Lung-derived innate cytokines: new epigenetic targets of allergen-specific sublingual immunotherapy

Document Type: Original Article


1 School of Medicine, Zabol University of Medical Sciences, Zabol, Iran

2 Student Research Committee, Immunology Research Center, Medical School, Mashhad University of Medical Sciences, Mashhad, Iran

3 Allergy Research Center, Medical School, Mashhad University of Medical Sciences, Mashhad, Iran

4 Division of Human Genetics, Immunology Research Center, Avicenna Research Institute, Mashhad University of Medical Sciences,Mashhad, Iran

5 Department of Food Science, Nour Branch, Islamic Azad University, Nour, Iran

6 Immunology Research Center, Medical School, Mashhad University of Medical Sciences, Mashhad, Iran


Objective(s):Sublingual allergen-specific immunotherapy is a safe and effective method for treatment of IgE-mediated respiratory allergies; however, the underlying mechanisms are not fully understood. This study was planned to test whether sublingual immunotherapy (SLIT) can exert epigenetic mechanisms through which the airway allergic responses can be extinguished.
Materials and Methods:BALB/c mice were sensitized intraperitoneally and challenged intranasally. Then, they received sublingual treatment with recombinant Che a 2 (rChe a 2), a major allergen of Chenopodium album. After SLIT, allergen-specific antibodies in sera, cytokine profiles of spleen cell cultures, mRNA and protein expression of lung-derived IL-33, IL-25, and TSLP (thymic stromal lymphopoietin), and histone modifications of these three genes were assessed.

Following Immunotherapy, systemic immune responses shifted from Th2 to Th1 profile as demonstrated by significant decrease in IgE and IL-4 and substantial increase in IgG2a and IFN-γ. At local site, mRNA and protein levels of lung-derived pro-inflammatory cytokines IL-33 and TSLP were markedly down-regulated following SLIT that was associated with marked enrichment of trimethylated lysine 27 of histone H3 at promoter regions of these two cytokines.
Conclusion:In our study, sublingual immunotherapy with recombinant allergen effectively attenuated allergic immune responses, at least partly, by induction of distinct histone modifications at specific loci. Additionally, the lung-derived pro-allergic cytokines IL-33 and TSLP could be promising mucosal candidates for either monitoring allergic conditions or therapeutic approaches.


1. Canonica GW, Cox L, Pawankar R, Baena-Cagnani CE, Blaiss M, Bonini S, et al. Sublingual immunotherapy: World Allergy Organization position paper 2013 update. World Allergy Organ J 2014; 7:6.

2. Swamy RS, Reshamwala N, Hunter T, Vissamsetti S, Santos CB, Baroody FM, et al. Epigenetic modifications and improved regulatory T-cell function in subjects undergoing dual sublingual immunotherapy. J Allergy Clin Immunol 2012; 130:215-224 e7.

3. Cheng RY, Shang Y, Limjunyawong N, Dao T, Das S, Rabold R, et al. Alterations of the lung methylome in allergic airway hyper-responsiveness. Environ Mol Mutagen 2014; 55:244-255.

4. Brand S, Kesper DA, Teich R, Kilic-Niebergall E, Pinkenburg O, Bothur E, et al. DNA methylation of TH1/TH2 cytokine genes affects sensitization and progress of experimental asthma. J Allergy Clin Immunol 2012; 129:1602-1610 e6.

5. Shang Y, Das S, Rabold R, Sham JS, Mitzner W, Tang WY. Epigenetic alterations by DNA methylation in house dust mite-induced airway hyperrespon-siveness. Am J Respir Cell Mol Biol 2013; 49:279-287.

6. Harb H, Renz H. Update on epigenetics in allergic disease. J Allergy Clin Immunol 2015; 135:15-24.

7. Holtzman MJ, Byers DE, Alexander-Brett J, Wang X. The role of airway epithelial cells and innate immune cells in chronic respiratory disease. Nat Rev Immunol. 2014; 14:686-698.

8. Bartemes KR, Kita H. Dynamic role of epithelium-derived cytokines in asthma. Clin Immunol 2012; 143:222-235.

9. Barderas R, Villalba M, Pascual CY, Batanero E, Rodriguez R. Profilin (Che a 2) and polcalcin (Che a 3) are relevant allergens of Chenopodium album pollen: isolation, amino acid sequences, and immunologic properties. J Allergy Clin Immunol 2004; 113:1192-1198.

10. Villalba M, Barderas R, Mas S, Colas C, Batanero E, Rodriguez R. Amaranthaceae pollens: review of an emerging allergy in the mediterranean area. J Investig Allergol Clin Immunol 2014; 24:371-381.

11. Fereidouni M, Hossini RF, Azad FJ, Assarehzadegan MA, Varasteh A. Skin prick test reactivity to common aeroallergens among allergic rhinitis patients in Iran. Allergol Immunopathol (Madr) 2009; 37:73-79.

12. Assarehzadegan MA, Shakurnia A, Amini A. The most common aeroallergens in a tropical region in Southwestern Iran. World Allergy Organ J 2013; 6:7.

