Research Center for Swine Disease, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu 611130, China.
Sichuan Science-Observation Experimental Station of Veterinary Drugs and Veterinary Diagnostic Technique, Ministry of Agriculture and Rural Affairs, Chengdu 611130, China.
Int J Mol Sci. 2024 May 14;25(10):5330. doi: 10.3390/ijms25105330.
, a zoonotic pathogen that produces a 146-kDa modular toxin (PMT), causes progressive atrophic rhinitis with severe turbinate bone degradation in pigs. However, its mechanism of cytotoxicity remains unclear. In this study, we expressed PMT, purified it in a prokaryotic expression system, and found that it killed PK15 cells. The host factor CXCL8 was significantly upregulated among the differentially expressed genes in a transcriptome sequencing analysis and qPCR verification. We constructed a CXCL8-knockout cell line with a CRISPR/Cas9 system and found that CXCL8 knockout significantly increased resistance to PMT-induced cell apoptosis. CXCL8 knockout impaired the cleavage efficiency of apoptosis-related proteins, including Caspase3, Caspase8, and PARP1, as demonstrated with Western blot. In conclusion, these findings establish that CXCL8 facilitates PMT-induced PK15 cell death, which involves apoptotic pathways; this observation documents that CXCL8 plays a key role in PMT-induced PK15 cell death.
，一种人畜共患病病原体，可产生 146kDa 模块化毒素（PMT），导致猪渐进性萎缩性鼻炎和严重鼻甲骨降解。然而，其细胞毒性机制尚不清楚。在本研究中，我们表达了 PMT，在原核表达系统中进行了纯化，并发现其可杀伤 PK15 细胞。在转录组测序分析和 qPCR 验证中，发现宿主因子 CXCL8 在差异表达基因中显著上调。我们使用 CRISPR/Cas9 系统构建了 CXCL8 敲除细胞系，发现 CXCL8 敲除显著增加了对 PMT 诱导的细胞凋亡的抗性。Western blot 结果表明，CXCL8 敲除可削弱凋亡相关蛋白（包括 Caspase3、Caspase8 和 PARP1）的切割效率。总之，这些发现证实了 CXCL8 促进 PMT 诱导的 PK15 细胞死亡，涉及凋亡途径；这一观察结果表明，CXCL8 在 PMT 诱导的 PK15 细胞死亡中发挥关键作用。
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