METHODS: Employing a human lepirudin whole-blood model, we analyzed complement and leukocyte activation by measuring the sC5b-9 complex and assessing CD11b expression. A 27-multiplex system was used for quantification of cytokines. Selective cell removal from whole blood and inhibition of C3, C5, and CD14 were also applied.
RESULTS: Our findings demonstrated a marked elevation in sC5b-9 and CD11b post-A. fumigatus incubation. Thirteen cytokines (TNF, IL-1β, IL-1ra, IL-4, IL-6, IL-8, IL-17, IFNγ, MCP-1, MIP-1α, MIP-1β, FGF-basic, and G-CSF) showed increased levels. A generally lower level of cytokine release and CD11b expression was observed with A. fumigatus conidia than with E. coli. Notably, monocytes were instrumental in releasing all cytokines except MCP-1. IL-1ra was found to be both monocyte and granulocyte-dependent. Pre-inhibiting with C3 and CD14 inhibitors resulted in decreased release patterns for six cytokines (TNF, IL-1β, IL-6, IL-8, MIP-1α, and MIP-1β), with minimal effects by C5-inhibition.
CONCLUSIONS: A. fumigatus conidia induced complement activation comparable to E. coli, whereas CD11b expression and cytokine release were lower, underscoring distinct inflammatory responses between these pathogens. Complement C3 inhibition attenuated cytokine release indicating a C3-level role of complement in A. fumigatus immunity.
方法:采用人类lepirudin全血模型,我们通过测量sC5b-9复合物和评估CD11b表达来分析补体和白细胞活化.27-多重系统用于细胞因子的定量。还应用了从全血中选择性去除细胞和抑制C3、C5和CD14。
结果:我们的发现表明sC5b-9和CD11b在A后明显升高。熏霉的孵化.13种细胞因子(TNF,IL-1β,IL-1ra,IL-4、IL-6、IL-8、IL-17、IFNγ、MCP-1,MIP-1α,MIP-1β,FGF-basic,和G-CSF)显示水平升高。与用大肠杆菌相比,用烟曲霉分生孢子观察到通常较低水平的细胞因子释放和CD11b表达。值得注意的是,单核细胞有助于释放除MCP-1以外的所有细胞因子。发现IL-1ra是单核细胞和粒细胞依赖性的。用C3和CD14抑制剂预抑制导致六种细胞因子的释放模式减少(TNF,IL-1β,IL-6,IL-8,MIP-1α和MIP-1β),C5抑制作用最小。
结论:A.烟曲霉分生孢子诱导的补体激活与大肠杆菌相当,而CD11b表达和细胞因子释放较低,强调这些病原体之间不同的炎症反应。补体C3抑制减弱细胞因子释放,表明补体在烟曲霉免疫中的C3水平作用。