Mature type-1 polarized dendritic cells with enhanced IL-12 production and methods of serum-free production and use

Abstract

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.

Description

[0001] This application claims priority from U.S. application Ser. No. 60 / 468,760, filed May 8, 2003, entitled “MATURE TYPE-1 POLARIZED DENDRITIC CELLS WITH ENHANCED IL-12 PRODUCTION AND METHODS OF SERUM-FREE PRODUCTION AND USE”.STATEMENT AS TO RIGHTS TO INVENTIONS MADE UNDER FEDERALLY SPONSORED RESEARCH AND DEVELOPMENT [0002] Portions of the present invention were made with support of the United States Government via a grant from the National Cancer Institute under grant number 1R01CA82016. The U.S. Government may therefore have certain rights in the invention.BACKGROUND [0003] 1. Field of the Invention [0004] The invention relates to methods of generating mature dendritic cells with enhanced IL-12 production and compositions and systems for such dendritic cells. [0005] 2. Discussion of the Background [0006] Dendritic cells (DCs), the most potent antigen presenting cells, are effective inducers of protective immunity against infectious diseases and cancer (Banchereau & Steinman 199...

AI Technical Summary

Benefits of technology

[0019] The present invention discloses novel dendritic cell (“DC”) maturation-inducing cytokine cocktails, and means for inducting type-1 polarized dendritic cells (“DC1s”) in serum-free conditions which enhance the desirable properties of DC1s generated in serum-supplemented cultures. The invention further discloses the use of IFNγ and other ligands of the IFNγ receptor, in combination with IFNα (or other type I interferons, such as IFNβ, known to bind to the same receptor), poly I:C, and other IFNα (and IFNβ) inducers to enhance the IL-12-producing properties of DCs. The invention also discloses the use of DC1s to induce Ag-specific T cells against tumors, intracellular pathogens, and atopic allergens for active and passive immunotherapy, immunomonitoring and research purposes. More specifically, the present invention discloses type-1 polarized DCs (DC1s) that have a unique combination of a fully-mature status and an elevated, instead of “exhausted”, ability to produce IL-12p70. These properties allow these DC1s to selectively induce high-intensity Th1-, CTL-, and NK cell-mediated type-1 immune responses, including those desirable in the treatment of cancer. Another preferred embodiment of the present invention shows that the inclusion of IFNα and / or poly-I:C to the “classical” DC1-inducing cocktail (TNFα / IL-1β / IFNγ) allows for the generation of fully-mature DC1s in serum- free AIM-V medium. In other preferred embodiments, the present invention discloses serum-free protocols of DC1 generation that facilitate the clinical application of DC1-based therapies and the identification of novel factors involved in the induction of Th1-, CTL-, and NK cell responses by DC1.

Problems solved by technology

Unfortunately, the maturation stage of DCs obtained in the currently-available protocols inversely correlates with their ability to produce IL-12p70 (Kalinski et al., 1999, Langenkamp et al., 2000), the cytokine with powerful anticancer Th1-and CTL-inducing properties (Trinchieri, 1998b); (Shurin et al., 1997).

However, use of IL-12 transduced DC in humans creates substantial logistic problems.

It also carries potential risks associated with the administration of genetically-manipulated material and the risks of direct IL-12 toxicity and of deregulating the immune system due to uncontrolled IL-12 production.

Specifically, none of the patents disclose or teach the combination of type I and type II interferons (such as IFNα and IFNγ), as a part of the cytokine cocktail used to produce fully mature DCs with high IL-12 producing capacity.

Thus, despite the efforts of many, the desirable combination of high immunostimulatory activity with a high capacity to produce IL-12p70 could not be attained by all previous DC-based vaccines which have employed either mature DCs exhibiting high stimulatory / low IL-12-secreting functions or immature DCs that display low stimulatory / high IL-12 secretion functions.

Unfortunately, the original DC1-inducing cytokine cocktail, composed of IL-1β, TNFα, and IFN-γ (Vieira et al 2000), does not allow for the induction of DC1s in serum-free media, which is desirable for clinical application.

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