Inhibition of dendritic cell-driven regulatory T cell activation and potentiation of tumor antigen-specific T cell responses by interleukin-15 and MAP kinase inhibitor
The invention involves the discovery that if dendritic cells loaded with a tumor antigen are cultured in interleukin-15 (IL-15), or if T cells activated by the dendritic cells are cultured in IL-15, Treg activity that is specific for the tumor antigen is reduced. This reduction in Treg activity results in an increase in anti-tumor immune response. Another embodiment of the invention involves the discovery that incubating dendritic cells with a MAP kinase inhibitor in combination with IL-15 gives synergistic benefits when the dendritic cells are used to activate T cells. Dendritic cell and T cell compositions incubated with IL-15 or a MAP kinase inhibitor are provided.
1. A method of preparing a therapeutic composition comprising antigen-specific T cells or dendritic cells loaded with an antigen, the method comprising:
(1) culturing adherent peripheral blood mononuclear cell (PBMC) dendritic cell recursors ex vivo in the presence of GM-CSF, IL-4, IL-15 and a p38 MAP kinase inhibitor in amounts and for a time effective to differentiate said precursors into dendritic cells that function to reduce Treg activity and FoxP3 expression in T cells, and further comprising stimulating antigen-specific T cells ex vivo with said dendritic cells that have been loaded with an antigen; or
(2) culturing adherent PBMC dendritic cell precursors ex vivo in the presence of GM-CSF, IL-4, IL-15 and a p38 MAP kinase inhibitor in amounts and for a time effective to differentiate said precursors into dendritic cells that function to reduce Treg activity and Foxp3 expression in T cells contacted with the dendritic cells, and loading the dendritic cells with an antigen ex vivo.
2. The method of claim 1 wherein the method comprises:
(1) culturing adherent peripheral blood mononuclear cell (PBMC) dendritic cell recursors ex vivo in the presence of GM-CSF IL-4, IL-15 and a p38 MAP kinase inhibitor in amounts and for a time effective to differentiate said precursors into dendritic cells that function to reduce Treg activity and Foxp3 expression in T cells, and further comprising stimulating antigen-specific T cells ex vivo with said dendritic cells that have been loaded with an antigen.
3. The method of claim 2 wherein the method comprises
(1) culturing adherent peripheral blood mononuclear cell (PBMC) dendritic cell recursors ex vivo in the presence of GM-CSF IL-4, IL-15 and a p38 MAP kinase inhibitor in amounts and for a time effective to differentiate said precursors into dendritic cells that function to reduce Treg activity and to reduce Foxp3 expression at least 2-fold in T cells, and further comprising stimulating antigen-specific T cells ex vivo with said dendritic cells that have been loaded with an antigen.
4. The method of claim 1 wherein the method comprises:
(2) culturing adherent PBMC dendritic cell precursors ex vivo in the presence of GM-CSF, IL-4, IL-15 and a p38 MAP kinase inhibitor in amounts and for a time effective to differentiate said precursors into dendritic cells that function to reduce Treg activity and Foxp3 expression in T cells contacted with the dendritic cells, and loading the dendritic cells with an antigen ex vivo.
5. A method of reducing Treg activity of T cells comprising:
(1) culturing adherent peripheral blood mononuclear cell (PBMC) dendritic cell precursors ex vivo in the presence of GM-CSF, IL-4, IL-15 and a p38 MAP kinase inhibitor in amounts and for a time effective to differentiate said precursors into dendritic cells that function to reduce Treg activity and FoxP3 expression in T cells, and further comprising stimulating antigen-specific T cells ex vivo with said dendritic cells that have been loaded with an antigen.
6. The method of claim 5 wherein the p38 MAP kinase inhibitor is (RS)-{4-[5-(4-fluorophenyl)-2-methylsulfanyl-3H-imidazol-4-yl]pyridin-2-yl}-(1-phenylethyl)amine]; 2-(4-chlorophenyl)-4-(4-fluorphenyl)-5-pyridin-4-yl-1,2-dihydropyrazol-3-one; 4-(3-(4-chlorophenyl)-5-(1-methylpiperidin-4-yl)-1H-pyrazol-4-yl)pyrimidine; or N-(2-methoxy-4-thiomethyl)benzoyl-4-benzylpiperidine.