A novel strategy for rapid and efficient isolation of human tumor-specific CD4(+) and CD8(+) T-cell clones.
Adoptive therapy with antigen-specific T cells is a promising approach for the treatment of infectious diseases and cancer. However, cloning of antigen-specific T cells by the traditional approach of limiting dilution is a time-consuming, laborious, and inefficient process. Here, we describe a novel flow cytometric strategy for rapid isolation of human tumor antigen-specific T-cell clones by using T-cell receptor (TCR) Vbeta antibodies in combination with carboxyfluorescein succinimidyl ester (CFSE)-based proliferation assay. The CFSE dilution following antigen stimulation identified proliferating antigen-specific T cells, and the TCRVbeta antibodies allowed distinguishing T cells at the clonal level from a heterogeneous T-cell population. This method of TCRVbeta/CFSE dilution was used for the isolation of four different human lymphoma and melanoma-specific CD4(+) and CD8(+) T-cell clones reactive against defined and undefined tumor antigens. Isolated tumor-specific T-cell clones could be expanded to large numbers ex vivo while maintaining phenotype, function, and tumor antigen specificity. The method was simple, efficient, and reproducible, and may have potential application for the development of adoptive immunotherapeutic strategies.
Lee
Seung-Tae
ST
Department of Lymphoma and Myeloma, The University of Texas M D Anderson Cancer Center, Houston, TX 77030, USA.
Liu
Shujuan
S
Radvanyi
Laszlo
L
Sukhumalchandra
Pariya
P
Molldrem
Jeffrey J
JJ
Wieder
Eric D
ED
Hwu
Patrick
P
Liu
Yong-Jun
YJ
Kwak
Larry W
LW
Lizée
Gregory
G
Neelapu
Sattva S
SS
D000951:Antigens, Neoplasm / Q000276:immunology*; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology*; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology*; D002460:Cell Line; D002469:Cell Separation / Q000379:methods*; D002999:Clone Cells / Q000276:immunology; D005434:Flow Cytometry / Q000379:methods*; D006801:Humans; D016219:Immunotherapy, Adoptive; D008223:Lymphoma / Q000276:immunology; D058965:MART-1 Antigen; D008545:Melanoma / Q000276:immunology; D009363:Neoplasm Proteins / Q000276:immunology; D016693:Receptors, Antigen, T-Cell, alpha-beta / Q000276:immunology*
D000951:Antigens, Neoplasm; D058965:MART-1 Antigen; C548667:MLANA protein, human; D009363:Neoplasm Proteins; D016693:Receptors, Antigen, T-Cell, alpha-beta