Cellular immunotherapy for follicular lymphoma using genetically modified CD20-specific CD8+ cytotoxic T lymphocytes

Mol Ther. 2004 Apr;9(4):577-86. doi: 10.1016/j.ymthe.2003.12.011.


Humoral immunotherapy using the monoclonal anti-CD20 antibody rituximab induces remissions in approximately 60% of patients with relapsed follicular lymphoma; however, most patients eventually relapse despite continued expression of CD20 on lymphoma cells. We have hypothesized that cellular immunotherapy targeting CD20(+) cells might provide a more effective mechanism for eliminating lymphoma cells than anti-CD20 antibodies and are therefore investigating the utility of cytotoxic T lymphocytes (CTL) genetically modified to target the CD20 antigen. Peripheral blood mononuclear cells were activated with anti-CD3 antibody (OKT3) and recombinant human interleukin-2 and electroporated with a plasmid containing a CD20-specific scFvFc:zeta chimeric T cell receptor gene and a neomycin phosphotransferase gene (neo(R)). Transfected cells were selected using the antibiotic G418 and cloned by limiting dilution. Using this approach, we have generated CD8(+) CTL clones with CD20-specific cytotoxicity, which specifically lysed CD20(+) target cells, including actual tumor cells from patients with follicular lymphoma, small lymphocytic lymphoma, splenic marginal zone lymphoma, diffuse large B cell lymphoma, and chronic lymphocytic leukemia. The CTL clones have been expanded to numbers sufficient for therapy ( approximately 10(9) cells). Our data indicate the feasibility of generating and expanding CD20-specific CTL and, for the first time, demonstrate that such CTL exhibit specific cytotoxicity against actual tumor cells isolated from patients with a variety of B lymphoid malignancies. In view of these promising findings, a Phase I clinical trial for relapsed follicular lymphoma is being initiated.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Antigens, CD20 / biosynthesis*
  • Antigens, CD20 / metabolism
  • Blotting, Southern
  • Blotting, Western
  • CD3 Complex / chemistry
  • CD8-Positive T-Lymphocytes / cytology*
  • CD8-Positive T-Lymphocytes / metabolism
  • Caspases / metabolism
  • Cell Line, Tumor
  • Cell Membrane / metabolism
  • Cells, Cultured
  • Chromium / metabolism
  • Cloning, Molecular
  • Electroporation
  • Flow Cytometry
  • Gene Transfer Techniques*
  • Genetic Therapy / methods*
  • Humans
  • Immunotherapy / methods*
  • Interleukin-2 / metabolism
  • Kanamycin Kinase / metabolism
  • Lymph Nodes / pathology
  • Lymphoma / metabolism
  • Lymphoma, Follicular / therapy*
  • Plasmids / metabolism
  • Receptors, Antigen, T-Cell / metabolism
  • T-Lymphocytes / metabolism
  • Time Factors
  • Transfection
  • Transgenes


  • Antigens, CD20
  • CD3 Complex
  • Interleukin-2
  • Receptors, Antigen, T-Cell
  • Chromium
  • Kanamycin Kinase
  • Caspases