Montero Jana, Coll Julio, Sevilla Noemí, Cuesta Alberto, Bols Niels C, Tafalla Carolina
Centro de Investigación en Sanidad Animal (CISA-INIA), Carretera de Algete a El Casar km. 8, 1. Valdeolmos, 28130 Madrid, Spain.
Dev Comp Immunol. 2008;32(11):1374-84. doi: 10.1016/j.dci.2008.05.004. Epub 2008 Jun 6.
In the current work, we have demonstrated that both rainbow trout (Oncorhynchus mykiss) interleukin 8 (IL-8), a CXC chemokine, and CK-6, a CC chemokine, are able of efficiently attracting RTS11, a rainbow trout established macrophage-monocyte-like cell line. Interestingly, two sub-populations of non-adherent cells are distinguishable by flow cytometry that could be identified as immature monocyte- and mature macrophage-like populations, respectively, and the two chemokines studied exert their effects on different populations. Although IL-8 specifically attracts the monocyte-like sub-population, CK-6 specifically attracts the macrophage-like cell sub-population. We have also determined the effects of both of these chemokines on RTS11 phagocytosis, respiratory burst and the expression of other immune-related genes. We found that IL-8 inhibited the phagocytosis capacity of RTS11 cells belonging to the macrophage-like profile. No effect was observed, however, on the respiratory burst, immune function that has been considerably affected throughout the establishment of the cell culture. Concerning the effect that IL-8 and CK-6 have on the expression of other immune genes, we found that IL-8 significantly induced the levels of expression of CK-6, IL-8, pro-inflammatory cytokines such as IL-1beta and tumour necrosis factor alpha (TNF-alpha) of RTS11 cells. On the other hand, CK-6 induced the levels of expression of IL-8, iNOS and the integrin CD-18, while it had very faint effect on pro-inflammatory cytokines. These results constitute one of the very few studies in which the effect of IL-8, a CXC chemokine, on monocyte-like cells is described. Moreover, it demonstrates that different monocyte-macrophage sub-populations have different reactivity to different chemokines.
在当前研究中,我们已证明虹鳟(Oncorhynchus mykiss)白细胞介素8(IL-8,一种CXC趋化因子)和CK-6(一种CC趋化因子)均能够有效吸引RTS11,一种已建立的虹鳟巨噬细胞-单核细胞样细胞系。有趣的是,通过流式细胞术可区分出两个非贴壁细胞亚群,它们分别可被鉴定为未成熟单核细胞样群体和成熟巨噬细胞样群体,并且所研究的两种趋化因子对不同群体发挥作用。尽管IL-8特异性吸引单核细胞样亚群,但CK-6特异性吸引巨噬细胞样细胞亚群。我们还确定了这两种趋化因子对RTS11吞噬作用、呼吸爆发以及其他免疫相关基因表达的影响。我们发现IL-8抑制了属于巨噬细胞样特征的RTS11细胞的吞噬能力。然而,对呼吸爆发未观察到影响,呼吸爆发这一免疫功能在细胞培养建立过程中受到了相当大的影响。关于IL-8和CK-6对其他免疫基因表达的影响,我们发现IL-8显著诱导了RTS11细胞中CK-6、IL-8、促炎细胞因子如IL-1β和肿瘤坏死因子α(TNF-α)的表达水平。另一方面,CK-6诱导了IL-8、诱导型一氧化氮合酶(iNOS)和整合素CD-18的表达水平,而对促炎细胞因子的影响非常微弱。这些结果构成了极少数描述CXC趋化因子IL-8对单核细胞样细胞作用的研究之一。此外,它表明不同的单核细胞-巨噬细胞亚群对不同趋化因子具有不同的反应性。