Retroviral transduction of human dendritic cells with a tumor-associated antigen gene.
Dendritic cells (DCs) are potent antigen-presenting cells that can activate quiescent T lymphocytes. When pulsed with tumor-associated antigen (TAA) peptide or protein, murine DCs can provide antitumor immunity. We reasoned that DCs retrovirally transduced with TAA genes might have important advantages over peptide- or protein-pulsed DCs, including long-term TAA presentation in vivo, and presentation of important but undefined epitopes. Therefore, we attempted to retrovirally transduce human DCs with a melanoma TAA gene (MART-1) and determine whether these transduced DCs could raise a specific antitumor response from quiescent autologous T lymphocytes. After retroviral transduction, human CD34+ cells were differentiated into DCs in vitro using granulocyte macrophage colony-stimulating factor, tumor necrosis factor alpha, and stem cell factor. This method consistently yielded a population of DCs as analyzed by morphology, phenotype, and MLR. Flow cytometric analysis revealed that 22-28% of cells expressing the DC phenotype also expressed a transduced marker gene. When DCs were transduced with the gene encoding MART-1, they stimulated much higher levels of cytokine release by MART-1-specific tumor-infiltrating lymphocytes than control DCs transduced with an irrelevant gene. In vitro stimulation using MART-1-transduced DCs but not control-transduced DCs raised specific antitumor CTLs from autologous quiescent T cells. These results provide evidence that human DCs can be retrovirally transduced with a TAA gene and that these transduced cells can raise a specific antitumor immune response in vitro. Transduced DCs may be useful for in vivo immunization against TAA.
Reeves
M E
ME
Surgery Branch, National Cancer Institute, NIH, Bethesda, Maryland 20892, USA.
Royal
R E
RE
Lam
J S
JS
Rosenberg
S A
SA
Hwu
P
P
D000951:Antigens, Neoplasm / Q000235:genetics* / Q000276:immunology; D003713:Dendritic Cells / Q000276:immunology*; D000939:Epitopes / Q000235:genetics* / Q000276:immunology; D005822:Genetic Vectors / Q000235:genetics* / Q000276:immunology; D006801:Humans; D007111:Immunity, Cellular; D016246:Lymphocytes, Tumor-Infiltrating / Q000276:immunology*; D008545:Melanoma / Q000276:immunology* / Q000628:therapy; D012190:Retroviridae / Q000235:genetics*; D014162:Transfection / Q000235:genetics / Q000379:methods*
D000951:Antigens, Neoplasm; D000939:Epitopes