13. Ahmadiafshar A, Sepehri S, Mousavinasab S, Torabi SZ. Recognition and frequency of common allergens in allergic patients of zanjan by skin prick test. Zanjan Univ Med Sci J 2008; 16:45-53.

14. Amini A, sankian M, Assarehzadegan MA, Vahedi F, Varasteh A. Chenopodium album pollen profilin (Che a 2): homology modeling and evaluation of cross-reactivity with allergenic profilins based on predicted potential IgE epitopes and IgE reactivity analysis. Mol Biol Rep. 2011; 38:2579-2587. Epub 2010/11/19.

15. Nouri HR, Sankian M, Afsharzadeh D, Varasteh A. Immunotherapy with a recombinant hybrid molecule alleviates allergic responses more efficiently than an allergenic cocktail or pollen extract in a model of Chenopodium album allergy. Int Arch Allergy Immunol 2013; 161:325-332.

16. Pfaffl MW. A new mathematical model for relative quantification in real-time RT–PCR. Nuclic Acids Res 2001; 29:e45.

17. Smith P, Krohn RI, Hermanson G, Mallia A, Gartner F, Provenzano M, et al. Measurement of protein using bicinchoninic acid. Anal Biochem 1985; 150:76-85.

18. Nouri HR, Varasteh A, Vahedi F, Chamani J, Afsharzadeh D, Sankian M. Constructing a hybrid molecule with low capacity of IgE binding from Chenopodium album pollen allergens. Immunol lett 2012; 144:67-77.

19. Taylor SC, Berkelman T, Yadav G, Hammond M. A defined methodology for reliable quantification of Western blot data. Mol Biotechnol 2013; 55:217-226.

20. Sailaja BS, Takizawa T, Meshorer E. Chromatin immunoprecipitation in mouse hippocampal cells and tissues. Methods Mol Biol 2012; 809:353-64.

21. Negishi H, Miki S, Sarashina H, Taguchi-Atarashi N, Nakajima A, Matsuki K, et al. Essential contribution of IRF3 to intestinal homeostasis and microbiota-mediated Tslp gene induction. Proc Natl Acad Sci U S A 2012; 109:21016-21021.

22. Wang Z, Zhang LJ, Guha G, Li S, Kyrylkova K, Kioussi C, et al. Selective ablation of Ctip2/Bcl11b in epidermal keratinocytes triggers atopic dermatitis-like skin inflammatory responses in adult mice. PLoS One 2012; 7:e51262.

23. Lefrançais E, Roga S, Gautier V, Gonzalez-de-Peredo A, Monsarrat B, Girard J-P, et al. IL-33 is processed into mature bioactive forms by neutrophil elastase and cathepsin G. Proc Natl Acad Sci USA 2012; 109:1673-1678.

24. Bernstein BE, Kamal M, Lindblad-Toh K, Bekiranov S, Bailey DK, Huebert DJ, et al. Genomic maps and comparative analysis of histone modifications in human and mouse. Cell 2005; 120:169-181.

25. Bernstein BE, Mikkelsen TS, Xie X, Kamal M, Huebert DJ, Cuff J, et al. A bivalent chromatin structure marks key developmental genes in embryonic stem cells. Cell 2006; 125:315-326.

26. Saenz SA, Taylor BC, Artis D. Welcome to the neighborhood: epithelial cell-derived cytokines license innate and adaptive immune responses at mucosal sites. Immunol Rev 2008; 226:172-190.

27. Nakanishi W, Yamaguchi S, Matsuda A, Suzukawa M, Shibui A, Nambu A, et al. IL-33, but not IL-25, is crucial for the development of house dust mite antigen-induced allergic rhinitis. PLoS One 2013; 8:e78099.

28. Byers DE. Defining the roles of IL-33, thymic stromal lymphopoietin, and IL-25 in human asthma. Am J Respir Crit Care Med 2014; 190:715-716.

29. Wallner M, Pichler U, Ferreira F. Recombinant allergens for pollen immunotherapy. Immunotherapy 2013; 5:1323-1338.

30. Wiedermann U, Jahn-Schmid B, Bohle B, Renz H, Kraft D, Ebner C. Suppression of antigen-specific               T-and B-cell responses by intranasal or oral administration of recombinant bet v 1, the major birch pollen allergen, in a murine model of type I allergy. J Allergy Clin Immunol 1999; 103:1202-1210.

31. Ahmadiafshar A, Taymourzadeh B, Shaikhi A, Mazloomzadeh S, Torabi Z. Evaluation of IL10, TGF-B and Specific IgE and IgG Levels during Sublingual Rye Grass Immunotherapy. J Allergy  Ther 2013; 4: 132.

32. Allam J-P, Novak N. Immunological mechanisms of sublingual immunotherapy. Curr Opin Allergy Clin Immunol 2014; 14:564-569.

33. Wei G, Wei L, Zhu J, Zang C, Hu-Li J, Yao Z, et al. Global mapping of H3K4me3 and H3K27me3 reveals specificity and plasticity in lineage fate determination of differentiating CD4+ T cells. Immunity 2009; 30:155-167.

34. Comer BS, Ba M, Singer CA, Gerthoffer WT. Epigenetic targets for novel therapies of lung diseases. Pharmacol Ther 2015; 147:91-110.