A rice-based edible vaccine expressing multiple T cell epitopes induces oral tolerance for inhibition of Th2-mediated IgE responses

Abstract

Peptide immunotherapy using multiple predominant allergen-specific T cell epitopes is a safe and promising strategy for the control of type I allergy. In this study, we developed transgenic rice plants expressing mouse dominant T cell epitope peptides of Cry j I and Cry j II allergens of Japanese cedar pollen as a fusion protein with the soybean seed storage protein glycinin. Under the control of the rice seed storage protein glutelin GluB-1 promoter, the fusion protein was specifically expressed and accumulated in seeds at a level of 0.5% of the total seed protein. Oral feeding to mice of transgenic rice seeds expressing the T cell epitope peptides of Cry j I and Cry j II before systemic challenge with total protein of cedar pollen inhibited the development of allergen-specific serum IgE and IgG antibody and CD4 ؉ T cell proliferative responses. The levels of allergen-specific CD4 ؉ T cell-derived allergy-associated T helper 2 cytokine production of IL-4, IL-5, and IL-13 and histamine release in serum were significantly decreased. Moreover, the development of pollen-induced clinical symptoms was inhibited in our experimental sneezing mouse model. These results indicate the potential of transgenic rice seeds in production and mucosal delivery of allergen-specific T cell epitope peptides for the induction of oral tolerance to pollen allergens.Japanese cedar pollinosis ͉ peptide immunotherapy ͉ seed-specific expression

Rice-based mucosal vaccine as a global strategy for cold-chain- and needle-free vaccination

“…The CTB gene of V. cholerae was modified to a suitable form for rice seed and inserted into a binary vector (pGPTV-35S-HPT) (23). The resulting plasmid (Fig.…”

“…Using the cetyltrimethylammonium bromide extraction method, we extracted genomic DNA from the leaf tissues of transgenic rice and analyzed the integration of the CTB gene in genomic DNA using PCR (23). The expression of mRNA encoding CTB in the rice seed was analyzed by Northern blot as previously described (23).…”

“…Using the cetyltrimethylammonium bromide extraction method, we extracted genomic DNA from the leaf tissues of transgenic rice and analyzed the integration of the CTB gene in genomic DNA using PCR (23). The expression of mRNA encoding CTB in the rice seed was analyzed by Northern blot as previously described (23). Briefly, total RNA (30 g) extracted from the developing seeds of rice plants using the phenol/ chloroform extraction method was separated on a 1.0% (wt/vol) formaldehyde/agarose gel and transferred to Hybond N ϩ membranes (GE Healthcare, Piscataway, NJ).…”

“…Total seed protein was extracted from seeds as described previously (23). Briefly, seeds of rice plants were ground to a fine powder using a Multibeads shocker (Yasui Kikai, Osaka, Japan) and extracted in 2% (wt/vol) SDS, 8 M urea, 5% (wt/vol) ␤-mercaptoethanol, 50 mM Tris⅐HCl (pH 6.8), and 20% (wt/vol) glycerol before being separated by SDS/PAGE with a 15% to 25% gradient polyacrylamide gel (Daiichi Pure Chemical, Tokyo, Japan).…”

Strategies of mucosal immunotherapy for allergic diseases

“…These results indicate the potential of transgenic rice seeds in the production and mucosal delivery of allergen-specific T-cell epitope peptides for the induction of oral tolerance to pollen allergens. 50 Adjuvants for mucosal immunotherapy Cholera toxin B (CTB). CTB is produced by Vibrio cholera.…”