Salmonella enterica serovar Typhi plasmid pRST98 enhances intracellular bacterial growth and S. typhi-induced macrophage cell death by suppressing autophagy

Objectives: Plasmid pRST98 is a hybrid resistance-virulence plasmid isolated from Salmonella enterica serovar Typhi (S. typhi). Previous studies demonstrated that pRST98 could enhance the virulence of its host bacteria. However, the mechanism of pRST98-increased bacterial virulence is still not full...

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Main Authors: Peiyan He, Shuyan Wu, Yuanyuan Chu, Yanru Yang, Yuanyuan Li, Rui Huang
Format: Article
Language:English
Published: Elsevier 2012-05-01
Series:Brazilian Journal of Infectious Diseases
Online Access:http://www.sciencedirect.com/science/article/pii/S1413867012703215
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author Peiyan He
Shuyan Wu
Yuanyuan Chu
Yanru Yang
Yuanyuan Li
Rui Huang
author_facet Peiyan He
Shuyan Wu
Yuanyuan Chu
Yanru Yang
Yuanyuan Li
Rui Huang
author_sort Peiyan He
collection DOAJ
description Objectives: Plasmid pRST98 is a hybrid resistance-virulence plasmid isolated from Salmonella enterica serovar Typhi (S. typhi). Previous studies demonstrated that pRST98 could enhance the virulence of its host bacteria. However, the mechanism of pRST98-increased bacterial virulence is still not fully elucidated. This study was designed to gain further insight into the roles of pRST98 in host responses. Methods: Human-derived macrophage-like cell line THP-1 was infected with wild-type (ST8), pRST98-deletion (ST8-ΔpRST98), and complemented (ST8-c-pRST98) S. typhi strains. Macrophage autophagy was performed by extracting the membrane-unbound LC3-I protein from cells, followed by flow cytometric detection of the membrane-associated fraction of LC3-II. Intracellular bacterial growth was determined by colony-forming units (cfu) assay. Macrophage cell death was measured by flow cytometry after propidium iodide (PI) staining. Autophagy activator rapamycin (RAPA) was added to the medium 2 h before infection to investigate the effect of autophagy on intracellular bacterial growth and macrophage cell death after S. typhi infection. Results: Plasmid pRST98 suppressed autophagy in infected macrophages and enhanced intracellular bacterial growth and S. typhi-induced macrophage cell death. Pretreatment with RAPA effectively restricted intracellular bacterial growth of ST8 and ST8-c-pRST98, and alleviated ST8 and ST8-c-pRST98-induced macrophage cell death, but had no significant effect on ST8-ΔpRST98. Conclusions: Plasmid pRST98 enhances intracellular bacterial growth and S. typhi-induced macrophage cell death by suppressing autophagy. Keywords: Plasmid pRST98, Macrophage, Autophagy, Intracellular bacterial growth, Cell death
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spelling doaj.art-30b3cf2ec7204cb382d42075dc6cb9462022-12-21T18:14:38ZengElsevierBrazilian Journal of Infectious Diseases1413-86702012-05-01163262266Salmonella enterica serovar Typhi plasmid pRST98 enhances intracellular bacterial growth and S. typhi-induced macrophage cell death by suppressing autophagyPeiyan He0Shuyan Wu1Yuanyuan Chu2Yanru Yang3Yuanyuan Li4Rui Huang5Microbiology Department, Medical College, Soochow University, Suzhou, Jiangsu, China; Jiaxing Center for Disease Control and Prevention, Jiaxing, Zhejiang, ChinaMicrobiology Department, Medical College, Soochow University, Suzhou, Jiangsu, ChinaMicrobiology Department, Medical College, Soochow University, Suzhou, Jiangsu, ChinaMicrobiology Department, Medical College, Soochow University, Suzhou, Jiangsu, ChinaMicrobiology Department, Medical College, Soochow University, Suzhou, Jiangsu, ChinaMicrobiology Department, Medical College, Soochow University, Suzhou, Jiangsu, China; Corresponding author at: Medical College of Soochow University, No. 199, Ren Ai Road, Suzhou, Jiangsu 215123, China.Objectives: Plasmid pRST98 is a hybrid resistance-virulence plasmid isolated from Salmonella enterica serovar Typhi (S. typhi). Previous studies demonstrated that pRST98 could enhance the virulence of its host bacteria. However, the mechanism of pRST98-increased bacterial virulence is still not fully elucidated. This study was designed to gain further insight into the roles of pRST98 in host responses. Methods: Human-derived macrophage-like cell line THP-1 was infected with wild-type (ST8), pRST98-deletion (ST8-ΔpRST98), and complemented (ST8-c-pRST98) S. typhi strains. Macrophage autophagy was performed by extracting the membrane-unbound LC3-I protein from cells, followed by flow cytometric detection of the membrane-associated fraction of LC3-II. Intracellular bacterial growth was determined by colony-forming units (cfu) assay. Macrophage cell death was measured by flow cytometry after propidium iodide (PI) staining. Autophagy activator rapamycin (RAPA) was added to the medium 2 h before infection to investigate the effect of autophagy on intracellular bacterial growth and macrophage cell death after S. typhi infection. Results: Plasmid pRST98 suppressed autophagy in infected macrophages and enhanced intracellular bacterial growth and S. typhi-induced macrophage cell death. Pretreatment with RAPA effectively restricted intracellular bacterial growth of ST8 and ST8-c-pRST98, and alleviated ST8 and ST8-c-pRST98-induced macrophage cell death, but had no significant effect on ST8-ΔpRST98. Conclusions: Plasmid pRST98 enhances intracellular bacterial growth and S. typhi-induced macrophage cell death by suppressing autophagy. Keywords: Plasmid pRST98, Macrophage, Autophagy, Intracellular bacterial growth, Cell deathhttp://www.sciencedirect.com/science/article/pii/S1413867012703215
spellingShingle Peiyan He
Shuyan Wu
Yuanyuan Chu
Yanru Yang
Yuanyuan Li
Rui Huang
Salmonella enterica serovar Typhi plasmid pRST98 enhances intracellular bacterial growth and S. typhi-induced macrophage cell death by suppressing autophagy
Brazilian Journal of Infectious Diseases
title Salmonella enterica serovar Typhi plasmid pRST98 enhances intracellular bacterial growth and S. typhi-induced macrophage cell death by suppressing autophagy
title_full Salmonella enterica serovar Typhi plasmid pRST98 enhances intracellular bacterial growth and S. typhi-induced macrophage cell death by suppressing autophagy
title_fullStr Salmonella enterica serovar Typhi plasmid pRST98 enhances intracellular bacterial growth and S. typhi-induced macrophage cell death by suppressing autophagy
title_full_unstemmed Salmonella enterica serovar Typhi plasmid pRST98 enhances intracellular bacterial growth and S. typhi-induced macrophage cell death by suppressing autophagy
title_short Salmonella enterica serovar Typhi plasmid pRST98 enhances intracellular bacterial growth and S. typhi-induced macrophage cell death by suppressing autophagy
title_sort salmonella enterica serovar typhi plasmid prst98 enhances intracellular bacterial growth and s typhi induced macrophage cell death by suppressing autophagy
url http://www.sciencedirect.com/science/article/pii/S1413867012703215
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