Huang Li-Hsin, Lin Hao-Yeh, Lyu Ying-Tzu, Gung Chiau-Ling, Huang Ching-Tsan
MycoMagic Biotechnology Co. Ltd., 8F-1, 12, Lane 270, Sec. 3, Beishen Road, New Taipei City, Taiwan.
Department of Biochemical Science and Technology, National Taiwan University, 1, Sec. 4, Roosevelt Road, Taipei, Taiwan.
Food Technol Biotechnol. 2019 Mar;57(1):105-112. doi: 10.17113/ftb.57.01.19.5865.
Orally administered fungal vaccines show promise for the prevention of infectious diseases. Edible mushrooms are deemed appropriate hosts to produce oral vaccines due to their low production cost and low risk of gene contamination. However, their low expression level of antigens has limited the potential development of oral vaccines using mushrooms. The low expression level might result from impurity of the transgenic mycelia since dikaryotic mycelia are commonly used as transformation materials. In this study, stable transgenic hepatitis B virus surface antigen (HBsAg) in transformants was obtained by -mediated transformation, followed by fruiting and basidiospore mating. The formation of HBsAg was detected by western blot analysis. The expression levels of HBsAg in transgenic fruiting bodies were (129.3±15.1), (110.9±1.7) and (161.1±8.5) ng/g total soluble protein. However, the values may be underestimated due to incomplete protein extraction. Two of the four pigs in the experimental group produced positive anti-HBsAg-specific IgG after being fed the HBsAg transgenic fruiting bodies for 20 weeks, while no anti-HBsAg antibody was detected in the control group. One of the positive pigs had HBsAg titres of 5.36 and 14.9 mIU/mL in weeks 10 and 14, respectively, but expression faded thereafter. The other positive pig displayed HBsAg titres of 9.75, 17.86 and 39.87 mIU/mL in weeks 14, 18 and 20, respectively. The successful immunogenicity in pigs fed transgenic fruiting bodies demonstrated the potential of using the fungus as an oral vaccine.
口服真菌疫苗在预防传染病方面显示出前景。食用蘑菇因其生产成本低和基因污染风险低，被认为是生产口服疫苗的合适宿主。然而，它们抗原的低表达水平限制了使用蘑菇的口服疫苗的潜在发展。低表达水平可能是由于转基因菌丝体的不纯导致的，因为双核菌丝体通常用作转化材料。在本研究中，通过介导转化获得转化体中稳定的转基因乙型肝炎病毒表面抗原（HBsAg），随后进行出菇和担孢子交配。通过蛋白质印迹分析检测HBsAg的形成。转基因子实体中HBsAg的表达水平为（129.3±15.1）、（110.9±1.7）和（161.1±8.5）ng/g总可溶性蛋白。然而，由于蛋白质提取不完全，这些值可能被低估。实验组的四只猪中有两只在喂食HBsAg转基因子实体20周后产生了阳性抗HBsAg特异性IgG，而对照组未检测到抗HBsAg抗体。其中一只阳性猪在第10周和第14周的HBsAg滴度分别为5.36和14.9 mIU/mL，但此后表达减弱。另一只阳性猪在第14周、第18周和第20周的HBsAg滴度分别为9.75、17.86和39.87 mIU/mL。在喂食转基因子实体的猪中成功诱导免疫原性证明了将这种真菌用作口服疫苗的潜力。
