After 1day, AHR to methacholine was assessed. blot analysis was applied with mice serum and mass spectrometry was used to identify the IgE-binding bands in birch pollen. Compared with PBS group, birch pollen sensitization and challenge BALB/c mice developed AHR, and IL4, IL5, IL6, IL10, and IL17 were significantly higher. Mice sensitized by birch pollen showed increased plasma levels of anti-BPE IgE, IgG1, and IgG2a. Histologic analyses showed that mice experienced peribranchial infiltration of inflammatory cells and mucosal hyperplasia. After SCIT, allergic symptoms effectively alleviated and kept for a long time. Interestingly, mice serum pool showed strong reactions to 70-kDa proteins. Mass spectrometry data suggests that the 70-kDa protein belongs to the HSP 70 family. SCIT inhibited the inflammatory response in the long term and a 70-kDa protein potentially belonging to the HSP 70 family plays a significant role in Chinese birch pollen-induced mice model. Key Words:Chinese birch pollen allergy, birch pollen allergen, mice model, asthma, 70-kDa allergen == INTRODUCTION == Birch pollen is one of the most important spring pollens in north of China. In Europe, 70% of birch pollen allergy patients have accompanying varying degrees of food allergy [1]. Their symptoms are mainly shown after ingesting birch pollen-related KL-1 foods [2], at least in part because of the cross-reaction between Bet v 1 homologous protein in food and the major allergen Bet v 1 in birch pollen, including apple (Mal d 1), cherry (Pru av. 1), pear (Pyr c 1), hazelnut (Cor a 1), celery (Api g 1), carrot (Dau c 1), soybean (Gly m 4), peanut (Ara h 8), jackfruit, and kiwi (Take action d 8) [3]. Subcutaneous immunotherapy (SCIT) is the only way to effectively control allergy symptoms KL-1 and it can change the natural courses from allergic rhinitis to allergic asthma [47]. They exhibited that the mechanism of SCIT is largely related to the reduction of Th2-related cytokines (IL4 and IL5) and Th17-related cytokines (IL17). It KL-1 is also thought that SCIT induces the production of protective antibodies IgG4 (IgG2a in mice) to inhibit mast cell activation [8,9]. Regulatory T cells (Treg) suppress the allergic inflammation by generating IL10 and TGF- in SCIT. IL-10 suppresses T cell responses, promotes IgG4 production, and suppresses IgE production [10]. TGF- is required for Treg differentiation. Moreover, IL-6 may regulate the balance between Th17 cells and Treg cells [11]. Adjuvants, especially the alum, play a central role in co-stimulating immune cells in KL-1 the SCIT, which also lead to local or systemic adverse effects. Seven allergens in birch pollen have been included in the recognized allergen Eledoisin Acetate list of the World Health Business and International Union of Immunological Societies (WHO/IUIS) allergen nomenclature subcommittee (www.allergen.org), named sequentially as Bet v 1 through Bet v 7. Research KL-1 is often biased toward a single major allergen (Bet v 1) [12]. Panallergens are ubiquitous with broad cross-sensitization and underlie certain pollen-food or plant-food syndromes [13]. However, there is little research conducted on birch pollen allergen in China. In this study, it is the first time to establish an allergy murine model with Chinese birch pollen and observe the long efficacy of SCIT in mice. We also hypothesized that SCIT without adjuvant (alum) remains efficient in mice. == MATERIALS AND METHODS == == Mice == Female BALB/c mice, at 56 weeks of age, were purchased from Beijing Vital River Laboratory Animal Technology Co, Beijing, China. All of the mice were given a 7-day period acclimate. The animals were kept under Specific Pathogen Free (SPF) laboratory conditions in the Center for Animal Experiments of Peking Union Medical College Hospital. The institutional animal ethics committee approved all experiments. == Allergens == Standard birch pollen extracts (BPE) were provided by the department of allergy, Peking Union Medical College Hospital, Beijing, China. These can be used for patients in clinics. The protein concentration was decided using the Pierce BCA Protein Assay Kit (Thermo Fisher Scientific, Waltham, Mass). The major protein Bet v1 of birch pollen extract was dependant on a Wager v.
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