Background: Allergen-specific subcutaneous immunotherapy (SCIT) is an antigen-specific therapy of IgE-mediated allergies. In the present study, we analyze the epitope specificities of antibody responses induced by SCIT with allergen extracts from pollen of trees belonging to the order Fagales (birch, alder, hazel) adsorbed onto aluminum hydroxide. Methods: The IgE, IgG1–4 and IgA responses to defined recombinant allergens (birch pollen: Bet v 1; alder pollen: Aln g 1; hazel pollen: Cor a 1; apple: Mal d 1) as well as to Bet v 1-derived recombinant fragments and synthetic peptides were analyzed in sera from patients who had undergone SCIT for different periods of time. Results: Long-term SCIT (>1 year; cumulative dose >1,000,000 SQ units) induced more pronounced IgG1, IgG2 and IgG4 responses to Bet v 1 and Bet v 1-related allergens according to the degree of sequence homology (Bet v 1>Aln g 1>Cor a 1>Mal d 1) than short-term SCIT (<1 year; cumulative dose <1,000,000 SQ units). In contrast to patients treated for <1 year, patients treated for >1 year mounted distinct IgG1, IgG2 and IgG4 responses against sequential Bet v 1 epitopes. No relevant allergen-specific IgA or IgG3 responses were induced by short- or long-term SCIT. Using a competitive ELISA assay, it could be shown that serum IgG from patients undergoing long-term SCIT inhibited IgE reactivity to Bet v 1 better than IgG from patients undergoing short-term SCIT. Conclusion: SCIT with allergen extracts adsorbed onto aluminum hydroxide induces IgG responses against new epitopes that block IgE binding and cross-react with structurally related allergens depending, among other factors, on duration of treatment and cumulative injected dose.

Keywords:
Epitopes, Immunotherapy, Peptides, Recombinant allergens
1.
Mothes N, Horak F, Valenta R: Transition from a botanical to a molecular classification in tree pollen allergy: implications for diagnosis and therapy. Int Arch Allergy Immunol 2004;135:357–373.
2.
Moverare R, Westritschnig K, Svensson M, Hayek B, Bende M, Pauli G, Sorva R, Haahtela T, Valenta R, Elfman L: Different IgE reactivity profiles in birch pollen-sensitive patients from six European populations revealed by recombinant allergens: an imprint of local sensitization. Int Arch Allergy Immunol 2002;128:325–335.
3.
Breiteneder H, Pettenburger K, Bito A, Valenta R, Kraft D, Rumpold H, Scheiner O, Breitenbach M: The gene coding for the major birch pollen allergen Bet v 1 is highly homologous to a pea disease resistance response gene. EMBO J 1989;8:1935–1938.
4.
Kazemi-Shirazi L, Pauli G, Purohit A, Spitzauer S, Froschl R, Hoffmann-Sommergruber K, Breiteneder H, Scheiner O, Kraft D, Valenta R: Quantitative IgE inhibition experiments with purified recombinant allergens indicate pollen-derived allergens as the sensitizing agents responsible for many forms of plant food allergy. J Allergy Clin Immunol 2000;105:116–125.
5.
Fritsch R, Bohle B, Vollmann U, Wiedermann U, Jahn-Schmid B, Krebitz M, Breiteneder H, Kraft D, Ebner C: Bet v 1, the major birch pollen allergen, and Mal d 1, the major apple allergen, cross-react at the level of allergen-specific T helper cells. J Allergy Clin Immunol 1998;102:679–686.
6.
Petersen BN, Janniche H, Munch EP, Wihl JA, Bowadt H, Ipsen H, Lowenstein H: Immunotherapy with partially purified and standardized tree pollen extracts. I. Clinical results from a three-year double-blind study of patients treated with pollen extracts either of birch or combinations of alder, birch and hazel. Allergy 1988;43:353–362.
7.
Wihl JA, Ipsen H, Petersen BN, Munch EP, Janniche H, Lowenstein H: Immunotherapy with partially purified and standardized tree pollen extracts. II. Results of skin prick tests and nasal provocation tests from a three-year double-blind study of patients treated with pollen extracts either of birch or combinations of alder, birch and hazel. Allergy 1988;43:363–369.
8.
Ipsen H, Schwartz B, Wihl JA, Petersen BN, Munch EP, Janniche H, Lowenstein H: Immunotherapy with partially purified and standardized tree pollen extracts. III. Specific IgE response to the major allergens of alder, birch and hazel pollen during immunotherapy. Allergy 1988;43:370–377.
9.
Möller C, Dreborg S: Cross-reactivity between deciduous trees during immunotherapy. I. In vivo results. Clin Allergy 1986;16:135–143.
