Elevated c-di-GMP Levels and Expression of the Type III Secretion System Promote Corneal Infection by Pseudomonas aeruginosa

Yam, Joey Kuok Hoong1; Aung, Thet Tun2,3; Chua, Song Lin4; Cheng, Yingying5; Kohli, Gurjeet Singh6; Zhou, Jianuan7; Constancias, Florentin8; Liu, Yang9; Cai, Zhao9; Salido, May Margarette Santillan1; Drautz-Moses, Daniela, I1; Rice, Scott A.1,10,11; Schuster, Stephan Christoph1,10; Boo, Zhao Zhi10; Wu, Bin10; Kjelleberg, Staffan1,10,12; Tolker-Nielsen, Tim13; Lakshminarayanan, Rajamani2,14,15; Beuerman, Roger W.2,16,17,18; Yang, Liang9; Givskov, Michael1,13

2022-08-01

10.1128/iai.00061-22

INFECTION AND IMMUNITY   Impact Factor & Quartile

0019-9567

1098-5522

Pseudomonas aeruginosa is generally believed to establish biofilm-associated infections under the regulation of the secondary messenger c-di-GMP. To evaluate P. aeruginosa biofilm physiology during ocular infections, comparative transcriptomic analysis was performed on wild-type P. aeruginosa PAO1, a Delta wspF mutant strain (high c-di-GMP levels), and a p(lac)-yhjH-containing strain (low c-di-GMP levels) from mouse corneal infection, as well as in vitro biofilm and planktonic cultures. The c-di-GMP content in P. aeruginosa during corneal infection was monitored using a fluorescent c-di-GMP reporter strain. Biofilm-related genes were induced in in vivo PAO1 compared to in vitro planktonic bacteria. Several diguanylate cyclases and phosphodiesterases were commonly regulated in in vivo PAO1 and in vitro biofilm compared to in vitro planktonic bacteria. Several exopolysaccharide genes and motility genes were induced and downregulated, respectively, in in vivo PAO1 and the in vivo Delta wspF mutant compared to the in vivo p(lac)-yhjH-containing strain. Elevation of c-di-GMP levels in P. aeruginosa began as early as 2 h postinfection. The Delta wspF mutant was less susceptible to host clearance than the p(lac)-yhjH-containing strain and could suppress host immune responses. The type III secretion system (T3SS) was induced in in vivo PAO1 compared to in vitro biofilm bacteria. Delta wspF mutant with a defective T3SS was more susceptible to host clearance than Delta wspF mutant with a functional T3SS. Our study suggests that elevated intracellular c-di-GMP levels and T3SS activity in P. aeruginosa are necessary for establishment of infection and modulation of host immune responses in mouse cornea.

Pseudomonas aeruginosa in vivo biofilms mouse corneal infection transcriptomics c-di-GMP type III secretion system immune response

SCI

English

Corresponding

AcRF Tier 2 from the Ministry of Education, Singapore[MOE2016-T2-1-010]

Immunology ; Infectious Diseases

Immunology ; Infectious Diseases

WOS:000835273900002

AMER SOC MICROBIOLOGY

IMMUNOLOGY

Web of Science

1.Nanyang Technol Univ, Singapore Ctr Environm Life Sci Engn SCELSE, Singapore, Singapore
2.Singapore Eye Res Inst, Ocular Infect & Antimicrobials Res Grp, Singapore, Singapore
3.Natl Univ Singapore, Natl Univ Hlth Syst, Yong Loo Lin Sch Med, Dept Microbiol & Immunol, Singapore, Singapore
4.Hong Kong Polytech Univ, Dept Appl Biol & Chem Technol, Hong Kong, Peoples R China
5.BGI Shenzhen, Forens Genom Int FGI, Shenzhen, Peoples R China
6.Alfred Wegener Inst, Helmholtz Zentrum Polar & Meeresforsch, Bremerhaven, Germany
7.South China Agr Univ, Integrat Microbiol Res Ctr, Guangdong Prov Key Lab Microbial Signals & Dis Co, Guangzhou, Peoples R China
8.Swiss Fed Inst Technol, Dept Hlth Sci & Technol, Zurich, Switzerland
9.Southern Univ Sci & Technol, Sch Med, Shenzhen, Guangdong, Peoples R China
10.Nanyang Technol Univ, Sch Biol Sci, Singapore, Singapore
11.CSIRO, Agr & Food Microbiomes Syst Hlth 1, Canberra, ACT, Australia
12.Univ New South Wales, Sch Biol Earth & Environm Sci, Sydney, NSW, Australia
13.Univ Copenhagen, Dept Immunol & Microbiol, Costerton Biofilm Ctr, Copenhagen, Denmark
14.Natl Univ Singapore, Dept Pharm, Singapore, Singapore
15.Duke NUS Med Sch, Acad Clin Program, Ophthalmol & Visual Sci Acad Clin Program, Singapore, Singapore
16.Duke NUS, SRP Neurosci & Behav Disorders & Emerging Infect, Singapore, Singapore
17.Natl Univ Singapore, Yong Loo Lin Sch Med, Ophthalmol, Singapore, Singapore
18.Univ Tennessee, Ctr Hlth Sci, Dept Ophthalmol, Memphis, TN 38163 USA

Yam, Joey Kuok Hoong,Aung, Thet Tun,Chua, Song Lin,et al. Elevated c-di-GMP Levels and Expression of the Type III Secretion System Promote Corneal Infection by Pseudomonas aeruginosa[J]. INFECTION AND IMMUNITY,2022.

Yam, Joey Kuok Hoong.,Aung, Thet Tun.,Chua, Song Lin.,Cheng, Yingying.,Kohli, Gurjeet Singh.,...&Givskov, Michael.(2022).Elevated c-di-GMP Levels and Expression of the Type III Secretion System Promote Corneal Infection by Pseudomonas aeruginosa.INFECTION AND IMMUNITY.

Yam, Joey Kuok Hoong,et al."Elevated c-di-GMP Levels and Expression of the Type III Secretion System Promote Corneal Infection by Pseudomonas aeruginosa".INFECTION AND IMMUNITY (2022).