Department of Tropical Pathology, Faculty of Tropical Medicine, Mahidol University, Ratchawithi Road, Ratchathewi, Bangkok, 10400, Thailand.
Department of Microbiology and Immunology, Faculty of Tropical Medicine, Mahidol University, Ratchawithi Road, Ratchathewi, Bangkok, 10400, Thailand.
Microb Ecol. 2022 Jan;83(1):216-235. doi: 10.1007/s00248-021-01750-7. Epub 2021 Apr 22.
Invasive aspergillosis and scedosporiosis are life-threatening fungal infections with similar clinical manifestations in immunocompromised patients. Contrarily, Scedosporium apiospermum is susceptible to some azole derivative but often resistant to amphotericin B. Histopathological examination alone cannot diagnose these two fungal species. Pathogenesis studies could contribute to explore candidate protein markers for new diagnosis and treatment methods leading to a decrease in mortality. In the present study, proteomics was conducted to identify significantly altered proteins in A549 cells infected with or without Aspergillus fumigatus and S. apiospermum as measured at initial invasion. Protein validation was performed with immunogold labelling alongside immunohistochemical techniques in infected A549 cells and lungs from murine models. Further, cytokine production was measured, using the Bio-Plex-Multiplex immunoassay. The cytoskeletal proteins HSPA9, PA2G4, VAT1, PSMA2, PEX1, PTGES3, KRT1, KRT9, CLIP1 and CLEC20A were mainly changed during A. fumigatus infection, while the immunologically activated proteins WNT7A, GAPDH and ANXA2 were principally altered during S. apiospermum infection. These proteins are involved in fungal internalisation and structural destruction leading to pulmonary disorders. Interleukin (IL)-21, IL-1α, IL-22, IL-2, IL-8, IL-12, IL-17A, interferon-γ and tumour necrosis factor-α were upregulated in both aspergillosis and scedosporiosis, although more predominately in the latter, in accordance with chitin synthase-1 and matrix metalloproteinase levels. Our results demonstrated that during invasion, A. fumigatus primarily altered host cellular integrity, whereas S. apiospermum chiefly induced and extensively modulated host immune responses.
侵袭性曲霉病和枝顶孢霉病是危及生命的真菌感染,在免疫功能低下的患者中具有相似的临床表现。相反,枝顶孢霉对一些唑类衍生物敏感,但通常对两性霉素 B 耐药。单纯的组织病理学检查无法诊断这两种真菌。发病机制研究有助于探索新的诊断和治疗方法的候选蛋白标志物,从而降低死亡率。在本研究中,进行了蛋白质组学研究,以鉴定初始侵袭时感染烟曲霉和枝顶孢霉的 A549 细胞中明显改变的蛋白质。使用免疫金标记和免疫组织化学技术在感染的 A549 细胞和鼠模型的肺部中对蛋白质进行验证。此外,使用 Bio-Plex-Multiplex 免疫分析测定细胞因子的产生。在烟曲霉感染过程中,主要改变的是细胞骨架蛋白 HSPA9、PA2G4、VAT1、PSMA2、PEX1、PTGES3、KRT1、KRT9、CLIP1 和 CLEC20A,而在枝顶孢霉感染过程中,主要改变的是免疫激活蛋白 WNT7A、GAPDH 和 ANXA2。这些蛋白质参与真菌内化和结构破坏,导致肺部疾病。白细胞介素(IL)-21、IL-1α、IL-22、IL-2、IL-8、IL-12、IL-17A、干扰素-γ和肿瘤坏死因子-α在曲霉病和枝顶孢霉病中均上调,尽管在后者中上调更为明显,与几丁质合酶-1 和基质金属蛋白酶水平一致。我们的结果表明,在侵袭过程中,烟曲霉主要改变宿主细胞的完整性,而枝顶孢霉主要诱导并广泛调节宿主免疫反应。