Journal
HUMAN GENE THERAPY
Volume 11, Issue 4, Pages 611-620Publisher
MARY ANN LIEBERT INC PUBL
DOI: 10.1089/10430340050015798
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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 hrIL-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 IgG(1)(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.
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