Jiang Wen-Zheng, Jin Ning-Yi, Li Zi-Jian, Zhang Li-Shu, Han Wen-Yu
Key Laboratory of Genetic Engineering of PLA, Quartermaster University of PLA, Changchun 130062, China.
Sheng Wu Gong Cheng Xue Bao. 2004 May;20(3):337-41.
To screening out Chinese vaccine candidate against HIV-1, Chinese vaccine strain 282E4 of fowlpox virus was used as the vector to construct the recombinant fowlpox virus (rFPV) coexpressing gp120 of Chinese HIV-1 strain and IL-18, and the recombinant virus was indentified by PCR and Western blot. The specific DNA fragment could be amplified by PCR from the genome of rFPV. Western blot analysis showed that gp120 and IL-18 could be expressed not only in chicken embryo fibroblast (CEF) cells infected by rFPV, but also in mammalian cells infected by rFPV. After the recombinant fowlpox virus was inoculated into BALB/c mice, the spleen specific CTL activities and serum antibodies in the immunized mice were detected, which demonstrated that the rFPV had good immunogenicity and could induce BALB/c mice to produce specific humoral and cellular immunity. IL-18 palyed the role of immunoadjuvant. The study lays the basis on the preparation of genetic engineering live vector vaccine against HIV-1.
为筛选抗HIV-1的中国疫苗候选株,以中国鸡痘病毒疫苗株282E4为载体构建共表达中国HIV-1株gp120和IL-18的重组鸡痘病毒(rFPV),并通过PCR和Western blot对重组病毒进行鉴定。PCR可从rFPV基因组中扩增出特异性DNA片段。Western blot分析表明,gp120和IL-18不仅能在感染rFPV的鸡胚成纤维细胞(CEF)中表达,也能在感染rFPV的哺乳动物细胞中表达。将重组鸡痘病毒接种到BALB/c小鼠后,检测免疫小鼠的脾脏特异性CTL活性和血清抗体,结果表明rFPV具有良好的免疫原性,能诱导BALB/c小鼠产生特异性体液免疫和细胞免疫。IL-18发挥了免疫佐剂的作用。该研究为抗HIV-1基因工程活载体疫苗的制备奠定了基础。