Nowadays, treatment of food allergy only considered the avoidance of the specific food. However, the possibility of cross-reactivity makes this practice not very effective. Immunotherapy may exhibit as a good alternative to food allergy treatment. The use of hypoallergenic molecules with reduced IgE binding capacity but with ability to stimulate the immune system is a promising tool which could be developed for immunotherapy. In this study, three mutants of Pru p 3, the principal allergen of peach, were produced based on the described mimotope and T cell epitopes, by changing the specific residues to alanine, named as Pru p 3.01, Pru p 3.02, and Pru p 3.03. Pru p 3.01 showed very similar allergenic activity as the wild type by in vitro assays. However, Pru p 3.02 and Pru p 3.03 presented reduced IgE binding with respect to the native form, by in vitro, ex vivo, and in vivo assays. In addition, Pru p 3.03 had affected the IgG4 binding capacity and presented a random circular dichroism, which was reflected in the nonrecognition by specific antibodies anti-Pru p 3. Nevertheless, both Pru p 3.02 and Pru p 3.03 maintained the binding to IgG1 and their ability to activate T lymphocytes. Thus, Pru p 3.02 and Pru p 3.03 could be good candidates for potential immunotherapy in peach-allergic patients. 1. Introduction IgE-mediated allergy is a hypersensitivity disease suffering from more than 25% of the population in industrialized countries. Currently, specific immunotherapy is the only allergen-specific approach for its treatment and for preventing its progression to severe manifestations [1, 2]. The administration of increasing doses of allergen extracts to patients is the method most commonly applied. However, the use of crude extracts has several disadvantages. It could induce severe anaphylactic side reactions [3] or lead to sensitization towards new allergens present in the mixture [4, 5]. Different strategies have been designed to try to overcome these negative effects [6], as the use of allergen-derived B cell peptides [7, 8], allergen-derived T cell epitope containing peptides [9, 10] or vaccination with allergen-encoding DNA [11, 12]. In food allergy, immunotherapy is not commonly used due to the negative side effects, although several studies have been performed [13, 14]. The use of hypoallergenic mutants would be a good strategy to avoid the nondesired side effects of immunotherapy. Hypoallergenic mutants have been developed for several pollens and foods allergens [15–18], and their utility for immunotherapy has been studied [19, 20]. These mutants
References
[1]
R. Valenta, “The future of antigen-specific immunotherapy of allergy,” Nature Reviews Immunology, vol. 2, no. 6, pp. 446–453, 2002.
[2]
R. Valenta, T. Ball, M. Focke et al., “Immunotherapy of allergic disease,” Advances in Immunology, vol. 82, pp. 105–153, 2004.
[3]
J. Bousquet, R. Lockey, H. J. Malling et al., “Allergen immunotherapy: therapeutic vaccines for allergic diseases,” Journal of Allergy and Clinical Immunology, vol. 102, no. 4 I, pp. 558–562, 1998.
[4]
T. Ball, W. R. Sperr, P. Valent et al., “Induction of antibody responses to new B cell epitopes indicates vaccination character of allergen immunotherapy,” European Journal of Immunology, vol. 29, no. 6, pp. 2026–2036, 1999.
[5]
R. Movérare, L. Elfman, E. Vesterinen, T. Metso, and T. Haahtela, “Development of new IgE specificities to allergenic components in birch pollen extract during specific immunotherapy studied with immunoblotting and pharmacia CAP System,” Allergy, vol. 57, no. 5, pp. 423–430, 2002.
[6]
R. Crameri, “Allergy vaccines: dreams and reality,” Expert Review of Vaccines, vol. 6, no. 6, pp. 991–999, 2007.
[7]
M. Focke, V. Mahler, T. Ball, D. Kraft, and R. Valenta, “Nonallergenic peptides from surface-exposed areas or B-cell epitopes of allergens for specific immunotherapy,” International Archives of Allergy and Immunology, vol. 124, no. 1–3, pp. 398–399, 2001.
[8]
M. Focke, V. Mahler, T. Ball et al., “Nonanaphylactic synthetic peptides derived from B cell epitopes of the major grass pollen allergen, Phl p 1, for allergy vaccination,” Federation of American Societies for Experimental Biology Journal, vol. 15, no. 11, pp. 2042–2044, 2001.
