Spatial Segregation of Virulence Gene Expression during Acute Enteric Infection with <named-content content-type="genus-species">Salmonella enterica</named-content> serovar Typhimurium
ABSTRACT To establish a replicative niche during its infectious cycle between the intestinal lumen and tissue, the enteric pathogen Salmonella enterica serovar Typhimurium requires numerous virulence genes, including genes for two type III secretion systems (T3SS) and their cognate effectors. To bet...
| Main Authors: | , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
American Society for Microbiology
2014-02-01
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| Series: | mBio |
| Online Access: | https://journals.asm.org/doi/10.1128/mBio.00946-13 |
| _version_ | 1819041479909179392 |
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| author | Richard C. Laughlin Leigh A. Knodler Roula Barhoumi H. Ross Payne Jing Wu Gabriel Gomez Roberta Pugh Sara D. Lawhon Andreas J. Bäumler Olivia Steele-Mortimer L. Garry Adams |
| author_facet | Richard C. Laughlin Leigh A. Knodler Roula Barhoumi H. Ross Payne Jing Wu Gabriel Gomez Roberta Pugh Sara D. Lawhon Andreas J. Bäumler Olivia Steele-Mortimer L. Garry Adams |
| author_sort | Richard C. Laughlin |
| collection | DOAJ |
| description | ABSTRACT To establish a replicative niche during its infectious cycle between the intestinal lumen and tissue, the enteric pathogen Salmonella enterica serovar Typhimurium requires numerous virulence genes, including genes for two type III secretion systems (T3SS) and their cognate effectors. To better understand the host-pathogen relationship, including early infection dynamics and induction kinetics of the bacterial virulence program in the context of a natural host, we monitored the subcellular localization and temporal expression of T3SS-1 and T3SS-2 using fluorescent single-cell reporters in a bovine, ligated ileal loop model of infection. We observed that the majority of bacteria at 2 h postinfection are flagellated, express T3SS-1 but not T3SS-2, and are associated with the epithelium or with extruding enterocytes. In epithelial cells, S. Typhimurium cells were surrounded by intact vacuolar membranes or present within membrane-compromised vacuoles that typically contained numerous vesicular structures. By 8 h postinfection, T3SS-2-expressing bacteria were detected in the lamina propria and in the underlying mucosa, while T3SS-1-expressing bacteria were in the lumen. Our work identifies for the first time the temporal and spatial regulation of T3SS-1 and -2 expression during an enteric infection in a natural host and provides further support for the concept of cytosolic S. Typhimurium in extruding epithelium as a mechanism for reseeding the lumen. IMPORTANCE The pathogenic bacterium Salmonella enterica serovar Typhimurium invades and persists within host cells using distinct sets of virulence genes. Genes from Salmonella pathogenicity island 1 (SPI-1) are used to initiate contact and facilitate uptake into nonphagocytic host cells, while genes within SPI-2 allow the pathogen to colonize host cells. While many studies have identified bacterial virulence determinants in animal models of infection, very few have focused on virulence gene expression at the single-cell level during an in vivo infection. To better understand when and where bacterial virulence factors are expressed during an acute enteric infection of a natural host, we infected bovine jejunal-ileal loops with S. Typhimurium cells harboring fluorescent transcriptional reporters for SPI-1 and -2 (PinvF and PssaG, respectively). After a prescribed time of infection, tissue and luminal fluid were collected and analyzed by microscopy. During early infection (≤2 h), bacteria within both intact and compromised membrane-bound vacuoles were observed within the epithelium, with the majority expressing SPI-1. As the infection progressed, S. Typhimurium displayed differential expression of the SPI-1 and SPI-2 regulons, with the majority of tissue-associated bacteria expressing SPI-2 and the majority of lumen-associated bacteria expressing SPI-1. This underscores the finding that Salmonella virulence gene expression changes as the pathogen transitions from one anatomical location to the next. |
| first_indexed | 2024-12-21T09:25:39Z |
| format | Article |
| id | doaj.art-94dff99ddb174ca8b1dc25b22ff70775 |
| institution | Directory Open Access Journal |
| issn | 2150-7511 |
| language | English |
| last_indexed | 2024-12-21T09:25:39Z |
| publishDate | 2014-02-01 |
| publisher | American Society for Microbiology |
| record_format | Article |
| series | mBio |
| spelling | doaj.art-94dff99ddb174ca8b1dc25b22ff707752022-12-21T19:08:54ZengAmerican Society for MicrobiologymBio2150-75112014-02-015110.1128/mBio.00946-13Spatial Segregation of Virulence Gene Expression during Acute Enteric Infection with <named-content content-type="genus-species">Salmonella enterica</named-content> serovar TyphimuriumRichard C. Laughlin0Leigh A. Knodler1Roula Barhoumi2H. Ross Payne3Jing Wu4Gabriel Gomez5Roberta Pugh6Sara D. Lawhon7Andreas J. Bäumler8Olivia Steele-Mortimer9L. Garry Adams10Department of Veterinary Pathobiology, College of Veterinary Medicine & Biomedical Sciences, Texas A&M University, College Station, Texas, USAPaul G. Allen School for Global Animal Health, College of Veterinary Medicine, Washington State University, Pullman, Washington, USADepartment of Veterinary Integrative Biosciences, College of Veterinary Medicine & Biomedical Sciences, Texas A&M University, College Station, Texas, USADepartment of Veterinary Pathobiology, College of Veterinary Medicine & Biomedical Sciences, Texas A&M University, College Station, Texas, USADepartment