Northwest Biotherapeutics, Inc.

Pacific Northwest Cancer Foundation

Benjamin A Tjoa

Marnix L Bosch

Gerald P Murphy

Alton L Boynton

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 CD14and expressed high levels of CD1. Upon maturation by contact with an agent such as, for example, BCG and IFN&#x3b3;, 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.

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

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 IFN&#x3b3;, 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.

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 CD14and expressed high levels of CD1a. Upon maturation by contact with an agent such as, for example, BCG and IFN&#x3b3;, 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.

Methods and compositions for use of human dendritic cells to activate T cells for immunotherapeutic responses against primary and metastatic prostate cancer are disclosed. In one embodiment, human dendritic cells, after exposure to a prostate cancer antigen or specific antigenic peptide, are administered to a prostate cancer patient to activate the relevant T cell responses in vivo. In an alternate embodiment, human dendritic cells are exposed to a prostate cancer antigen or specific antigenic peptide in vitro and incubated or cultured with primed or unprimed T cells to activate the relevant T cell responses in vitro. The activated T cells are then administered to a prostate cancer patient. Methods and compositions for human dendritic cells with extended life span and cryopreserved dendritic cells are disclosed.

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Seattle, WA, United States of America

Benjamin A Tjoa