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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

Effect of GM-CSF on P. aeruginosa PAO1 persister cells was abolished by heat inactivation and anti-GM-CSF.(a) The persister cells isolated from stationary phase cultures underwent treatment with active 0.17 pM GM-CSF or heat-inactivated 0.17 pM GM-CSF in the presence of 5 μg/mL ciprofloxacin for 3.5 h. (b) The persister cells were isolated from exponential phase cultures. All samples underwent the same incubation time. The figure shows the viability of persister cells treated with GM-CSF alone, anti-GM-CSF alone, or GM-CSF neutralized by anti-GM-CSF (2 h incubation) followed by addition of 5 μg/mL ciprofloxacin and incubation for 3.5 h. The amount of BSA (0.1%) was adjusted to be the same for all samples. Following the treatment, the viability of persister cells was determined by counting CFU. The samples were tested in triplicate (n = 3). Error bars represent SD; * p < 0.05, ** p < 0.01,**** p < 0.0001, one-way ANOVA followed by Tukey test.
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f2: Effect of GM-CSF on P. aeruginosa PAO1 persister cells was abolished by heat inactivation and anti-GM-CSF.(a) The persister cells isolated from stationary phase cultures underwent treatment with active 0.17 pM GM-CSF or heat-inactivated 0.17 pM GM-CSF in the presence of 5 μg/mL ciprofloxacin for 3.5 h. (b) The persister cells were isolated from exponential phase cultures. All samples underwent the same incubation time. The figure shows the viability of persister cells treated with GM-CSF alone, anti-GM-CSF alone, or GM-CSF neutralized by anti-GM-CSF (2 h incubation) followed by addition of 5 μg/mL ciprofloxacin and incubation for 3.5 h. The amount of BSA (0.1%) was adjusted to be the same for all samples. Following the treatment, the viability of persister cells was determined by counting CFU. The samples were tested in triplicate (n = 3). Error bars represent SD; * p < 0.05, ** p < 0.01,**** p < 0.0001, one-way ANOVA followed by Tukey test.

Mentions: To confirm if the observed effects were caused by GM-CSF rather than any contaminant, we inactivated the GM-CSF stock by heating at 75 °C overnight. This treatment abolished the effects of GM-CSF on P. aeruginosa PAO1 persister cells during treatment with ciprofloxacin (Fig. 2a). This finding is consistent with the heat sensitive nature of proteins. To further confirm that the observed effects were caused by GM-CSF specifically, we also tested the effects in the presence of anti-GM-CSF antibody. As shown in Fig. 2b, pretreatment of GM-CSF with anti-GM-CSF (2 h incubation) abolished the effects of GM-CSF. Thus, the observed effects on persister cells were indeed caused by GM-CSF.


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)

Effect of GM-CSF on P. aeruginosa PAO1 persister cells was abolished by heat inactivation and anti-GM-CSF.(a) The persister cells isolated from stationary phase cultures underwent treatment with active 0.17 pM GM-CSF or heat-inactivated 0.17 pM GM-CSF in the presence of 5 μg/mL ciprofloxacin for 3.5 h. (b) The persister cells were isolated from exponential phase cultures. All samples underwent the same incubation time. The figure shows the viability of persister cells treated with GM-CSF alone, anti-GM-CSF alone, or GM-CSF neutralized by anti-GM-CSF (2 h incubation) followed by addition of 5 μg/mL ciprofloxacin and incubation for 3.5 h. The amount of BSA (0.1%) was adjusted to be the same for all samples. Following the treatment, the viability of persister cells was determined by counting CFU. The samples were tested in triplicate (n = 3). Error bars represent SD; * p < 0.05, ** p < 0.01,**** p < 0.0001, 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

f2: Effect of GM-CSF on P. aeruginosa PAO1 persister cells was abolished by heat inactivation and anti-GM-CSF.(a) The persister cells isolated from stationary phase cultures underwent treatment with active 0.17 pM GM-CSF or heat-inactivated 0.17 pM GM-CSF in the presence of 5 μg/mL ciprofloxacin for 3.5 h. (b) The persister cells were isolated from exponential phase cultures. All samples underwent the same incubation time. The figure shows the viability of persister cells treated with GM-CSF alone, anti-GM-CSF alone, or GM-CSF neutralized by anti-GM-CSF (2 h incubation) followed by addition of 5 μg/mL ciprofloxacin and incubation for 3.5 h. The amount of BSA (0.1%) was adjusted to be the same for all samples. Following the treatment, the viability of persister cells was determined by counting CFU. The samples were tested in triplicate (n = 3). Error bars represent SD; * p < 0.05, ** p < 0.01,**** p < 0.0001, one-way ANOVA followed by Tukey test.
Mentions: To confirm if the observed effects were caused by GM-CSF rather than any contaminant, we inactivated the GM-CSF stock by heating at 75 °C overnight. This treatment abolished the effects of GM-CSF on P. aeruginosa PAO1 persister cells during treatment with ciprofloxacin (Fig. 2a). This finding is consistent with the heat sensitive nature of proteins. To further confirm that the observed effects were caused by GM-CSF specifically, we also tested the effects in the presence of anti-GM-CSF antibody. As shown in Fig. 2b, pretreatment of GM-CSF with anti-GM-CSF (2 h incubation) abolished the effects of GM-CSF. Thus, the observed effects on persister cells were indeed caused by GM-CSF.

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