呼吸道合胞病毒重组蛋白候选疫苗的质粒构建、表达及免疫原性和保护性研究

PCR扩增呼吸道合胞病毒（respiratory syncytial virus,RSV）M2 蛋白的CD8＋T细胞表位F/M2:81-95和RSV-G蛋白的B细胞表位片段G:125～225（简称G1）,以一个Linker连接,插入质粒pET-DsbA中构建原核表达重组质粒, 转染E.coli BL21(DE3)后成功表达了融合蛋白DsbA-G1-Linker-F/M2:81-95（简称D-G1LF/M2）,Western-blot结果表明该融合蛋白是RSV特异性的,采用Ni+螯合亲和层析法纯化变性的包涵体溶液,经梯度透析法复性,用该蛋白免疫BALB/c小鼠,结果表明被免疫小鼠肺部及血清中产生了高滴度的抗D-G1LF/M2及抗RSV IgG抗体和中和抗体,同时还诱导产生了RSV特异性的CTL应答;IgG的亚型IgG1/IgG2a的比值为2.66;用RSV攻击免疫后的小鼠,病毒滴定法检测肺部RSV滴度,结果表明D-G1LF/M2对小鼠肺部具有保护作用。     

Daphnane diterpenoids isolated from Trigonostemon thyrsoideum as HIV-1 antivirals

Four highly oxygenated daphnane diterpenoids, trigonothyrins D-G (1-4), were isolated from the stems of Trigonostemon thyrsoideum, and their structures were elucidated on the basis of extensive spectroscopic studies. Inhibitory activity against HIV-1 was assessed for compounds 1, 3 and 4, wherein, 3 showed activity with an EC₅₀ value of 0.13 μg/mL and a therapeutic index (TI) of 75.1.