[9]
B. M. Haselden, A. B. Kay, and M. Larché, “Immunoglobulin E-independent major histocompatibility complex- restricted T cell peptide epitope-induced late asthmatic reactions,” Journal of Experimental Medicine, vol. 189, no. 12, pp. 1885–1894, 1999.
[10]
D. Moldaver and M. Larché, “Immunotherapy with peptides,” Allergy, vol. 66, no. 6, pp. 784–791, 2011.
[11]
R. Hochreiter, T. Stepanoska, F. Ferreira et al., “Prevention of allergen-specific IgE production and suppression of an established Th2-type response by immunization with DNA encoding hypoallergenic allergen derivatives of Bet v 1, the major birch-pollen allergen,” European Journal of Immunology, vol. 33, no. 6, pp. 1667–1676, 2003.
[12]
Y. Darcan, A. Petersen, W. M. Becker et al., “Immunoprophylaxis with DNA vaccination inhibits Th2-mediated responses and airway inflammation in mice sensitised with the major timothy grass pollen allergen Phl p 5b,” Vaccine, vol. 23, no. 33, pp. 4203–4211, 2005.
[13]
H. S. Nelson, J. Lahr, R. Rule, A. Bock, and D. Leung, “Treatment of anaphylactic sensitivity to peanuts by immunotherapy with injections of aqueous peanut extract,” Journal of Allergy and Clinical Immunology, vol. 99, no. 6, pp. 744–751, 1997.
[14]
M. Fernández-Rivas, S. Garrido Fernández, J. A. Nadal et al., “Randomized double-blind, placebo-controlled trial of sublingual immunotherapy with a Pru p 3 quantified peach extract,” Allergy, vol. 64, no. 6, pp. 876–883, 2009.
[15]
S. Vrtala, M. Focke, J. Kopec et al., “Genetic engineering of the major timothy grass pollen allergen, Phl p 6, to reduce allergenic activity and preserve immunogenicity,” Journal of Immunology, vol. 179, no. 3, pp. 1730–1739, 2007.
[16]
A. Orlandi, F. Grasso, S. Corinti et al., “The recombinant major allergen of Parietaria judaica and its hypoallergenic variant: in vivo evaluation in a murine model of allergic sensitization,” Clinical and Experimental Allergy, vol. 34, no. 3, pp. 470–477, 2004.
[17]
M. Toda, G. Reese, G. Gadermaier et al., “Protein unfolding strongly modulates the allergenicity and immunogenicity of Pru p 3, the major peach allergen,” Journal of Allergy and Clinical Immunology, vol. 128, no. 5, pp. 1022–1030.e7, 2011.
[18]
K. Westritschnig, B. Linhart, M. Focke-Tejkl et al., “A hypoallergenic vaccine obtained by tail-to-head restructuring of timothy grass pollen profilin, Phl p 12, for the treatment of cross-sensitization to profilin,” Journal of Immunology, vol. 179, no. 11, pp. 7624–7634, 2007.
[19]
S. T. H. P. Bolhaar, L. Zuidmeer, Y. Ma et al., “A mutant of the major apple allergen, Mal d 1, demonstrating hypo-allergenicity in the target organ by double-blind placebo-controlled food challenge,” Clinical and Experimental Allergy, vol. 35, no. 12, pp. 1638–1644, 2005.
[20]
V. Niederberger, F. Horak, S. Vrtala et al., “Vaccination with genetically engineered allergens prevents progression of allergic disease,” Proceedings of the National Academy of Sciences of the United States of America, vol. 101, no. 2, pp. 14677–14682, 2004.
[21]
G. Reese, B. K. Ballmer-Weber, A. Wangorsch, S. Randow, and S. Vieths, “Allergenicity and antigenicity of wild-type and mutant, monomeric, and dimeric carrot major allergen Dau c 1: destruction of conformation, not oligomerization, is the roadmap to save allergen vaccines,” Journal of Allergy and Clinical Immunology, vol. 119, no. 4, pp. 944–951, 2007.
