Grayfer Leon, Belosevic Miodrag
Department of Biological Sciences, University of Alberta, Edmonton, Alberta, Canada.
Dev Comp Immunol. 2009 Feb;33(2):235-46. doi: 10.1016/j.dci.2008.09.001. Epub 2008 Oct 1.
We report on the characterization and functional analysis of IFNgamma of the goldfish. Quantitative analysis indicated the highest expression of goldfish IFNgamma in the spleen, with lower expressions in brain, gill, kidney, heart, intestine and muscle. An increase in IFNgamma expression was observed in kidney leukocytes following stimulation with PHA, PolyI:C and during mixed leukocyte reaction. IFNgamma was expressed in different goldfish immune cell populations and this expression was upregulated in several of these populations following treatment of cells with recombinant goldfish TNFalpha-2. A recombinant form of goldfish IFNgamma (rgIFNgamma) was produced and functionally analyzed. The rgIFNgamma primed goldfish macrophages and neutrophils for enhanced respiratory burst responses and increased the phagocytic and nitric oxide responses of macrophages. Furthermore, treatment of goldfish macrophages with rgIFNgamma induced increased expression of several proinflammatory genes including TNFalpha-1, TNFalpha-2, IL-beta-1, IL-beta-2, IL-12-p35, IL-12-p40, RSAD-2 (=viparin), CXCL-8 (=IL-8), CCL-1, iNOS A and B and IFNgamma, and decreased the expression of TLR-3, while the expression of TGFbeta remained unchanged. Our results indicate that IFNgamma is a central proinflammatory cytokine of teleosts.
我们报告了金鱼γ干扰素（IFNγ）的特性及功能分析。定量分析表明，金鱼IFNγ在脾脏中的表达最高，在脑、鳃、肾、心脏、肠道和肌肉中的表达较低。在用PHA、聚肌胞苷酸（PolyI:C）刺激后以及在混合淋巴细胞反应过程中，肾白细胞中的IFNγ表达增加。IFNγ在不同的金鱼免疫细胞群体中表达，在用重组金鱼肿瘤坏死因子α-2（TNFα-2）处理细胞后，其中几个群体的这种表达上调。制备了重组形式的金鱼IFNγ（rgIFNγ）并进行了功能分析。rgIFNγ使金鱼巨噬细胞和中性粒细胞致敏，以增强呼吸爆发反应，并增加巨噬细胞的吞噬和一氧化氮反应。此外，用rgIFNγ处理金鱼巨噬细胞可诱导包括TNFα-1、TNFα-2、白细胞介素β-1（IL-β-1）、白细胞介素β-2（IL-β-2）、白细胞介素12-p35、白细胞介素12-p40、RSAD-2（=维帕菌素）、CXCL-8（=白细胞介素8）、CCL-1、诱导型一氧化氮合酶A和B以及IFNγ在内的几种促炎基因的表达增加，并降低Toll样受体3（TLR-3）的表达，而转化生长因子β（TGFβ）的表达保持不变。我们的结果表明，IFNγ是硬骨鱼的一种核心促炎细胞因子。
