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

J Immunol Methods. 2007 Dec 1;328(1-2):169-81. doi: 10.1016/j.jim.2007.08.013. Epub 2007 Sep 29.

Abstract

Adoptive cell transfer immunotherapy has been utilized to treat EBV related human malignancies including post-transplant lymphoproliferative diseases, Hodgkin's lymphoma and nasopharyngeal carcinoma. However, there are limited options available for tumor antigen-specific T cell purification. Here we describe a novel solid phase T cell selection system, in which monocytes or EBV transformed B-lymphocytes are immobilized on solid support for antigen-specific T cell purification. We hypothesize and prove that antigen-specific T cells recognize their cognate antigens and bind to them faster than non-antigen specific T cells. Therefore antigen-specific T cells can be concentrated on the surface after removing the non-adherent cells by washing. The optimal selection time for both EBV-specific T cells and LMP2-specific T cells is studied. Our data demonstrate that the frequency of antigen-specific T cells can be increased by >20-fold after selection. Moreover, activated antigen-specific T cells proliferate more rapidly than non-specific T cells, further increasing the frequency and purity of antigen-specific T cells. This new T cell selection system is superior to traditional repeated stimulation methods in generating tumor antigen-specific T cells. We are able to generate large quantities of highly purified T cells of subdominant antigens LMP2 within 2 weeks after T cell activation for adoptive cell transfer immunotherapy with this simple, rapid and inexpensive T cell selection system.

MeSH terms

  • Antigens, Neoplasm / immunology*
  • Cell Separation / methods*
  • Herpesvirus 4, Human / immunology*
  • Humans
  • Immunotherapy / methods*
  • In Vitro Techniques
  • Lymphocyte Activation
  • T-Lymphocytes, Cytotoxic / immunology*
  • Time
  • Viral Matrix Proteins / immunology

Substances

  • Antigens, Neoplasm
  • EBV-associated membrane antigen, Epstein-Barr virus
  • Viral Matrix Proteins