Cytotoxic T Lymphocyte Therapy for Epstein-Barr Virus+ Hodgkin's Disease

Abstract

Epstein Barr virus (EBV)+ Hodgkin's disease (HD) expresses clearly identified tumor antigens derived from the virus and could, in principle, be a target for adoptive immunotherapy with viral antigen–specific T cells. However, like most tumor-associated antigens in immunocompetent hosts, these potential targets are only weakly immunogenic, consisting primarily of the latent membrane protein (LMP)1 and LMP2 antigens. Moreover, Hodgkin tumors possess a range of tumor evasion strategies. Therefore, the likely value of immunotherapy with EBV-specific cytotoxic effector cells has been questioned. We have now used a combination of gene marking, tetramer, and functional analyses to track the fate and assess the activity of EBV cytotoxic T lymphocyte (CTL) lines administered to 14 patients treated for relapsed EBV+ HD. Gene marking studies showed that infused effector cells could further expand by several logs in vivo, contribute to the memory pool (persisting up to 12 mo), and traffic to tumor sites. Tetramer and functional analyses showed that T cells reactive with the tumor-associated antigen LMP2 were present in the infused lines, expanded in peripheral blood after infusion, and also entered tumor. Viral load decreased, demonstrating the biologic activity of the infused CTLs. Clinically, EBV CTLs were well tolerated, could control type B symptoms (fever, night sweats, and weight loss), and had antitumor activity. After CTL infusion, five patients were in complete remission at up to 40 mo, two of whom had clearly measurable tumor at the time of treatment. One additional patient had a partial response, and five had stable disease. The performance and fate of these human tumor antigen–specific T cells in vivo suggests that they might be of value for the treatment of EBV+ Hodgkin lymphoma.

In vitro methods for generating highly purified EBV associated tumor antigen-specific T cells by using solid phase T cell selection system for immunotherapy

“…In this approach, the quantities of LMP2-specific T cells represent only minority of EBV-specific T cells, which is consistent with the fact that LMP2 are EBV subdominant antigens. Even so, the quantities of LMP2-specific T cells generated from this new T cell selection method appears to be much more than that in EBV-specific T cells generated through traditional repeated stimulation method, in which on average only about 1% of LMP2-specific T cells was generated using PBMC from both normal individual and Hodgkin's Lymphoma patients (Bollard et al, 2004b). In patients with known HLA restricted LMP2 epitope peptide, another approach is to select and expand activated LMP2-specific T cells directly.…”

“…In addition, Results from several clinical trials have proved autologous EBV-targeted T cells infusions effective in promoting clinical tumor response, including clinical remissions in some instances in patients with HL and NPC (Bollard et al, 2004a;Straathof et al, 2005;Bollard et al 2004b;Comoli et al, 2005). In these clinical trials, EBV-specific cytotoxic T lymphocytes (CTLs) are generated with peripheral blood mononuclear cells (PBMC) by repeated stimulation using patient-derived EBV transformed B lymphocyte (LCL) as antigen presenting cells.…”

Epstein‐Barr virus (EBV) latent membrane protein‐1‐specific cytotoxic T lymphocytes targeting EBV‐carrying natural killer cell malignancies

“…Extension of a similar strategy to other EBV-associated malignancies, such as HD [14] and NPC [15], has been reported to be efficacious in some patients. However, the majority of lymphoblastoid cell line (LCL)-activated CTL used in the reported studies were directed to immunodominant EBNA3A, EBNA3B, and EBNA3C Ag, which are not expressed in the malignant cells of HD and NPC cases.…”

“…A subdominant portion of LCL-activated CTL may recognize peptides derived from LMP2 [14,15], which would contribute to immunotherapeutic effects in treated patients. However, T cells directing LMP1 peptides are rare [15], reflecting a low CTL precursor frequency [16].…”

Immunotherapies for Hodgkin's lymphoma

“…Peripheral blood from HL patients was initially used to create both an EBV-specific CTL line and B-lymphoblastoid cell line (LCL, an EBVtransformed cell line). The EBV-specific CTL were activated in vitro using LCLs as antigen presenters, then expanded, genetically marked, and reinfused into patients [32,33]. Patientderived CTL expanded less easily and had lower T cell receptor zeta chain expression than CTL from healthy donors, but had comparable cytotoxicity against EBV in vitro [33].…”