Limits...
Killing of Staphylococcus aureus in murine macrophages by chloroquine used alone and in combination with ciprofloxacin or azithromycin.

Dey S, Bishayi B - J Inflamm Res (2015)

Bottom Line: We present here data obtained with a model of S. aureus-infected mouse peritoneal macrophages in which the intracellular growth of the bacteria and the influence of antibiotics was monitored for 30, 60, and 90 minutes in the presence or absence of CQ along with the production of ROS and alteration in levels of antioxidant enzymes and cytokines.It can be suggested that action of AZM in mediating bacterial killing is enhanced by the presence of CQ, indicating enhanced uptake of AZM during early infection that may be essential for bacteria killing by AZM.Based on these observations, one may speculate that in an inflammatory milieu, CQ loaded with AZM elicits a stronger proinflammatory response by increasing the intracellular uptake of AZM or CIP, thus enabling the immune system to mount a more robust and prolonged response against intracellular pathogens.

View Article: PubMed Central - PubMed

Affiliation: Department of Physiology, Immunology laboratory, University of Calcutta, University Colleges of Science and Technology, Calcutta, India.

ABSTRACT
This study aimed to determine any alteration in the killing of Staphylococcus aureus in murine peritoneal macrophages when chloroquine (CQ) is used alone compared with when it is used in combination with ciprofloxacin (CIP) or azithromycin (AZM). The study also aimed to find out the implication of reactive oxygen species (ROS) production and cytokine release in the intracellular killing of S. aureus in macrophages. We present here data obtained with a model of S. aureus-infected mouse peritoneal macrophages in which the intracellular growth of the bacteria and the influence of antibiotics was monitored for 30, 60, and 90 minutes in the presence or absence of CQ along with the production of ROS and alteration in levels of antioxidant enzymes and cytokines. It was observed that S. aureus-triggered cytokine response was regulated when macrophages were co-cultured with CQ and AZM as compared with CQ stimulation only. It can be suggested that action of AZM in mediating bacterial killing is enhanced by the presence of CQ, indicating enhanced uptake of AZM during early infection that may be essential for bacteria killing by AZM. Reduction of oxidative stress burden on the S. aureus-infected macrophages may pave the way for better killing of internalized S. aureus by CQ plus ciprofloxacin (CIP) or CQ plus AZM. Based on these observations, one may speculate that in an inflammatory milieu, CQ loaded with AZM elicits a stronger proinflammatory response by increasing the intracellular uptake of AZM or CIP, thus enabling the immune system to mount a more robust and prolonged response against intracellular pathogens.

No MeSH data available.


Related in: MedlinePlus

Effect of chloroquine (CQ) and antibiotic treatment on Staphylococcus aureus infection induced NO release in the lysate and supernatant in murine peritoneal macrophages. The lysate (A) and supernatant (B) recovered after time-dependent phagocytosis in the presence or absence of CQ or antibiotics were used to determine the nitric-oxide release by Griess assay.Notes: nitric-oxide release is expressed in terms of mM/106 cells. Results are presented as mean ± standard deviation of three independent experiments. *Significant difference with respect to CM; #significant difference with respect to SAM; ^significant difference with respect to CQ plus S. aureus-infected macrophages at the P<0.05 level of significance; $significant difference with respect to either ciprofloxacin (CIP) or azithromycin (AZM) plus S. aureus-infected macrophages.Abbreviations: CM, control macrophages; CQ + SAM, chloroquine-pretreated plus S. aureus-infected macrophages; CQ + SAM + AZM, chloroquine-pretreated macrophages infected with S. aureus and then treated with azithromycin; CQ + SAM + CIP, chloroquine-pretreated macrophages infected with S. aureus and then treated with ciprofloxacin; SAM, S. aureus-infected macrophages; SAM + AZM, S. aureus-infected macrophages treated with azithromycin; SAM + CIP, S. aureus-infected macrophages treated with ciprofloxacin.
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4309780&req=5

