Abstract：
OBJECTIVE: Animals exhibit physiological changes designed to eliminate the perceived danger, provoking similar symptoms of fever. However, a high-grade fever indicates poor clinical outcomes. Caspase11 (Casp11) is involved in many inflammatory diseases. Whether Casp11 leads to fever remains unclear. In this study, we investigate the role of the preoptic area of the hypothalamus (PO/AH) microglia Casp11 in fever.
METHODS: We perform experiments using a rat model of LPS-induced fever. We measure body temperature and explore the functions of peripheral macrophages and PO/AH microglia in fever signaling by ELISA, immunohistochemistry, immunofluorescence, flow cytometry, macrophage depletion, protein blotting, and RNA-seq. Then, the effects of macrophages on microglia in a hyperthermic environment are observed in vitro. Finally, adeno-associated viruses are used to knockdown or overexpress microglia Casp11 in PO/AH to determine the role of Casp11 in fever.
RESULTS: We find peripheral macrophages and PO/AH microglia play important roles in the process of fever, which is proved by macrophage and microglia depletion. By RNA-seq analysis, we find Casp11 expression in PO/AH is significantly increased during fever. Co-culture and conditioned-culture simulate the induction of microglia Casp11 activation by macrophages in a non-contact manner. Microglia Casp11 knockdown decreases body temperature, pyrogenic factors, and inflammasome, and vice versa.
CONCLUSIONS: We report that Casp11 drives fever. Mechanistically, peripheral macrophages transmit immune signals via cytokines to microglia in PO/AH, which activate the Casp11 non-canonical inflammasome. Our findings identify a novel player, the microglia Casp11, in the control of fever, providing an explanation for the transmission and amplification of fever immune signaling.
METHODS: We perform experiments using a rat model of LPS-induced fever. We measure body temperature and explore the functions of peripheral macrophages and PO/AH microglia in fever signaling by ELISA, immunohistochemistry, immunofluorescence, flow cytometry, macrophage depletion, protein blotting, and RNA-seq. Then, the effects of macrophages on microglia in a hyperthermic environment are observed in vitro. Finally, adeno-associated viruses are used to knockdown or overexpress microglia Casp11 in PO/AH to determine the role of Casp11 in fever.
RESULTS: We find peripheral macrophages and PO/AH microglia play important roles in the process of fever, which is proved by macrophage and microglia depletion. By RNA-seq analysis, we find Casp11 expression in PO/AH is significantly increased during fever. Co-culture and conditioned-culture simulate the induction of microglia Casp11 activation by macrophages in a non-contact manner. Microglia Casp11 knockdown decreases body temperature, pyrogenic factors, and inflammasome, and vice versa.
CONCLUSIONS: We report that Casp11 drives fever. Mechanistically, peripheral macrophages transmit immune signals via cytokines to microglia in PO/AH, which activate the Casp11 non-canonical inflammasome. Our findings identify a novel player, the microglia Casp11, in the control of fever, providing an explanation for the transmission and amplification of fever immune signaling.
摘要：
目的：动物表现出旨在消除感知危险的生理变化，引起类似的发烧症状。然而,高热表明临床结局较差.Caspase11(Casp11)与许多炎症性疾病有关。Casp11是否会导致发烧尚不清楚。在这项研究中,我们研究了下丘脑视前区（PO/AH）小胶质细胞Casp11在发热中的作用。
方法：我们使用LPS诱导的发热大鼠模型进行实验。我们通过ELISA测量体温并探讨外周巨噬细胞和PO/AH小胶质细胞在发热信号中的功能，免疫组织化学，免疫荧光，流式细胞术,巨噬细胞耗竭，蛋白质印迹，和RNA-seq。然后，在体外观察到巨噬细胞在高温环境中对小胶质细胞的影响。最后,腺相关病毒用于敲除或过表达PO/AH中的小胶质细胞Casp11，以确定Casp11在发热中的作用。
结果：我们发现外周巨噬细胞和PO/AH小胶质细胞在发热过程中起重要作用，巨噬细胞和小胶质细胞耗竭证明了这一点。通过RNA-seq分析,我们发现发热期间PO/AH中Casp11的表达显著增加。共培养和条件培养以非接触方式模拟巨噬细胞对小胶质细胞Casp11活化的诱导。小胶质细胞Casp11敲低降低体温，致热因素，和炎性体，反之亦然。
结论：我们报告Casp11引起发烧。机械上,外周巨噬细胞通过细胞因子传递免疫信号给PO/AH中的小胶质细胞，激活Casp11非规范炎性体。我们的发现发现了一个新颖的玩家，小胶质细胞Casp11，在控制发烧中，为发热免疫信号的传递和放大提供解释。
方法：我们使用LPS诱导的发热大鼠模型进行实验。我们通过ELISA测量体温并探讨外周巨噬细胞和PO/AH小胶质细胞在发热信号中的功能，免疫组织化学，免疫荧光，流式细胞术,巨噬细胞耗竭，蛋白质印迹，和RNA-seq。然后，在体外观察到巨噬细胞在高温环境中对小胶质细胞的影响。最后,腺相关病毒用于敲除或过表达PO/AH中的小胶质细胞Casp11，以确定Casp11在发热中的作用。
结果：我们发现外周巨噬细胞和PO/AH小胶质细胞在发热过程中起重要作用，巨噬细胞和小胶质细胞耗竭证明了这一点。通过RNA-seq分析,我们发现发热期间PO/AH中Casp11的表达显著增加。共培养和条件培养以非接触方式模拟巨噬细胞对小胶质细胞Casp11活化的诱导。小胶质细胞Casp11敲低降低体温，致热因素，和炎性体，反之亦然。
结论：我们报告Casp11引起发烧。机械上,外周巨噬细胞通过细胞因子传递免疫信号给PO/AH中的小胶质细胞，激活Casp11非规范炎性体。我们的发现发现了一个新颖的玩家，小胶质细胞Casp11，在控制发烧中，为发热免疫信号的传递和放大提供解释。