10.
Bodtger U, Poulsen LK, Jacobi HH, Malling HJ: The safety and efficacy of subcutaneous birch pollen immunotherapy – a one-year, randomised, double-blind, placebo-controlled study. Allergy 2002;57:297–305.
11.
Arvidsson MB, Löwhagen O, Rak S: Effect of 2-year placebo-controlled immunotherapy on airway symptoms and medication in patients with birch pollen allergy. J Allergy Clin Immunol 2002;109:777–783.
12.
Pauli G, Larsen TH, Rak S, Horak F, Pastorello E, Valenta R, Purohit A, Arvidsson M, Kavina A, Schroeder JW, Mothes N, Spitzauer S, Montagut A, Galvain S, Melac M, Andre C, Poulsen LK, Malling HJ: Efficacy of recombinant birch pollen vaccine for the treatment of birch-allergic rhinoconjunctivitis. J Allergy Clin Immunol 2008;122:951–960.
13.
Reisinger J, Horak F, Pauli G, van Hage M, Cromwell O, Konig F, Valenta R, Niederberger V: Allergen-specific nasal IgG antibodies induced by vaccination with genetically modified allergens are associated with reduced nasal allergen sensitivity. J Allergy Clin Immunol 2005;116:347–354.
14.
Niederberger V, Horak F, Vrtala S, Spitzauer S, Krauth MT, Valent P, Reisinger J, Pelzmann M, Hayek B, Kronqvist M, Gafvelin G, Gronlund H, Purohit A, Suck R, Fiebig H, Cromwell O, Pauli G, van Hage-Hamsten M, Valenta R: Vaccination with genetically engineered allergens prevents progression of allergic disease. Proc Natl Acad Sci USA 2004;101(suppl 2):14677–14682.
15.
Focke M, Linhart B, Hartl A, Wiedermann U, Sperr WR, Valent P, Thalhamer J, Kraft D, Valenta R: Non-anaphylactic surface-exposed peptides of the major birch pollen allergen, Bet v 1, for preventive vaccination. Clin Exp Allergy 2004;34:1525–1533.
16.
Vrtala S, Hirtenlehner K, Vangelista L, Pastore A, Eichler HG, Sperr WR, Valent P, Ebner C, Kraft D, Valenta R: Conversion of the major birch pollen allergen, Bet v 1, into two nonanaphylactic T cell epitope-containing fragments: candidates for a novel form of specific immunotherapy. J Clin Invest 1997;99:1673–1681.
17.
Vrtala S, Susani M, Sperr WR, Valent P, Laffer S, Dolecek C, Kraft D, Valenta R: Immunologic characterization of purified recombinant timothy grass pollen (Phleum pratense) allergens (Phl p 1, Phl p 2, Phl p 5). J Allergy Clin Immunol 1996;97:781–787.
18.
Loveless MH: Immunological studies of pollinosis. I. The presence of two antibodies related to the same pollen-antigen in the serum of treated hay-fever patients. J Immunol 1940;38:25–50.
19.
Breiteneder H, Ferreira F, Reikerstorfer A, Duchene M, Valenta R, Hoffmann-Sommergruber K, Ebner C, Breitenbach M, Kraft D, Scheiner O: Complementary DNA cloning and expression in Escherichia coli of Aln g 1, the major allergen in pollen of alder (Alnus glutinosa). J Allergy Clin Immunol 1992;90:909–917.
20.
Breiteneder H, Ferreira F, Hoffmann-Sommergruber K, Ebner C, Breitenbach M, Rumpold H, Kraft D, Scheiner O: Four recombinant isoforms of Cor a 1, the major allergen of hazel pollen, show different IgE-binding properties. Eur J Biochem 1993;212:355–362.
21.
Vanek-Krebitz M, Hoffmann-Sommergruber K, Laimer da Camara Machado M, Susani M, Ebner C, Kraft D, Scheiner O, Breiteneder H: Cloning and sequencing of Mal d 1, the major allergen from apple (Malus domestica), and its immunological relationship to Bet v 1, the major birch pollen allergen. Biochem Biophys Res Commun 1995;214:538–551.
22.
Mothes N, Heinzkill M, Drachenberg KJ, Sperr WR, Krauth MT, Majlesi Y, Semper H, Valent P, Niederberger V, Kraft D, Valenta R: Allergen-specific immunotherapy with a monophosphoryl lipid A-adjuvanted vaccine: reduced seasonally boosted immunoglobulin E production and inhibition of basophil histamine release by therapy-induced blocking antibodies. Clin Exp Allergy 2003;33:1198–1208.
23.
