Genetic modification of human T cells with CD20: a strategy to purify and lyse transduced cells with anti-CD20 antibodies.
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.
Introna
M
M
Istituto di Ricerche Farmacologiche Mario Negri, Milan, Italy. martino@IRFMN.MNEGRI.IT
Barbui
A M
AM
Bambacioni
F
F
Casati
C
C
Gaipa
G
G
Borleri
G
G
Bernasconi
S
S
Barbui
T
T
Golay
J
J
Biondi
A
A
Rambaldi
A
A
D000911:Antibodies, Monoclonal / Q000276:immunology*; D058846:Antibodies, Monoclonal, Murine-Derived; D018951:Antigens, CD20 / Q000235:genetics / Q000276:immunology*; D001483:Base Sequence; D002455:Cell Division; D002460:Cell Line; D002469:Cell Separation; D017931:DNA Primers; D018076:DNA, Complementary; D005434:Flow Cytometry; D005822:Genetic Vectors; D006801:Humans; D012190:Retroviridae / Q000235:genetics; D000069283:Rituximab; D013601:T-Lymphocytes / Q000276:immunology*; D014161:Transduction, Genetic
D000911:Antibodies, Monoclonal; D058846:Antibodies, Monoclonal, Murine-Derived; D018951:Antigens, CD20; D017931:DNA Primers; D018076:DNA, Complementary; D000069283:Rituximab