title: Modulation of immunogenicity and allergenicity by controlling the number of immunostimulatory oligonucleotides linked to Amb a 1.
abstract: BACKGROUND
  Immunostimulatory DNA sequences (ISS) are potent immunomodulators that can drive T(H)1 responses to antigens or allergens. This effect can be dramatically enhanced by direct linkage of ISS to the protein.
  
  
  OBJECTIVE
  Evaluate the effects of the number of ISS bound to the major ragweed allergen Amb a 1 on immunogenicity and allergenicity.
  
  
  METHODS
  Immunogenicity in mice and allergenicity using PBMC or sera from subjects with ragweed allergy were assayed.
  
  
  RESULTS
  Both antibody induction in vivo and antibody recognition in vitro were highly sensitive to the number of ISSs linked. IgE recognition of Amb a 1 in competitive ELISA or histamine release assays was inhibited by ISS linkage and showed an inverse relationship to the number of ISSs bound. Type and magnitude of antibody induction in mice was also highly dependent on the number of ISS bound. At the highest ISS to protein ratios, antibody induction was very low. Moderate ISS to protein ratios induced high antibody responses in which IgG(2a) generally predominated. Low ISS to protein ratios produced the highest overall antibody responses in which IgG(1) predominated. In contrast, varied ISS to protein ratios did not affect T-cell responses. In both in vivo mouse studies and in vitro human PBMC studies, all ISS to protein ratios evaluated induced similar responses represented by high levels of IFN-gamma and low levels of T(H)2 cytokines.
  
  
  CONCLUSIONS
  Controlling the number of ISS bound to a protein allows manipulation of antibody recognition and induction while retaining the potent T(H)1 properties of an ISS-linked protein.
  
  
  CONCLUSIONS
  Immunostimulatory DNA sequence-linked Amb a 1 conjugate represents a safe, novel therapeutic approach for treating ragweed allergy.
authors.affiliation: Dynavax Technologies Corp., 2929 Seventh Street, Berkeley, CA 94710, USA. dhiggins@dvax.com
authors.forename: Deborah
authors.initials: D
authors.lastname: Higgins
authors.forename: Roberto
authors.initials: R
authors.lastname: Rodriguez
authors.forename: Robert
authors.initials: R
authors.lastname: Milley
authors.forename: Jason
authors.initials: J
authors.lastname: Marshall
authors.forename: Christi
authors.initials: C
authors.lastname: Abbate
authors.forename: Tracy
authors.initials: T
authors.lastname: dela Cruz
authors.forename: Kathryn
authors.initials: K
authors.lastname: Patton
authors.forename: Fiona
authors.initials: F
authors.lastname: Walker
authors.forename: Kristin
authors.initials: K
authors.lastname: Chichester
authors.forename: Joseph
authors.initials: J
authors.lastname: Eiden
authors.forename: Stephen
authors.initials: S
authors.lastname: Tuck
authors.forename: Gary
authors.initials: G
authors.lastname: Van Nest
mesh_terms: D000276:Adjuvants, Immunologic / Q000494:pharmacology; D000485:Allergens / Q000276:immunology / Q000494:pharmacology*; D000818:Animals; D000906:Antibodies / Q000276:immunology*; D052179:Antigens, Plant; D002478:Cells, Cultured; D016207:Cytokines / Q000096:biosynthesis / Q000276:immunology; D005260:Female; D006636:Histamine Release; D006801:Humans; D006967:Hypersensitivity / Q000276:immunology / Q000628:therapy; D007114:Immunization; D007073:Immunoglobulin E / Q000276:immunology; D007963:Leukocytes, Mononuclear / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D009841:Oligonucleotides / Q000494:pharmacology*; D010940:Plant Proteins / Q000276:immunology / Q000494:pharmacology*
chemical_list: D000276:Adjuvants, Immunologic; D000485:Allergens; C026967:Amb a I protein, Ambrosia artemisiifolia; D000906:Antibodies; D052179:Antigens, Plant; D016207:Cytokines; D009841:Oligonucleotides; D010940:Plant Proteins; D007073:Immunoglobulin E