Generation of dendritic cells from monocytic dendritic precursor cells with GM-CSF in the absence of additional cytokines

摘要

The present invention it was determined that dendritic cells could be derived from various sources including peripheral blood monocytes in the presence of only GM-CSF without other cytokines if the monocytes were not activated. By preventing activation, such as by preventing binding of the cells to the surface of the culture vessel, the monocytes do not require the presence of additional cytokines, such as IL-4 or IL-13, to prevent differentiation into a non-dendritic cell lineage. The immature DCs generated and maintained in this manner were CD14− and expressed high levels of CD1a. Upon maturation by contact with an agent such as, for example, BCG and IFNγ, the cells were determined to express surface molecules typical of mature dendritic cells purified by prior methods and cultured in the presence of GM-CSF and IL-4. The mature dendritic cells produced from monocytes without activation and cultured in GM-CSF alone are suitable for use in dendritic cell-based immunotherapy methods, such as for use in the treatment of disease, including cancer.

主权项

1. A method for differentiating human monocytic dendritic cell precursors (CD14+CD1a−) into immature dendritic cells having increased CD1a and having no CD14 on their cell surface, comprising:
a) providing a cell population comprising non-activated human monocytic dendritic cell precursors (CD 14+CD1a−); b) contacting the non-activated human dendritic cell precursors in a culture vessel with a dendritic cell culture media supplemented with at least 1% animal or human protein and an effective amount of granulocyte-macrophage colony stimulating factor in the absence of additional cytokines for a time period sufficient for the human monocytic dendritic cell precursors to differentiate into immature dendritic cells having no expression of CD14 and having increased expression of CD1a on their cell surface.