Generation of antigen-presenting cells using cultured dendritic cells and amplified autologous tumor mRNA.
Novel antigen-presenting cells (APCs) were generated using cultured dendritic cells (DCs) and amplified tumor mRNA, and the potential of tumor antigen-reactive T cell induction by the tumor RNA-introduced DCs (DC/tumor RNA) was analyzed in a patient with melanoma antigen-encoding gene (MAGE3)-positive malignant melanoma of the esophagus. DCs were generated from an adherent fraction of peripheral blood mononuclear cells in the presence of granulocyte macrophage colony-stimulating factor and interleukin-4. Tumor mRNA was purified from tumor tissue, amplified in vitro using a T7 RNA polymerase system, and then introduced into DCs by electroporation (150 V/150 microF or 100 V/200 microF). The gene introduction efficiency was 44-55% as measured by enhanced green fluorescent protein reporter gene expression, and the viability of RNA-introduced DCs was approximately 80%. DC/tumor RNA could induce tumor antigen-reactive cytotoxic T lymphocytes (CTLs) in an mRNA-specific manner, but had no effect on the self-antigen-reactive T cells. DC/tumor RNA could induce the polyspecific antigen-reactive CTL responses mediated by both human leukocyte antigen class I and class II molecules, whereas MAGE3 peptide-pulsed DCs induced only the monospecific MAGE3-reactive CTL responses mediated by human leukocyte antigen class I molecules, showing the superiority of the DC/tumor RNA over the DC/peptide. It is suggested that the use of DC/tumor RNA as antigen-presenting cells may be more effective, convenient and practical for the DC-based anti-cancer immunotherapy.
Minami
Kazuhito
K
Department of Surgical Oncology, Research Institute for Radiation Biology and Medicine, Hiroshima University, Japan.
Yamaguchi
Yoshiyuki
Y
Ohshita
Akiko
A
Kawabuchi
Yoshiharu
Y
Ohta
Koji
K
Hihara
Jun
J
Toge
Tetsuya
T
D000938:Antigen-Presenting Cells / Q000166:cytology*; D000951:Antigens, Neoplasm / Q000737:chemistry / Q000378:metabolism; D002448:Cell Adhesion; D018929:Cell Culture Techniques / Q000379:methods*; D012321:DNA-Directed RNA Polymerases / Q000378:metabolism; D003713:Dendritic Cells / Q000166:cytology*; D018274:Electroporation; D004797:Enzyme-Linked Immunosorbent Assay; D004938:Esophageal Neoplasms / Q000276:immunology* / Q000378:metabolism; D005434:Flow Cytometry; D017930:Genes, Reporter; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006801:Humans; D007371:Interferon-gamma / Q000378:metabolism; D015847:Interleukin-4 / Q000378:metabolism; D007963:Leukocytes, Mononuclear / Q000378:metabolism; D008545:Melanoma / Q000276:immunology* / Q000378:metabolism; D009363:Neoplasm Proteins / Q000378:metabolism; D010455:Peptides / Q000737:chemistry; D012313:RNA / Q000737:chemistry / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D012334:RNA, Neoplasm / Q000378:metabolism*; D013601:T-Lymphocytes / Q000378:metabolism; D014158:Transcription, Genetic; D014764:Viral Proteins / Q000378:metabolism
D000951:Antigens, Neoplasm; C072013:MAGEA3 protein, human; D009363:Neoplasm Proteins; D010455:Peptides; D012333:RNA, Messenger; D012334:RNA, Neoplasm; D014764:Viral Proteins; D049452:Green Fluorescent Proteins; D015847:Interleukin-4; D012313:RNA; D007371:Interferon-gamma; D016178:Granulocyte-Macrophage Colony-Stimulating Factor; C062210:bacteriophage T7 RNA polymerase; D012321:DNA-Directed RNA Polymerases