title: The influence of antigen targeting to sub-cellular compartments on the anti-allergic potential of a DNA vaccine.
abstract: BACKGROUND
  Gene vaccines offer attractive rationales for prophylactic as well as therapeutic treatments of type I allergies. DNA and mRNA vaccines have been shown to prevent from allergic sensitization and to counterbalance established allergic immune reactions. Recent advances in gene vaccine manipulation offer additional opportunities for modulation of T helper cell profiles by specific targeting of cellular compartments.
  
  
  METHODS
  DNA vaccines encoding the major birch pollen allergen Bet v 1.0101 were equipped with different leader sequences to shuttle the antigen to lysosomes (LIMP-II), to trigger cellular secretion (hTPA), or to induce proteasomal degradation via forced ubiquitination (ubi). Mice were pre-vaccinated with these constructs and the protective efficacy was tested by subcutaneous Th2-promoting challenges, followed by allergen inhalation. IgG antibody subclass distribution and allergen-specific IgE as well as cytokine profiles from re-stimulated splenocytes and from BALFs were assessed. The cellular composition of BALFs, and lung resistance and compliance were determined.
  
  
  RESULTS
  Immunization with all targeting variants protected from allergic sensitization, i.e. IgE induction, airway hyperresponsiveness, lung inflammation, and systemic and local Th2 cytokine expression. Surprisingly, protection did not clearly correlate with the induction of a systemic Th1 cytokine profile, but rather with proliferating CD4+ CD25+ FoxP3+ T regulatory cells in splenocyte cultures. Targeting the allergen to proteasomal or lysosomal degradation severely down-regulated antibody induction after vaccination, while T cell responses remained unaffected. Although secretion of antigen promoted the highest numbers of Th1 cells, this vaccine type was the least efficient in suppressing the establishment of an allergic immune response.
  
