Transgene expression in dendritic cells to induce antigen-specific cytotoxic T cells in healthy donors.
Immunization with specific tumor-associated antigen (Ag) (TAA)-pulsed dendritic cells (DC) has proven to be efficacious in a variety of animal models and is being investigated for the treatment of cancer patients. Use of DC pulsed with specific peptides or transfected with TAA genes has been a focused area of investigation for the induction of potent tumor and viral immune responses. In this study we demonstrate transgene expression, including expression of the MART-1 gene, in DC transfected with plasmid DNA and cationic liposome complexes. These transiently transfected DC, derived from healthy donor monocytes cultured with granulocyte macrophage colony-stimulating factor and interleukin-4, express the transgene and can stimulate naive CD8+ T cells to elicit an antitumor immune response. These cytotoxic T lymphocytes (CTL) were capable of recognizing both known and unknown TAA epitopes and were able to exhibit cytolytic activity against human histocompatibility leukocyte Ag-matched tumor cells expressing the Ag. In addition to their cytolytic function, the CTL displayed an oligoclonal T-cell receptor repertoire, indicating that the presented Ag induced alterations in the T-cell population. The ability to induce tumor-specific CTL in vitro using gene-modified DC transiently expressing TAAs demonstrates the potential use of these Ag-presenting cells to generate future in vivo cancer vaccine strategies.
Philip
R
R
RPR Gencell, Santa Clara, California 95054, USA. ramila.philip@rp-rorer.com
Brunette
E
E
Ashton
J
J
Alters
S
S
Gadea
J
J
Sorich
M
M
Yau
J
J
O'Donoghue
G
G
Lebkowski
J
J
Okarma
T
T
Philip
M
M
D000951:Antigens, Neoplasm; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology / Q000378:metabolism; D002272:Carcinoembryonic Antigen / Q000235:genetics* / Q000378:metabolism; D002478:Cells, Cultured; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics / Q000378:metabolism; D003713:Dendritic Cells / Q000166:cytology / Q000187:drug effects / Q000502:physiology*; D000229:Dependovirus / Q000235:genetics; D005805:Genes, MHC Class I; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000494:pharmacology; D006801:Humans; D015847:Interleukin-4 / Q000494:pharmacology; D008081:Liposomes; D058965:MART-1 Antigen; D008285:Major Histocompatibility Complex; D009000:Monocytes; D009363:Neoplasm Proteins / Q000235:genetics* / Q000378:metabolism; D011948:Receptors, Antigen, T-Cell / Q000187:drug effects / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology*; D014162:Transfection; D019076:Transgenes
D000951:Antigens, Neoplasm; D002272:Carcinoembryonic Antigen; D008081:Liposomes; D058965:MART-1 Antigen; C548667:MLANA protein, human; D009363:Neoplasm Proteins; D011948:Receptors, Antigen, T-Cell; D011994:Recombinant Proteins; D015847:Interleukin-4; D016178:Granulocyte-Macrophage Colony-Stimulating Factor; D015500:Chloramphenicol O-Acetyltransferase