Background: The efficacy and safety of allergen-specific immunotherapy (AIT) are highly dose-dependent. Methods: We investigated the dosage effects of AIT and the underlying mechanisms in a murine model of shrimp hypersensitivity. BALB/c mice were sensitized with recombinant shrimp allergen rMet e 1 and challenged orally with a high dose of rMet e 1 to elicit an allergic response. These sensitized mice were then treated with a low (0.01 mg), medium (0.05 mg), or high dosage (0.1 mg) of rMet e 1 intraperitoneally before receiving a second oral challenge. The allergic responses and immunological changes in the gut were compared between animals receiving different dosages. Results: We found that all sensitized mice that received rMet e 1 immunotherapy were desensitized, regardless of the dosage, and protected at the second oral challenge. Nevertheless, the mice in the high-dosage group experienced severe systemic reactions during the treatment phase. In contrast, regulatory T (Treg) cell-associated genes were upregulated only in the low- and medium-dosage groups, and Foxp3+ cells were more abundant in the gut lymphoid tissues than in the high-dosage group. Conclusions: Our results demonstrate that low-dosage immunotherapy favors the induction of local Foxp3+ Treg cells and the upregulation of regulatory cytokines. The safety advantages and long-term efficacy of low-dosage immunotherapy should be taken into consideration when developing immunotherapy dose schedules.

Keywords:
Tropomyosin, Desensitization, Dosage, Regulatory T cells, Foxp3
1.
Sicherer SH, Sampson HA: Food allergy: epidemiology, pathogenesis, diagnosis, and treatment. J Allergy Clin Immunol 2014;133:291-307.
2.
Bacal LR: The impact of food allergies on quality of life. Pediatr Ann 2013;42:141-145.
3.
Gupta R, Holdford D, Bilaver L, Dyer A, Holl JL, Meltzer D: The economic impact of childhood food allergy in the United States. JAMA Pediatr 2013;167:1026-1031.
4.
Demoly P, Calderon MA: Dosing and efficacy in specific immunotherapy. Allergy 2011;66(suppl 95):38-40.
5.
Didier A, Malling HJ, Worm M, Horak F, Jäger S, Montagut A, André C, de Beaumont O, Melac M: Optimal dose, efficacy, and safety of once-daily sublingual immunotherapy with a 5-grass pollen tablet for seasonal allergic rhinitis. J Allergy Clin Immunol 2007;120:1338-1345.
6.
Akdis M, Akdis CA: Mechanisms of allergen-specific immunotherapy: multiple suppressor factors at work in immune tolerance to allergens. J Allergy Clin Immunol 2014;133:621-631.
7.
Dioszeghy V, Mondoulet L, Dhelft V, Ligouis M, Puteaux E, Dupont C, Benhamou PH: The regulatory T cells induction by epicutaneous immunotherapy is sustained and mediates long-term protection from eosinophilic disorders in peanut-sensitized mice. Clin Exp Allergy 2014;44:867-881.
8.
Akdis CA, Akdis M: Mechanisms of allergen-specific immunotherapy. J Allergy Clin Immunol 2011;127:18-27.
9.
Thalayasingam M, Gerez IF, Yap GC, Llanora GV, Chia IP, Chua L, Lee CJ, Ta LD, Cheng YK, Thong BY, Tang CY, Van Bever HP, Shek LP, Curotto de Lafaille MA, Lee BW: Clinical and immunochemical profiles of food challenge proven or anaphylactic shrimp allergy in tropical Singapore. Clin Exp Allergy 2015;45:687-697.
10.
Leung PS, Lee YS, Tang CY, Kung WY, Chuang YH, Chiang BL, Fung MC, Chu KH: Induction of shrimp tropomyosin-specific hypersensitivity in mice. Int Arch Allergy Immunol 2008;147:305-314.
11.
Lam YF, Tong KK, Kwan KM, Tsuneyama K, Shu SA, Leung PS, Chu KH: Gastrointestinal immune response to the shrimp allergen tropomyosin: histological and immunological analysis in an animal model of shrimp tropomyosin hypersensitivity. Int Arch Allergy Immunol 2015;167:29-40.