  
  CONCLUSIONS
  This comparative analysis highlights the modulatory effect of antigen targeting on the resulting immune response, with a special emphasis on prophylactic anti-allergy DNA vaccination. Targeting the antigen to proteasomal or lysosomal degradation reduces the availability of native allergen, thereby rendering the vaccine hypoallergenic without compromising efficacy, an important feature for a therapeutic setting.
authors: Department of Molecular Biology, University of Salzburg, 5020 Salzburg, Austria.
authors: Esther E
authors: EE
authors: Weinberger
authors: Almedina
authors: A
authors: Isakovic
authors: Sandra
authors: S
authors: Scheiblhofer
authors: Christina
authors: C
authors: Ramsauer
authors: Katrin
authors: K
authors: Reiter
authors: Cornelia
authors: C
authors: Hauser-Kronberger
authors: Josef
authors: J
authors: Thalhamer
authors: Richard
authors: R
authors: Weiss
mesh_terms: D000485:Allergens / Q000235:genetics / Q000276:immunology; D000818:Animals; D018926:Anti-Allergic Agents / Q000276:immunology* / Q000493:pharmacokinetics*; D029662:Betula; D016207:Cytokines / Q000378:metabolism; D005260:Female; D006967:Hypersensitivity / Q000276:immunology / Q000517:prevention & control*; D007073:Immunoglobulin E / Q000097:blood; D007074:Immunoglobulin G / Q000097:blood; D007963:Leukocytes, Mononuclear / Q000276:immunology; D008168:Lung / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D021381:Protein Transport; D013154:Spleen / Q000276:immunology; D019444:Vaccines, DNA / Q000276:immunology* / Q000493:pharmacokinetics*
chemical_list: D000485:Allergens; D018926:Anti-Allergic Agents; D016207:Cytokines; D007074:Immunoglobulin G; D019444:Vaccines, DNA; D007073:Immunoglobulin E
keywords: 20-amino-acid C-terminal tail of lysosomal integral membrane protein-II; AA; AHR; APC; BAL; BALF; DNA vaccine; ER-targeting; Immunotherapy; LIMP-II; LIMPII; SIT; T helper; T regulatory; Th; Treg; Type I allergy; Ubi; Ubiquitination; airway hyperreactivity; amino acid; antigen presenting cell; bronchoalveolar lavage; bronchoalveolar lavage fluid; human tissue plasminogen activator leader peptide; specific immunotherapy; tPA; ubiquitin
references: J Infect Dis. 2006 Jan 15;193(2):205-13
references: 16362884
references: Curr Drug Targets Inflamm Allergy. 2005 Oct;4(5):585-97
references: 16248827
references: Int Arch Allergy Immunol. 1999 Sep;120(1):17-29
references: 10529585
references: J Virol. 1998 Jun;72(6):5174-81
references: 9573289
references: J Neuroinflammation. 2012 Jun 22;9:139
references: 22727044
references: Eur J Immunol. 2013 Sep;43(9):2421-9
references: 23722868
references: Int Arch Allergy Immunol. 2006;139(4):332-45
references: 16534215
references: Infect Immun. 2000 Jun;68(6):3097-102
references: 10816449
references: Immunol Lett. 2009 Jan 29;122(1):68-75
references: 19111573
references: J Allergy Clin Immunol. 2009 Nov;124(5):1070-7.e1-11
references: 19665781
references: Clin Exp Allergy. 2003 Sep;33(9):1198-208
references: 12956739
references: J Immunol. 2012 Sep 15;189(6):2995-3006
references: 22908332
references: Infect Immun. 2000 Oct;68(10):5914-9
references: 10992502
references: Vaccine. 1999 Dec 10;18(9-10):815-24
references: 10580194
references: J Clin Invest. 2003 Jul;112(1):22-4
references: 12840054
references: Allergy. 2006 Jul;61(7):828-35
references: 16792580
references: J Allergy Clin Immunol. 2011 Feb;127(2):509-516.e1-5
references: 21281875
references: Nat Med. 1996 May;2(5):540-4
references: 8616712
references: Adv Exp Med Biol. 2009;647:156-73
references: 19760073
references: J Biol Chem. 1994 Mar 4;269(9):6622-31
references: 7509809
references: J Allergy Clin Immunol. 2009 Apr;123(4):911-6
references: 19348926
references: Vaccine. 2011 Mar 3;29(11):2140-8
references: 21215346
references: J Virol. 2001 Nov;75(21):10421-30
references: 11581410
references: Eur J Immunol. 2004 Sep;34(9):2480-8
references: 15307180
references: J Cell Biochem. 2006 May 15;98(2):235-42
references: 16440328
references: J Biol Chem. 1994 Feb 18;269(7):5210-7
references: 8106503
references: J Intern Med. 2012 Aug;272(2):144-57
references: 22640224
references: J Allergy Clin Immunol. 2006 Jul;118(1):269-76
references: 16815165
references: ScientificWorldJournal. 2007 Jun 12;7:987-99
references: 17619780
references: Methods. 2004 Mar;32(3):328-39
references: 14962768
references: Nat Methods. 2011 Sep 29;8(10):785-6
references: 21959131
references: Am J Respir Crit Care Med. 2011 May 1;183(9):1153-63
references: 21297073
references: J Mol Cell Biol. 2012 Feb;4(1):3-10
references: 22147847
references: J Immunol. 1997 Dec 15;159(12):6037-43
references: 9550402
references: Ann Allergy Asthma Immunol. 2008 May;100(5):463-8
references: 18517079
references: PLoS One. 2007 Oct 03;2(10):e984
references: 17912361
references: Allergy. 2012 Dec;67(12):1511-8
references: 23004934
references: Mol Immunol. 2010 Jun;47(10):1914-22
references: 20392496
references: Proc Natl Acad Sci U S A. 2010 Oct 5;107(40):17292-7
references: 20855629
references: Immunity. 2007 Jan;26(1):79-92
references: 17182262
references: Hum Gene Ther. 1999 Nov 20;10(17):2727-40
references: 10584920
references: J Allergy Clin Immunol. 2006 Sep;118(3):734-41
references: 16950295
references: Curr Opin Allergy Clin Immunol. 2009 Feb;9(1):50-4
references: 19532092
references: Clin Exp Allergy. 2003 Feb;33(2):218-25
references: 12580915
references: J Exp Med. 2002 Jul 15;196(2):237-46
references: 12119348
references: J Allergy Clin Immunol. 1999 Jan;103(1 Pt 1):107-13
references: 9893193
references: PLoS One. 2012;7(9):e45916
references: 23029313
references: Recent Pat DNA Gene Seq. 2010 Jun;4(2):122-31
references: 20550512
references: J Biol Chem. 1989 May 5;264(13):7715-9
references: 2540202
references: Nat Immunol. 2005 Nov;6(11):1123-32
references: 16200070