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

[0046] Alpha type-1 DC (αDC1) of the present invention have a superior ability to induce melanoma-specific CTL responses. To analyze their CTL-inducing activity, individual populations of DC1 (generated under serum-free, or serum-supplemented conditions), or serum-free control IL-1β/TNFα/IL-6/PGE2-matured cDC, were pulsed with melanoma-associated antigenic peptides, and used to sensitize autologous CD8+ T cells from HLA-A2+ melanoma patients. The long-term CD8+ T cell lines obtained by further expansion with autologous PBMCs were harvested and used as responder cells against T2 cells pulsed with individual peptides, or their combination. As shown in FIG. 8, each of the DC1s obtained in each of the protocols tested induced significantly higher numbers of melanoma specific (MART-1-, gp100-, and tyrosinase-specific) CD8+ T cells, when compared to the DCs matured in the presence of IL-1β/TNFα/IL-6/TNFα/IL-6 and PGE2 (“control DC”: Jonuleit et al, 1997). In addition, to being superior to control DCs in the induction high numbers of melanoma-specific CD8+ T cells, alpha-type-1 DC also proved superior in the induction of high cytotoxic activity against target cells pulsed with MAA peptides (FIG. 9).

[0047] In addition to being superior inducers of the cytolytic activity of tumor-specific CD8+ T cells (CTLs), αDC1 also proved superior in their ability to induce similar, cytolytic functions in CD4+ Th cells, and in isolated NK cells, allowing them to efficiently kill transformed cells (FIG. 10).

[0048] Of particular importance for their ability to function in vivo, as carriers of anticancer vaccines, αDC1 can effectively migrate in response to CCR7 ligands (FIG. 11A), known to be produced in the T cell areas of the lymph nodes and to be responsible for the local accumulation of immu

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

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