Platform of dendritic cell (DC)-based vaccination
View Patent ↗The present invention discloses novel dendritic cell maturation-inducing cytokine cocktails, and methods for inducting type-1 polarized dendritic cells in serum-free conditions which enhance the desirable properties of DC1s generated in serum-supplemented cultures. The invention further discloses methods and systems using IFNγ and other ligands of the IFNγ receptor, in combination with IFNα (or other type I interferons), poly I:C, and other IFNα (and IFNβ) inducers to enhance the IL-12-producing properties of dendritic cells. More specifically, the present invention discloses type-1 polarized dendritic cells that have a unique combination of a fully-mature status and an elevated, instead of “exhausted”, ability to produce IL-12p70. allows for the generation of fully-mature DC1s in serum-free AIM-V medium. The invention discloses systems that use the foregoing products and methods to facilitate the clinical application of DC1-based vaccines and the identification of novel factors involved in the induction of Th1 and CTL responses by DC1.
1. A method for producing activated T lymphocytes or NK cells, comprising
producing mature dendritic cells; and
contacting the mature dendritic cells with T lymphocytes or NK cells, thereby producing activated T lymphocytes or NK cells, respectively,
wherein producing dendritic cells comprises adding, in vitro, a) an effective amount of interferon (IFN)-α or IFN-β; and b) an effective amount of IFN-γ to a population of immature dendritic cells from a subject of interest; and culturing the immature dendritic cells in vitro to produce mature dendritic cells, wherein after subsequent stimulation of the mature dendritic cells with CD40 ligand, said mature dendritic cells produce more IL-12p70 as compared to immature dendritic cells stimulated with CD40 ligand.
2. The method of claim 1 , wherein the method comprises adding a) 0.1 pg/ml to 10 mg/ml of IFN-α or IFN-β and b) 0.1 pg/ml to 10 mg/ml of IFN-γ to the population of immature dendritic cells.
3. The method of claim 1 , wherein the method comprises adding a) 0.1 pg/ml to 10 mg/ml of IFN-α and b) 0.1 pg/ml to 10 mg/ml of IFN-γ to the population of immature dendritic cells.
4. The method of claim 1 , wherein the method further comprises adding an effective amount of lipopolysaccharide (LPS) or LPS analog to the immature dendritic cells in vitro.
5. The method of claim 1 , wherein the method further comprises adding an effective amount of interleukin-1β, tumor necrosis factor (TNF)-α or both to the immature dendritic cells in vitro.
6. The method of claim 1 , comprising adding an effective amount of a) IFN-α or IFN-β b) an effective amount of IFN-γ, and c) an effective amount of poly-I:C to the immature dendritic cells in vitro.
7. The method of claim 1 , wherein the T lymphocytes are CD4+ T cells.
8. The method of claim 1 , wherein the immature dendritic cells are cultured in a serum-free medium.
9. The method of claim 1 , wherein the dendritic cells produce IL-12p70 at least 10-fold higher than dendritic cells from the subject of interest matured in the presence of IL-1β, TNF-α, IL-6 and prostaglandin (PG) E 2 .
10. The method of claim 1 , wherein the method comprises adding 25 ng/ml IL-1β, 50 ng/ml TNF-α, 1000 U/ml IFN-γ, 20 μg/ml poly-I:C, and 3000 U/ml IFN-α to the immature dendritic cells.
11. The method of claim 1 , comprising adding an effective amount of a) IFN-α or IFN-β b) an effective amount of IFN-γ, and c) an effective amount of poly-I:C to the immature dendritic cells in vitro.
12. The method of claim 1 , wherein the method produces activated T lymphocytes and wherein the method further comprises adding an effective amount of lipopolysaccharide (LPS) or LPS analog to the immature dendritic cells in vitro.