Expansion of EBV latent membrane protein 2a specific cytotoxic T cells for the adoptive immunotherapy of EBV latency type 2 malignancies: influence of recombinant IL12 and IL15

Cytotherapy. 2003;5(3):231-40. doi: 10.1080/14653240310001262.


Background: EBV-associated malignancies with a Type II latency gene expression pattern, such as EBV-positive HD, or nasopharyngeal carcinoma, frequently express the EBV latency Ag LMP2a. Hence, they provide a potential target for adoptive immunotherapy using in vitro-generated LMP2a-specific cytotoxic T lymphocytes (CTL). In this study, LMP2a-specific CTL were specifically amplified and the influence of rIL12 and rIL15 on the culture outcome was tested.

Methods: PBMC from donors were stimulated twice with autologous DC transduced with an adenovirus vector expressing LMP2a. This led to a significant expansion of LMP2a-tetramer-specific CTL, which were subsequently further expanded with autologous EBV-transformed B-lymphoblastoid cells (LCL). The addition of rIL12 and rIL15 to the standard IL2-containing culture medium enhanced the proliferation of LMP2a-specific CTL.

Results: While rIL15 did not change the pattern of cytokines secreted by LMP2a-CTL, rIL12 enhanced the production of Th1/Tc1 cytokines, such as IFN-n, while suppressing the production of the Th2/Tc2 cytokine IL5.

Discussion: Stimulation of CTL cultures with rIL12 or rIL15 will generate CTL more rapidly, facilitating the application of this approach for patients with these EBV-associated disorders.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenoviridae / genetics
  • Cell Line
  • Cytokines / metabolism
  • Dendritic Cells / immunology
  • Epstein-Barr Virus Infections / immunology*
  • Epstein-Barr Virus Infections / therapy
  • Genetic Vectors
  • Humans
  • Immunotherapy, Adoptive
  • Interleukin-12 / genetics
  • Interleukin-12 / pharmacology*
  • Interleukin-15 / genetics
  • Interleukin-15 / pharmacology*
  • Leukocytes, Mononuclear / immunology
  • Lymphocyte Activation
  • Recombinant Proteins / pharmacology
  • T-Lymphocytes, Cytotoxic / immunology*
  • T-Lymphocytes, Cytotoxic / transplantation
  • Viral Matrix Proteins / genetics
  • Viral Matrix Proteins / immunology*


  • Cytokines
  • EBV-associated membrane antigen, Epstein-Barr virus
  • Interleukin-15
  • Recombinant Proteins
  • Viral Matrix Proteins
  • Interleukin-12