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Salmonella-Infected Aortic Aneurysm: Investigating Pathogenesis Using Salmonella Serotypes

Polish Journal of Microbiology
Volume 68 (2019): Issue 4 (January 2019)
By:
Open Access
|Oct 2019

Figures & Tables

Fig. 1.

Autophagy and inflammasome induced by Salmonella infection.(A, C) Western blotting was performed with anti-LC3-I/II and anti-ASC antibodies. β-Actin Western blots were used as loading controls. LC3 was identified as a double band (i.e., LC3-I and LC3-II). (A) THP-1 macrophages and (C) THP-1 macrophage-derived foam cells were infected by different serotypes of Salmonella with or without virulence plasmid for 0.5 and 2 h. Uninfected macrophages and foam cells were the negative controls. (B, D) The LC3 I/II and ASC bands were quantified, and the ratios of autophagy and inflammasome were calculated from the ratios of infected to uninfected LC3-II/I cells and of infected to uninfected ASC, respectively. All values are represented as means ± standard error (n = 3). a–e indicate significant differences of autophagy formation between strains in the 0.5 and 2 h post-infection (p < 0.05); x, y, z indicate significant differences of inflammasome formation between strains in the 0.5 and 2 h post-infection. (p < 0.05). nc: uninfected cells.
Autophagy and inflammasome induced by Salmonella infection.(A, C) Western blotting was performed with anti-LC3-I/II and anti-ASC antibodies. β-Actin Western blots were used as loading controls. LC3 was identified as a double band (i.e., LC3-I and LC3-II). (A) THP-1 macrophages and (C) THP-1 macrophage-derived foam cells were infected by different serotypes of Salmonella with or without virulence plasmid for 0.5 and 2 h. Uninfected macrophages and foam cells were the negative controls. (B, D) The LC3 I/II and ASC bands were quantified, and the ratios of autophagy and inflammasome were calculated from the ratios of infected to uninfected LC3-II/I cells and of infected to uninfected ASC, respectively. All values are represented as means ± standard error (n = 3). a–e indicate significant differences of autophagy formation between strains in the 0.5 and 2 h post-infection (p < 0.05); x, y, z indicate significant differences of inflammasome formation between strains in the 0.5 and 2 h post-infection. (p < 0.05). nc: uninfected cells.

Fig. 2.

CD36 expression in THP-1 macrophage-derived foam cells after different serotypes Salmonella infection. After treated with ox-LDL, THP-1 macrophage-derived foam cells were infected with plasmid-bearing and -less S. Typhimurium, Enteritidis, and Cholerae suis, respectively. CD36 expression was analyzed through flow cytometry.
CD36 expression in THP-1 macrophage-derived foam cells after different serotypes Salmonella infection. After treated with ox-LDL, THP-1 macrophage-derived foam cells were infected with plasmid-bearing and -less S. Typhimurium, Enteritidis, and Cholerae suis, respectively. CD36 expression was analyzed through flow cytometry.

Fig. 3.

IL-1β production by THP-1 macrophage-derived foam cells after Salmonella infection.ELISA was performed for IL-1β produced after infection by different Salmonella serotypes. Foam cells were infected by plasmid-bearing S. Typhimurium OU5045, plasmid-less S. Typhimurium OU5046, plasmid-bearing S. Enteriditis OU7130, plasmid-less S. Enteriditis OU7067, and plasmid-bearing S. Choleraesuis OU7085 and plasmid-less S. Choleraesuis OU7266 for 0.5 and 2 h, and the supernatants were harvested and used for experiments. The experiments were performed in triplicate and presented as mean ± SD. (***p < 0.005, one-way ANOVA). NC: uninfected cells; ST: S. Typhimurium; SE: S. Enteritidis; SC: S. Choleraesuis.
IL-1β production by THP-1 macrophage-derived foam cells after Salmonella infection.ELISA was performed for IL-1β produced after infection by different Salmonella serotypes. Foam cells were infected by plasmid-bearing S. Typhimurium OU5045, plasmid-less S. Typhimurium OU5046, plasmid-bearing S. Enteriditis OU7130, plasmid-less S. Enteriditis OU7067, and plasmid-bearing S. Choleraesuis OU7085 and plasmid-less S. Choleraesuis OU7266 for 0.5 and 2 h, and the supernatants were harvested and used for experiments. The experiments were performed in triplicate and presented as mean ± SD. (***p < 0.005, one-way ANOVA). NC: uninfected cells; ST: S. Typhimurium; SE: S. Enteritidis; SC: S. Choleraesuis.

Fig. 4.

Cytokines expression in response to Salmonella infection.ELISA for (A) interleukin (IL)-12p40, (B) IL-12p35, and (C) IFN-α produced after infection by different Salmonella serotypes. THP-1 macrophage-derived foam cells were infected by Salmonella with or without virulence plasmids for 0.5 and 2 h, and the supernatants were harvested and used for experiments. All values are presented as means ± standard error (n = 3). a-e indicate significant differences between strains 0.5 h after infection (p < 0.05); w-z indicate significant differences between strains 2 h after infection (p < 0.05). nc: uninfected cells.
Cytokines expression in response to Salmonella infection.ELISA for (A) interleukin (IL)-12p40, (B) IL-12p35, and (C) IFN-α produced after infection by different Salmonella serotypes. THP-1 macrophage-derived foam cells were infected by Salmonella with or without virulence plasmids for 0.5 and 2 h, and the supernatants were harvested and used for experiments. All values are presented as means ± standard error (n = 3). a-e indicate significant differences between strains 0.5 h after infection (p < 0.05); w-z indicate significant differences between strains 2 h after infection (p < 0.05). nc: uninfected cells.

CD36 expression based on fluorescence density and gate (%) on foam cell interaction among different Salmonella serotypes_

Sample%ParentMean
Foam NC4.52 594
Foam NC-CD36 FITC5.48 796
Foam 5045-CD36 FITC5.27 204
Foam 5046-CD36 FITC7.711 194
Foam 7130-CD36 FITC1.84 807
Foam 7067-CD36 FITC6.33 204
Foam 7085-CD36 FITC7.25 341
Foam 7266-CD36 FITC9.611 067

Characteristics of S_ Typhimurium, S_ Enteritidis, and S_ Choleraesuis strains_

SerovarsStrainsCharacteristics of virulence plasmid
S. TyphimuriumOU5045With a 90-kb pSTV as a wild type
OU5046Without pSTV from wild type
S. EnteritidisOU7130With a 60-kb pSEV as a wild type
OU7067Without pSEV from wild type
S. CholeraesuisOU7085With a 50-kb pSCV as a wild type
OU7266Without pSCV from wild type
DOI: https://doi.org/10.33073/pjm-2019-043 | Journal eISSN: 2544-4646 | Journal ISSN: 1733-1331
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Language: English
Page range: 439 - 447
Submitted on: Apr 17, 2019
Accepted on: Aug 19, 2019
Published on: Oct 22, 2019
Published by: Polish Society of Microbiologists
In partnership with: Paradigm Publishing Services
Publication frequency: 4 times per year
Keywords:
serotype,
virulence plasmid,
foam cell,
autophagy,
inflammasome
Related subjects:
Life sciences,
Microbiology and virology

© 2019 CHISHIH CHU, MIN YI WONG, CHENG-HSUN CHIU, YUAN-HSI TSENG, CHYI-LIANG CHEN, YAO-KUANG HUANG, published by Polish Society of Microbiologists
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 License.

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