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Early Host Cell Targets of Yersinia pestis during Primary Pneumonic Plague

Pechous, Roger D., Sivaraman, Vijay, Price, Paul A., Stasulli, Nikolas M., Goldman, William E.·2013·PLoS Pathogens
DOI 10.1371/journal.ppat.1003679PMC PMC3789773
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Figure 1 YopE-TEM translocation in the lung during pneumonic plague. (A) Groups of mice were inoculated intranasally with 106 CFU of Y. pestis expressing YopE-TEM or PBS alone (mock), followed by staining with CCF2-AM. Lungs were harvested at 12 hpi, and live cells were gated to evaluate blue/green fluorescence by flow cytometry. Cells showing blue fluorescence have been injected with YopE-TEM. Histograms show representative data from a single mouse; (B) Total injection events (cells exhibiting blue fluorescence) in the lungs of mice inoculated with 106 CFU Y. pestis YopE-TEM; (C) The percentage of total cells in the lung injected with YopE-TEM was evaluated for samples shown in (B) at 6, 12, and 24 hpi; (D) Lung bacterial burden was evaluated in mice inoculated with 106 CFU Y. pestis YopE-TEM. (E) Live injected cells in lungs of mice inoculated with 106 CFU Y. pestis YopE-TEM were gated for identification of alveolar macrophages (F480+CD11blow/midCD11chigh), interstitial macrophages (F480+CD11bhighCD11cmid/low), F480+CD11bhighCD11chigh macrophages, monocytes (F480−CD11bhighLy-6G−), neutrophils (F480−CD11bhighLy-6G+), and CD11b high and low dendritic cells (F480− CD11c+); All data are representative of at least three independent experiments with three to five mice at each time point. Error bars represent SEM.
Figure 1 YopE-TEM translocation in the lung during pneumonic plague. (A) Groups of mice were inoculated intranasally with 106 CFU of Y. pestis expressing YopE-TEM or PBS alone (mock), followed by staining with CCF2-AM. Lungs were harvested at 12 hpi, and live cells were gated to evaluate blue/green fluorescence by flow cytometry. Cells showing blue fluorescence have been injected with YopE-TEM. Histograms show representative data from a single mouse; (B) Total injection events (cells exhibiting blue fluorescence) in the lungs of mice inoculated with 106 CFU Y. pestis YopE-TEM; (C) The percentage of total cells in the lung injected with YopE-TEM was evaluated for samples shown in (B) at 6, 12, and 24 hpi; (D) Lung bacterial burden was evaluated in mice inoculated with 106 CFU Y. pestis YopE-TEM. (E) Live injected cells in lungs of mice inoculated with 106 CFU Y. pestis YopE-TEM were gated for identification of alveolar macrophages (F480+CD11blow/midCD11chigh), interstitial macrophages (F480+CD11bhighCD11cmid/low), F480+CD11bhighCD11chigh macrophages, monocytes (F480−CD11bhighLy-6G−), neutrophils (F480−CD11bhighLy-6G+), and CD11b high and low dendritic cells (F480− CD11c+); All data are representative of at least three independent experiments with three to five mice at each time point. Error bars represent SEM.CC BY · PubMed Central
Figure 2 Lung host cell repertoire of mice inoculated intranasally with Y. pestis. (A) Total cell numbers of innate immune cell populations in the lungs of mice inoculated with 106 CFU Y. pestis YopE-TEM or PBS (mock) at 6, 12, and 24 hpi. (B) Y. pestis causes severe inflammation in the lungs that is visible by 48 hpi. Mice were inoculated with 106 CFU Y. pestis and lungs were inflated with 10% formalin at 24 and 48 hpi prior to H and E staining. Images are representative of experiments done a minimum of three times. Data are representative of at least three independent experiments with five mice at each time point. Error bars represent SEM.
Figure 2 Lung host cell repertoire of mice inoculated intranasally with Y. pestis. (A) Total cell numbers of innate immune cell populations in the lungs of mice inoculated with 106 CFU Y. pestis YopE-TEM or PBS (mock) at 6, 12, and 24 hpi. (B) Y. pestis causes severe inflammation in the lungs that is visible by 48 hpi. Mice were inoculated with 106 CFU Y. pestis and lungs were inflated with 10% formalin at 24 and 48 hpi prior to H and E staining. Images are representative of experiments done a minimum of three times. Data are representative of at least three independent experiments with five mice at each time point. Error bars represent SEM.CC BY · PubMed Central
Figure 3 Pre-treatment of mice with clodrosome or neutrophil-depleting antibody α-Ly-6G. (A) Total YopE-TEM injection events 6 hpi in mice inoculated with 106 CFU Y. pestis YopE-TEM with and without prior alveolar macrophage depletion with clodrosome. (B) Total YopE-TEM injection events 24 hpi in Y. pestis-infected mice with and without pre-treatment with neutrophil-depleting antibody. (C) Identity of injected cells 6 hpi in mice with and without prior clodrosome treatment. (D) Identity of injected cells 24 hpi with and without pre-treatment with neutrophil-depleting antibody. Data are representative of at least three independent experiments with three to five mice at each time point. Error bars represent SEM for all graphs.
Figure 3 Pre-treatment of mice with clodrosome or neutrophil-depleting antibody α-Ly-6G. (A) Total YopE-TEM injection events 6 hpi in mice inoculated with 106 CFU Y. pestis YopE-TEM with and without prior alveolar macrophage depletion with clodrosome. (B) Total YopE-TEM injection events 24 hpi in Y. pestis-infected mice with and without pre-treatment with neutrophil-depleting antibody. (C) Identity of injected cells 6 hpi in mice with and without prior clodrosome treatment. (D) Identity of injected cells 24 hpi with and without pre-treatment with neutrophil-depleting antibody. Data are representative of at least three independent experiments with three to five mice at each time point. Error bars represent SEM for all graphs.CC BY · PubMed Central
Figure 4 Evaluating the progression of pneumonic plague in mice pre-treated with clodrosome or neutrophil-depleting antibody α-Ly-6G. (A) Lung bacterial burden in mice with and without pre-treatment with clodrosome or neutrophil-depleting antibody and after inoculation with 104 CFU Y. pestis; (B) Lung histology 48 hpi in mice inoculated with 104 CFU Y. pestis with and without prior treatment with clodrosome or neutrophil-depleting antibody. Bar graph represents experiments repeated at least three times with three to five mice. Error bars represent SEM. Histopathology shows representative images from experiments repeated at least twice.
Figure 4 Evaluating the progression of pneumonic plague in mice pre-treated with clodrosome or neutrophil-depleting antibody α-Ly-6G. (A) Lung bacterial burden in mice with and without pre-treatment with clodrosome or neutrophil-depleting antibody and after inoculation with 104 CFU Y. pestis; (B) Lung histology 48 hpi in mice inoculated with 104 CFU Y. pestis with and without prior treatment with clodrosome or neutrophil-depleting antibody. Bar graph represents experiments repeated at least three times with three to five mice. Error bars represent SEM. Histopathology shows representative images from experiments repeated at least twice.CC BY · PubMed Central

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Research works printed but not linked: 34

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Metadata

Authors
Pechous, R. D., Sivaraman, V., Price, P. A., Stasulli, N. M., & Goldman, W. E.
Venue
PLoS Pathogens
Publisher
Public Library of Science (PLoS)
Published
2013-10-03
Volume
9
Issue
10
Pages
e1003679
ISSN
1553-7374
DOI
10.1371/journal.ppat.1003679
PMC
PMC3789773
Cited by
90
References
0
Language
en
Licence
creativecommons.org/licenses/by/4.0/ (unspecified)

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