Persistent protective effect of heat-killed Escherichia coli producing “engineered,” recombinant peanut proteins in a murine model of peanut allergy

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Persistent protective effect of heat-killed Escherichia coli producing “engineered,” recombinant peanut proteins in a murine model of peanut allergy Xiu-Min Li, MDa, Kamal Srivastava, MPhila, Alexander Grishin, PhDa, Chih-Kang Huang, MSa, Brian Schofield, JDb, Wesley Burks, MDc, Hugh A. Sampson, MDa Journal of Allergy and Clinical Immunology Volume 112, Issue 1, Pages 159-167 (July 2003) DOI: 10.1067/mai.2003.1622 Copyright © 2003 Mosby, Inc. Terms and Conditions

Fig. 1 Experimental protocol. C3H/HeJ mice were sensitized intragastrically with freshly ground, whole roasted peanut together with cholera toxin over an 8-week period. Treatment was initiated at week 10. Six groups (n = 12/group) were involved: Group 1 (sham) received methylcellulose vehicle only; group 2 received HKE-MP123 0.9 μg (low dose); group 3 received HKE-MP123, 9 μg (medium dose); group 4 received HKE-MP123, 90 μg (high dose); group 5 received HKE-vector; and group 6 was naive. The treatments were administered rectally in methylcellulose as vehicle 3 times at weekly intervals. Mice were challenged 2, 6, and 10 weeks after the termination of therapy (week 14, 18, and 22, respectively, after desensitization). After each challenge, 4 mice were killed for collection of blood and tissue samples. Journal of Allergy and Clinical Immunology 2003 112, 159-167DOI: (10.1067/mai.2003.1622) Copyright © 2003 Mosby, Inc. Terms and Conditions

Fig. 2 Mouse experiencing anaphylaxis. A, Normal C3H/HeJ mouse. B, Peanut-sensitized C3H/HeJ mouse experiencing an anaphylactic reaction after intragastric feeding of peanut; note swelling about the eyes and snout, pilar erecti, and cyanosis of the ears and feet. Journal of Allergy and Clinical Immunology 2003 112, 159-167DOI: (10.1067/mai.2003.1622) Copyright © 2003 Mosby, Inc. Terms and Conditions

Fig. 3 Persistent protection against peanut-induced anaphylactic reactions by HKE-MP123. Mice were challenged at weeks 2 (A) , 6 (B) , and 10 (C) after the last HKE-MP123 treatment. Anaphylactic symptom scores were determined 30 minutes after challenge. Each point indicates an individual mouse. Bars indicate the median of 12 mice (A) , 8 mice (B) , and 4 mice (C) in each group. *P < .05, and **P < .01 vs sham. Journal of Allergy and Clinical Immunology 2003 112, 159-167DOI: (10.1067/mai.2003.1622) Copyright © 2003 Mosby, Inc. Terms and Conditions

Fig. 4 Effect of therapy on plasma histamine levels. Thirty to 40 minutes after challenge, blood was collected and plasma was obtained. Histamine levels were measured by use of an enzyme immunoassay kit. Data are means ± SEM for each group of 12 mice (A) , 8 mice (B) , and 4 mice (C) in each group. *P < .05, and **P < .01 vs sham. Journal of Allergy and Clinical Immunology 2003 112, 159-167DOI: (10.1067/mai.2003.1622) Copyright © 2003 Mosby, Inc. Terms and Conditions

Fig. 5 Effect of therapy on peanut-specific IgE levels. Sera from all groups of mice were obtained during sensitization (3, 6, and 8 weeks), before treatment, and 1 day before each challenge (weeks 14, 18, and 22). Peanut-specific IgE (A) and IgG2a (B) levels were determined by ELISA. Data are mean ± SEM for each group as in Fig 2. *P < .05 vs sham; ***P < .001 vs sham. Journal of Allergy and Clinical Immunology 2003 112, 159-167DOI: (10.1067/mai.2003.1622) Copyright © 2003 Mosby, Inc. Terms and Conditions

Fig. 6 Effect of therapy on cytokine production. Cell suspensions were cultured in complete culture medium in the presence of crude peanut extract, medium alone, or concanavalin A (data not shown). Supernatants were collected 72 hours later, and cytokine levels were determined by ELISA. Results are expressed as mean ± SEM of 2 duplicate cultures of 4 mice. *P < .05; ** P < .01,***P < .001 vs sham. Journal of Allergy and Clinical Immunology 2003 112, 159-167DOI: (10.1067/mai.2003.1622) Copyright © 2003 Mosby, Inc. Terms and Conditions