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利用非复制型痘苗病毒载体，构建了能同时表达乙型肝炎（乙肝）病毒ＳＳ１、麻疹病毒ＨＡ和Ｆ及白细胞介素２（ＩＬ２）的非复制型重组痘苗病毒ＶＨＦＩＬ２ΔＣＫＳＳ１，及其相应的复制型重组痘苗病毒ＶＨＦＩＬ２ＳＳ１。这两株重组病毒在ＣＥＦ细胞上连续传至第２５代，经Ｓｏｕｔｈｅｒｎｂｌｏｔ证实，两株病毒均在Ａ２４、Ａ２７间稳定整合有ＳＳ１基因，非复制型重组病毒Ｃ、Ｋ间基因稳定缺失。经ＲＩＡ、ＥＬＩＳＡ及Ｗｅｓｔｅｒｎｂｌｏｔ方法检测，这两株病毒均可稳定分泌表达ＳＳ１蛋白，其分泌表达的ＳＳ１蛋白与ＣＨＯ细胞表达的ＳＳ１蛋白性质相近，接近于天然。经免疫打点检测，两株病毒均可稳定表达麻疹病毒的ＨＡ与Ｆ蛋白以及ＩＬ２。而且ＶＨＦＩＬ２ΔＣＫＳＳ１与ＶＨＦＩＬ２ＳＳ１表达多种外源抗原的水平无显著性差异。ＶＨＦＩＬ２ΔＣＫＳＳ１经皮内注射免疫家兔，４周后可诱发抗乙肝病毒ＳＳ１、抗麻疹病毒及抗痘苗病毒的特异性ＩｇＧ抗体；加强免疫２周后，特异性抗体滴度可上升４～１６倍，与其初始病毒ＶＨＦＩＬ２ΔＣＫ诱发的抗体水平相同，比ＶＨＦＩＬ２ＳＳ１诱发的抗体滴度低０～４倍。非复制型多价重组痘苗病毒的构建，为评价非复制型痘苗病毒载体作为多价疫苗载?
A non-replicating recombinant vaccinia virus VHFIL2ΔCKSS1 that can express hepatitis B virus (HBV) SS1, measles virus HA and F and interleukin 2 (IL2) at the same time was constructed by using non-replicating vaccinia virus vector and its corresponding replication Recombinant vaccinia virus VHFIL2SS1. The two recombinant viruses were continuously transmitted to the 25th passage on CEF cells. Southern blot confirmed that both viruses were stably integrated with SS1 gene in A24 and A27, and non-replicating recombinant viruses C and K were stably deleted. The results of RIA, ELISA and Western blot showed that both of the two viruses could stably secrete SS1 protein, and the secreted SS1 protein was similar to the SS1 protein expressed by CHO cells, close to the natural one. The immunization RBI test, two viruses can be stable expression of measles virus HA and F protein and IL2. Moreover, there was no significant difference in the levels of VHFIL2ΔCKSS1 and VHFIL2SS1 expressing multiple foreign antigens. VHFIL2ΔCKSS1 was immunized intradermally in rabbits. After 4 weeks, specific IgG antibodies against hepatitis B virus SS1, anti-measles virus and anti-vaccinia virus could be induced. After 2 weeks of booster immunization, the specific antibody titer increased 4 ~ 16 times, The level of antibody induced by its original virus VHFIL2 [Delta] CK is 0 to 4 times lower than the antibody titer induced by VHFIL2SS1. Construction of Non-replicating Multivalent Recombinant Vaccinia Virus To Assess Non-replicating Vaccinia Virus Vectors as Multivalent Vaccines