囊性包虫重组14-3-3脂质体疫苗的制备及理化性质的检测

Preparation and Property of rEg14-3-3 Liposome Vaccine

笔者制备了囊性包虫重组14-3-3脂质体疫苗(Recombinant Echinoccocus granulosus 14-3-3,r Eg14-3-3),观察并分析脂质体疫苗的脂质体形态、疫苗抗原包封率及动物使用的安全性,确定囊性包虫重组14-3-3脂质体疫苗的制备工艺。笔者以DOPC、DOPG、MPBL等磷脂物质为原料,采用薄膜分散法制成均匀脂质薄膜,脂膜与14-3-3和MPLA水合、超声后制备出包载14-3-3蛋白和MPLA的多层交联脂质体疫苗。采用电子显微镜测定脂质体的形态。采用BCA蛋白定量法检测r Eg14-3-3脂质体疫苗的抗原包封率。将r Eg14-3-3脂质体疫苗和空脂质体分别注射入5只6周龄ICR小鼠皮下,观察其生物安全性,并在0周、2周、4周、6周和10周从尾部静脉采血,收集血清,用ELISA法检测14-3-3脂质体疫苗免疫后特异性Ig G抗体水平。制备的脂质体疫苗为乳白色形态较均一的球形,疫苗包封率为72.45%,体外释放缓慢,性状稳定,对小鼠无明显毒副作用。14-3-3脂质体免疫组小鼠Ig G水平在免疫后2周后迅速升高,与空白脂质体组相比差异显著。薄膜分散-超声法制备囊性包虫多层交联脂质体疫苗,抗原包封率高、性状稳定、安全无毒。该脂质体疫苗可有效诱导机体特异性免疫应答。

To prepare the r Eg14-3-3 liposome vaccine and investigate its physical and chemical properties.Prepare liposome with phospholipid,cholesterol and steary lamine by reverse evaporation,then add purified recombinant Echinoccocos granulosus 14-3-3 protein and Monophosphoryl lipid A(MPLA)for preparation of r Eg14-3-3 liposome vaccine by ultrosound.Observe the prepared vaccine formorphology under electron microscope,determine the envelope rate and release rate in vitro,and observe the biological safety of the liposome vaccine used in mice.Then r Eg14-3-3 liposome and the blank liposome were injected into 5 six weeks old mice sepratively to test IgG level with ELISA.The prepared r Eg14-3-3 liposome vaccine under electron microscope were round particles distributed.The envelope rate of the vaccine was more than 72%.The vaccine showed no stimulation or abnormal toxicity and was released slowly.The mice injected with r Eg14-3-3 liposome could produce high level of IgG antibody after 2 weeks,which showed significant difference compared mice injected with blank liposome.The prepared r Eg14-3-3 liposome vaccine showed stable property and safe.r Eg14-3-3 liposome vaccine could induce effective specific immunity response.Which shows that the r Eg14-3-3 liposome vaccine can be a promising anti-echinoccocosis vaccine.