Cellular immunotherapy for follicular lymphoma using genetically modified CD20-specific CD8+ cytotoxic T lymphocytes.
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.
Wang
Jinjuan
J
Fred Hutchinson Cancer Research Center, 1100 Fairview Avenue North, Seattle, WA 98109, USA.
Press
Oliver W
OW
Lindgren
Catherine G
CG
Greenberg
Philip
P
Riddell
Stanley
S
Qian
Xiaojun
X
Laugen
Christian
C
Raubitschek
Andrew
A
Forman
Stephen J
SJ
Jensen
Michael C
MC
D018951:Antigens, CD20 / Q000096:biosynthesis* / Q000378:metabolism; D015139:Blotting, Southern; D015153:Blotting, Western; D017252:CD3 Complex / Q000737:chemistry; D018414:CD8-Positive T-Lymphocytes / Q000166:cytology* / Q000378:metabolism; D020169:Caspases / Q000378:metabolism; D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000378:metabolism; D002478:Cells, Cultured; D002857:Chromium / Q000378:metabolism; D003001:Cloning, Molecular; D018274:Electroporation; D005434:Flow Cytometry; D018014:Gene Transfer Techniques; D015316:Genetic Therapy / Q000379:methods*; D006801:Humans; D007167:Immunotherapy / Q000379:methods*; D007376:Interleukin-2 / Q000378:metabolism; D019868:Kanamycin Kinase / Q000378:metabolism; D008198:Lymph Nodes / Q000473:pathology; D008223:Lymphoma / Q000378:metabolism; D008224:Lymphoma, Follicular / Q000628:therapy*; D010957:Plasmids / Q000378:metabolism; D011948:Receptors, Antigen, T-Cell / Q000378:metabolism; D013601:T-Lymphocytes / Q000378:metabolism; D013997:Time Factors; D014162:Transfection; D019076:Transgenes
D018951:Antigens, CD20; D017252:CD3 Complex; D007376:Interleukin-2; D011948:Receptors, Antigen, T-Cell; D002857:Chromium; D019868:Kanamycin Kinase; D020169:Caspases