Pretreatment of Mice with Streptomycin Provides a<i>Salmonella enterica</i>Serovar Typhimurium Colitis Model That Allows Analysis of Both Pathogen and Host

Barthel, Manja; Hapfelmeier, Siegfried; Quintanilla-Martı́nez, Leticia; Kremer, Marcus; Rohde, Manfred; Hogardt, Michael; Pfeffer, Klaus; Rüssmann, Holger; Hardt, Wolf‐Dietrich

doi:10.1128/iai.71.5.2839-2858.2003

Cited by 909 publications

(1,279 citation statements)

References 81 publications

Supporting

Mentioning

1,221

Contrasting

Unclassified

Order By: Relevance

“…Future attempts should also include the transfer of the Dual RNA-seq technique to three-dimensional tissue models (e.g. Pusch et al, 2011) or to animal models such as the streptomycin-treated mouse (Barthel et al, 2003).…”

Section: Towards In Vivo Dual Rna-seqmentioning

confidence: 99%

Dual RNA-seq of pathogen and host

2012

View full text Add to dashboard Cite

SummaryThe infection of a eukaryotic host cell by a bacterial pathogen is one of the most intimate examples of cross-kingdom interactions in biology. Infection processes are highly relevant from both a basic research as well as a clinical point of view. Sophisticated mechanisms have evolved in the pathogen to manipulate the host response and vice versa host cells have developed a wide range of anti-microbial defense strategies to combat bacterial invasion and clear infections.However, it is this diversity and complexity that makes infection research so challenging to technically address as common approaches have either been optimized for bacterial or eukaryotic organisms. Instead, methods are required that are able to deal with the often dramatic discrepancy between host and pathogen with respect to various cellular properties and processes. One class of cellular macromolecules that exemplify this host-pathogen heterogeneity is given by their transcriptomes: Bacterial transcripts differ from their eukaryotic counterparts in many aspects that involve both quantitative and qualitative traits. The entity of RNA transcripts present in a cell is of paramount interest as it reflects the cell's physiological state under the given condition. Genome-wide transcriptomic techniques such as RNA-seq have therefore been used for single-organism analyses for several years, but their applicability has been limited for infection studies.The present work describes the establishment of a novel transcriptomic approach for infection biology which we have termed "Dual RNA-seq". Using this technology, it was intended to shed light particularly on the contribution of non-protein-encoding transcripts to virulence, as these classes have mostly evaded previous infection studies due to the lack of suitable methods. The performance of Dual RNA-seq was evaluated in an in vitro infection model based on the important facultative intracellular pathogen Salmonella enterica serovar Typhimurium and different human cell lines. Dual RNA-seq was found to be capable of capturing all major bacterial and human transcript classes and proved reproducible. During the course of these experiments, a previously largely uncharacterized bacterial small non-coding RNA (sRNA), referred to as STnc440, was identified as one of the most strongly induced genes in intracellular Salmonella.Interestingly, while inhibition of STnc440 expression has been previously shown to cause a virulence defect in different animal models of Salmonellosis, the underlying molecular mechanisms have remained obscure. Here, classical genetics, transcriptomics and biochemical assays proposed a complex model of Salmonella gene expression control that is orchestrated by this sRNA. In particular, STnc440 was found to be involved in the regulation of multiple bacterial target mRNAs by direct base pair interaction with consequences for Salmonella virulence and

show abstract

Section: Towards In Vivo Dual Rna-seqmentioning

confidence: 99%

Dual RNA-seq of pathogen and host

2012

View full text Add to dashboard Cite

show abstract

“…5 A mouse model of typhlocolitis was developed to investigate the interactions occurring between Salmonella and the host in the gastrointestinal tract by pretreating mice with streptomycin sulfate to deplete commensal bacteria before infection with streptomycin-resistant Salmonella strains. 6,7 Intestinal inflammation in this model is observed mainly in the cecum (typhlitis) and to a lesser extent in the colon (colitis) and is characterized by loss of epithelial integrity (erosion and exfoliation), submucosal edema, infiltration of polymorphonuclear cells to the lamina propria and loss of goblet cells. The model allows the identification of the impact of host genes such as Nramp1 (Nramp1) and Tlr5 on the severity of intestinal inflammation during infection.…”

Section: Introductionmentioning

confidence: 99%

Impact of Usp18 and IFN signaling in Salmonella-induced typhlitis

et al. 2011

View full text Add to dashboard Cite

In humans, Salmonella infection causes two major clinical diseases, typhoid fever and a self-limiting gastro-enteritidis. Salmonella transmission occurs by the fecal-oral route and the interactions between the bacteria and the digestive tract epithelium are central to the outcome of the infection. Using a mouse model of typhoid fever, we previously identified a mutation in USP18 affecting type I interferon (IFN) signaling resulting in increased susceptibility to systemic Salmonella infection. In this study, we demonstrate the effects of this mutation during the early response to Salmonella using a model of typhlitis. Mutant Usp18 mice showed a minimal inflammatory response early after Salmonella Typhimurium infection that was associated with low pathologic scores and low IFN-g production. This resulted in an increased interaction of Salmonella with the cecal epithelium and earlier systemic dissemination of the bacteria. The global transcriptional signature in the cecum of mouse during Salmonella infection showed normal expression of tissue specific genes and upregulation of type I IFN pathway in mutant mice. In control mice, there was a significant over-representation of genes involved in cellular recruitment and antibacterial activity paralleling the histopathological features. These results show the impact of USP18 in the development of Salmonella-induced typhlitis.

show abstract

“…Animals were then supplied with water and food ad libitum. As previously described (2), at 20 h after streptomycin treatment, water and food were withdrawn again for 4 h and the mice were gavaged with either 1 ϫ 10 7 colony-forming units of wild-type S. enterica Serovar Typhimurium (50-l suspension in HBSS), the flagellindeficient mutant strain fliC Ϫ /fljB Ϫ , or with sterile HBSS. Six hours after gavage, mice were killed and intestinal tissue was harvested for analysis.…”

Section: Methodsmentioning

confidence: 99%

“…In vivo experiments of Salmonella infection were performed by using a previously described streptomycin-pretreated mouse model (2). The protocol was approved by the University of Chicago Animal Care (IACUC) committee.…”

Section: Methodsmentioning

confidence: 99%

“…To determine the contribution of flagellin to the induction of intestinal epithelial Hsp25 in vivo, the streptomycin animal model of Salmonella infection was used (2). Mice are intrinsically resistant to developing enterocolitis from Salmonella infection, but this model allows development of a murine intestinal salmonellosis that closely resembles the enterocolitis caused by S. typhimurium in humans.…”

Section: Flagellin Is a Major Heat Shock Protein-inducing Effector Anmentioning

confidence: 99%

See 1 more Smart Citation