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Human Granulocyte Macrophage Colony-Stimulating Factor Enhances Antibiotic Susceptibility of Pseudomonas aeruginosa Persister Cells.

Choudhary GS, Yao X, Wang J, Peng B, Bader RA, Ren D - Sci Rep (2015)

Bottom Line: The DNA microarray and qPCR results indicated that GM-CSF induced the genes for flagellar motility and pyocin production in the persister cells, but not the normal cells of P. aeruginosa PAO1.Consistently, the supernatants from GM-CSF treated P. aeruginosa PAO1 persister cell suspensions were found cidal to the pyocin sensitive strain P. aeruginosa PAK.Collectively, these findings suggest that host immune factors and bacterial persisters may directly interact, leading to enhanced susceptibility of persister cells to antibiotics.

View Article: PubMed Central - PubMed

Affiliation: Department of Biomedical and Chemical Engineering, Syracuse University, Syracuse, NY 13244, USA.

ABSTRACT
Bacterial persister cells are highly tolerant to antibiotics and cause chronic infections. However, little is known about the interaction between host immune systems with this subpopulation of metabolically inactive cells, and direct effects of host immune factors (in the absence of immune cells) on persister cells have not been studied. Here we report that human granulocyte macrophage-colony stimulating factor (GM-CSF) can sensitize the persister cells of Pseudomonas aeruginosa PAO1 and PDO300 to multiple antibiotics including ciprofloxacin, tobramycin, tetracycline, and gentamicin. GM-CSF also sensitized the biofilm cells of P. aeruginosa PAO1 and PDO300 to tobramycin in the presence of biofilm matrix degrading enzymes. The DNA microarray and qPCR results indicated that GM-CSF induced the genes for flagellar motility and pyocin production in the persister cells, but not the normal cells of P. aeruginosa PAO1. Consistently, the supernatants from GM-CSF treated P. aeruginosa PAO1 persister cell suspensions were found cidal to the pyocin sensitive strain P. aeruginosa PAK. Collectively, these findings suggest that host immune factors and bacterial persisters may directly interact, leading to enhanced susceptibility of persister cells to antibiotics.

No MeSH data available.


Related in: MedlinePlus

Effects of supernatants of GM-CSF treated P. aeruginosa PAO1 persister cell suspensions.The normal cells harvested from stationary phase cultures of P. aeruginosa PAO1 and PAK were treated with the supernatants collected from the persister cell suspensions of P. aeruginosa PAO1 (a), and PA0620::phoA (b) after treatment with 0.17 pM or 0.17 nM GM-CSF for 2 h. The amount of BSA (0.1%) was adjusted to be the same for all samples during GM-CSF treatment. Following the treatment, the viability of PAO1 and PAK cells was determined by counting CFU. The samples were tested in triplicate (n = 3). Error bars represent SD; *p < 0.05, one-way ANOVA followed by Tukey test.
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f7: Effects of supernatants of GM-CSF treated P. aeruginosa PAO1 persister cell suspensions.The normal cells harvested from stationary phase cultures of P. aeruginosa PAO1 and PAK were treated with the supernatants collected from the persister cell suspensions of P. aeruginosa PAO1 (a), and PA0620::phoA (b) after treatment with 0.17 pM or 0.17 nM GM-CSF for 2 h. The amount of BSA (0.1%) was adjusted to be the same for all samples during GM-CSF treatment. Following the treatment, the viability of PAO1 and PAK cells was determined by counting CFU. The samples were tested in triplicate (n = 3). Error bars represent SD; *p < 0.05, one-way ANOVA followed by Tukey test.

Mentions: R-type pyocins produced by P. aeruginosa strains can be categorized into five types termed R1 to R542. Kohler et al.39 showed that the R1-pyocin producing P. aeruginosa PAK strain is susceptible to the R2-pyocins produced by P. aeruginosa PAO1. We found that treatment with GM-CSF induced R-pyocin related genes in P. aeruginosa PAO1 persister cells including PA0617, PA0619–22, and PA0625–30. Further test showed that, after P. aeruginosa PAO1 persister cells were treated with GM-CSF at 0.17 pM or 0.17 nM, the supernatants from the treated cell samples killed 65.3 ± 14.5% (p = 0.0201) and 67.8 ± 9.7% (p = 0.0132) of normal cells of P. aeruginosa PAK, respectively, compared to the GM-CSF free control (Fig. 7a). In contrast, no significant difference (p > 0.1) was observed for the same treatment of the normal cells of R2-pyocin resistant P. aeruginosa PAO1 (Fig. 7a). Moreover, when the persister cells of PA0620::phoA, an isogenic deletion mutant of PA0620 encoding the R2-pyocin tail fiber protein, were treated with 0.17 pM or 0.17 nM GM-CSF, the supernatants did not change the viability of P. aeruginosa PAO1 and PAK (Fig. 7b). Collectively, these results indicate that pyocin production in PAO1 might have been induced by GM-CSF. To our best knowledge, the interaction between P. aeruginosa and GM-CSF in pyocin production has not been explored to date.


