Development of Recombinant Vaccinia Virus Vaccine Against Adult T-cell Leukemia

1)为构建高水平表达外源基因的牛痘病毒载体，克隆了牛痘病毒a型包涵体（ATI）的编码基因。ATI基因的启动子比p7.5基因的启动子强数倍。为了构造更好的启动子，我们将ATI启动子与p7.5启动子的几组早期区域结合，得到了在vv感染的早期和晚期都能发挥良好作用的启动子。2)检测了表达env、gag和px基因的重组VV_s的性质。它们都产生了真正的htlc - 1蛋白。3)用表达env基因的重组VV免疫家兔，对htlv - 1产生细胞（MT-2）的攻击具有保护作用。在对野食蟹进行类似实验时，发现除了抗嫉妒抗体外，T细胞的启动也起着重要的保护作用。4) LC16mO在不同VV毒株中诱导的抗enb抗体水平很高，但其神经毒力很低。因此，该菌株是人类疫苗接种载体的良好候选者。5)为了检测JTLV-I的哪个成分是细胞毒性T细胞（CTL）的靶点，我们将产生HTLV-I成分的大鼠细胞感染到同基因大鼠体内，然后用各种重组vv感染的大鼠细胞检测其淋巴样细胞的CTL活性。结果表明，gag蛋白和pX蛋白是主要的靶点。

1) To develop the vaccinia virus vectors(VV) that express foreign genes at higher level, we cloned the gene encoding A-type inclusion body (ATI) of cowpox virus. The promoter of ATI gene was seceral times stronger than that of p7.5 gene. To constract better promoter, we combined the ATI promoter and the several sets of early region of p7.5 promoter so as to obtain the promoter which can act very well at both early and late times of vv infection.2) We examined the properties of the recombinant VV_s that express the env, gag or px gene.They all produced authentic HTLC-I proteins.3) The rabbits which had been immunized by the recombinant VV expressing the env gene were protected from challenge of the HTLV-I producing cells (MT-2). When the similar experiment using cynomolgus nomkeys was done, it was rebealed that not only anti env antibodies but also priming of T cells were important for the protection.4) LC16mO among various VV strains induced anti enb antibody at quite high level although it had very low neurovirulence. Thus, this strain is a good candidate as a vector for vaccination of humans.5) To examine which component of JTLV-I is the target of cytotoxic T cells (CTL), rat cells producing HTLV-I components were inhected into the syngenic rats and then the CTL activity of their lymphoid cells were measured using the various recombinant VV-infected rat cells. The results showed that gag and pX proteins were the major targets.