Department of Veterinary and Biomedical Sciences, South Dakota State University, Brookings, SD 57007, USA.
Virus Res. 2012 Feb;163(2):461-8. doi: 10.1016/j.virusres.2011.11.007. Epub 2011 Nov 20.
The cytokine interleukin-1 beta (IL-1β) is a potent inflammatory mediator in response to infection, and can be used as an immunological adjuvant. In this study, we constructed a recombinant porcine reproductive and respiratory syndrome virus (vP129/swIL1β) expressing swine IL-1β from the separate subgenomic mRNA inserted between the ORF1b and ORF2 genome region. MARC-145 cells infected with vP129/swIL1β secreted 1947 pg of IL-1β per 2 × 10(5)cells at 36 h post-infection. In vitro growth kinetics analysis in MARC-145 cells showed that the vP129/swIL1β virus had a similar replication rate as that of parental virus. We further performed in vivo characterization of the vP129/swIL1β virus in a nursery pig disease model. The vP129/swIL1β infected pigs did not show visible clinical signs, while respiratory distress and lethargy were evident in pigs infected with the parental virus. The expression of various cytokines from peripheral blood mononuclear cells measured by fluorescent microsphere immunoassay showed that IL-1β, IL-4 and IFN-γ expression levels were up-regulated in pigs infected with vP129/swIL1β at 7 and 14 days post-infection. However, no detectable level of IL-1β was found in serum samples from pigs infected with either vP129/swIL1β or parental virus. In summary, this study demonstrated a recombinant PRRSV as a useful tool to study the role of different cytokines in disease progression and immune responses, which represents a new strategy for future therapeutic application and vaccine development.
细胞因子白细胞介素-1β(IL-1β)是一种针对感染的有效炎症介质,可用作免疫佐剂。在这项研究中,我们构建了一种表达猪白细胞介素-1β的重组猪繁殖与呼吸综合征病毒(vP129/swIL1β),该病毒的 swIL1β 基因由单独的亚基因组 mRNA 插入 ORF1b 和 ORF2 基因组区域之间构建。感染 vP129/swIL1β 的 MARC-145 细胞在感染后 36 小时分泌 1947pg 的 IL-1β/2×10(5)细胞。在 MARC-145 细胞中的体外生长动力学分析表明,vP129/swIL1β 病毒的复制率与亲本病毒相似。我们进一步在仔猪疾病模型中对 vP129/swIL1β 病毒进行了体内特性研究。vP129/swIL1β 感染的猪没有表现出明显的临床症状,而感染亲本病毒的猪则表现出呼吸窘迫和昏睡。通过荧光微球免疫分析测量外周血单核细胞中的各种细胞因子表达,结果表明在感染 vP129/swIL1β 后的第 7 和 14 天,猪的 IL-1β、IL-4 和 IFN-γ 表达水平上调。然而,在感染 vP129/swIL1β 或亲本病毒的猪的血清样本中未检测到 IL-1β。总之,这项研究表明,重组 PRRSV 是研究不同细胞因子在疾病进展和免疫反应中的作用的有用工具,这代表了未来治疗应用和疫苗开发的新策略。