Abstract
Abstract Rotavirus-induced diarrhea is a life-threatening disease in children in developing countries. We have developed a novel system for prophylaxis and therapy against rotavirus using transgenic rice expressing the neutralizing variable domain of llama heavy-chain antibody fragments against rhesus-monkey rotavirus serotype G3P (MucoRice-ARP1). MucoRice-ARP1 was produced at high levels in rice seeds using an overexpression system and RNAi technology to suppress the production of major rice endogenous storage proteins. MucoRice-ARP1 was water-soluble and expressed in protein body II as a rice storage organelle and cytoplasm between protein bodies of in rice. We confirmed the full sequence of 123 amino acids of ARP1 by mass spectrometry. MucoRice-ARP1 neutralized human RV strains (ST-3 G4P[6], 69M G8P[10], F45 G9P[8], P G3P[8] and Va70 G4P[8]) in vitro. When administered orally, MucoRice-ARP1 markedly decreased the prevalence of diarrhea and viral load in a murine pup model. Long-term storage (>1yr) at room temperature of MucoRice-ARP1 did not impair its in vivo neutralizing activity. Oral MucoRice-ARP1 thus offer a novel cost-effective approach to prevent and treat rotavirus-induced diarrhea not only in children but also in adults who would not be expected to respond to the live attenuated vaccine.
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