title: Protamine nanoparticles with CpG-oligodeoxynucleotide prevent an allergen-induced Th2-response in BALB/c mice.
abstract: The currently applied immunotherapy of type I allergy with aluminum hydroxide (alum) as adjuvant elicits - among other side effects - an initial IgE-boost. In contrast, CpG-oligodeoxynucleotides (ODNs) drive the immune response toward Th1. The biodegradable material protamine can spontaneously form nanoparticles together with such ODNs. Our aim was to investigate the immune response induced by protamine-based nanoparticles (proticles) with CpG-ODN as an allergen delivery system. Proticles complexed with Ara h 2 extracted from raw peanuts as model allergen were injected subcutaneously into naïve BALB/c mice. Ara h 2-specific antibodies were analyzed by ELISA and rat basophilic leukemia (RBL) cell assay. Cytokine levels were investigated in supernatants of stimulated splenocytes. The in vivo distribution after subcutaneous injection was examined via fluorescence imaging. BMDCs were stimulated with proticles, and expression of stimulation and maturation markers as well as cytokines in supernatants was investigated. A favorable increase in Ara h 2-specific IgG2a antibodies was found after immunization with proticles-Ara h 2, whereas Ara h 2-specific IgE was not detectable. Accordingly, the ratio of IL-5/IFN-gamma was low in this group. Granuloma formation was completely absent at injection sites of proticles. The distribution of Ara h 2 after subcutaneous injection was markedly decelerated when complexed to proticles. Stimulation of BMDCs with proticles-Ara h 2 caused upregulation of CD11c and CD80 as well as an increased IL-6 production. Our data suggest that biodegradable protamine-based nanoparticles with CpG-ODN counteract the Th2-dominated immune response induced by an allergen and therefore are suitable as novel carrier system for immunotherapy of allergy.
authors.affiliation: Messerli Research Institute of the University of Veterinary Medicine Vienna, Medical University of Vienna, University of Vienna, Vienna, Austria; Department of Pathophysiology and Allergy Research, Medical University of Vienna, Vienna, Austria. Electronic address: isabella.pali@meduniwien.ac.at.
authors.forename: Isabella
authors.initials: I
authors.lastname: Pali-Schöll
authors.forename: Helen
authors.initials: H
authors.lastname: Szöllösi
authors.forename: Philipp
authors.initials: P
authors.lastname: Starkl
authors.forename: Bernhard
authors.initials: B
authors.lastname: Scheicher
authors.forename: Caroline
authors.initials: C
authors.lastname: Stremnitzer
authors.forename: Alexander
authors.initials: A
authors.lastname: Hofmeister
authors.forename: Franziska
authors.initials: F
authors.lastname: Roth-Walter
authors.forename: Anna
authors.initials: A
authors.lastname: Lukschal
authors.forename: Susanne C
authors.initials: SC
authors.lastname: Diesner
authors.forename: Andreas
authors.initials: A
authors.lastname: Zimmer
authors.forename: Erika
authors.initials: E
authors.lastname: Jensen-Jarolim
mesh_terms: D000485:Allergens / Q000276:immunology; D000536:Aluminum Hydroxide / Q000276:immunology; D000818:Animals; D001491:Basophils / Q000276:immunology; D016207:Cytokines / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D007074:Immunoglobulin G / Q000276:immunology; D007167:Immunotherapy / Q000379:methods; D007279:Injections, Subcutaneous; D015850:Interleukin-6 / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D053758:Nanoparticles; D009838:Oligodeoxyribonucleotides / Q000008:administration & dosage / Q000494:pharmacology*; D011479:Protamines / Q000737:chemistry*; D051381:Rats; D018418:Th2 Cells / Q000276:immunology*
chemical_list: D000485:Allergens; C408982:CPG-oligonucleotide; D016207:Cytokines; D007074:Immunoglobulin G; D015850:Interleukin-6; D009838:Oligodeoxyribonucleotides; D011479:Protamines; D000536:Aluminum Hydroxide
keywords: 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid); ABTS; Allergen delivery system; Allergy; Alum; BMDCs; BSA; CpG; DCs; DMSO; EDTA; ELISA; FDA; IFN; IL; Immunotherapy; ODNs; Oligodeoxynucleotides; PBS; Protamine-based nanoparticles; Proticles; RBL; RT; SDS–PAGE; STDV; T helper; TBST; TLR; TNF; Th; aluminum hydroxide; bone marrow derived dendritic cells; bovine serum albumin; cytosine phosphate guanine; dendritic cells; dimethyl sulfoxide; enzyme-linked immunosorbent assay; ethylene-diamine tetra-acetic acid; food and drug administration; interferon; interleukin; o.n.; oligodeoxynucleotides; overnight; phosphate-buffered saline; rat basophilic leukemia; room temperature; sodium dodecyl sulfate polyacrylamide gel electrophoresis; standard deviation; toll-like receptor; tris-buffered saline with 0*05% Tween®; tumor necrosis factor