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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

GM-CSF did not sensitize the persister cells of E. coli K12 and E. coli ATCC 53505 to antibiotics.The persister cells were isolated from exponential phase cultures of E. coli K12 and E. coli ATCC 53505 by killing the normal cells with 100 μg/mL ampicillin and 20 μg/mL ofloxacin (ATTCC 53505 is resistant to ampicillin, data not shown) respectively for 3.5 h, and then treated with or without 0.17 pM GM-CSF for 1 h, followed by adding an antibiotic as indicated and incubating for 3.5 h. The amount of BSA (contained in GM-CSF stocks) was adjusted to be the same in all samples. Following the treatment, the cell viability was determined by counting CFU. Cip: 2 μg/mL ciprofloxacin. Tob: 70 μg/mL tobramycin. Tet: 20 μg/mL tetracycline. Gen: 200 μg/mL gentamicin. The samples were tested in triplicate (n = 3). The number of CFU in the GM-CSF free sample (antibiotic only) varied depending on the antibiotic used, so the CFU of corresponding GM-CSF free control for each antibiotic was normalized as 100% for the convenience of comparison.
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f5: GM-CSF did not sensitize the persister cells of E. coli K12 and E. coli ATCC 53505 to antibiotics.The persister cells were isolated from exponential phase cultures of E. coli K12 and E. coli ATCC 53505 by killing the normal cells with 100 μg/mL ampicillin and 20 μg/mL ofloxacin (ATTCC 53505 is resistant to ampicillin, data not shown) respectively for 3.5 h, and then treated with or without 0.17 pM GM-CSF for 1 h, followed by adding an antibiotic as indicated and incubating for 3.5 h. The amount of BSA (contained in GM-CSF stocks) was adjusted to be the same in all samples. Following the treatment, the cell viability was determined by counting CFU. Cip: 2 μg/mL ciprofloxacin. Tob: 70 μg/mL tobramycin. Tet: 20 μg/mL tetracycline. Gen: 200 μg/mL gentamicin. The samples were tested in triplicate (n = 3). The number of CFU in the GM-CSF free sample (antibiotic only) varied depending on the antibiotic used, so the CFU of corresponding GM-CSF free control for each antibiotic was normalized as 100% for the convenience of comparison.

Mentions: To understand if the observed activities of GM-CSF are species specific, we also tested it on E. coli strains including the lab strain E. coli K1236 and a pathogenic strain ATCC 5350537 associated with human urinary tract infection. The concentration of antibiotics which showed 2 log reduction in CFU of normal cells was selected for the experiments. Unlike P. aeruginosa PAO1 and PDO300, GM-CSF did not exhibit significant effect on the planktonic persister cells of E. coli. For example, treatment with 0.17 pM GM-CSF did not change the susceptibility of E. coli K12 persister cells to 2 μg/mL ciprofloxacin (p = 0.93) or 70 μg/mL tobramycin (p = 0.95) (Fig. 5). Similar results were found for the pathogenic E. coli ATCC 53505. As shown in Fig. 5, GM-CSF did not sensitize the persister cells isolated from exponential phase cultures to 2 μg/mL ciprofloxacin (p = 0.55) or 70 μg/mL tobramycin (p = 0.31). These results indicate that GM-CSF is not effective on the persister cells of E. coli K12 and ATCC 53505.


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)

GM-CSF did not sensitize the persister cells of E. coli K12 and E. coli ATCC 53505 to antibiotics.The persister cells were isolated from exponential phase cultures of E. coli K12 and E. coli ATCC 53505 by killing the normal cells with 100 μg/mL ampicillin and 20 μg/mL ofloxacin (ATTCC 53505 is resistant to ampicillin, data not shown) respectively for 3.5 h, and then treated with or without 0.17 pM GM-CSF for 1 h, followed by adding an antibiotic as indicated and incubating for 3.5 h. The amount of BSA (contained in GM-CSF stocks) was adjusted to be the same in all samples. Following the treatment, the cell viability was determined by counting CFU. Cip: 2 μg/mL ciprofloxacin. Tob: 70 μg/mL tobramycin. Tet: 20 μg/mL tetracycline. Gen: 200 μg/mL gentamicin. The samples were tested in triplicate (n = 3). The number of CFU in the GM-CSF free sample (antibiotic only) varied depending on the antibiotic used, so the CFU of corresponding GM-CSF free control for each antibiotic was normalized as 100% for the convenience of comparison.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

f5: GM-CSF did not sensitize the persister cells of E. coli K12 and E. coli ATCC 53505 to antibiotics.The persister cells were isolated from exponential phase cultures of E. coli K12 and E. coli ATCC 53505 by killing the normal cells with 100 μg/mL ampicillin and 20 μg/mL ofloxacin (ATTCC 53505 is resistant to ampicillin, data not shown) respectively for 3.5 h, and then treated with or without 0.17 pM GM-CSF for 1 h, followed by adding an antibiotic as indicated and incubating for 3.5 h. The amount of BSA (contained in GM-CSF stocks) was adjusted to be the same in all samples. Following the treatment, the cell viability was determined by counting CFU. Cip: 2 μg/mL ciprofloxacin. Tob: 70 μg/mL tobramycin. Tet: 20 μg/mL tetracycline. Gen: 200 μg/mL gentamicin. The samples were tested in triplicate (n = 3). The number of CFU in the GM-CSF free sample (antibiotic only) varied depending on the antibiotic used, so the CFU of corresponding GM-CSF free control for each antibiotic was normalized as 100% for the convenience of comparison.
Mentions: To understand if the observed activities of GM-CSF are species specific, we also tested it on E. coli strains including the lab strain E. coli K1236 and a pathogenic strain ATCC 5350537 associated with human urinary tract infection. The concentration of antibiotics which showed 2 log reduction in CFU of normal cells was selected for the experiments. Unlike P. aeruginosa PAO1 and PDO300, GM-CSF did not exhibit significant effect on the planktonic persister cells of E. coli. For example, treatment with 0.17 pM GM-CSF did not change the susceptibility of E. coli K12 persister cells to 2 μg/mL ciprofloxacin (p = 0.93) or 70 μg/mL tobramycin (p = 0.95) (Fig. 5). Similar results were found for the pathogenic E. coli ATCC 53505. As shown in Fig. 5, GM-CSF did not sensitize the persister cells isolated from exponential phase cultures to 2 μg/mL ciprofloxacin (p = 0.55) or 70 μg/mL tobramycin (p = 0.31). These results indicate that GM-CSF is not effective on the persister cells of E. coli K12 and ATCC 53505.

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