12.
Wai CY, Leung NY, Ho MH, Gershwin LJ, Shu SA, Leung PS, Chu KH: Immunization with hypoallergens of shrimp allergen tropomyosin inhibits shrimp tropomyosin specific IgE reactivity. PLoS One 2014;9:e111649.
13.
Wai CY, Leung NY, Leung PS, Chu KH: T cell epitope immunotherapy ameliorates allergic responses in a murine model of shrimp allergy. Clin Exp Allergy 2016;46:491-503.
14.
Li XM, Serebrisky D, Lee SY, Huang CK, Bardina L, Schofield BH, Stanley JS, Burks AW, Bannon GA, Sampson HA: A murine model of peanut anaphylaxis: T- and B-cell responses to a major peanut allergen mimic human responses. J Allergy Clin Immunol 2000;106:150-158.
15.
Fox EM, Torrero MN, Evans H, Mitre E: Immunologic characterization of 3 murine regimens of allergen-specific immunotherapy. J Allergy Clin Immunol 2015;135:1341-1351.
16.
Leung NY, Wai CY, Ho MH, Liu R, Lam KS, Wang JJ, Shu SA, Chu KH, Leung PS: Screening and identification of mimotopes of the major shrimp allergen tropomyosin using one-bead-one-compound peptide libraries. Cell Mol Immunol 2017;14:308-318.
17.
Moolenbeek C, Ruitenberg EJ: The “Swiss roll”: a simple technique for histological studies of the rodent intestine. Lab Anim 1981;15:57-59.
18.
Schmittgen TD, Livak KJ: Analyzing real-time PCR data by the comparative C(T) method. Nat Protoc 2008;3:1101-1108.
19.
Lin XP, Magnusson J, Ahlstedt S, Dahlman-Höglund A, Hanson LL, Magnusson O, Bengtsson U, Telemo E: Local allergic reaction in food-hypersensitive adults despite a lack of systemic food-specific IgE. J Allergy Clin Immunol 2002;109:879-887.
20.
Oppenheimer JJ, Nelson HS, Bock SA, Christensen F, Leung DY: Treatment of peanut allergy with rush immunotherapy. J Allergy Clin Immunol 1992;90:256-262.
21.
Nelson HS, Lahr J, Rule R, Bock A, Leung D: Treatment of anaphylactic sensitivity to peanuts by immunotherapy with injections of aqueous peanut extract. J Allergy Clin Immunol 1997;99:744-751.
22.
Sampson HA: Peanut oral immunotherapy: is it ready for clinical practice? J Allergy Clin Immunol Pract 2013;1:15-21.
23.
Moran TP, Burks AW: Is clinical tolerance possible after allergen immunotherapy? Curr Allergy Asthma Rep 2015;15:23.
24.
Calderon MA, Larenas D, Kleine-Tebbe J, Jacobsen L, Passalacqua G, Eng PA, Varga EM, Valovirta E, Moreno C, Malling HJ, Alvarez-Cuesta E, Durham S, Demoly P: European Academy of Allergy and Clinical Immunology Task Force report on “dose-response relationship in allergen-specific immunotherapy.” Allergy 2011;66:1345-1359.
25.
Daul CB, Slattery M, Reese G, Lehrer SB: Identification of the major brown shrimp (Penaeus aztecus) allergen as the muscle protein tropomyosin. Int Arch Allergy Immunol 1994;105:49-55.
26.
Ayuso R, Lehrer SB, Reese G: Identification of continuous, allergenic regions of the major shrimp allergen Pen a 1 (tropomyosin). Int Arch Allergy Immunol 2002;127:27-37.
27.
Yang AC, Arruda LK, Santos AB, Barbosa MC, Chapman MD, Galvao CE, Kalil J, Morato-Castro FF: Measurement of IgE antibodies to shrimp tropomyosin is superior to skin prick testing with commercial extract and measurement of IgE to shrimp for predicting clinically relevant allergic reactions after shrimp ingestion. J Allergy Clin Immunol 2010;125:872-878.
