Construction of vectors expressing bioactive heterodimeric and single-chain murine interleukin-12 for gene therapy.
Construction of vectors expressing bioactive heterodimeric and single-chain murine interleukin-12 for gene therapy.
复制标题
用于基因治疗的表达生物活性异二聚体和单链鼠白细胞介素 12 的载体的构建。
DOI：
10.1089/hum.1998.9.4-457
复制
发表时间：
1998
影响因子：
4.2
通讯作者：
Bor
中科院分区：
文献类型：
--
作者：
Yueh Lun Lee;Mi;Yen;Bor
白细胞介素-12（IL-12）可以用于防御多种病原体，抑制肿瘤生长和转移，甚至可以用作疫苗的佐剂以增强有益的1型T辅助细胞（Th 1）应答而不是有害的2型T辅助细胞（Th 2）应答。为了将IL-12基因应用于基因治疗如DNA疫苗，构建了含有两个巨细胞病毒（CMV）启动子的pIL-12载体，以分别驱动p35和p40亚基的表达。此外，设计了一个pscIL-12载体，该载体带有一个连接子，将p35 cDNA与p40 cDNA融合，以产生单链IL-12蛋白，不仅确保了p35和p40亚基的表达相等，而且没有游离的p40亚基干扰IL-12活性。结果表明，pIL-12转染COS细胞系和C2 C12肌细胞系后，可产生较高水平的具有生物学活性的IL-12。有趣的是，pscIL-12载体也可以在体外表达具有生物活性的鼠IL-12融合蛋白。此外，体内功能研究还表明，用表达B肝炎病毒（HBV）主要包膜蛋白的pS载体和编码天然IL-12或单链IL-12融合蛋白的IL-12载体共免疫小鼠，引起更高水平的IgG 2a抗-HBs抗体和Th 1相关细胞因子。因为p35和p40基因可以通过使用单个启动子在载体中表达，所以pscIL-12在将来用于核酸疫苗接种或用于针对疾病的基因治疗中应该是有用的。
It has been well demonstrated that interleukin-12 (IL-12) could be useful to defend against a variety of pathogens, to suppress tumor growth and metastasis, and even to be employed as an adjuvant of vaccines to enhance beneficial type 1 T helper (Th1) cell response over detrimental type 2 T helper (Th2) cell responses. To apply IL-12 genes in gene therapy such as a DNA vaccine, a pIL-12 vector was constructed that contained two cytomegalovirus (CMV) promoters to drive the expression of p35 and p40 subunits, respectively. In addition, a pscIL-12 vector was designed with a linker to fuse p35 cDNA with p40 cDNA to produce a single-chain IL-12 protein, ensuring not only that the expression of p35 and p40 subunits was equally expressed, but also that no free p40 subunits interfered with IL-12 activity. The data suggested pIL-12 could produce a rather high level of biologically active IL-12 after transfection of COS cell lines as well as C2C12 muscle cell lines, as measured by both concanavalin A blast proliferation assay and enzyme-linked immunosorbent assay. Interestingly, the pscIL-12 vector could also express a bioactive murine IL-12 fusion protein in vitro. Furthermore, in vivo functional studies also demonstrated that mice co-immunized with a pS vector expressing the major envelope protein of hepatitis B virus (HBV) and IL-12 vectors encoding native IL-12 or single-chain IL-12 fusion protein elicited higher levels of IgG2a anti-HBs antibody and of Th1-related cytokine. Because p35 and p40 genes can be expressed in a vector by using a single promoter, pscIL-12 should be useful in future applications for nucleic acid vaccination or for gene therapy against diseases.
登录
查看更多内容
影响因子：
20.3
作者：
M. Kubin;JM Chow;G. Trinchieri
通讯作者：
M. Kubin;JM Chow;G. Trinchieri
影响因子：
56.9
作者：
HSIEH, CS;MACATONIA, SE;MURPHY, KM
通讯作者：
MURPHY, KM
影响因子：
56.9
作者：
BIRD, RE;HARDMAN, KD;WHITLOW, M
通讯作者：
WHITLOW, M
DOI：
10.1073/pnas.85.16.5879
发表时间：
1988-08-01
影响因子：
11.1
作者：
HUSTON, JS;LEVINSON, D;OPPERMANN, H
通讯作者：
OPPERMANN, H
表达生物活性人白细胞介素 12 的逆转录病毒载体的构建和表征。
影响因子：
4.2
作者：
Zitvogel,L;Tahara,H;Cai,Q;Storkus,WJ;Muller,G;Wolf,SF;Gately,M;Robbins,PD;Lotze,MT
通讯作者：
Lotze,MT