Center for Genome Regulation, Facultad de Ciencias, Universidad de Chile, Santiago, Chile.
BMC Biol. 2010 Dec 22;8:151. doi: 10.1186/1741-7007-8-151.
Studies on innate immunity have benefited from the introduction of zebrafish as a model system. Transgenic fish expressing fluorescent proteins in leukocyte populations allow direct, quantitative visualization of an inflammatory response in vivo. It has been proposed that this animal model can be used for high-throughput screens aimed at the identification of novel immunomodulatory lead compounds. However, current assays require invasive manipulation of fish individually, thus preventing high-content screening.
Here we show that specific, noninvasive damage to lateral line neuromast cells can induce a robust acute inflammatory response. Exposure of fish larvae to sublethal concentrations of copper sulfate selectively damages the sensory hair cell population inducing infiltration of leukocytes to neuromasts within 20 minutes. Inflammation can be assayed in real time using transgenic fish expressing fluorescent proteins in leukocytes or by histochemical assays in fixed larvae. We demonstrate the usefulness of this method for chemical and genetic screens to detect the effect of immunomodulatory compounds and mutations affecting the leukocyte response. Moreover, we transformed the assay into a high-throughput screening method by using a customized automated imaging and processing system that quantifies the magnitude of the inflammatory reaction.
This approach allows rapid screening of thousands of compounds or mutagenized zebrafish for effects on inflammation and enables the identification of novel players in the regulation of innate immunity and potential lead compounds toward new immunomodulatory therapies. We have called this method the chemically induced inflammation assay, or ChIn assay. See Commentary article: http://www.biomedcentral.com/1741-7007/8/148.
先天免疫研究受益于斑马鱼模型系统的引入。在白细胞群体中表达荧光蛋白的转基因鱼可直接、定量地观察体内炎症反应。有人提出，这种动物模型可用于高通量筛选，以鉴定新型免疫调节先导化合物。然而，目前的检测方法需要对鱼进行单独的侵入性操作，因此无法进行高内涵筛选。
我们发现，对侧线神经丘细胞的特定、非侵入性损伤可诱导强烈的急性炎症反应。亚致死浓度硫酸铜暴露于幼鱼中可选择性地破坏感觉毛细胞群，在 20 分钟内诱导白细胞浸润神经丘。通过在白细胞中表达荧光蛋白的转基因鱼或通过固定幼虫的组织化学检测，可实时检测炎症。我们证明了该方法在化学和遗传筛选中的有效性，以检测免疫调节化合物和影响白细胞反应的突变的作用。此外，我们通过使用定制的自动成像和处理系统将该检测转化为高通量筛选方法，该系统可定量炎症反应的程度。
该方法可快速筛选数千种化合物或诱变斑马鱼对炎症的影响，并可识别先天免疫调节中的新型参与者和潜在的免疫调节治疗新先导化合物。我们将这种方法称为化学诱导炎症检测，或 ChIn 检测。参见评论文章：http://www.biomedcentral.com/1741-7007/8/148。
