Development of combination vaccine by using cytokine expressing transgenic plants.

轮状病毒是引起哺乳动物腹泻的最重要病原体之一。我们已经培育出表达轮状病毒VP6 cDNA基因的转基因马铃薯植株，该基因编码一种外壳蛋白。此外，我们还在小鼠体内证实了转基因马铃薯的抗原性。在本研究中，我们培育了引入人干扰素基因的转基因草莓。利用共转化技术制备了同时表达牛IL-1和轮状病毒VP6蛋白的转基因草莓。在这里，我们研究了使用上述转基因草莓和/或转基因VP6土豆口服疫苗和细胞因子组合给鼠的效果。BALB/c（8周龄）小鼠每周自发饲喂转基因植物1次，共3次。最后一次（第三次）给药1周后，用10^5 TCID50剂量的轮状病毒EW株（小鼠组）攻毒。检测攻毒小鼠血液和粪便中的病毒抗原和抗病毒抗体。在给药小鼠粪便中检测到分泌性IgA。另一方面，hlFN草莓组、IL-1草莓组和非转基因草莓组的免疫标记物差异不显著；

Rotavirus is one of the most significant pathogens causing diarrhea in mammals. We have already produced the transgenic potato plants expressing rotavirus VP6 cDNA gene which encodes a coat protein. In addition, we have confirmed the antigenicity of the transgenic potato in mice.In this study we produced the transgenic strawberries which were introduced human interferon(hIFN) gene. Also we have produced the transgenic strawberry expressing both bovine IL-1 and rotavirus VP6 protein by using co-transformation technique. And here we examined the effect of oral administration to mice with combinations of vaccine and cytokine, using the transgenic strawberries and/or the transgenic VP6 potato mentioned above.BALB/c (8 week-old) mice were spontaneously fed with the transgenic plants once in a week, three times. After 1 week of the last(3rd) administration, those mice were challenged using rotavirus EW strain (murine group A) with dose of 10^5 TCID50.Virus antigen and anti-virus antibody in blood and feces of the challenged mice were examined. Secretory IgA was detected in feces of VP6 administrated mice. On the other hand, there was no significant difference in immune-markers among the hlFN strawberry, the IL-1 strawberry and non lransgenic strawberry administrated groups ;