Provincial Key Laboratory for Transfusion-Transmitted Infectious Diseases, Institute of Blood Transfusion, Chinese Academy of Medical Sciences and Peking Union Medical College, Chengdu, Sichuan 610052, China.
Provincial Key Laboratory for Transfusion-Transmitted Infectious Diseases, Institute of Blood Transfusion, Chinese Academy of Medical Sciences and Peking Union Medical College, Chengdu, Sichuan 610052, China.
Life Sci. 2020 Sep 15;257:118064. doi: 10.1016/j.lfs.2020.118064. Epub 2020 Jul 8.
Exosomes contain functional molecules from their cells of origin and can enter recipient cells for intercellular communication. Interferon β (IFNβ) has been shown to induce some lncRNAs to regulate host immune response and play a major role in the positive regulation of the activity of natural killer (NK) cells. We aim to clarify whether IFNβ induced exosomes can regulate the cytotoxicity of NK cells by transferring specific lncRNAs into NK cells.
Exosomes were isolated from the supernatants of A549 cells with or without IFNβ treatment. Co-culture and ELISA assay were used to analyze the effect of exosomes on the cytotoxicity of NK cells. Human transcriptome array (HTA) was performed to analyze the profiling of RNAs wrapped in exosomes. Then subcellular location, qPCR, western blotting, dual-luciferase reporter assay and ELISA were used to determine long noncoding RNAs (lcnRNAs) location, sponge absorb effects, the expression of NKp46 and cytotoxicity of NK cells.
ELISA assay showed IFNβ induced exosomes can strengthen the cytotoxicity of NK cells. Through HTA we found the expression levels of 69 lncRNAs were significantly changed within IFNβ induced exosomes. Additionally, we found a specific exosomal cargo, linc-EPHA6-1, acted as a competing endogenous RNA (ceRNA) for hsa-miR-4485-5p which subsequently up-regulate one of the natural cytotoxicity receptors (NKp46) expression. Furthermore, we verified over-expression of linc-EPHA6-1 significantly enhances the cytotoxicity of NK cells against A549 cells and Zika virus infected A549 cells.
Our results demonstrated that IFNβ-induced exosomal linc-EPHA6-1 can regulate the cytotoxicity of NK cells.
外泌体包含其来源细胞的功能分子，并可进入受体细胞进行细胞间通讯。干扰素β（IFNβ）已被证明可诱导一些长链非编码 RNA（lncRNA）来调节宿主免疫反应，并在自然杀伤（NK）细胞活性的正向调节中发挥主要作用。我们旨在阐明 IFNβ 诱导的外泌体是否可以通过将特定的 lncRNA 转移到 NK 细胞中来调节 NK 细胞的细胞毒性。
用或不用 IFNβ 处理 A549 细胞，从其上清液中分离出外泌体。共培养和 ELISA 测定用于分析外泌体对 NK 细胞细胞毒性的影响。进行人类转录组芯片（HTA）分析以分析包裹在外泌体中的 RNA 的谱图。然后使用亚细胞定位、qPCR、western blot、双荧光素酶报告基因测定和 ELISA 来确定长链非编码 RNA（lncRNA）的位置、海绵吸收效应、NKp46 的表达和 NK 细胞的细胞毒性。
ELISA 测定表明 IFNβ 诱导的外泌体可以增强 NK 细胞的细胞毒性。通过 HTA，我们发现 IFNβ 诱导的外泌体中 69 个 lncRNA 的表达水平显著改变。此外，我们发现一种特定的外泌体 cargo，linc-EPHA6-1，作为 hsa-miR-4485-5p 的竞争内源性 RNA（ceRNA），随后上调一种自然细胞毒性受体（NKp46）的表达。此外，我们验证了 linc-EPHA6-1 的过表达显著增强了 NK 细胞对 A549 细胞和 Zika 病毒感染的 A549 细胞的细胞毒性。
我们的结果表明，IFNβ 诱导的外泌体 linc-EPHA6-1 可以调节 NK 细胞的细胞毒性。
