Genetic modification of human T cells with CD20: a strategy to purify and lyse transduced cells with anti-CD20 antibodies

Hum Gene Ther. 2000 Mar 1;11(4):611-20. doi: 10.1089/10430340050015798.

Abstract

A retroviral vector has been constructed that contains the human CD20 cDNA under the control of the Moloney murine leukemia virus (Mo-MuLV) LTR. Freshly isolated mononuclear cells are infected for three consecutive days in the presence of PHA and hrlL-2, and a mean 15.9% of the cells (range, 6.5 to 31.7%) acquire a CD3+CD20+ phenotype. Transduced T lymphocytes grow and expand in vitro for up to 3 weeks like mock-infected cells and, as observed for the T lymphoblastoid CEM cell line, CD20 expression is maintained for several months with no change in the growth curve of the cells. CD20-expressing CEM and fresh T lymphocytes can be positively immunoselected on columns using different anti-CD20 antibodies. Exposure to monoclonal chimeric anti-CD20 IgG1(kappa) Rituximab antibody (Roche), in the presence of complement, results in effective and rapid killing of the transduced CD3+CD20+ human T cells in vitro. This approach represents a new and alternative method to gene manipulation with "suicide" genes for the production of drug-responsive T cell populations, a crucial step for the future management of graft-versus-host disease in bone marrow transplant patients.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antibodies, Monoclonal / immunology*
  • Antibodies, Monoclonal, Murine-Derived
  • Antigens, CD20 / genetics
  • Antigens, CD20 / immunology*
  • Base Sequence
  • Cell Division
  • Cell Line
  • Cell Separation
  • DNA Primers
  • DNA, Complementary
  • Flow Cytometry
  • Genetic Vectors
  • Humans
  • Retroviridae / genetics
  • Rituximab
  • T-Lymphocytes / immunology*
  • Transduction, Genetic*

Substances

  • Antibodies, Monoclonal
  • Antibodies, Monoclonal, Murine-Derived
  • Antigens, CD20
  • DNA Primers
  • DNA, Complementary
  • Rituximab