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Salmonella-Infected Aortic Aneurysm: Investigating Pathogenesis Using Salmonella Serotypes

CHISHIH CHU
CHISHIH CHU
, 
MIN YI WONG
MIN YI WONG
, 
CHENG-HSUN CHIU
CHENG-HSUN CHIU
, 
YUAN-HSI TSENG
YUAN-HSI TSENG
, 
CHYI-LIANG CHEN
CHYI-LIANG CHEN
 et 
YAO-KUANG HUANG
YAO-KUANG HUANG
   | 22 oct. 2019
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Article Category: original-paper
Publié en ligne: 22 oct. 2019
Pages: 439 - 447
Reçu: 17 avr. 2019
Accepté: 19 août 2019
DOI: https://doi.org/10.33073/pjm-2019-043
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© 2019 Chishih Chu et al., published by Sciendo
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.

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
eISSN:
2544-4646
Langue:
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Life Sciences, Microbiology and Virology
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