MOI对PK-15细胞感染PPV后细胞因子mRNA转录时相的影响

Effect of MOI on transcriptional profiles of cytokines mRNA after PK-15 cell cultures infected with porcine parvovirus

摘要:

【目的】更好地了解感染复数(MOI)对猪细小病毒(PPV)感染PK-15细胞后引起细胞因子的反应, 探讨宿主-病毒之间的作用关系。【方法】运用荧光定量PCR技术, 测定和分析3种感染复数对PPV感染PK-15细胞引起的病毒DNA量的变化和细胞因子IFN-β、IRF-3、TNF-α和IL-18分泌水平。【结果】PPV感染PK-15细胞12 h病毒开始大量迅速增殖, 感染48 h且MOI为1.0时达到最高峰; PPV感染后可引起PK-15细胞中IFN-β、IRF-3、TNF-α和IL-18的表达量显著增加, 其中IRF-3基因在感染后1 h且MOI为10.0时表达量达到967倍。【结论】细胞因子的分泌水平显著依赖感染复数和感染时间的交互作用。

Abstract:

[Objective] Porcine Kidney-15 cells (PK-15) were infected by Porcine Parvovirus (PPV) to survey the host inflammatory responses, particularly multiplicity of infection (MOI) for the cytokines responses of PPV infection, and the host-PPV interaction. [Methods] Then the viral DNA was measured and analyzed using real-time PCR. The transcript level of cytokines (IFN-β, IRF-3, TNF-α and IL-18) were detected by real-time PCR too. [Results] We found that PPV proliferated rapidly in PK-15 cell 12 hours after infection, and reached peak after 48 hours infection and MOI=1.0; the transcription levels of IFN-β, IRF-3, TNF-α and IL-18 increased obviously, and the transcription level of IRF-3 gene increased to 967 times in 1 hour and MOI=10.0 after infection. [Conclusion] The results revealed that the level of cytokines significantly dependent on the interaction of MOI and infection time.