Method for stimulating the immune, inflammatory or neuroprotective response

#1. Field of the Invention

#The present invention relates to the stimulation and enhancement of the immune or inflammatory response, including the use of adjuvants to enhance immune response to a vaccine. The present invention also relates to treatment of injuries, diseases, disorders and conditions that result in neurodegeneration.

#2. Description of the Related Art

#FIGS. 1A-1H are graphs of the surface phenotype of DCs exposed to c-di-GMP. DCs were left untreated (Untr) or were treated with NaCl, c-di-GMP or LPS for 24 h. DCs were then analysed for expression of the indicated markers, HLA-DR (FIG. 1A), CD83 (FIG. 1B), CXCR4 (FIG. 1C), CCR7 (FIG. 1D), CD80 (FIG. 1E), CD86 (FIG. 1F), MR (FIG. 1G), and CD32 (FIG. 1H), by staining with PE- or FITC-conjugated mAbs. Isotype controls for direct stains exhibited mean fluorescence <5. Results are expresses as percentage of positive cells (□) and as mean fluorescence intensity (MFI, ▪).

#FIG. 2 is a graph showing endocytic activity of DCs exposed to c-di-GMP. DCs were left untreated (Untr) or were treated with NaCl, c-di-GMP or LPS for 24 h. Mannose receptor-mediated endocytosis was evaluated as the cellular uptake of FITC-dextran (DX) and measured using FACS. Results are expressed as percentage of positive cells.

#FIGS. 3A and 3B are graphs showing the effect of c-di-GMP on cytokine production. Analysis of cytokine supernatant concentration in NaCl (used as control) or c-di-GMP-treated immature (FIG. 3A) or mature (LPS-treated; FIG. 3B) DCs was determined by ELISA. Supernatants were harvested after 24 h of treatment and tested for TNF-α, IL-6, IL-1β, IL-10 and IL-12 Results are expressed as pg/ml. IL-12 concentration was undetectable in immature DC culture supernatants. iDC=immature DC; mDC=mature DC.

#The results shown in the Example presented hereinbelow show that c-di-GMP stimulates and activates DCs, T cells and the Th-1 response, up-regulates the expression of important costimulatory molecules and the proinflammatory response and show that c-di-GMP has neuroactivity and is neuroprotective against nerve damage.

#Stimulation or enhancement (immunostimulation) of the immune and inflammatory response can be achieved by exogenous cyclic di-GMP or cyclic dinucleotides and cyclic dinucleotide analogs. Accordingly, cyclic di-GMP and cyclic dinucleotide analogues thereof can also be used in the development of a drug platform technology against a variety of diseases and immunological and inflammatory diseases including but not limited to infectious disease, cancer, HIV and AIDS, rheumatoid arthritis, and Hodgkin's disease. Cyclic di-GMP and cyclic dinucleotide analogues thereof are also useful as immunotherapeutic agents against cancers and allergic reactions, and as a vaccine adjuvant (e.g. in DNA vaccines, live attenuated vaccines, killed vaccines). Cyclic di-GMP and cyclic dinucleotide analogues thereof are also useful in affecting neuroactivity and in the inhibition or treatment of various brain, nervous and neural disorders.

#Several chemotactic cytokines, or chemokines, inhibit HIV replication by blocking or down regulating chemokine receptors that serve as entry cofactors for the virus. The role of chemokine receptors in HIV pathogenesis has been the subject of intense interest.