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脂多糖刺激下虹鳟幼鱼中白细胞介素-8和CXC趋化因子受体1的检测与定位

Detection and Localization of IL-8 and CXCR1 in Rainbow Trout Larvae in Response to Lipopolysaccharide.

作者信息

Santana Paula A, Forero Juan C, Guzmán Fanny, Gaete Sandra, Acosta Félix, Mercado Luis A, Álvarez Claudio A

机构信息

Instituto de Ciencias Aplicadas, Facultad de Ingeniería, Universidad Autónoma de Chile, Santiago 8910060, Chile.

Laboratorio de Bioingeniería de Tejidos e Innovación Odontológica, Facultad de Odontología, Universidad de Valparaíso, Valparaíso 2360004, Chile.

出版信息

Animals (Basel). 2024 Oct 6;14(19):2878. doi: 10.3390/ani14192878.

Abstract

The salmonid industry faces challenges due to the susceptibility of fish to opportunistic pathogens, particularly in early developmental stages. Understanding the immunological capacity during these stages is crucial for developing effective disease control strategies. IL-8R, a member of the G-protein-coupled receptor family, acts as a receptor for Interleukin 8 (IL-8). The binding of IL-8 to IL-8R plays a major role in the pathophysiology of a wide spectrum of inflammatory conditions. This study focused on the immune response capacity of rainbow trout () larvae by analyzing IL-8/CXCR1 response to lipopolysaccharide (LPS) from . Previous research demonstrated that LPS from acts as a potent immunostimulant in teleost, enhancing pro-inflammatory cytokines. The methodology included in silico analysis and the synthesis and characterization of an omCXCR1-derived epitope peptide, which was used to produce omCXCR1-specific anti98 serum in mice. The research revealed that rainbow trout larvae 19 days post-hatching (dph) exhibited pronounced immune responses post-stimulation with 1 µg/mL of LPS. This was evidenced by the upregulated protein expression of IL-8 and omCXCR1 in trout larvae 2 and 8 h after LPS challenge, as analyzed by ELISA and immunohistochemistry. Furthermore, fluorescence microscopy successfully revealed the colocalization of IL-8 and its receptor in cells from mucosal tissues after LPS challenge in larvae 19 dph. These findings underscore the efficacy of LPS immersion as a method to activate the innate immune system in trout larvae. Furthermore, we propose IL-8 and its receptor as molecular markers for evaluating immunostimulation in the early developmental stages of salmonids.

摘要

由于鱼类,特别是在早期发育阶段,对机会性病原体敏感,鲑鱼养殖业面临挑战。了解这些阶段的免疫能力对于制定有效的疾病控制策略至关重要。白细胞介素8受体(IL-8R)是G蛋白偶联受体家族的成员,作为白细胞介素8(IL-8)的受体。IL-8与IL-8R的结合在多种炎症性疾病的病理生理学中起主要作用。本研究通过分析虹鳟()幼体对嗜水气单胞菌脂多糖(LPS)的IL-8/CXCR1反应,聚焦于其免疫反应能力。先前的研究表明,嗜水气单胞菌的LPS在硬骨鱼中作为一种有效的免疫刺激剂,可增强促炎细胞因子。该方法包括计算机分析以及omCXCR1衍生表位肽的合成与表征,该表位肽用于在小鼠中产生omCXCR1特异性抗血清。研究表明,孵化后19天(dph)的虹鳟幼体在受到1μg/mL LPS刺激后表现出明显的免疫反应。通过ELISA和免疫组织化学分析,LPS攻击后2小时和8小时的虹鳟幼体中IL-8和omCXCR1的蛋白表达上调证明了这一点。此外,荧光显微镜成功揭示了19 dph幼体LPS攻击后,IL-8及其受体在黏膜组织细胞中的共定位。这些发现强调了LPS浸泡作为激活虹鳟幼体先天免疫系统方法的有效性。此外,我们提出IL-8及其受体作为评估鲑鱼早期发育阶段免疫刺激的分子标志物。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/daa5/11475925/1042681a7232/animals-14-02878-g001.jpg

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