of Veterinary Pathobiology, College of Veterinary Medicine & Biomedical Sciences, Texas A&M University, College Station, Texas, USADepartment of Veterinary Pathobiology, College of Veterinary Medicine & Biomedical Sciences, Texas A&M University, College Station, Texas, USADepartment of Veterinary Pathobiology, College of Veterinary Medicine & Biomedical Sciences, Texas A&M University, College Station, Texas, USADepartment of Veterinary Pathobiology, College of Veterinary Medicine & Biomedical Sciences, Texas A&M University, College Station, Texas, USADepartment of Medical Microbiology and Immunology, School of Medicine, University of California at Davis, Davis, California, USALaboratory of Intracellular Parasites, Rocky Mountain Laboratories, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Hamilton, Montana, USADepartment of Veterinary Pathobiology, College of Veterinary Medicine & Biomedical Sciences, Texas A&M University, College Station, Texas, USAABSTRACT To establish a replicative niche during its infectious cycle between the intestinal lumen and tissue, the enteric pathogen Salmonella enterica serovar Typhimurium requires numerous virulence genes, including genes for two type III secretion systems (T3SS) and their cognate effectors. To better understand the host-pathogen relationship, including early infection dynamics and induction kinetics of the bacterial virulence program in the context of a natural host, we monitored the subcellular localization and temporal expression of T3SS-1 and T3SS-2 using fluorescent single-cell reporters in a bovine, ligated ileal loop model of infection. We observed that the majority of bacteria at 2 h postinfection are flagellated, express T3SS-1 but not T3SS-2, and are associated with the epithelium or with extruding enterocytes. In epithelial cells, S. Typhimurium cells were surrounded by intact vacuolar membranes or present within membrane-compromised vacuoles that typically contained numerous vesicular structures. By 8 h postinfection, T3SS-2-expressing bacteria were detected in the lamina propria and in the underlying mucosa, while T3SS-1-expressing bacteria were in the lumen. Our work identifies for the first time the temporal and spatial regulation of T3SS-1 and -2 expression during an enteric infection in a natural host and provides further support for the concept of cytosolic S. Typhimurium in extruding epithelium as a mechanism for reseeding the lumen. IMPORTANCE The pathogenic bacterium Salmonella enterica serovar Typhimurium invades and persists within host cells using distinct sets of virulence genes. Genes from Salmonella pathogenicity island 1 (SPI-1) are used to initiate contact and facilitate uptake into nonphagocytic host cells, while genes within SPI-2 allow the pathogen to colonize host cells. While many studies have identified bacterial virulence determinants in animal models of infection, very few have focused on virulence gene expression at the single-cell level during an in vivo infection. To better understand when and where bacterial virulence factors are expressed during an acute enteric infection of a natural host, we infected bovine jejunal-ileal loops with S. Typhimurium cells harboring fluorescent transcriptional reporters for SPI-1 and -2 (PinvF and PssaG, respectively). After a prescribed time of infection, tissue and luminal fluid were collected and analyzed by microscopy. During early infection (≤2 h), bacteria within both intact and compromised membrane-bound vacuoles were observed within the epithelium, with the majority expressing SPI-1. As the infection progressed, S. Typhimurium displayed differential expression of the SPI-1 and SPI-2 regulons, with the majority of tissue-associated bacteria expressing SPI-2 and the majority of lumen-associated bacteria expressing SPI-1. This underscores the finding that Salmonella virulence gene expression changes as the pathogen transitions from one anatomical location to the next.https://journals.asm.org/doi/10.1128/mBio.00946-13 |
| spellingShingle | Richard C. Laughlin Leigh A. Knodler Roula Barhoumi H. Ross Payne Jing Wu Gabriel Gomez Roberta Pugh Sara D. Lawhon Andreas J. Bäumler Olivia Steele-Mortimer L. Garry Adams Spatial Segregation of Virulence Gene Expression during Acute Enteric Infection with <named-content content-type="genus-species">Salmonella enterica</named-content> serovar Typhimurium mBio |
| title | Spatial Segregation of Virulence Gene Expression during Acute Enteric Infection with <named-content content-type="genus-species">Salmonella enterica</named-content> serovar Typhimurium |
| title_full | Spatial Segregation of Virulence Gene Expression during Acute Enteric Infection with <named-content content-type="genus-species">Salmonella enterica</named-content> serovar Typhimurium |
| title_fullStr | Spatial Segregation of Virulence Gene Expression during Acute Enteric Infection with <named-content content-type="genus-species">Salmonella enterica</named-content> serovar Typhimurium |
| title_full_unstemmed | Spatial Segregation of Virulence Gene Expression during Acute Enteric Infection with <named-content content-type="genus-species">Salmonella enterica</named-content> serovar Typhimurium |
| title_short | Spatial Segregation of Virulence Gene Expression during Acute Enteric Infection with <named-content content-type="genus-species">Salmonella enterica</named-content> serovar Typhimurium |
| title_sort | spatial segregation of virulence gene expression during acute enteric infection with named content content type genus species salmonella enterica named content serovar typhimurium |
| url | https://journals.asm.org/doi/10.1128/mBio.00946-13 |
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