Generation of tumor-specific T lymphocytes using dendritic cell/tumor fusions and anti-CD3/CD28.
Adoptive immunotherapy with tumor-specific T cells represents a promising treatment strategy for patients with malignancy. However, the efficacy of T-cell therapy has been limited by the ability to expand tumor-reactive cells with an activated phenotype that effectively target malignant cells. We have developed an anticancer vaccine in which patient-derived tumor cells are fused with autologous dendritic cells (DCs), such that a wide array of tumor antigens are presented in the context of DC-mediated costimulation. In this study, we demonstrate that DC/tumor fusions induce T cells that react with tumor and are dramatically expanded by subsequent ligation of the CD3/CD28 costimulatory complex. These T cells exhibit a predominantly activated phenotype as manifested by an increase in the percentage of cells expressing CD69 and interferon gamma. In addition, the T cells upregulate granzyme B expression and are highly effective in lysing autologous tumor targets. Targeting of tumor-specific antigen was demonstrated by the expansion of T cells with specificity for the MUC1 tetramer. Stimulation with anti-CD3/CD28 followed by DC/tumor fusions or either agent alone failed to result in a similar expansion of tumor-reactive T cells. Consistent with these findings, spectratyping analysis demonstrates selective expansion of T-cell clones as manifested by considerable skewing of the Vbeta repertoire following sequential stimulation with DC/tumor fusions and anti-CD3/CD28. Gene expression analysis was notable for the upregulation of inflammatory pathways. These findings indicate that stimulation with DC/tumor fusions provides a unique platform for subsequent expansion with anti-CD3/CD28 in adoptive T-cell therapy of cancer.
Beth Israel Deaconess Medical Center, Boston, MA 02215, USA. jrosenb1@bidmc.harvard.edu
Jacalyn
J
Rosenblatt
Zekui
Z
Wu
Baldev
B
Vasir
Corrine
C
Zarwan
Richard
R
Stone
Heidi
H
Mills
Thea
T
Friedman
Panagiotis A
PA
Konstantinopoulos
Dimitrios
D
Spentzos
Musie
M
Ghebremichael
Kristen
K
Stevenson
Donna
D
Neuberg
James D
JD
Levine
Robin
R
Joyce
Dimitrios
D
Tzachanis
Vassiliki
V
Boussiotis
Donald
D
Kufe
David
D
Avigan
D017951:Antigen Presentation / Q000276:immunology; D015703:Antigens, CD / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology; D000943:Antigens, Differentiation / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology; D019496:Cancer Vaccines; D002459:Cell Fusion; D049109:Cell Proliferation; D002999:Clone Cells; D003602:Cytotoxicity, Immunologic; D003713:Dendritic Cells / Q000276:immunology / Q000378:metabolism* / Q000473:pathology; D019674:Genes, T-Cell Receptor beta; D053804:Granzymes / Q000096:biosynthesis* / Q000235:genetics; D006801:Humans; D007371:Interferon-gamma / Q000378:metabolism; D008213:Lymphocyte Activation; D018396:Mucin-1 / Q000276:immunology / Q000378:metabolism; D013601:T-Lymphocytes / Q000276:immunology / Q000378:metabolism* / Q000473:pathology
D015703:Antigens, CD; D000943:Antigens, Differentiation; D019496:Cancer Vaccines; C060192:MUC1 protein, human; D018396:Mucin-1; D007371:Interferon-gamma; D053804:Granzymes