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Imaging InlC Secretion to Investigate Cellular Infection by the Bacterial Pathogen Listeria monocytogenes
Published on: September 19, 2013
TMT-Based Quantitative Proteomic Analysis of Intestinal Organoids Infected by Listeria monocytogenes Strains with
Cong Zhou1, Yafang Zou1, Jie Huang1
1Jiangsu Collaborative Innovation Center of Meat Production and Processing, Quality and Safety Control, College of Food Science and Technology, Nanjing Agricultural University, Nanjing 210095, China.
Abstract:
L. monocytogenes, consisting of 13 serotypes, is an opportunistic food-borne pathogen that causes different host reactions depending on its serotypes. In this study, highly toxic L. monocytogenes 10403s resulted in more severe infections and lower survival rates. Additionally, to investigate the remodeling of the host proteome by strains exhibiting differential toxicity, the cellular protein responses of intestinal organoids were analyzed using tandem mass tag (TMT) labeling and high-performance liquid chromatography−mass spectrometry. The virulent strain 10403s caused 102 up-regulated and 52 down-regulated proteins, while the low virulent strain M7 caused 188 up-regulated and 25 down-regulated proteins. Based on the analysis of gene ontology (GO) and KEGG databases, the expressions of differential proteins in organoids infected by L. monocytogenes 10403s (virulent strain) or M7 (low virulent strain) were involved in regulating essential processes such as the biological metabolism, the energy metabolism, and immune system processes. The results showed that the immune system process, as the primary host defense response to L. monocytogenes, comprised five pathways, including ECM−receptor interaction, the complement and coagulation cascades, HIF-1, ferroptosis, and NOD-like receptor signaling pathways. As for the L. monocytogenes 10403s vs. M7 group, the expression of differential proteins was involved in two pathways: systemic lupus erythematosus and transcriptional mis-regulation in cancer. All in all, these results revealed that L. monocytogenes strains with different toxicity induced similar biological functions and immune responses while having different regulations on differential proteins in the pathway.
Insights
Different Listeria monocytogenes strains cause varying host responses. Proteomic analysis revealed distinct protein regulations in intestinal organoids, impacting metabolism and immune pathways, despite overall similar biological functions.
Area of Science:
- Microbiology
- Proteomics
- Host-Pathogen Interactions
Background:
- Listeria monocytogenes is an opportunistic foodborne pathogen with 13 serotypes, causing varied host reactions.
- Strain toxicity influences infection severity and host survival rates.
Purpose of the Study:
- To investigate host proteome remodeling by Listeria monocytogenes strains with differential toxicity.
- To analyze cellular protein responses in intestinal organoids upon infection.
Main Methods:
- Tandem mass tag (TMT) labeling coupled with high-performance liquid chromatography-mass spectrometry (HPLC-MS) for proteomic analysis.
- Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis.
Main Results:
- The virulent strain 10403s and low-virulent strain M7 induced differential protein expression in intestinal organoids.
- Infected organoids showed altered biological metabolism, energy metabolism, and immune system processes.
- Immune responses involved pathways like ECM-receptor interaction and complement cascades; strain comparison highlighted differences in systemic lupus erythematosus and cancer-related pathways.
Conclusions:
- Listeria monocytogenes strains with differing toxicity induce comparable biological functions and immune responses.
- Differential protein regulation in specific pathways occurs despite overall functional similarity.