Human Granulocyte Macrophage Colony-Stimulating Factor Enhances Antibiotic Susceptibility of Pseudomonas aeruginosa Persister Cells.

Choudhary GS, Yao X, Wang J, Peng B, Bader RA, Ren D - Sci Rep (2015)

Effects of supernatants of GM-CSF treated P. aeruginosa PAO1 persister cell suspensions.The normal cells harvested from stationary phase cultures of P. aeruginosa PAO1 and PAK were treated with the supernatants collected from the persister cell suspensions of P. aeruginosa PAO1 (a), and PA0620::phoA (b) after treatment with 0.17 pM or 0.17 nM GM-CSF for 2 h. The amount of BSA (0.1%) was adjusted to be the same for all samples during GM-CSF treatment. Following the treatment, the viability of PAO1 and PAK cells was determined by counting CFU. The samples were tested in triplicate (n = 3). Error bars represent SD; *p < 0.05, one-way ANOVA followed by Tukey test.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4663479&req=5

f7: Effects of supernatants of GM-CSF treated P. aeruginosa PAO1 persister cell suspensions.The normal cells harvested from stationary phase cultures of P. aeruginosa PAO1 and PAK were treated with the supernatants collected from the persister cell suspensions of P. aeruginosa PAO1 (a), and PA0620::phoA (b) after treatment with 0.17 pM or 0.17 nM GM-CSF for 2 h. The amount of BSA (0.1%) was adjusted to be the same for all samples during GM-CSF treatment. Following the treatment, the viability of PAO1 and PAK cells was determined by counting CFU. The samples were tested in triplicate (n = 3). Error bars represent SD; *p < 0.05, one-way ANOVA followed by Tukey test.
Mentions: R-type pyocins produced by P. aeruginosa strains can be categorized into five types termed R1 to R542. Kohler et al.39 showed that the R1-pyocin producing P. aeruginosa PAK strain is susceptible to the R2-pyocins produced by P. aeruginosa PAO1. We found that treatment with GM-CSF induced R-pyocin related genes in P. aeruginosa PAO1 persister cells including PA0617, PA0619–22, and PA0625–30. Further test showed that, after P. aeruginosa PAO1 persister cells were treated with GM-CSF at 0.17 pM or 0.17 nM, the supernatants from the treated cell samples killed 65.3 ± 14.5% (p = 0.0201) and 67.8 ± 9.7% (p = 0.0132) of normal cells of P. aeruginosa PAK, respectively, compared to the GM-CSF free control (Fig. 7a). In contrast, no significant difference (p > 0.1) was observed for the same treatment of the normal cells of R2-pyocin resistant P. aeruginosa PAO1 (Fig. 7a). Moreover, when the persister cells of PA0620::phoA, an isogenic deletion mutant of PA0620 encoding the R2-pyocin tail fiber protein, were treated with 0.17 pM or 0.17 nM GM-CSF, the supernatants did not change the viability of P. aeruginosa PAO1 and PAK (Fig. 7b). Collectively, these results indicate that pyocin production in PAO1 might have been induced by GM-CSF. To our best knowledge, the interaction between P. aeruginosa and GM-CSF in pyocin production has not been explored to date.

Bottom Line: The DNA microarray and qPCR results indicated that GM-CSF induced the genes for flagellar motility and pyocin production in the persister cells, but not the normal cells of P. aeruginosa PAO1.Consistently, the supernatants from GM-CSF treated P. aeruginosa PAO1 persister cell suspensions were found cidal to the pyocin sensitive strain P. aeruginosa PAK.Collectively, these findings suggest that host immune factors and bacterial persisters may directly interact, leading to enhanced susceptibility of persister cells to antibiotics.

View Article: PubMed Central - PubMed

Affiliation: Department of Biomedical and Chemical Engineering, Syracuse University, Syracuse, NY 13244, USA.

ABSTRACT
Bacterial persister cells are highly tolerant to antibiotics and cause chronic infections. However, little is known about the interaction between host immune systems with this subpopulation of metabolically inactive cells, and direct effects of host immune factors (in the absence of immune cells) on persister cells have not been studied. Here we report that human granulocyte macrophage-colony stimulating factor (GM-CSF) can sensitize the persister cells of Pseudomonas aeruginosa PAO1 and PDO300 to multiple antibiotics including ciprofloxacin, tobramycin, tetracycline, and gentamicin. GM-CSF also sensitized the biofilm cells of P. aeruginosa PAO1 and PDO300 to tobramycin in the presence of biofilm matrix degrading enzymes. The DNA microarray and qPCR results indicated that GM-CSF induced the genes for flagellar motility and pyocin production in the persister cells, but not the normal cells of P. aeruginosa PAO1. Consistently, the supernatants from GM-CSF treated P. aeruginosa PAO1 persister cell suspensions were found cidal to the pyocin sensitive strain P. aeruginosa PAK. Collectively, these findings suggest that host immune factors and bacterial persisters may directly interact, leading to enhanced susceptibility of persister cells to antibiotics.

No MeSH data available.


Related in: MedlinePlus