The Generation and Characterization of LMP2-Specific CTLs for Use as Adoptive Transfer From Patients With Relapsed EBV-Positive Hodgkin Disease

Abstract

Cellular adoptive immunotherapy for virus-associated malignant disease is an attractive strategy, since viral antigens provide targets for specific T lymphocytes. In Epstein-Barr virus (EBV)-positive Hodgkin disease (HD), a limited number of EBV-encoded antigens such as the latent membrane antigens (LMP) 1 and 2 are expressed on the malignant Reed-Sternberg cells. The authors aimed to generate cytotoxic T lymphocytes (CTLs) from patients with relapsed HD by specifically targeting LMP2A. Patients with relapsed HD have highly immunosuppressive tumors and have been heavily pretreated with cytotoxic agents. As a result, monocytes and lymphocytes are numerically reduced and functionally impaired. Approaches using dendritic cells (DCs) as the sole antigen-presenting cell to expand LMP2-specific CTL lines in vitro have proved impractical. The authors now show how small amounts of patient peripheral blood can be used to produce DCs expressing LMP2 after Ad5F35 transduction, and how an initial reactivation of LMP2-specific CTLs can be followed by stimulation with lymphoblastoid cell lines overexpressing LMP2 from the same vector. Large numbers of LMP2-specific cytotoxic lymphocytes are produced that contain both CD4+ and CD8+ T cells (favoring long-term persistence in vivo) and recognize multiple LMP2 epitopes (minimizing the risk of tumor antigen loss variants). This approach is being used in a current clinical trial.

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 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.…”

Computational Prediction and Identification of Epstein-Barr Virus Latent Membrane Protein 2A Antigen-Specific CD8+ T-Cell Epitopes

“…Recently, there have been many attempts to use genetransfected DCs to sensitize T cells to recognize target cells expressing relative antigen in research of EBV-relative malignancies (23)(24)(25)(26). In a variety of vectors, vaccinia virus has more recently been used as a versatile eukaryotic expression vector and is important in tumor gene transfecting.…”

Immunotherapies for Hodgkin's lymphoma

“…A potential limitation of using autologous LCLs for antigen presentation is that T cells specific for immunodominant EBV antigens or early lytic cycle antigens (which are not expressed in HL) are preferentially amplified [34]. Nevertheless, the CTL lines generated with this approach contained small populations of LMP2-specific clones [33].…”

“…CTL specific for LMP1 [36] or for LMP2 [37,38] have been successfully generated. One approach is to stimulate patient T cells using autologous dendritic cells and LCLs, both overexpressing LMP2A through an adenoviral vector [34]. This can generate large numbers of LMP2-specific CD4+ and CD8+ T cells [34].…”

“…One approach is to stimulate patient T cells using autologous dendritic cells and LCLs, both overexpressing LMP2A through an adenoviral vector [34]. This can generate large numbers of LMP2-specific CD4+ and CD8+ T cells [34]. In a study of 16 patients with EBV+HL or non-Hodgkin's lymphoma who had relapsed or were at high risk of relapse, infusion of such autologous LMP2-CTL was well tolerated and was associated with an increased frequency of LMP2-CTL in the blood [39].…”