[22]
K. W. Chen, G. Fuchs, K. Sonneck et al., “Reduction of the in vivo allergenicity of Der p 2, the major house-dust mite allergen, by genetic engineering,” Molecular Immunology, vol. 45, no. 9, pp. 2486–2498, 2008.
[23]
M. Wallner, A. St？cklinger, T. Thalhamer et al., “Allergy multivaccines created by DNA shuffling of tree pollen allergens,” Journal of Allergy and Clinical Immunology, vol. 120, no. 2, pp. 374–380, 2007.
[24]
J. Cuesta-Herranz, D. Barber, C. Blanco et al., “Differences among pollen-allergic patients with and without plant food allergy,” International Archives of Allergy and Immunology, vol. 153, no. 2, pp. 182–192, 2010.
[25]
G. Salcedo, R. Sánchez-Monge, D. Barber, and A. Díaz-Perales, “Plant non-specific lipid transfer proteins: an interface between plant defence and human allergy,” Biochimica et Biophysica Acta, vol. 1771, no. 6, pp. 781–791, 2007.
[26]
O. A. Duffort, F. Polo, M. Lombardero et al., “Immunoassay to quantify the major peach allergen Pru p 3 in foodstuffs. Differential allergen release and stability under physiological conditions,” Journal of Agricultural and Food Chemistry, vol. 50, no. 26, pp. 7738–7741, 2002.
[27]
M. Fernández-Rivas, E. González-Mancebo, R. Rodríguez-Pérez et al., “Clinically relevant peach allergy is related to peach lipid transfer protein, Pru p 3, in the Spanish population,” Journal of Allergy and Clinical Immunology, vol. 112, no. 4, pp. 789–795, 2003.
[28]
G. García-Casado, L. F. Pacios, A. Díaz-Perales et al., “Identification of IgE-binding epitopes of the major peach allergen Pru p 3,” Journal of Allergy and Clinical Immunology, vol. 112, no. 3, pp. 599–605, 2003.
[29]
L. F. Pacios, L. Tordesillas, J. Cuesta-Herranz et al., “Mimotope mapping as a complementary strategy to define allergen IgE-epitopes: peach Pru p 3 allergen as a model,” Molecular Immunology, vol. 45, no. 8, pp. 2269–2276, 2008.
[30]
L. Tordesillas, J. Cuesta-Herranz, M. Gonzalez-Mu？oz et al., “T-cell epitopes of the major peach allergen, Pru p 3: identification and differential T-cell response of peach-allergic and non-allergic subjects,” Molecular Immunology, vol. 46, no. 4, pp. 722–728, 2009.
[31]
L. Tordesillas, L. F. Pacios, A. Palacin et al., “Molecular basis of allergen cross-reactivity: non-specific lipid transfer proteins from wheat flour and peach fruit as models,” Molecular Immunology, vol. 47, no. 2-3, pp. 534–540, 2009.
[32]
B. Linhart and R. Valenta, “Molecular design of allergy vaccines,” Current Opinion in Immunology, vol. 17, no. 6, pp. 646–655, 2005.
[33]
E. M. Shakir, D. S. Cheung, and M. H. Grayson, “Mechanisms of immunotherapy: a historical perspective,” Annals of Allergy, Asthma and Immunology, vol. 105, no. 5, pp. 340–347, 2010.
[34]
A. Bonura, R. Passantino, M. A. Costa et al., “Characterization of a Par j 1/Par j 2 mutant hybrid with reduced allergenicity for immunotherapy of Parietaria allergy,” Clinical and Experimental Allergy, vol. 42, no. 3, pp. 471–480, 2012.
[35]
J. Cuesta-Herranz, M. Lazaro, E. Figueredo, J. M. Igea, A. Umpierrez, and M. De-Las-Heras, “Allergy to plant-derived fresh foods in a birch- and ragweed-free area,” Clinical and Experimental Allergy, vol. 30, no. 10, pp. 1411–1416, 2000.
[36]
Subcommittee on Skin Tests of the European Academy of Allergology and Clinical Immunology, “Skin tests for diagnosis of IgE-mediated allergy,” Allergy, vol. 44, supplement s10, pp. 31–37, 1989.
