Dendritic cells generated from acute myeloid leukemia (AML) blasts maintain the expression of immunogenic leukemia associated antigens.
Recently, the focus is on new specific immunotherapies for AML such as cellular therapies employing dendritic cells (DCs) generated from AML blasts. AML-DCs express constitutionally leukemia-associated antigens (LAAs) present in AML blasts they are generated from. Here we investigated whether the generation of AML-DCs would alter the expression level of LAAs. Moreover, we evaluated the presence of HLA and costimulatory molecules on AML blasts versus AML-DCs. Quantitative real-time polymerase chain reaction (PCR) was performed for the following LAAs: preferentially expressed antigen in melanoma (PRAME), the receptor for hyaluronic acid mediated motility (RHAMM/CD168), Wilms' tumor gene 1 (WT-1) and proteinase 3. The expression of HLA-ABC, HLA-DR, CD40, CD80, CD83 and CD86 was evaluated by flow cytometry. RHAMM protein expression was evaluated by immunocytochemistry, recognition of AML-DCs by PRAME epitope-specific T cells was evaluated in a chromium-release assay. Quantitative real-time PCR for AML-DCs versus AML blasts showed an alteration in mRNA expression of LAAs. An elevated PCR signal for PRAME was detected in 7/12 AML-DC preparations. 6/12 AML-DC preparations showed a significant upregulation of the PCR signal for RHAMM. A stronger WT-1 and proteinase-3 signal was observed in PCR for only 2/12 and 1/12 AML-DCs , respectively. All preparations showed a strong expression of at least one of the LAAs examined. As demonstrated by flow cytometry, AML-DCs strongly upregulated costimulatory molecules like CD40 and CD80 in comparison with AML blasts. AML-DCs tested positive for RHAMM protein. PRAME positive AML-DCs were recognized by specific T cells. AML-DCs might constitute a powerful tool in immunotherapy for AML. Real-time PCR allows a quick and quantitative assessment of immunologically relevant LAA expression with only 10(5) DCs and might be helpful for the decision whether the AML-DC vaccination strategy is favourable or not.
Li
Li
L
Third Department of Internal Medicine, University of Ulm, Robert-Koch-Street 8, 89081 Ulm, Germany.
Reinhardt
Peter
P
Schmitt
Anita
A
Barth
Thomas F E
TF
Greiner
Jochen
J
Ringhoffer
Mark
M
Döhner
Hartmut
H
Wiesneth
Markus
M
Schmitt
Michael
M
D000208:Acute Disease; D015703:Antigens, CD / Q000378:metabolism; D000951:Antigens, Neoplasm / Q000276:immunology* / Q000378:metabolism; D001752:Blast Crisis; D002470:Cell Survival / Q000276:immunology; D003713:Dendritic Cells / Q000276:immunology*; D016326:Extracellular Matrix Proteins / Q000378:metabolism; D006684:HLA-DR Antigens / Q000378:metabolism; D006801:Humans; D018960:Hyaluronan Receptors / Q000378:metabolism; D007951:Leukemia, Myeloid / Q000276:immunology* / Q000473:pathology; D053146:Myeloblastin; D012334:RNA, Neoplasm / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012697:Serine Endopeptidases / Q000378:metabolism; D013601:T-Lymphocytes / Q000276:immunology / Q000378:metabolism; D014407:Tumor Cells, Cultured; D025721:WT1 Proteins / Q000378:metabolism
D015703:Antigens, CD; D000951:Antigens, Neoplasm; D016326:Extracellular Matrix Proteins; D006684:HLA-DR Antigens; D018960:Hyaluronan Receptors; C404944:PRAME protein, human; D012334:RNA, Neoplasm; D025721:WT1 Proteins; C098946:hyaluronan-mediated motility receptor; D012697:Serine Endopeptidases; D053146:Myeloblastin