Work

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

Year: 2024

Type: article

Abstract: 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 • 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.

Source: STAR Protocols

DOI: 10.1016/j.xpro.2024.103301

Authors: Jiahui Huang, Yao Zhang, Shuo Sheng, Lian-Hui Zhang, Zeling Xu

Institutions: South China Agricultural University, Zunyi Medical University

Language: English

Topic: Bacterial biofilms and quorum sensing

Subfield: Molecular Biology

Field: Biochemistry, Genetics and Molecular Biology

Domain: Life Sciences

SDG: Good health and well-being

Open Access status: gold

Estimated APC paid: $1,500