28.
Gamez C, Sanchez-Garcia S, Ibanez MD, Lopez R, Aguado E, Lopez E, Sastre B, Sastre J, del Pozo V: Tropomyosin IgE-positive results are a good predictor of shrimp allergy. Allergy 2011;66:1375-1383.
29.
Herz U, Renz H, Wiedermann U: Animal models of type I allergy using recombinant allergens. Methods 2004;32:271-280.
30.
Parungo CP, Soybel DI, Colson YL, Kim SW, Ohnishi S, DeGrand AM, Laurence RG, Soltesz EG, Chen FY, Cohn LH, Bawendi MG, Frangioni JV: Lymphatic drainage of the peritoneal space: a pattern dependent on bowel lymphatics. Ann Surg Oncol 2007;14:286-298.
31.
Spahn TW, Weiner HL, Rennert PD, Lügering N, Fontana A, Domschke W, Kucharzik T: Mesenteric lymph nodes are critical for the induction of high-dose oral tolerance in the absence of Peyer's patches. Eur J Immunol 2002;32:1109-1113.
32.
Cassani B, Villablanca EJ, Quintana FJ, Love PE, Lacy-Hulbert A, Blaner WS, Sparwasser T, Snapper SB, Weiner HL, Mora JR: Gut-tropic T cells that express integrin α4β7 and CCR9 are required for induction of oral immune tolerance in mice. Gastroenterology 2011;141:2109-2118.
33.
Esterhazy D, Loschko J, London M, Jove V, Oliveira TY, Mucida D: Classical dendritic cells are required for dietary antigen-mediated induction of peripheral T(reg) cells and tolerance. Nat Immunol 2016;17:545-555.
34.
Syed A, Garcia MA, Lyu SC, Bucayu R, Kohli A, Ishida S, Berglund JP, Tsai M, Maecker H, O'Riordan G, Galli SJ, Nadeau KC: Peanut oral immunotherapy results in increased antigen-induced regulatory T-cell function and hypomethylation of forkhead box protein 3 (FOXP3). J Allergy Clin Immunol 2014;133:500-510.
35.
Rauter I, Krauth MT, Westritschnig K, Horak F, Flicker S, Gieras A, Repa A, Balic N, Spitzauer S, Huss-Marp J, Brockow K, Darsow U, Behrendt H, Ring J, Kricek F, Valent P, Valenta R: Mast cell-derived proteases control allergic inflammation through cleavage of IgE. J Allergy Clin Immunol 2008;121:197-202.
36.
Lu LF, Lind EF, Gondek DC, Bennett KA, Gleeson MW, Pino-Lagos K, Scott ZA, Coyle AJ, Reed JL, Van Snick J, Strom TB, Zheng XX, Noelle RJ: Mast cells are essential intermediaries in regulatory T-cell tolerance. Nature 2006;442:997-1002.
37.
Hosken NA, Shibuya K, Heath AW, Murphy KM, O'Garra A: The effect of antigen dose on CD4+ T helper cell phenotype development in a T cell receptor-alpha beta-transgenic model. J Exp Med 1995;182:1579-1584.
38.
Hashiguchi M, Hachimura S, Ametani A, Kaminogawa S: Th2 polarization enhanced by oral administration of higher doses of antigen. Cytotechnology 2000;33:237-245.
39.
Stamm C, Barthelmann J, Kunz N, Toellner KM, Westermann J, Kalies K: Dose-dependent induction of murine Th1/Th2 responses to sheep red blood cells occurs in two steps: antigen presentation during second encounter is decisive. PLoS One 2013;8:e67746.
40.
Kretschmer K, Apostolou I, Hawiger D, Khazaie K, Nussenzweig MC, von Boehmer H: Inducing and expanding regulatory T cell populations by foreign antigen. Nat Immunol 2005;6:1219-1227.
41.
Bedoret D, Singh AK, Shaw V, Hoyte EG, Hamilton R, DeKruyff RH, Schneider LC, Nadeau KC, Umetsu DT: Changes in antigen-specific T-cell number and function during oral desensitization in cow's milk allergy enabled with omalizumab. Mucosal Immunol 2012;5:267-276.
