)

EBV-associated Hodgkin's Disease (HD) and some non-Hodgkins lymphoma (NHL) show type II latency expressing the subdominant EBV antigens EBNA1, LMP1 and LMP2, which may serve as targets for immunotherapy approaches. In previous studies, we used polyclonal EBV-specific CTL in patients with relapsed EBV +ve HD and saw 2 complete and 1 partial response in 11 patients. Analyses of EBV-CTL lines showed that small populations of T cells reactive against the tumor-associated antigen LMP2 were present in the majority of the infused lines, with some expansion in the peripheral blood following infusion. We therefore hypothesized that CTL specifically targeting LMP2 might have greater efficacy in these patients. LMP2-CTL were generated from 10 patients using Dendritic Cells and Lymphoblastoid Cell Lines (LCL) that had been genetically modified to overexpress LMP2 by transduction with an Ad5f35LMP2 vector. Polyclonal LMP2-CTL lines recognized 2-6 (median 4) LMP2 epitopes, as determined using overlapping LMP2 peptide pools in ELISPOT assays. A mean of 22.8% (5-42.1%) of CD8+ T cells bound HLA-restricted LMP2 tetramers, compared to a mean of 0.11% (0.01-0.24%) of LMP2-tetramer positive CD8+ T cells found in CTL generated with genetically unmodified LCL from the same patients. So far, 10 patients have been treated on this dose escalation study. No toxicity has been seen |[ndash]| 6 patients have been treated on dose level 1 (2 doses of CTL at 2x10e7/m2/dose given 2 weeks apart in the outpatient clinic) and 4 patients on dose level 2 (1 dose of 2|[times]|10e7/m2 and second dose of 1|[times]|10e8/m2). No immediate toxicity was observed. In patients with identified LMP2-epitopes, measurement of IFN|[gamma]| secretion by CD8+ T cells after stimulation with appropriate LMP2-peptides in ELISPOT assays showed a 4-25-fold increase in spot forming cells after infusions. In contrast, frequencies of CMV and superantigen-specific T cells did not increase. Four of five patients who received LMP2-CTL as adjuvant therapy post stem cell transplant or chemotherapy remain in remission up to 12 months post LMP2-CTL. One patient presented with progressive disease 8 weeks post CTL therapy. Five patients had detectable disease at the time of CTL therapy. 1 patient had a very good partial response and for 2 patients it is too early to assess. The two remaining patients had stable disease 8 weeks post LMP2-CTL and were eligible to receive further doses. Both patients received 2 further doses of LMP2-CTL. One patient continues with stable disease 6 months post CTL therapy and the other patient had a complete radiological response. This patient had a supraclavicular lymph node resection, which showed selective accumulation of LMP2-tetramer positive cells (0.3% compared to 0.01% in the peripheral blood) with few residual tumor cells. Immunotherapy with autologous LMP2-CTL is therefore well tolerated in patients with relapsed EBV+ve HD/NHL and infused LMP2-CTL cells can localize to the tumor and induce a clinical response.