Roles of Host Phospholipase D during Aspergillus fumigatus Infection in Mice
Roles of Host Phospholipase D during Aspergillus fumigatus Infection in Mice
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DOI：
10.15212/zoonoses-2022-0044
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发表时间：
2023-05
期刊：
影响因子：
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通讯作者：
Fangyan Chen;Xiaoyu Liu;Rui Zhao;Jingya Zhao;Dingchen Li;Li Han
中科院分区：
文献类型：
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作者：
Fangyan Chen;Xiaoyu Liu;Rui Zhao;Jingya Zhao;Dingchen Li;Li Han
肺中的烟曲霉感染伴随着先天免疫细胞的募集、吞噬作用和炎症因子的释放。磷脂酶D（PLD）是细胞迁移和吞噬的关键调节因子，但PLD缺乏对动物抗真菌感染的影响尚不清楚。本研究旨在探讨PLD对宿主免疫应答的影响。在免疫活性或免疫抑制状态下的烟曲霉感染。侵袭性肺曲霉菌病小鼠模型的建立采用改良的免疫抑制方案。为了从小鼠收集支气管肺泡灌洗液（BALF），用0.5ml PBS洗涤肺8次。使用库尔特计数器测定BALF中的总细胞计数。采用流式细胞术检测支气管肺泡灌洗液中肺泡巨噬细胞、中性粒细胞和单核细胞的含量，并采用FlowJo V10软件进行分析。采用多重免疫分析法测定BALF中炎性细胞因子的浓度。在免疫功能正常的小鼠中，肺泡巨噬细胞是A.烟曲霉感染后，肺泡中招募了大量中性粒细胞和单核细胞。pld 1和pld 2基因的缺失不影响BALF中肺泡巨噬细胞、中性粒细胞或单核细胞的含量。在醋酸氢化可的松诱导的免疫抑制下，pld 1-/-pld 2-/-小鼠在A.与野生型小鼠相比，烟曲霉感染的小鼠具有更高的真菌负荷和更低数量的肺组织中致密炎性浸润的突出局灶性区域。感染后pld 1-/-pld 2-/-小鼠BALF中IL-12 p40显著降低，IL-10显著升高。我们的研究结果表明，在A. pld 1和pld 2的缺乏不利于炎性细胞向肺组织的浸润，但促进了IL-10的释放并阻断了IL-12的释放，从而增加了真菌负荷和死亡率。
Aspergillus fumigatus infection in the lungs is accompanied by the recruitment of innate immune cells, phagocytosis, and the release of inflammatory factors. Phospholipase D (PLD) is a key regulator of cell migration and phagocytosis, but the effect of PLD deficiency on antifungal infection in animals is unknown. This study aims to investigate the impact of PLD on the host immune response to A. fumigatus infection under either immunocompetent or immunosuppressed status. The invasive pulmonary aspergillosis mouse model was created using a modified protocol with immunosuppression by steroids. For collection of bronchoalveolar lavage fluid (BALF) from mice, the lungs were washed eight times with 0.5 ml of PBS. Total cell counts in BALF were determined using a Coulter Counter. The content of alveolar macrophages, neutrophils, and monocytes in BALF was examined by flow cytometry and analyzed by FlowJo V10 software. Multiplex immunoassays were used to determine the concentrations of inflammatory cytokines in BALF. In immunocompetent mice, alveolar macrophages were the major cell population in BALF after A. fumigatus infection, and a number of neutrophils and monocytes were recruited in the alveoli. Loss of both pld1 and pld2 genes did not affect the content of alveolar macrophages, neutrophils, or monocytes in BALF. Under immunosuppression induced by hydrocortisone acetate, pld1-/-pld2-/- mice showed higher mortality after A. fumigatus infection and had a higher fungal burden and much lower number of prominent focal areas of dense inflammatory infiltrates in lung tissue than wild type mice. Moreover, interleukin (IL)-12p40 significantly decreased, and IL-10 markedly increased, in BALF from pld1 -/- pld2 -/- mice after infection. Our findings revealed that, during A. fumigatus infection, deficiency in both pld1 and pld2 in mice was not conducive to the infiltration of inflammatory cells into lung tissue but promoted the release of IL-10 and blocked the release of IL-12, thereby increasing fungal burden and mortality.