[37]
A. Diaz-Perales, G. Garcia-Casado, R. Sanchez-Monge, F. J. Garcia-Selles, D. Barber, and G. Salcedo, “cDNA cloning and heterologous expression of the major allergens from peach and apple belonging to the lipid-transfer protein family,” Clinical and Experimental Allergy, vol. 32, no. 1, pp. 87–92, 2002.
[38]
A. Diaz-Perales, M. Lombardero, R. Sanchez-Monge et al., “Lipid-transfer proteins as potential plant panallergens: cross-reactivity among proteins of Artemisia pollen, Castanea nut and Rosaceae fruits, with different IgE-binding capacities,” Clinical and Experimental Allergy, vol. 30, no. 10, pp. 1403–1410, 2000.
[39]
A. Palacin, J. Varela, S. Quirce et al., “Recombinant lipid transfer protein Tri a 14: a novel heat and proteolytic resistant tool for the diagnosis of baker's asthma,” Clinical and Experimental Allergy, vol. 39, no. 8, pp. 1267–1276, 2009.
[40]
P. Gamboa, R. Sanchez-Monge, M. L. Sanz, A. Palacín, G. Salcedo, and A. Diaz-Perales, “Sensitization to Cannabis sativa caused by a novel allergenic lipid transfer protein, Can s 3,” Journal of Allergy and Clinical Immunology, vol. 120, no. 6, pp. 1459–1460, 2007.
[41]
M. L. Sanz, P. M. Gamboa, I. Antépara et al., “Flow cytometric basophil activation test by detection of CD63 expression in patients with immediate-type reactions to betalactam antibiotics,” Clinical and Experimental Allergy, vol. 32, no. 2, pp. 277–286, 2002.
[42]
L. Tordesillas, P. Gamboa, M. L. Sanz et al., “A mutant of the major melon allergen, Cuc m 2, with reduced IgE binding capacity is a good candidate for specific immunotherapy,” Molecular Immunology, vol. 49, no. 3, pp. 504–511, 2011.
[43]
G. López-Torrejón, J. F. Crespo, R. Sánchez-Monge et al., “Allergenic reactivity of the melon profilin Cuc m 2 and its identification as major allergen,” Clinical and Experimental Allergy, vol. 35, no. 8, pp. 1065–1072, 2005.
[44]
S. Vale-Pereira, A. Todo-Bom, L. Geraldes, C. Schmidt-Weber, C. A. Akdis, and A. Mota-Pinto, “FoxP3, GATA-3 and T-bet expression in elderly asthma,” Clinical and Experimental Allergy, vol. 41, no. 4, pp. 490–496, 2011.
[45]
M. W. Lipscomb, L. Chen, J. L. Taylor et al., “Ectopic T-bet expression licenses dendritic cells for IL-12-independent priming of type 1 T cells in vitro,” Journal of Immunology, vol. 183, no. 11, pp. 7250–7258, 2009.
[46]
K. J. Livak and T. D. Schmittgen, “Analysis of relative gene expression data using real-time quantitative PCR and the method,” Methods, vol. 25, no. 4, pp. 402–408, 2001.
[47]
F. Ferreira, K. Hirtenlehner, A. Jilek et al., “Dissection of immunoglobulin E and T lymphocyte reactivity of isoforms of the major birch pollen allergen Bet v 1: potential use of hypoallergenic isoforms for immunotherapy,” Journal of Experimental Medicine, vol. 183, no. 2, pp. 599–609, 1996.
[48]
F. Ferreira, C. Ebner, B. Kramer et al., “Modulation of IgE reactivity of allergens by site-directed mutagenesis: potential use of hypoallergenic variants for immunotherapy,” The FASEB Journal, vol. 12, no. 2, pp. 231–242, 1998.
[49]
C. A. Akdis and M. Akdis, “Mechanisms of allergen-specific immunotherapy,” Journal of Allergy and Clinical Immunology, vol. 127, no. 1, pp. 18–27, 2011.
[50]
V. Schulten, A. Radakovics, C. Hartz et al., “Characterization of the allergic T-cell response to Pru p 3, the nonspecific lipid transfer protein in peach,” Journal of Allergy and Clinical Immunology, vol. 124, no. 1, pp. 100–107, 2009.
