International Medical University, 57000 Kuala Lumpur, Malaysia.
Department of Medical Microbiology, Faculty of Medicine, University of Malaya, 50603 Kuala Lumpur, Malaysia.
Med Mycol. 2020 Apr 1;58(3):333-340. doi: 10.1093/mmy/myz061.
Cladosporium is one of the most abundant spore. Fungi of this genus can cause respiratory allergy and intrabronchial lesion. We studied the differential expression of host genes after the interaction of Cladosporium sphaerospermum conidia with Human Bronchial Epithelial Cells (BEAS-2B) and Human Pulmonary Alveolar Epithelial Cells (HPAEpiC). C. sphaerospermum conidia were harvested and co-cultured with BEAS-2B cells or HPAEpiC cells for 48 hours respectively. This culture duration was chosen as it was associated with high germination rate. RNA was extracted from two biological replicates per treatment. RNA of BEAS-2B cells was used to assess changes in gene expression using AffymetrixGeneChip® Human Transcriptome Array 2.0. After co-culture with Cladosporium spores, 68 individual genes were found differentially expressed (P ≤ 0.05) and up-regulated ≥ 1.5 folds while 75 genes were found differentially expressed at ≤ -1.5 folds compared with controls. Reverse transcription and qPCR were performed on the RNA collected from both BEAS-2B cells and HPAEpiC cells to validate the microarray results with 7 genes. Based on the findings, infected pulmonary epithelial cells exhibited an increase in cell death-related genes and genes associated with innate immunity.
枝孢菌是最丰富的孢子真菌之一。该属真菌可引起呼吸道过敏和支气管内病变。我们研究了枝孢菌分生孢子与人体支气管上皮细胞(BEAS-2B)和人肺泡上皮细胞(HPAEpiC)相互作用后宿主基因的差异表达。收集枝孢菌分生孢子并分别与 BEAS-2B 细胞或 HPAEpiC 细胞共培养 48 小时。选择此培养时间是因为它与高发芽率相关。每个处理提取两份生物重复的 RNA。使用 AffymetrixGeneChip® Human Transcriptome Array 2.0 评估 BEAS-2B 细胞的 RNA 中基因表达的变化。与对照相比,与枝孢菌孢子共培养后,有 68 个个体基因被发现差异表达(P≤0.05)且上调≥1.5 倍,而有 75 个基因被发现差异表达≤-1.5 倍。对来自 BEAS-2B 细胞和 HPAEpiC 细胞的 RNA 进行逆转录和 qPCR,用 7 个基因验证微阵列结果。根据研究结果,感染的肺上皮细胞表现出与细胞死亡相关的基因和与先天免疫相关的基因增加。