Biological activity of dendritic cells generated from cord blood CD34+ hematopoietic progenitors in IL-7- and IL-13-conditioned cultures.
BACKGROUND
Dendritic cells (DCs) are required for initiation of the immune response and may therefore be used for the production of cancer vaccines. As mature DCs (mDCs) are the most potent antigen-presenting cells, there is increasing interest in generating them ex vivo. The present study was designed to obtain mDCs from CD34+ hematopoietic progenitors by culturing them in different media.


METHODS
Cord blood CD34+ hematopoietic progenitors were expanded for 7 days in FST medium containing fms-related tyrosine kinase 3 ligand (Flt3-L), stem cell factor (SCF), and thrombopoietin (TPO). Then the cells were divided into three parts and cultured for 21 days in different media: FST medium or FST enriched in interleukin (IL)-3 (FST3 medium) or supplemented with IL-7 and IL-13 (FST713 medium). At the end of culture part of the cells was harvested, counted, and analyzed while the other part was matured with proinflammatory cytokines for 2 days. The cells' phenotypes, ability to induce proliferation of allogeneic lymphocytes in the mixed lymphocyte reaction (allo-MLR), chemotaxis, phagocytosis, and O2(-) production were determined.


RESULTS
The average fold increase of DCs at the end of culture in FST medium was 127, in FST3 1043, and in FST713 71. In comparison with the other media, FST713 medium supported the generation of mDCs that were characterized by higher expression of CD83, costimulatory molecules, and HLA-DR, enhanced ability to induce allo-MLR and migration to -macrophage inflammatory protein (MIP) 3beta poor phagocytosis, and O2(-) production.


CONCLUSIONS
This study indicates that FST713 medium allows the generation of limited numbers of more mature DCs, while FST3 medium leads to the production of immature DCs in high numbers.
Mytar
Bozenna
B
Department of Clinical Immunology, Polish-American Institute of Pediatrics, Jagiellonian University Medical College, Cracow, Poland.
Stec
Małgorzata
M
Weglarczyk
Kazimierz
K
Zembala
Marek
M
D002454:Cell Differentiation / Q000187:drug effects; D002478:Cells, Cultured / Q000187:drug effects; D054415:Chemokine CCL19 / Q000494:pharmacology; D002633:Chemotaxis / Q000187:drug effects; D003470:Culture Media / Q000494:pharmacology; D003713:Dendritic Cells / Q000276:immunology*; D005312:Fetal Blood / Q000166:cytology*; D006684:HLA-DR Antigens / Q000096:biosynthesis; D006412:Hematopoietic Stem Cells / Q000166:cytology* / Q000187:drug effects; D006801:Humans; D016130:Immunophenotyping; D007231:Infant, Newborn; D007377:Interleukin-3 / Q000494:pharmacology*; D015851:Interleukin-7 / Q000494:pharmacology*; D008213:Lymphocyte Activation; D007959:Lymphocyte Culture Test, Mixed; D008565:Membrane Proteins / Q000494:pharmacology; D010587:Phagocytosis; D054400:Receptors, CCR7 / Q000096:biosynthesis / Q000235:genetics; D019089:Stem Cell Factor / Q000494:pharmacology; D013481:Superoxides / Q000378:metabolism; D013926:Thrombopoietin / Q000494:pharmacology
C515370:CCR7 protein, human; D054415:Chemokine CCL19; D003470:Culture Media; D006684:HLA-DR Antigens; D007377:Interleukin-3; D015851:Interleukin-7; D008565:Membrane Proteins; D054400:Receptors, CCR7; D019089:Stem Cell Factor; C084881:flt3 ligand protein; D013481:Superoxides; D013926:Thrombopoietin