Serum IgE response to orally ingested antigen: A novel IgE response model with allergen-specific T-cell receptor transgenic mice Kan Shida a,b, Satoshi.

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Serum IgE response to orally ingested antigen: A novel IgE response model with allergen-specific T-cell receptor transgenic mice Kan Shida a,b, Satoshi Hachimura, PhDa, Akio Ametani, PhDa, Mina Ishimori a, Mei Ling a, Masaaki Hashiguchi a, Yoshihiro Ueda a, Takehito Sato, PhDc, Yoshihiro Kumagai, PhDd, Kotaro Takamizawa, PhDb, Sonoko Habu, MD, PhDc, Shuichi Kaminogawa, PhDa Journal of Allergy and Clinical Immunology Volume 105, Issue 4, Pages 788-795 (April 2000) DOI: 10.1067/mai.2000.104934 Copyright © 2000 Mosby, Inc. Terms and Conditions

Fig. 1 Levels of OVA-specific IgE and total IgE in the sera of transgenic (Tg) mice fed the OVA diet. Age-matched transgenic and BALB/c mice were fed the OVA diet for the indicated period, and the levels of OVA-specific IgE (A) and total IgE (B) in sera were determined by using ELISA. OVA specific IgE level was expressed as relative units compared with hyperimmunized mouse serum (1000 U/mL). Data are expressed as means ± SD of 8 (transgenic) or 6 (BALB/c) mice. Points not sharing a common letter are significantly different. In BALB/c mice ANOVA did not show a significant difference between levels of IgE at each time point. Experiments were repeated 3 times with similar results. Journal of Allergy and Clinical Immunology 2000 105, 788-795DOI: (10.1067/mai.2000.104934) Copyright © 2000 Mosby, Inc. Terms and Conditions

Fig. 2 OVA-specific antibody responses in terms of IgG1, IgG2a, IgA, and IgM classes in transgenic (Tg) mice fed the OVA diet. Transgenic and BALB/c mice were fed the OVA diet for the indicated period, and OVA-specific antibody levels in sera were determined by using ELISA. OVA-specific antibody level was expressed as optical density at 405 nm. Data are expressed as means ± SD of 8 (transgenic) or 6 (BALB/c) mice. Points not sharing a common letter are significantly different. NS, Not significant. Letters in parentheses are for BALB/c mice. Experiments were repeated 3 times with similar results. Journal of Allergy and Clinical Immunology 2000 105, 788-795DOI: (10.1067/mai.2000.104934) Copyright © 2000 Mosby, Inc. Terms and Conditions

Fig. 3 Comparison of the antibody response elicited by oral feeding of OVA and that elicited by intraperitoneal injection of OVA. Transgenic mice were either fed the OVA diet for 5 weeks (Fed) or immunized with OVA adsorbed onto alum intraperitoneally twice with a 2-week interval (ip/alum) . The sera were collected and assayed for OVA-specific antibody by using ELISA. OVA-specific antibody level was expressed as optical density at 405 nm. Data are expressed as means ± SD of 4 mice. Experiments were repeated twice with similar results. NT, not treated; NS, not significant. *P < .05; **P < .01. Journal of Allergy and Clinical Immunology 2000 105, 788-795DOI: (10.1067/mai.2000.104934) Copyright © 2000 Mosby, Inc. Terms and Conditions

Fig. 4 Correlation between anaphylactic reactions and serum OVA-specific IgE level. Anaphylactic scores and serum OVA-specific IgE levels of individual mice (n = 30) in experiment 2 of Table I are plotted. The Spearmen R value is shown (P < .001). Journal of Allergy and Clinical Immunology 2000 105, 788-795DOI: (10.1067/mai.2000.104934) Copyright © 2000 Mosby, Inc. Terms and Conditions

Fig. 5 Cytokine production by splenocytes from transgenic mice fed the OVA diet. Splenocytes from transgenic mice fed the OVA diet for various periods were cultured with OVA for 44 hours, and the levels of cytokines in the culture supernatants were determined by using ELISA. Data are expressed as means ± SD of 3 independent spleens of transgenic mice. Bars not sharing a common letter are significantly different. Experiments were repeated 3 times with similar results. Journal of Allergy and Clinical Immunology 2000 105, 788-795DOI: (10.1067/mai.2000.104934) Copyright © 2000 Mosby, Inc. Terms and Conditions

Fig. 6 Helper activity of splenic T cells from OVA-fed transgenic mice in promoting IgE production. CD4+ T cells were prepared from pooled splenocytes of 3 transgenic mice fed the OVA diet for various periods, and the isolated cells were cultured with naive B cells from unsensitized BALB/c mice in the presence of OVA for 7 days. The supernatants were assayed for total IgE levels by using ELISA. Data are expressed as means ± SD of 3 independent cultures. Experiments were repeated twice with similar results. Journal of Allergy and Clinical Immunology 2000 105, 788-795DOI: (10.1067/mai.2000.104934) Copyright © 2000 Mosby, Inc. Terms and Conditions

Fig. 7 Cytokine production by Peyer’s patch cells from transgenic mice fed the OVA diet. Peyer’s patch cells were prepared from pooled Peyer’s patches of 3 transgenic mice fed the OVA diet for various periods, and the isolated cells were cultured with OVA for 44 hours. The supernatants were assayed for cytokine levels by using ELISA. Data are expressed as means ± SD of 3 independent cultures. Bars not sharing a common letter are significantly different. Experiments were repeated 3 times with similar results. Journal of Allergy and Clinical Immunology 2000 105, 788-795DOI: (10.1067/mai.2000.104934) Copyright © 2000 Mosby, Inc. Terms and Conditions