Singleton Helen, Graham Simon P, Bodman-Smith Katherine B, Frossard Jean-Pierre, Steinbach Falko
Virology Department, Animal and Plant Health AgencySurrey, UK; Faculty of Health and Medical Sciences, University of SurreySurrey, UK.
Faculty of Health and Medical Sciences, University of Surrey Surrey, UK.
Front Microbiol. 2016 Jun 2;7:832. doi: 10.3389/fmicb.2016.00832. eCollection 2016.
Monocyte-derived macrophages (MoMØ) and monocyte-derived dendritic cells (MoDC) are two model systems well established in human and rodent systems that can be used to study the interaction of pathogens with host cells. Porcine reproductive and respiratory syndrome virus (PRRSV) is known to infect myeloid cells, such as macrophages (MØ) and dendritic cells (DC). Therefore, this study aimed to establish systems for the differentiation and characterization of MoMØ and MoDC for subsequent infection with PRRSV-1. M-CSF differentiated MoMØ were stimulated with activators for classical (M1) or alternative (M2) activation. GM-CSF and IL-4 generated MoDC were activated with the well established maturation cocktail containing PAMPs and cytokines. In addition, MoMØ and MoDC were treated with dexamethasone and IL-10, which are known immuno-suppressive reagents. Cells were characterized by morphology, phenotype, and function and porcine MØ subsets highlighted some divergence from described human counterparts, while MoDC, appeared more similar to mouse and human DCs. The infection with PRRSV-1 strain Lena demonstrated different replication kinetics between MoMØ and MoDC and within subsets of each cell type. While MoMØ susceptibility was significantly increased by dexamethasone and IL-10 with an accompanying increase in CD163/CD169 expression, MoDC supported only a minimal replication of PRRSV These findings underline the high variability in the susceptibility of porcine myeloid cells toward PRRSV-1 infection.
单核细胞衍生的巨噬细胞(MoMØ)和单核细胞衍生的树突状细胞(MoDC)是在人类和啮齿动物系统中建立得很好的两种模型系统,可用于研究病原体与宿主细胞的相互作用。已知猪繁殖与呼吸综合征病毒(PRRSV)可感染髓样细胞,如巨噬细胞(MØ)和树突状细胞(DC)。因此,本研究旨在建立MoMØ和MoDC的分化和表征系统,以便随后用PRRSV-1进行感染。用激活剂刺激经M-CSF分化的MoMØ进行经典(M1)或替代(M2)激活。用含有病原体相关分子模式(PAMPs)和细胞因子的成熟鸡尾酒激活经GM-CSF和IL-4生成的MoDC。此外,用已知的免疫抑制试剂地塞米松和IL-10处理MoMØ和MoDC。通过形态、表型和功能对细胞进行表征,猪MØ亚群与所描述的人类对应物存在一些差异,而MoDC似乎与小鼠和人类DC更相似。用PRRSV-1毒株Lena感染显示MoMØ和MoDC之间以及每种细胞类型的亚群内具有不同的复制动力学。虽然地塞米松和IL-10显著增加了MoMØ的易感性,并伴随着CD163/CD169表达的增加,但MoDC仅支持PRRSV的最小复制。这些发现强调了猪髓样细胞对PRRSV-1感染易感性的高度变异性。