Generation of dendritic cells from patients with chronic myelogenous leukemia.
Dendritic cells (DCs) are professional antigen-presenting cells (APCs) specialized to internalize, process, and present antigen. They have the capacity to stimulate the primary immune response of resting T-cells. We generated DCs from the adherent cell fraction of peripheral blood, as well as from purified CD34+ cells from CML patients. Characterizing DCs from ten CML patients by flow cytometry, we found that these cells are highly positive for HLA-DR, CD1a, CD23, and CD80 and negative for CD14, CD15, and CD16. The yield of DCs ranged from 19.5 to 68%. In addition, we used a functional test of FITC-dextran uptake to verify that early DCs take up large particles (0.5-3 microm) by macropinocytosis while monocytes do not. FITC-dextran uptake was detected by flow cytometry, showing that DCs had accumulated these fluorescent particles. Electron-microscopic analysis showed no major morphological differences between normal and CML-derived DCs. Furthermore, cultured DCs were isolated by FAC sorting for CD1a and HLA-DR expression. In these highly purified cells the Ph chromosome was detected by interphase fluorescence in situ hybridization (FISH) and by fluorescence immunophenotyping and interphase cytogenetics as a tool for the investigation of neoplasms (FICTION); 30-85% of DCs generated were Ph-chromosome positive. It might therefore be possible not only to prime T-cells with bcr/abl-specific synthetic peptides, but also to stimulate T-cells directly with Ph-positive DCs. Use of DCs might serve as a novel therapeutic approach in CML patients, due to their ability to induce highly specific T-cell responses in an autologous system.
Heinzinger
M
M
Department of Hematology/Oncology, University Medical Center Freiburg, Germany.
Waller
C F
CF
von den Berg
A
A
Rosenstiel
A
A
Lange
W
W
D000328:Adult; D015703:Antigens, CD / Q000032:analysis; D002452:Cell Count; D002455:Cell Division; D003713:Dendritic Cells / Q000166:cytology* / Q000276:immunology / Q000378:metabolism; D003911:Dextrans; D004705:Endocytosis; D005260:Female; D005434:Flow Cytometry; D016650:Fluorescein-5-isothiocyanate; D006684:HLA-DR Antigens / Q000032:analysis; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D015464:Leukemia, Myelogenous, Chronic, BCR-ABL Positive / Q000097:blood / Q000473:pathology*; D007959:Lymphocyte Culture Test, Mixed; D008297:Male; D008854:Microscopy, Electron; D008875:Middle Aged; D010641:Phenotype
D015703:Antigens, CD; D003911:Dextrans; D006684:HLA-DR Antigens; D016650:Fluorescein-5-isothiocyanate