Interferon is an important cytokine that plays a critical role in the initial host defense against viral infection. Recombinant human adenoviruses expressing human interferon-alpha (Ad-HIFNalpha) or pig interferon-beta fused with interleukin-18 (Ad-PIFNbeta-IL18) were constructed and used to induce an early protective response against foot and mouth disease (FMD). To analyze the antiviral effect, bovine thyroid and porcine kidney IBRS-2 cells and ICR mice were treated with Ad-HIFNalpha, Ad-PIFNbeta-IL18, and cocktail of Ad-HIFNalpha and Ad-PIFNbeta-IL18. The survival rate of suckling mice was monitored after foot and mouth disease virus (FMDV) challenge following intra-peritoneal (IP) administration of appropriate adenovirus. Indirect antigen ELISA was performed to evaluate inhibition of FMDV replication following challenge with the FMDV O, A, or Asia 1 serotypes in vitro. These recombinant adenoviruses reduced the replication of FMDV in susceptible cells, thereby decreasing the fatality in mice, suggesting that they can be a useful control method for the early protection against FMD infection in livestock after field trial.
Adenoviridae ; Adenoviruses, Human ; Animals ; Asia ; Enzyme-Linked Immunosorbent Assay ; Foot ; Foot-and-Mouth Disease ; Foot-and-Mouth Disease Virus ; Humans ; Interferon-alpha ; Interferon-beta ; Interferons ; Interleukin-18 ; Kidney ; Livestock ; Mice ; Mice, Inbred ICR ; Survival Rate ; Thyroid Gland

[摘 要] 目的：探讨DJ-1基因过表达对人胃癌MGC803细胞增殖、迁移、侵袭与上皮间质转化（EMT）的影响及其机制。方法：利用基因转染技术构建DJ-1基因过表达MGC803细胞，实验分为MGC803、空载体和DJ-1过表达组。采用MTT、平板克隆形成、细胞划痕和Transwell实验分别检测DJ-1过表达对MGC803细胞增殖、克隆形成、迁移与侵袭的影响；qPCR和WB法检测DJ-1过表达对各组细胞DJ-1、PTEN、Akt、p-Akt、Snail、vimentin、E-cadherin、MMP-9与TIMP-3表达的影响，相差显微镜下观察MGC803细胞形态学的变化。裸鼠荷瘤实验检测DJ-1过表达对MGC803细胞移植瘤体内生长的影响。结果：成功构建DJ-1基因稳定过表达的MGC803细胞。与MGC803组和空载体组比较，DJ-1过表达组细胞的增殖能力与克隆形成数均显著增加（均P<0.05），细胞迁移距离明显增加、划痕距离明显缩短（均P<0.05），迁移与侵袭细胞数显著增多（均P<0.05），DJ-1 mRNA与蛋白表达明显上调、PTEN mRNA与蛋白表达下调（均P<0.05），Akt总蛋白各组比较无明显差异（均P>0.05），p-Akt蛋白表达明显上调（P<0.05），Snail、vimentin与MMP-9表达上调、E-cadherin与TIMP-3表达下调（均P<0.05）。相差显微镜下见长梭形细胞数目增多，圆形与椭圆形细胞减少，异型性更为明显。荷瘤裸鼠体内实验结果表明，与MGC803组相比较，DJ-1过表达组MGC803细胞移植瘤生长速度明显加快、移植瘤质量显著增加（均P<0.05）。结论：DJ-1过表达可通过PTEN/Akt通路在体内外抑制MGC803细胞的增殖、迁移、侵袭与EMT。