Bioactive murine and human interleukin-12 fusion proteins which retain antitumor activity in vivo
Bioactive murine and human interleukin-12 fusion proteins which retain antitumor activity in vivo
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DOI：
10.1038/nbt0197-35
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发表时间：
1997-01-01
影响因子：
46.9
通讯作者：
Mulligan, RC
中科院分区：
文献类型：
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作者：
Lieschke, GJ;Rao, PK;Mulligan, RC
白细胞介素 12 (IL-12) 在细胞因子中是独特的，因为它是两个单独编码的亚基（p35 和 p40）的二硫键连接的异二聚体。我们从逆转录病毒构建体表达单链IL-12蛋白，其中两个IL-12亚基通过6-15个氨基酸的多肽接头连接，并删除了尾随亚基的22个氨基酸前导序列。含有(Gly(4)Ser)(3)接头的鼠融合蛋白IL-12.p40.L.Delta p35稳定表达，具有体外生物活性，并且具有与天然和重组IL-12相当的表观比活性。蛋白质印迹证实鼠IL-12.p40.L.Delta p35保留了连接多肽序列。含有Gly(6)Ser接头的类似人IL-12.p40.L.Delta p35融合蛋白具有与重组人IL-12相当的表观比活性的生物活性。在预先存在的 CMS-5 肿瘤模型中，分泌天然或融合蛋白形式的 IL-12 的 CMS-5 细胞延长了存活时间并导致肿瘤完全消退。
Interleukin-12 (IL-12) is unique amongst cytokines in being a disulfide-linked heterodimer of two separately encoded subunits (p35 and p40). We expressed single chain IL-12 proteins from retroviral constructs in which the two IL-12 subunits were linked by a 6-15 amino acid polypeptide linker, with deletion of the 22 amino acid leader sequence of the,trailing subunit. The murine fusion protein IL-12.p40.L.Delta p35 containing a (Gly(4)Ser)(3) linker was stably expressed, bioactive in vitro, and had an apparent specific activity comparable to that of native and recombinant IL-12. Western blotting confirmed that murine IL-12.p40.L.Delta p35 retained the linking polypeptide sequences. The analogous human IL-12.p40.L.Delta p35 fusion protein containing a Gly(6)Ser linker was bioactive with an apparent specific activity comparable to recombinant human IL-12. In a preexisting CMS-5 tumor model, CMS-5 cells secreting either native or fusion protein forms of IL-12 prolonged survival and led to complete tumor regression.