| Summary: | 碩士 === 國立中興大學 === 食品暨應用生物科技學系所 === 97 === Enterovirus 71 (EV71) is the main causative agent of Hand, Foot and Mouth Disease (HFMD) and is associated with severe neurological diseases resulting in high mortalities. Currently, there is neither vaccine nor therapeutic treatment available. The significant increase in recent EV71 epidemic throughout the Asia-Pacific region was clinically observed recently.
Bacillus subtilis is a Gram-positive, facultative anaerobic rod-shaped endospore-forming bacterium that occurs naturally in soil and water. It has been used in the production of fermented food, extracellular enzymes as well as special chemicals. Some strains of B. subtilis are exploited as probiotics or biological control agents. In the field of genetic engineering, B. subtilis has been developed as a host for the production of homologous and heterologous proteins.
This study developed an effective vaccine by transforming the partial genes of VP1, VP1e and VP1f into Bacillus subtilis. VP1 is a coat protein of EV71 and considered as potent epitope; VP1e is the N-terminal sequence of VP1 (amino acid is 1-81) and VP1f is the hydrophilic sequences fusion (amino acid are 95-109, 162-170, 210-221 and 266-274). In this study, we first generated a Bacillus subtilis secretory production system to produce the VP1e and VP1f epitope protein, high level of recombinant protein was expressed and secreted. Then, vaccination with recombinated protein VP1e, VP1f, or combined VP1e, VP1f and rLZ8 who administered to Balb/c mice. The VP1-specific serum IgG1 and IgG2a were examined. The VP1-specific serum IgG antibody induced by VP1e triggered both IgG subtype responses, indicating that Bacillus subtilis expressing VP1e was successful and high efficiency as vaccine. This study not only demonstrates the feasibility of using Bacillus subtilis secetory system as vaccine producer, but also suggests the probability of enterovirus vaccine development using VP1e as potent epitope.
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