Thoracic Surgery, The First Hospital of China Medical University, Shenyang, China.
Pathology Department, The 202 Hospital of the People's Liberation Army, China.
Microb Pathog. 2019 Jun;131:98-105. doi: 10.1016/j.micpath.2019.03.038. Epub 2019 Apr 3.
With the widespread use of invasive surgery, immunosuppressive therapy and broad-spectrum antibiotics, there has resulted a corresponding increase in severe systemic infections as produced by Candida albicans (C.albicans), as it combines with bacterial infections. Such infections often result in high rates of mortality. In this report, we examined the effects of the C. albicans cell wall mannoprotein (MP) on macrophage immunity. The MTS assay was used to detect cell proliferation activity and neutral red staining to observe cell phagocytosis. The Griess method was used to detect NO secretion in culture supernatants and apoptosis of macrophages were determined with use of FITC-Annexin V and PI staining. mRNA and protein expressions of JAK2, STAT3, IL-1β, IL-6, TNF-α and iNOS in RAW264.7 cells were determined with use of RT-PCR and western blot. MP significantly promoted the proliferation of RAW264.7 cells, inhibited their phagocytic capacity, but exerted no significant effects on apoptosis of macrophages. In addition, MP not only up-regulated the expression of cytokines, but also the expressions of p-stat3 and p-jak2. Interestingly, when MP was combined with lipopolysaccharide (LPS) a markedly accentuated release of inflammatory cytokines was observed. MP promotes macrophage inflammation induced by LPS and participates in the inflammatory response. One of the potential mechanisms of this effect involves MP activation of the JAK2/STAT3 signaling pathway in RAW264.7 cells, which enables macrophages to transform from M0 to M1 and promote the occurrence of inflammation.
随着侵袭性手术、免疫抑制治疗和广谱抗生素的广泛应用,白色念珠菌(C.albicans)与细菌感染相结合导致的严重全身感染相应增加。此类感染常导致高死亡率。在本报告中,我们研究了白色念珠菌细胞壁甘露糖蛋白(MP)对巨噬细胞免疫的影响。MTS 法检测细胞增殖活性,中性红染色观察细胞吞噬作用。Griess 法检测培养上清液中 NO 的分泌,FITC-Annexin V 和 PI 染色检测巨噬细胞凋亡。用 RT-PCR 和 Western blot 检测 RAW264.7 细胞中 JAK2、STAT3、IL-1β、IL-6、TNF-α和 iNOS 的 mRNA 和蛋白表达。MP 显著促进 RAW264.7 细胞的增殖,抑制其吞噬能力,但对巨噬细胞凋亡无明显影响。此外,MP 不仅上调细胞因子的表达,还上调 p-stat3 和 p-jak2 的表达。有趣的是,当 MP 与脂多糖(LPS)结合时,观察到炎症细胞因子的释放明显增强。MP 促进 LPS 诱导的巨噬细胞炎症,并参与炎症反应。这种作用的潜在机制之一涉及 MP 激活 RAW264.7 细胞中的 JAK2/STAT3 信号通路,使巨噬细胞从 M0 转化为 M1,并促进炎症的发生。