title: A minimal human immunostimulatory CpG motif that potently induces IFN-gamma and IFN-alpha production.
abstract: Recent reports have shown that immunostimulatory sequences (ISS) containing CpG motifs have minimal length requirements (>/=12 bases) for the exertion of immune-enhancing function upon mammalian cells. Herein we demonstrate that short ISS (5-7 bases), which exhibit no activity on their own, induce IFN-gamma and IFN-alpha secretion from human peripheral blood mononuclear cells when adsorbed to the surface of cationic poly(D,L-lactide-co-glycolide) microparticles (cPLGA). Utilizing this technique, we discovered a minimal ISS sequence for induction of IFN-gamma and IFN-alpha from human cells: 5'-TCGXX-3'. These short ISS/cPLGA formulations targeted PDC in similar fashion to longer ISS ODN, the activity of which does not require (but is enhanced by) cPLGA. PDC stimulated with short ISS/cPLGA responded with enhanced uptake of ISS and elevated production of cytokines, including IFN-alpha. However, ISS-responsive B cells did not respond to short ISS/cPLGA, underlining the plasmacytoid dendritic cell selectivity of this formulation. These results describe a novel technique for formulating active, but very short, ISS oligodeoxynucleotide that allows for the dissection and characterization of minimal immunostimulatory CpG motifs.
authors.affiliation: Dynavax Technologies Corporation, Berkeley, CA 94710, USA.
authors.forename: Karen
authors.initials: K
authors.lastname: Fearon
authors.forename: Jason D
authors.initials: JD
authors.lastname: Marshall
authors.forename: Christi
authors.initials: C
authors.lastname: Abbate
authors.forename: Sandhya
authors.initials: S
authors.lastname: Subramanian
authors.forename: Priscilla
authors.initials: P
authors.lastname: Yee
authors.forename: Josh
authors.initials: J
authors.lastname: Gregorio
authors.forename: Glen
authors.initials: G
authors.lastname: Teshima
authors.forename: Gary
authors.initials: G
authors.lastname: Ott
authors.forename: Stephen
authors.initials: S
authors.lastname: Tuck
authors.forename: Gary
authors.initials: G
authors.lastname: Van Nest
authors.forename: Robert L
authors.initials: RL
authors.lastname: Coffman
mesh_terms: D000276:Adjuvants, Immunologic / Q000494:pharmacology*; D001402:B-Lymphocytes / Q000187:drug effects / Q000276:immunology; D001483:Base Sequence; D003713:Dendritic Cells / Q000187:drug effects / Q000276:immunology; D005786:Gene Expression Regulation / Q000187:drug effects; D006801:Humans; D066298:In Vitro Techniques; D016898:Interferon-alpha / Q000096:biosynthesis* / Q000235:genetics; D007371:Interferon-gamma / Q000096:biosynthesis* / Q000235:genetics; D019344:Lactic Acid; D007963:Leukocytes, Mononuclear / Q000187:drug effects / Q000276:immunology; D009838:Oligodeoxyribonucleotides / Q000235:genetics / Q000494:pharmacology*; D011100:Polyglycolic Acid; D000077182:Polylactic Acid-Polyglycolic Acid Copolymer; D011108:Polymers
chemical_list: D000276:Adjuvants, Immunologic; C408982:CPG-oligonucleotide; D016898:Interferon-alpha; D009838:Oligodeoxyribonucleotides; D011108:Polymers; D000077182:Polylactic Acid-Polyglycolic Acid Copolymer; D011100:Polyglycolic Acid; D019344:Lactic Acid; D007371:Interferon-gamma