Institute of Animal Health (IUSA), University of Las Palmas de Gran Canaria, Arucas 35413, Spain.
Servicio de Microbiología, Hospital Universitario Marqués de Valdecilla e Instituto IDIVAL, Santander, Spain.
Microbes Infect. 2015 Apr;17(4):258-65. doi: 10.1016/j.micinf.2014.06.006. Epub 2014 Jun 21.
Streptococcus iniae has become one of the most serious aquatic pathogens in the last decade, causing large losses in wild and farmed fish worldwide. There is clear evidence that this pathogen is capable not only of causing serious disease in fish but also of being transferred to and infecting humans. In this study, we investigate the interaction of S. iniae with two murine macrophage cell lines, J774-A1 and RAW 264.7. Cytotoxicity assay demonstrated significant differences between live and UV-light killed IUSA-1 strains. The burst respiratory activity decreased to baseline after 1 and 4 h of exposure for J774-A1 and RAW 264.7, respectively. Immunofluorescent and ultrastructural study of infected cells confirmed the intracellular localization of bacteria at 1 h and 24 h post-infection. Using qRT-PCR arrays, we investigated the changes in the gene expression of immune relevant genes associated with macrophage activation. In this screening, we identified 11 of 84 genes up-regulated, we observed over-expression of pro-inflammatory response as IL-1α, IL-1β, and TNF-α, without a good anti-inflammatory response. Present findings suggest a capacity of S. iniae to modulate a mammalian macrophages cell lines to their survival and replication intracellular, which makes this cell type as a reservoir for continued infection.
无乳链球菌在过去十年中已成为最严重的水生病原体之一,在世界范围内给野生和养殖鱼类造成了巨大损失。有明确的证据表明,这种病原体不仅能够导致鱼类严重疾病,还能够转移并感染人类。在本研究中,我们研究了无乳链球菌与两种鼠源巨噬细胞系 J774-A1 和 RAW 264.7 的相互作用。细胞毒性测定表明,活菌株和紫外线灭活菌株之间存在显著差异。J774-A1 和 RAW 264.7 的呼吸爆发活性分别在暴露 1 和 4 小时后降至基线。感染细胞的免疫荧光和超微结构研究证实了细菌在感染后 1 小时和 24 小时的细胞内定位。使用 qRT-PCR 芯片,我们研究了与巨噬细胞激活相关的免疫相关基因的表达变化。在这次筛选中,我们鉴定了 84 个基因中上调的 11 个基因,我们观察到促炎反应的过度表达,如 IL-1α、IL-1β 和 TNF-α,而没有良好的抗炎反应。目前的研究结果表明,无乳链球菌能够调节哺乳动物巨噬细胞系的生存和复制,使其成为持续感染的储存库。