Protocol for examining the T3SS-mediated cytotoxicity of Pseudomonas aeruginosa using the A549 cell line

• Published in: STAR protocols Vol. 5; no. 3; p. 103301
• Main Authors: Huang, Jiahui, Zhang, Yao, Sheng, Shuo, Zhang, Lian-Hui, Xu, Zeling
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 20-09-2024; Elsevier
• Subjects: A549 Cells; Cell Biology; Cell culture; Humans; Microbiology; Molecular Biology; Protocol; Pseudomonas aeruginosa; Pseudomonas Infections - microbiology; Type III Secretion Systems - metabolism; Cell culture; Microbiology; Molecular Biology; Cell Biology
• ISSN: 2666-1667; 2666-1667
• DOI: 10.1016/j.xpro.2024.103301

Pseudomonas aeruginosa utilizes a type III secretion system (T3SS) to directly inject effector proteins into host cells, leading to severe acute infections. Here, we present a protocol for detecting the T3SS-mediated cytotoxicity of P. aeruginosa using the A549 cell line. We describe the steps for the preparation of the A549 cell line and P. aeruginosa strains, cell seeding, bacterial culture, infection, and cytotoxicity assay. Additionally, we provide detailed procedures for data analysis.
For complete details on the use and execution of this protocol, please refer to Huang et al.1
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•Steps for culture, passage, and seeding of the A549 cell line•Guidance for preparation and collection of P. aeruginosa•Instructions for cytotoxicity assay and data analysis
Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.
Pseudomonas aeruginosa utilizes a type III secretion system (T3SS) to directly inject effector proteins into host cells, leading to severe acute infections. Here, we present a protocol for detecting the T3SS-mediated cytotoxicity of P. aeruginosa using the A549 cell line. We describe the steps for the preparation of the A549 cell line and P. aeruginosa strains, cell seeding, bacterial culture, infection, and cytotoxicity assay. Additionally, we provide detailed procedures for data analysis.