Rompato G, Ling E, Chen Z, Van Kruiningen H, Garmendia A E
Department of Pathobioloy and Veterinary Sciences, University of Connecticut, 61 N. Eagleville Rd, Storrs, CT, USA.
Vet Immunol Immunopathol. 2006 Jan 15;109(1-2):151-60. doi: 10.1016/j.vetimm.2005.08.015. Epub 2005 Sep 12.
This study investigated the effect of swine interleukin 2 (IL-2) and swine interleukin 4 (IL-4) on the development of immune responses induced by a PRRSV-ORF7 DNA vaccine (phCMV-ORF7). The two cytokines were cloned separately in the eukaryotic expression vector phCMV, and delivered via gene gun as adjuvants for the DNA vaccine. Groups of 3-week-old certified PRRSV-free, castrated male, Yorkshire crossbred pigs, were vaccinated with or without the IL-2 or IL-4. The ensuing humoral and cellular immune responses were analyzed by a PRRSV-specific ELISA, and by an in vitro blastogenic response of peripheral blood mononuclear cells (PBMC) stimulated by viral antigen, respectively. The animals were boosted 21 days post-vaccination and challenged 28 days afterward. The virus loads post-challenge were measured by real time PCR. The group of swine receiving the vaccine plus IL-2 had significant virus-specific blastogenic responses 3 weeks after the vaccine-cytokine boost, when compared to those of the experimental pigs that received the vaccine plus IL-4, vaccine alone, unvaccinated controls or the pigs vaccinated with the DNA vaccine cloned in the reverse orientation (phCMV-ORF7(Rev)). None of the experimental swine had detectable specific antibodies against the virus during the vaccination phase. The virus load peak in vaccinated animals was delayed by about 72h as compared to that of the control pigs (unvaccinated and vaccinated with the phCMV-ORF7(Rev) construct). Interestingly, animals that received the phCMV-ORF7 vaccine alone consistently had low virus loads throughout the study. These results demonstrate that IL-2 has a positive inductive effect on the activation of vaccine-induced virus-specific cellular immunity, while IL-4 appeared to have a suppressive effect. Our data also suggest that ORF7 may play a role in reducing the virus load in PRRSV infected animals.
本研究调查了猪白细胞介素2(IL-2)和猪白细胞介素4(IL-4)对猪繁殖与呼吸综合征病毒(PRRSV)开放阅读框7(ORF7)DNA疫苗(phCMV-ORF7)诱导的免疫反应发展的影响。这两种细胞因子分别克隆于真核表达载体phCMV中,并通过基因枪作为DNA疫苗的佐剂进行递送。将3周龄经认证无PRRSV的去势雄性约克夏杂交猪分组,分别接种或不接种IL-2或IL-4。随后分别通过PRRSV特异性酶联免疫吸附测定(ELISA)以及病毒抗原刺激外周血单个核细胞(PBMC)的体外增殖反应,分析体液免疫和细胞免疫反应。动物在接种疫苗21天后进行加强免疫,并在之后28天进行攻毒。通过实时聚合酶链反应(PCR)测定攻毒后的病毒载量。与接受疫苗加IL-4、单独疫苗、未接种对照或接种反向克隆的DNA疫苗(phCMV-ORF7(Rev))的实验猪相比,接受疫苗加IL-2的猪群在疫苗-细胞因子加强免疫3周后具有显著的病毒特异性增殖反应。在接种阶段,没有实验猪检测到针对该病毒的特异性抗体。与对照猪(未接种和接种phCMV-ORF7(Rev)构建体)相比,接种疫苗动物的病毒载量峰值延迟了约72小时。有趣的是,在整个研究过程中,单独接受phCMV-ORF7疫苗的动物病毒载量始终较低。这些结果表明,IL-2对疫苗诱导的病毒特异性细胞免疫激活具有正向诱导作用,而IL-4似乎具有抑制作用。我们的数据还表明,ORF7可能在降低PRRSV感染动物的病毒载量中发挥作用。