title: PMA induces vaccine adjuvant activity by the modulation of TLR signaling pathway.
abstract: Toll-like receptor (TLR) ligands are being developed for use as vaccine adjuvants and as immunomodulators because of their ability to stimulate innate and adaptive immune responses. Flagellin, a TLR5 ligand, was reported to show potent mucosal vaccine adjuvant activity. To identify ligands that potentiate the adjuvant activity of flagellin, we screened a plant library using HEK293T cells transiently cotransfected with phTLR5 and pNF- κ B-SEAP plasmids. The 90% EtOH extract from Croton tiglium showed significant NF- κ B transactivation in a TLR5-independent manner along with the increase of a flagellin activity. We have studied to characterize an active component from Croton tiglium and to elucidate the action mechanisms. Phorbol 12-myristate 13-acetate (PMA) was isolated as an active component of Croton tiglium by activity-guided fractionation, column chromatography, HPLC, NMR, and MS. PMA at a range of nM induced PKC-dependent NF- κ B activation and IL-8 production in both TLR5- and TLR5+ assay systems. In in vivo mouse vaccination model, PMA induced antigen-specific IgG and IgA antibody responses and increased IL-12 production corresponding to T cell responses in spleen lymphocytes. These results suggest that PMA would serve as an efficacious mucosal vaccine adjuvant.
authors.affiliation: Jeollanamdo Institute of Natural Resources Research, Jangheung, Jeollanamdo 529-851, Republic of Korea.
authors.forename: Dool-Ri
authors.initials: DR
authors.lastname: Oh
authors.affiliation: Jeollanamdo Institute of Natural Resources Research, Jangheung, Jeollanamdo 529-851, Republic of Korea.
authors.forename: Hu Won
authors.initials: HW
authors.lastname: Kang
authors.affiliation: Korea INSPharm Inc. 1065, Dong-myeon, Hwasun, Jeollanamdo 519-882, Republic of Korea.
authors.forename: Jong-Ro
authors.initials: JR
authors.lastname: Kim
authors.affiliation: Jeollanamdo Institute of Natural Resources Research, Jangheung, Jeollanamdo 529-851, Republic of Korea.
authors.forename: Sunoh
authors.initials: S
authors.lastname: Kim
authors.affiliation: Department of Biomedical Science and BK21 PLUS Center for Creative Biomedical Scientists, Chonnam National University Medical School, Gwangju 501-757, Republic of Korea.
authors.forename: In-Kyu
authors.initials: IK
authors.lastname: Park
authors.affiliation: Clinical Vaccine R&D Center, Chonnam National University Medical School, Gwangju 501-757, Republic of Korea.
authors.forename: Joon Haeng
authors.initials: JH
authors.lastname: Rhee
authors.affiliation: Korea Bioactive Natural Material Bank, Research Institute of Pharmaceutical Sciences, College of Pharmacy, Seoul National University, Seoul 151-742, Republic of Korea.
authors.forename: Won Keun
authors.initials: WK
authors.lastname: Oh
authors.affiliation: Department of Pharmaceutical Engineering, Dongshin University, Naju, Jeollanamdo 520-724, Republic of Korea.
authors.forename: Young Ran
authors.initials: YR
authors.lastname: Kim
mesh_terms: D000276:Adjuvants, Immunologic / Q000737:chemistry*; D000818:Animals; D018938:Caco-2 Cells; D002467:Cell Nucleus / Q000378:metabolism; D029802:Croton / Q000737:chemistry; D003600:Cytosol / Q000378:metabolism; D005408:Flagellin / Q000737:chemistry; D057809:HEK293 Cells; D006801:Humans; D016209:Interleukin-8 / Q000378:metabolism; D008024:Ligands; D008214:Lymphocytes / Q000166:cytology; D051379:Mice; D008807:Mice, Inbred BALB C; D016328:NF-kappa B / Q000378:metabolism; D010936:Plant Extracts / Q000737:chemistry; D015398:Signal Transduction; D013154:Spleen / Q000166:cytology; D013601:T-Lymphocytes / Q000276:immunology; D013755:Tetradecanoylphorbol Acetate / Q000737:chemistry*; D051198:Toll-Like Receptor 5 / Q000378:metabolism; D051193:Toll-Like Receptors / Q000378:metabolism*; D015533:Transcriptional Activation; D014612:Vaccines
chemical_list: D000276:Adjuvants, Immunologic; D016209:Interleukin-8; D008024:Ligands; D016328:NF-kappa B; D010936:Plant Extracts; C495346:TLR5 protein, human; D051198:Toll-Like Receptor 5; D051193:Toll-Like Receptors; D014612:Vaccines; D005408:Flagellin; D013755:Tetradecanoylphorbol Acetate