U.S. Army Research Institute of Environmental Medicine, Natick, MA, USA.
Military Operational Medicine Research Program, Fort Detrick, MA, USA.
FEBS Lett. 2023 May;597(9):1225-1232. doi: 10.1002/1873-3468.14616. Epub 2023 Apr 4.
Macrophages play an important role in the response to infection and/or repair of injury in tissues. To examine the NF-κB pathway in response to an inflammatory stimulus, we used wild-type bone-marrow-derived macrophages (BMDMs) or BMDMs with knockout (KO) of myeloid differentiation primary response 88 (MyD88) and/or Toll/interleukin-1 receptor domain-containing adapter-inducing interferon-β (TRIF) via CRISPR/Cas9. Following treatment of BMDMs with lipopolysaccharide (LPS) to induce an inflammatory response, translational signalling of NF-κB was quantified via immunoblot and cytokines were measured. Our findings reveal that MyD88 KO, but not TRIF KO, decreased LPS-induced NF-κB signalling, and 10% expression of basal MyD88 expression was sufficient to partially rescue the abolished inflammatory cytokine secretion observed upon MyD88 KO.
巨噬细胞在组织对感染和/或损伤的反应中发挥重要作用。为了研究 NF-κB 途径对炎症刺激的反应，我们使用了野生型骨髓来源的巨噬细胞（BMDM）或通过 CRISPR/Cas9 敲除髓样分化初级反应 88（MyD88）和/或 Toll/白细胞介素-1 受体域包含衔接诱导干扰素-β（TRIF）的 BMDM。在用脂多糖（LPS）处理 BMDM 诱导炎症反应后，通过免疫印迹定量 NF-κB 的翻译信号，测量细胞因子。我们的研究结果表明，MyD88 KO，但不是 TRIF KO，降低了 LPS 诱导的 NF-κB 信号，并且基础 MyD88 表达的 10%表达足以部分挽救在 MyD88 KO 时观察到的被废除的炎症细胞因子分泌。
