Kovacevic Nikolina, Belosevic Miodrag
Department of Biological Sciences, University of Alberta, Edmonton, Alberta, Canada.
Department of Biological Sciences, University of Alberta, Edmonton, Alberta, Canada; Department of Microbiology and Immunology, University of Alberta, Edmonton, Alberta, Canada.
Fish Shellfish Immunol. 2015 Dec;47(2):942-53. doi: 10.1016/j.fsi.2015.10.041. Epub 2015 Oct 31.
Quantitative expression analysis of goldfish SAA revealed the highest mRNA levels in the kidney, spleen and intestine with lower mRNA levels in muscle and liver. Goldfish SAA was differentially expressed in goldfish immune cells with highest mRNA levels observed in neutrophils. To functionally assess goldfish SAA, recombinant protein (rgSAA) was generated by prokaryotic expression and functionally characterized. Monocytes and macrophages treated with rgSAA exhibited differential gene expression of pro-inflammatory and anti-inflammatory cytokines. rgSAA induced gene expression of both pro-inflammatory (TNFα1, TNFα2) and anti-inflammatory cytokines (IL-10, TGFβ) in monocytes. rgSAA induced IL-1β1 and SAA gene expression in macrophages. rgSAA was chemotactic to macrophages and neutrophils, but not monocytes. rgSAA did not affect respiratory burst induced by heat-killed Aeromonas salmonicida. rgSAA treatment of macrophages down-regulated their production of nitric oxide. rgSAA exhibited antibacterial properties against Escherichia coli in a concentration dependent manner.
金鱼SAA的定量表达分析显示，肾脏、脾脏和肠道中的mRNA水平最高，而肌肉和肝脏中的mRNA水平较低。金鱼SAA在金鱼免疫细胞中差异表达，在中性粒细胞中观察到最高的mRNA水平。为了从功能上评估金鱼SAA，通过原核表达产生了重组蛋白（rgSAA）并对其进行了功能表征。用rgSAA处理的单核细胞和巨噬细胞表现出促炎和抗炎细胞因子的差异基因表达。rgSAA诱导单核细胞中促炎细胞因子（TNFα1、TNFα2）和抗炎细胞因子（IL-10、TGFβ）的基因表达。rgSAA诱导巨噬细胞中IL-1β1和SAA基因表达。rgSAA对巨噬细胞和中性粒细胞具有趋化作用，但对单核细胞没有趋化作用。rgSAA不影响热灭活的杀鲑气单胞菌诱导的呼吸爆发。rgSAA处理巨噬细胞会下调其一氧化氮的产生。rgSAA对大肠杆菌具有浓度依赖性的抗菌特性。
