Rodríguez-Gómez Irene M, Käser Tobias, Gómez-Laguna Jaime, Lamp Benjamin, Sinn Leonie, Rümenapf Till, Carrasco Librado, Saalmüller Armin, Gerner Wilhelm
Institute of Immunology, Department of Pathobiology, University of Veterinary Medicine, Vienna, Veterinärplatz 1, 1210, Vienna, Austria.
Department of Anatomy and Comparative Pathology, Faculty of Veterinary Medicine, University of Córdoba, "International Excellence Agrifood Campus - CeiA3", 14014, Córdoba, Spain.
Vet Res. 2015 May 20;46(1):54. doi: 10.1186/s13567-015-0186-z.
In vitro generated monocyte-derived dendritic cells (moDCs) have frequently been used to study the influence of porcine reproductive and respiratory syndrome virus (PRRSV) infection on antigen presenting cells. However, obtained results have often been conflicting in regard to expression of co-stimulatory molecules and interaction with T cells. In this study we performed a detailed phenotypic characterisation of PRRSV-infected moDCs and non-infected moDCs. For CD163 and CD169, which are involved in PRRSV-entry into host cells, our results show that prior to infection porcine moDCs express high levels of CD163 but only very low levels for CD169. Following infection with either PRRSV-1 or PRRSV-2 strains after 24 h, PRRSV-nucleoprotein (N-protein)(+) and N-protein(-) moDCs derived from the same microculture were analyzed for expression of swine leukocyte antigen-DR (SLA-DR) and CD80/86. N-protein(+) moDCs consistently expressed higher levels of SLA-DR and CD80/86 compared to N-protein(-) moDCs. We also investigated the influence of PRRSV-infected moDCs on proliferation and frequency of Foxp3(+) regulatory T cells present within CD4(+) T cells in in vitro co-cultures. Neither CD3-stimulated nor unstimulated CD4(+) T cells showed differences in regard to proliferation and frequency of Foxp3(+) T cells following co-cultivation with either PRRSV-1 or PRRSV-2 infected moDCs. Our results suggest that a more detailed characterisation of PRRSV-infected moDCs will lead to more consistent results across different laboratories and PRRSV strains as indicated by the major differences in SLA-DR and CD80/86 expression between PRRSV-infected and non-infected moDCs present in the same microculture.
体外生成的单核细胞来源的树突状细胞(moDCs)经常被用于研究猪繁殖与呼吸综合征病毒(PRRSV)感染对抗抗原呈递细胞的影响。然而,关于共刺激分子的表达以及与T细胞的相互作用,所获得的结果常常相互矛盾。在本研究中,我们对感染PRRSV的moDCs和未感染的moDCs进行了详细的表型特征分析。对于参与PRRSV进入宿主细胞过程的CD163和CD169,我们的结果表明,在感染前,猪moDCs表达高水平的CD163,但CD169的表达水平极低。在用PRRSV -1或PRRSV -2毒株感染24小时后,对来自同一微量培养物的PRRSV核蛋白(N蛋白)阳性(N-protein(+))和阴性(N-protein(-))moDCs进行猪白细胞抗原-DR(SLA-DR)和CD80/86表达分析。与N蛋白阴性(N-protein(-))moDCs相比,N蛋白阳性(N-protein(+))moDCs始终表达更高水平的SLA-DR和CD80/86。我们还研究了感染PRRSV的moDCs对体外共培养中CD4(+) T细胞内Foxp3(+)调节性T细胞增殖和频率的影响。在用感染PRRSV -1或PRRSV -2的moDCs共培养后,无论是CD3刺激的还是未刺激的CD4(+) T细胞,在Foxp3(+) T细胞的增殖和频率方面均未显示出差异。我们的结果表明,如同一微量培养物中感染PRRSV与未感染PRRSV的moDCs之间SLA-DR和CD80/86表达的主要差异所示,对感染PRRSV的moDCs进行更详细的特征分析将在不同实验室和PRRSV毒株中产生更一致的结果。