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Development of Inducible Pattern Recognition Receptors and A Unified Dendritic Cell Enhancement Vector
Author(s) -
Narayanan Priyadharshini,
Spencer David M
Publication year - 2008
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.22.1_supplement.1076.17
Subject(s) - microbiology and biotechnology , receptor , pattern recognition receptor , dendritic cell , trif , chemistry , toll like receptor , cd40 , biology , innate immune system , in vitro , immunology , immune system , biochemistry , cytotoxic t cell
Recognition of pathogen‐associated molecular patterns (PAMPS) via pattern recognition receptors (PRRs) on the surface of dendritic cells (DCs), results in their activation and maturation. PRRs include cell surface Toll‐like receptors (TLRs) and intracellular receptors including RNA helicases and NOD‐like receptors. Stimulation with a TLR4 ligand, in conjunction with CD40 co‐stimulation, leads to potent activation, maturation and migration of human monocyte‐derived DCs. To circumvent drawbacks associated with the use and clinical availability of synthetic TLR4 ligands, we set out to develop a simplified, unified vector consisting of an adjuvant‐responsive domain, namely, a TIR domain from a TLR and a CD40 costimulatory receptor domain. This chimeric receptor will enable optimal DC activation in vitro and in vivo using the chemically induced dimerization (CID) system, thus replacing the panel of activation reagents, currently in use to activate DCs in vitro . Toward our goal, we have developed several inducible PRRs. We are now comparing the efficacy of iPRRs using reporter assays that monitor downstream activation of transcription factors such as NF‐κB, IRF3 and IRF7 in various cell lines. We observe that iRIG‐I and iNOD2 activate NF‐κB several fold, while iTLRs show weak induction and require additional optimization. In conclusion, iRIG‐I and iNOD2 are potent activators of NF‐κB making them ideal candidates for fusion with the CD40 domain to make the chimeric receptor. The chimeric receptor will be tested in mouse bone marrow‐derived dendritic cells and used to generate “enhanced DCs” tested for their efficacy in mouse tumor models. Funding: R01‐ CA120411