42.
Galli SJ, Tsai M, Piliponsky AM: The development of allergic inflammation. Nature 2008;454:445-454.
43.
Long SA, Rieck M, Tatum M, Bollyky PL, Wu RP, Muller I, Ho JC, Shilling HG, Buckner JH: Low-dose antigen promotes induction of FOXP3 in human CD4+ T cells. J Immunol 2011;187:3511-3520.
44.
Maggi E: T-cell responses induced by allergen-specific immunotherapy. Clin Exp Immunol 2010;161:10-18.
45.
Palomares O, Martin-Fontecha M, Lauener R, Traidl-Hoffmann C, Cavkaytar O, Akdis M, Akdis CA: Regulatory T cells and immune regulation of allergic diseases: roles of IL-10 and TGF-β. Genes Immun 2014;15:511-520.
46.
Carrier Y, Yuan J, Kuchroo VK, Weiner HL: Th3 cells in peripheral tolerance. II. TGF-β-transgenic Th3 cells rescue IL-2-deficient mice from autoimmunity. J Immunol 2007;178:172-178.
47.
Pabst O, Mowat AM: Oral tolerance to food protein. Mucosal Immunol 2012;5:232-239.
48.
Allam JP, Würtzen PA, Reinartz M, Winter J, Vrtala S, Chen KW, Valenta R, Wenghoefer M, Appel T, Gros E, Niederhagen B, Bieber T, Lund K, Novak N: Phl p 5 resorption in human oral mucosa leads to dose-dependent and time-dependent allergen binding by oral mucosal Langerhans cells, attenuates their maturation, and enhances their migratory and TGF-β1 and IL-10-producing properties. J Allergy Clin Immunol 2010;126:638-645.
49.
Uermösi C, Zabel F, Manolova V, Bauer M, Beerli RR, Senti G, Kündig TM, Saudan P, Bachmann MF: IgG-mediated down-regulation of IgE bound to mast cells: a potential novel mechanism of allergen-specific desensitization. Allergy 2014;69:338-347.
50.
Cheng LE, Wang ZE, Locksley RM: Murine B cells regulate serum IgE levels in a CD23-dependent manner. J Immunol 2010;185:5040-5047.
51.
Oefner CM, Winkler A, Hess C, Lorenz AK, Holecska V, et al: Tolerance induction with T cell-dependent protein antigens induces regulatory sialylated IgGs. J Allergy Clin Immunol 2012;129:1647-1655.
52.
Calderon MA, Casale TB, Togias A, Bousquet J, Durham SR, Demoly P: Allergen-specific immunotherapy for respiratory allergies: from meta-analysis to registration and beyond. J Allergy Clin Immunol 2011;127:30-38.
53.
Kleine-Tebbe J, Ribel M, Herold DA: Safety of a SQ-standardised grass allergen tablet for sublingual immunotherapy: a randomized, placebo-controlled trial. Allergy 2006;61:181-184.
54.
Larsen TH, Poulsen LK, Melac M, Combebias A, André C, Malling HJ: Safety and tolerability of grass pollen tablets in sublingual immunotherapy - a phase-1 study. Allergy 2006;61:1173-1176.
55.
Pocobelli D, Del Bono A, Venuti L, Falagiani P, Venuti A: Nasal immunotherapy at constant dosage: a double-blind, placebo-controlled study in grass-allergic rhinoconjunctivitis. J Investig Allergol Clin Immunol 2001;11:79-88.
56.
Marcucci F, Sensi LG, Caffarelli C, Cavagni G, Bernardini R, Tiri A, Riva G, Novembre E: Low-dose local nasal immunotherapy in children with perennial allergic rhinitis due to Dermatophagoides. Allergy 2002;57:23-28.
57.
Yanagida N, Sato S, Asaumi T, Okada Y, Ogura K, Ebisawa M: A single-center, case-control study of low-dose-induction oral immunotherapy with cow's milk. Int Arch Allergy Immunol 2015;168:131-137.
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