title: Cellular immunotherapy for follicular lymphoma using genetically modified CD20-specific CD8+ cytotoxic T lymphocytes.
abstract: 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.
authors.affiliation: Fred Hutchinson Cancer Research Center, 1100 Fairview Avenue North, Seattle, WA 98109, USA.
authors.forename: Jinjuan
authors.initials: J
authors.lastname: Wang
authors.forename: Oliver W
authors.initials: OW
authors.lastname: Press
authors.forename: Catherine G
authors.initials: CG
authors.lastname: Lindgren
authors.forename: Philip
authors.initials: P
authors.lastname: Greenberg
authors.forename: Stanley
authors.initials: S
authors.lastname: Riddell
authors.forename: Xiaojun
authors.initials: X
authors.lastname: Qian
authors.forename: Christian
authors.initials: C
authors.lastname: Laugen
authors.forename: Andrew
authors.initials: A
authors.lastname: Raubitschek
authors.forename: Stephen J
authors.initials: SJ
authors.lastname: Forman
authors.forename: Michael C
authors.initials: MC
authors.lastname: Jensen
mesh_terms: 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
chemical_list: D018951:Antigens, CD20; D017252:CD3 Complex; D007376:Interleukin-2; D011948:Receptors, Antigen, T-Cell; D002857:Chromium; D019868:Kanamycin Kinase; D020169:Caspases