Zwollo P, Ehrlich-Kautzky E, Scharf SJ, Ansari AA, Erlich HA, Marsh DG: Sequencing of HLA-D in responders and nonresponders to short ragweed allergen, Amb a V. Immunogenetics 1991;33:141–151.
24.
Mari A, Wallner M, Ferreira F: Fagales pollen sensitization in a birch-free area: a respiratory cohort survey using Fagales pollen extracts and birch recombinant allergens (rBet v 1, rBet v 2, rBet v 4). Clin Exp Allergy 2003;33:1419–1428.
25.
Ferreira F, Hawranek T, Gruber P, Wopfner N, Mari A: Allergic cross-reactivity: from gene to the clinic. Allergy 2004;59:243–267.
26.
Gastaminza G, Algorta J, Audicana M, Etxenagusia M, Fernandez E, Munoz D: Systemic reactions to immunotherapy: influence of composition and manufacturer. Clin Exp Allergy 2003;33:470–474.
27.
Grier TJ, Hazelhurst DM, Duncan EA, West TK, Esch RE: Major allergen measurements: sources of variability, validation, quality assurance, and utility for laboratories, manufacturers, and clinics. Allergy Asthma Proc 2002;23:125–131.
28.
Focke M, Marth K, Flicker S, Valenta R: Heterogeneity of commercial timothy grass pollen extracts. Clin Exp Allergy 2008;38:1400–1408.
29.
Bolhaar ST, Tiemessen MM, Zuidmeer L, van Leeuwen A, Hoffmann-Sommergruber K, Bruijnzeel-Koomen CA, Taams LS, Knol EF, van Hoffen E, van Ree R, Knulst AC: Efficacy of birch-pollen immunotherapy on cross-reactive food allergy confirmed by skin tests and double-blind food challenges. Clin Exp Allergy 2004;34:761–769.
30.
Bucher X, Pichler WJ, Dahinden CA, Helbling A: Effect of tree pollen specific, subcutaneous immunotherapy on the oral allergy syndrome to apple and hazelnut. Allergy 2004;59:1272–1276.
31.
Niederberger V, Reisinger J, Valent P, Krauth MT, Pauli G, van Hage M, Cromwell O, Horak F, Valenta R: Vaccination with genetically modified birch pollen allergens: immune and clinical effects on oral allergy syndrome. J Allergy Clin Immunol 2007;119:1013–1016.
32.
Pree I, Reisinger J, Focke M, Vrtala S, Pauli G, van Hage M, Cromwell O, Gadermaier E, Egger C, Reider N, Horak F, Valenta R, Niederberger V: Analysis of epitope-specific immune responses induced by vaccination with structurally folded and unfolded recombinant Bet v 1 allergen derivatives in man. J Immunol 2007;179:5309–5316.
33.
Focke M, Marth K, Valenta R: Molecular composition and biological activity of commercial birch pollen allergen extracts. Eur J Clin Invest 2009;39:429-436.
34.
Vrtala S, Ball T, Spitzauer S, Pandjaitan B, Suphioglu C, Knox B, Sperr WR, Valent P, Kraft D, Valenta R: Immunization with purified natural and recombinant allergens induces mouse IgG1 antibodies that recognize similar epitopes as human IgE and inhibit the human IgE-allergen interaction and allergen-induced basophil degranulation. J Immunol 1998;160:6137–6144.
35.
Orlandi A, Grasso F, Corinti S, Marinaro M, Bonura A, Boirivant M, Colombo P, Di Felice G: The recombinant major allergen of Parietaria judaica and its hypoallergenic variant: in vivo evaluation in a murine model of allergic sensitization. Clin Exp Allergy 2004;34:470–477.
36.
Jutel M, Jaeger L, Suck R, Meyer H, Fiebig H, Cromwell O: Allergen-specific immunotherapy with recombinant grass pollen allergens. J Allergy Clin Immunol 2005;116:608–613.
37.
Flicker S, Valenta R: Renaissance of the blocking antibody concept in type I allergy. Int Arch Allergy Immunol 2003;132:13–24.
38.
van Neerven RJ, Wikborg T, Lund G, Jacobsen B, Brinch-Nielsen A, Arnved J, Ipsen H: Blocking antibodies induced by specific allergy vaccination prevent the activation of CD4+ T cells by inhibiting serum-IgE-facilitated allergen presentation. J Immunol 1999;163:2944–2952.
39.
Valenta R, Ball T, Focke M, Linhart B, Mothes N, Niederberger V, Spitzauer S, Swoboda I, Vrtala S, Westritschnig K, Kraft D: Immunotherapy of allergic disease. Adv Immunol 2004;82:105–153.
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2009
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