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

1)为了研制高水平表达外源基因的痘苗病毒载体，我们克隆了牛痘病毒A型包涵体（ATI）基因。ATI基因的启动子比p7.5基因的启动子强数倍。为了获得更好的启动子，我们将ATI启动子与p7.5启动子的几组早期序列相结合，以获得在vv感染的早期和晚期都能发挥作用的启动子。3）表达env基因的重组VV免疫家兔后，对HTLC-I的攻击有保护作用。I产生细胞（MT-2）。用食蟹猴做类似的实验，结果表明，不仅抗env抗体，而且T细胞的启动也是重要的。4）不同VV毒株中的LC 16 mO虽然具有很低的神经毒力，但仍能诱导相当高水平的抗enb抗体。5）为了检测JTLV-I的哪种成分是细胞毒性T细胞（CTL）的靶，将产生HTLV-I成分的大鼠细胞注射到同系大鼠中，然后使用各种重组VV感染的大鼠细胞测量其淋巴细胞的CTL活性。结果表明，gag和pX蛋白是主要的靶蛋白。

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.