Stably Integrated <i>luxCDABE</i> for Assessment of <i>Salmonella</i> Invasion Kinetics

Flentie, Kelly; Qi, Min; Gammon, Seth T.; Razia, Yasmin; Lui, Felix H.; Marpegan, Luciano; Manglik, Aashish; Piwnica‐Worms, David; McKinney, Jeffrey S.

doi:10.2310/7290.2008.00024

Cited by 17 publications

(14 citation statements)

References 40 publications

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“…Consistent with previous studies, 26,56,[64][65][66][67] we observed that an InvC deficiency caused a significant impairment of bacterial entrance to non-phagocytic cells, as compared with WT Salmonella. Lack of InvC is thought to render Salmonella unable to translocate the SPI-1-derived effector molecules required to induce phagocytosis by epithelial cells.…”

Section: Discussionsupporting

confidence: 92%

Salmonella pathogenicity island 1 differentially modulates bacterial entry to dendritic and non‐phagocytic cells

et al. 2010

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SummarySalmonella enterica serovar Typhimurium can enter non-phagocytic cells, such as intestinal epithelial cells, by virtue of a Type Three Secretion System (TTSS) encoded in the Salmonella Pathogenicity Island 1 (SPI-1), which translocates bacterial effector molecules into the host cell. Salmonella can also be taken up by dendritic cells (DCs). Although the role of SPI-1 in non-phagocytic cell invasion is well established, its contribution to invasion of phagocytic cells has not been evaluated. Here, we have tested the invasive capacity of a S. Typhimurium strain lacking a key component of its TTSS-1 (DInvC) leading to defective translocation of SPI-1-encoded effectors. Whereas this mutant Salmonella strain was impaired for invasion of non-phagocytic cells, it was taken up by DCs at a significantly higher rate than wild-type Salmonella. Similar to wild-type Salmonella, the DInvC mutant strain retained the capacity to avoid antigen presentation to T cells. However, mice infected with the DInvC mutant strain showed higher survival rate and reduced organ colonization. Our data suggest that, besides promoting phagocytosis by non-phagocytic cells, SPI-1 modulates the number of bacteria that enters DCs. The SPI-1 could be considered not only as an inducer of epithelial cell invasion but as a controller of DC entry.

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Section: Discussionsupporting

confidence: 92%

Salmonella pathogenicity island 1 differentially modulates bacterial entry to dendritic and non‐phagocytic cells

et al. 2010

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“…The second strategy involves the use of the entire lux operon from a bacterium such as Photorhabdus luminescens (formerly Xenorhabdus luminescens ). A transposon insertion or plasmid vector containing the luciferase enzyme, as well as the components for substrate production can be engineered with a transposon insertion (Flentie et al, 2008). This system will emit light in the range of 495–500 nm.…”

Section: Introductionmentioning

confidence: 99%

In vivo bioluminescence imaging of Burkholderia mallei respiratory infection and treatment in the mouse model

Massey¹

2011

Front. Microbio.

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Bioluminescent imaging (BLI) technology is a powerful tool for monitoring infectious disease progression and treatment approaches. BLI is particularly useful for tracking fastidious intracellular pathogens that might be difficult to recover from certain organs. Burkholderia mallei, the causative agent of glanders, is a facultative intracellular pathogen and has been classified by the CDC as a Category B select agent due to its highly infectious nature and potential use as a biological weapon. Very little is known regarding pathogenesis or treatment of glanders. We investigated the use of bioluminescent reporter constructs to monitor the dynamics of infection as well as the efficacy of therapeutics for B. mallei in real-time. A stable luminescent reporter B. mallei strain was created using the pUTmini-Tn5::luxKm2 plasmid and used to monitor glanders in the BALB/c murine model. Mice were infected via the intranasal route with 5 × 103 bacteria and monitored by BLI at 24, 48, and 72 h. We verified that our reporter construct maintained similar virulence and growth kinetics compared to wild-type B. mallei and confirmed that it maintains luminescent stability in the presence or absence of antibiotic selection. The luminescent signal was initially seen in the lungs, and progressed to the liver and spleen over the course of infection. We demonstrated that antibiotic treatment 24 h post-infection resulted in reduction of bioluminescence that can be attributed to decreased bacterial burden in target organs. These findings suggest that BLI can be used to monitor disease progression and efficacy of therapeutics during glanders infections. Finally, we report an alternative method to mini-Tn5::luxKm2 transposon using mini-Tn7-lux elements that insert site-specifically at known genomic attachment sites and that can also be used to tag bacteria.

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“…The lack of PhoP-PhoQ two-component system has always been linked to major intramacrophage survival defects and marked attenuation in the murine typhoid model. Hence, human cancer cell lines are commonly used to study invasion of Salmonella to eukaryotic cells [19]. The attenuated Salmonella used in this study still maintained its ability to invade MCF-7 and MCF-10A mammalian cells, where 1.2% and 0.5% of the cells were infected, respectively.…”

Section: Discussionmentioning

confidence: 87%

AttenuatedSalmonella typhimuriumSV4089 as a Potential Carrier of Oral DNA Vaccine in Chickens

Jazayeri

Ideris

Zakaria

et al. 2012

Journal of Biomedicine and Biotechnology

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Attenuated Salmonella has been used as a carrier for DNA vaccine. However, in vitro and in vivo studies on the bacteria following transfection of plasmid DNA were poorly studied. In this paper, eukaryotic expression plasmids encoding avian influenza virus (AIV) subtype H5N1 genes, pcDNA3.1/HA, NA, and NP, were transfected into an attenuated Salmonella enteric typhimurium SV4089. In vitro stability of the transfected plasmids into Salmonella were over 90% after 100 generations. The attenuated Salmonella were able to invade MCF-7 (1.2%) and MCF-10A (0.5%) human breast cancer cells. Newly hatched specific-pathogen-free (SPF) chicks were inoculated once by oral gavage with 109 colony-forming unit (CFU) of the attenuated Salmonella. No abnormal clinical signs or deaths were recorded after inoculation. Viable bacteria were detected 3 days after inoculation by plating from spleen, liver, and cecum. Fluorescent in situ hybridization (FISH) and polymerase chain reaction (PCR) were carried out for confirmation. Salmonella was not detected in blood cultures although serum antibody immune responses to Salmonella O antiserum group D1 factor 1, 9, and 12 antigens were observed in all the inoculated chickens after 7 days up to 35 days. Our results showed that live attenuated S. typhimurium SV4089 harboring pcDNA3.1/HA, NA, and NP may provide a unique alternative as a carrier for DNA oral vaccine in chickens.

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Stably Integrated luxCDABE for Assessment of Salmonella Invasion Kinetics

Cited by 17 publications

References 40 publications

Salmonella pathogenicity island 1 differentially modulates bacterial entry to dendritic and non‐phagocytic cells

Salmonella pathogenicity island 1 differentially modulates bacterial entry to dendritic and non‐phagocytic cells

In vivo bioluminescence imaging of Burkholderia mallei respiratory infection and treatment in the mouse model

AttenuatedSalmonella typhimuriumSV4089 as a Potential Carrier of Oral DNA Vaccine in Chickens

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