背景:在炎症反应的早期阶段使用后剩余的多余细胞因子即使在反应结束后也会刺激免疫细胞,引起继发性炎症反应并最终损害宿主,这被称为细胞因子风暴。抑制热休克蛋白90(Hsp90),最近被证明在调节各种细胞类型的炎症中起重要作用,可能有助于控制过度的炎症反应和细胞因子风暴。
方法:我们通过使用Hsp90抑制剂的化学化合物文库测量CD86表达对脾DC(sDC)的抑制作用,发现了抗炎化合物。随后,选择命中的化合物,在骨髓来源的DC(BMDC)和腹膜巨噬细胞上测量细胞因子的产生和表面分子的表达.然后,我们分析了抗原特异性Th1细胞的反应。最后,我们使用急性肺损伤(ALI)和迟发型超敏反应(DTH)模型证实了该化合物的作用。
结果:我们确定Be01是命中化合物,其中CD86在sDC中的表达降低最多。用Be01治疗减少了促炎细胞因子的产生(IL-6,TNF-α,和IL-1β)在LPS刺激的BMDC和腹膜巨噬细胞中。在DTH模型下,Be01治疗减少了耳朵肿胀和脾脏中的促炎细胞因子。同样,在ALI模型中的Be01治疗减少了中性粒细胞浸润和较低水平的分泌的细胞因子(IL-6、TNF-α)。
结论:Be01减少DCs上CD80和CD86的表达表明Th1细胞的继发性炎症反应减少,腹膜巨噬细胞释放促炎细胞因子的减少可能最初控制细胞因子风暴。
BACKGROUND: The surplus cytokines remaining after use in the early stages of the inflammatory response stimulate immune cells even after the response is over, causing a secondary inflammatory response and ultimately damaging the host, which is called a cytokine storm. Inhibiting heat shock protein 90 (Hsp90), which has recently been shown to play an important role in regulating inflammation in various cell types, may help control excessive inflammatory responses and cytokine storms.
METHODS: We discovered an anti-inflammatory compound by measuring the inhibitory effect of CD86 expression on spleen DCs (sDCs) using the chemical compounds library of Hsp90 inhibitors. Subsequently, to select the hit compound, the production of cytokines and expression of surface molecules were measured on the bone marrow-derived DCs (BMDCs) and peritoneal macrophages. Then, we analyzed the response of antigen-specific Th1 cells. Finally, we confirmed the effect of the compound using acute lung injury (ALI) and delayed-type hypersensitivity (DTH) models.
RESULTS: We identified Be01 as the hit compound, which reduced CD86 expression the most in sDCs. Treatment with Be01 decreased the production of pro-inflammatory cytokines (IL-6, TNF-α, and IL-1β) in BMDC and peritoneal macrophages stimulated by LPS. Under the DTH model, Be01 treatment reduced ear swelling and pro-inflammatory cytokines in the spleen. Similarly, Be01 treatment in the ALI model decreased neutrophil infiltration and lower levels of secreted cytokines (IL-6, TNF-α).
CONCLUSIONS: Reduction of CD80 and CD86 expression on DCs by Be01 indicates reduced secondary inflammatory response by Th1 cells, and reduced release of pro-inflammatory cytokines by peritoneal macrophages may initially control the cytokine storm.