PLATFORM OF DENDRITIC CELL (DC)-BASED VACCINATION
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 . An isolated population of mature dendritic cells, wherein the mature dendritic cells in the population express CD83, CD56 and CCR7 on their cell surface and produce IL-12p70 after stimulation with CD40 ligand.
2 . The isolated population of mature dendritic cells of claim 1 , produced by the process of:
adding in vitro a) an effective amount of interferon (IFN)-α, IFN-β, poly-I:C or combination thereof and b) an effective amount of IFN-γ or both to a population of immature dendritic cells from a subject of interest; and
culturing the immature dendritic cells in vitro,
thereby producing the population of mature dendritic cells.
3 . The isolated population of mature dendritic cells of claim 1 , wherein the process 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.
4 . The isolated population of mature dendritic cells of claim 1 , wherein the process comprises 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.
5 . The isolated population of mature dendritic cells of claim 1 , wherein the process further comprises adding an effective amount of lippopolysaccharide (LPS).
6 . The isolated population of mature dendritic cells of claim 1 , wherein the process further comprises adding an effective amount of interleukin-1β, tumor necrosis factor (TNF)-α or both to the immature dendritic cells.
7 . The isolated population of mature dendritic cells of claim 1 , wherein the mature dendritic cells produce more IL-12p70 after stimulation with CD40 ligand compared to immature dendritic cells from a subject of interest.
8 . The isolated population of mature dendritic cells of claim 1 , wherein the mature dendritic cells produce IL-12p70 at least 10-fold higher than dendritic cells from a subject of interest matured in the presence of IL-1β, TNF-α, IL-6 and prostaglandin (PG) E 2 .
9 . The isolated population of mature dendritic cells of claim 2 , wherein the process comprises 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.
10 . An isolated population of mature dendritic cells produced by a process comprising
obtaining a sample of peripheral blood from a subject; and
isolating immature dendritic cells from the peripheral blood of the subject;
contacting the immature dendritic cells with a) an effective amount of exogenous interferon (IFN)-α or IFN-β and b) an effective amount of exogenous IFN-γ, poly-I:C or both; and
contacting the immature dendritic cells with an antigen of interest that binds the Major Histocompatibility Complex (MHC),
thereby producing mature antigen presenting cells that present the antigen of interest.
11 . The isolated population of mature dendritic cells of claim 10 , wherein the process comprises wherein the antigen is tumor-specific or specific for an infectious agent.
12 . The isolated population of mature dendritic cells of claim 10 , wherein the process comprises wherein the method comprises contacting immature dendritic cells with 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.
13 . The isolated population of mature dendritic cells of claim 10 , wherein the process comprises contacting mononuclear cells with 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.
14 . The isolated population of mature dendritic cells of claim 1 , wherein the mature dendritic cells migrate in response to a CCR7 ligand.