f4-jir-8-029: Effect of chloroquine (CQ) and antibiotic treatment on Staphylococcus aureus infection induced NO release in the lysate and supernatant in murine peritoneal macrophages. The lysate (A) and supernatant (B) recovered after time-dependent phagocytosis in the presence or absence of CQ or antibiotics were used to determine the nitric-oxide release by Griess assay.Notes: nitric-oxide release is expressed in terms of mM/106 cells. Results are presented as mean ± standard deviation of three independent experiments. *Significant difference with respect to CM; #significant difference with respect to SAM; ^significant difference with respect to CQ plus S. aureus-infected macrophages at the P<0.05 level of significance; $significant difference with respect to either ciprofloxacin (CIP) or azithromycin (AZM) plus S. aureus-infected macrophages.Abbreviations: CM, control macrophages; CQ + SAM, chloroquine-pretreated plus S. aureus-infected macrophages; CQ + SAM + AZM, chloroquine-pretreated macrophages infected with S. aureus and then treated with azithromycin; CQ + SAM + CIP, chloroquine-pretreated macrophages infected with S. aureus and then treated with ciprofloxacin; SAM, S. aureus-infected macrophages; SAM + AZM, S. aureus-infected macrophages treated with azithromycin; SAM + CIP, S. aureus-infected macrophages treated with ciprofloxacin.

Mentions: CQ inhibits NO production when added during the stimulation of peritoneal macrophages with S. aureus. A statistically significant decrease in the amount of NO released both in the lysate (Figure 4A) and supernatant (Figure 4B) was observed (P<0.05) in CQ-pretreated macrophages before infection, compared with untreated S. aureus-infected macrophages at 30 and 60 minutes post-incubation. The amount of NO release decreased in the case of macrophages treated with AZM or CIP alone before S. aureus infection compared with untreated S. aureus-infected macrophages (Figure 4A and B). The NO release was significantly lower at 30 and 60 minutes in the supernatant and at 60 minutes in the cell lysate when CQ-pretreated macrophages were incubated with CIP or AZM before S. aureus infection, compared with CIP- or AZM-treated S. aureus-infected macrophages.


Killing of Staphylococcus aureus in murine macrophages by chloroquine used alone and in combination with ciprofloxacin or azithromycin.

Dey S, Bishayi B - J Inflamm Res (2015)

Effect of chloroquine (CQ) and antibiotic treatment on Staphylococcus aureus infection induced NO release in the lysate and supernatant in murine peritoneal macrophages. The lysate (A) and supernatant (B) recovered after time-dependent phagocytosis in the presence or absence of CQ or antibiotics were used to determine the nitric-oxide release by Griess assay.Notes: nitric-oxide release is expressed in terms of mM/106 cells. Results are presented as mean ± standard deviation of three independent experiments. *Significant difference with respect to CM; #significant difference with respect to SAM; ^significant difference with respect to CQ plus S. aureus-infected macrophages at the P<0.05 level of significance; $significant difference with respect to either ciprofloxacin (CIP) or azithromycin (AZM) plus S. aureus-infected macrophages.Abbreviations: CM, control macrophages; CQ + SAM, chloroquine-pretreated plus S. aureus-infected macrophages; CQ + SAM + AZM, chloroquine-pretreated macrophages infected with S. aureus and then treated with azithromycin; CQ + SAM + CIP, chloroquine-pretreated macrophages infected with S. aureus and then treated with ciprofloxacin; SAM, S. aureus-infected macrophages; SAM + AZM, S. aureus-infected macrophages treated with azithromycin; SAM + CIP, S. aureus-infected macrophages treated with ciprofloxacin.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4309780&req=5

f4-jir-8-029: Effect of chloroquine (CQ) and antibiotic treatment on Staphylococcus aureus infection induced NO release in the lysate and supernatant in murine peritoneal macrophages. The lysate (A) and supernatant (B) recovered after time-dependent phagocytosis in the presence or absence of CQ or antibiotics were used to determine the nitric-oxide release by Griess assay.Notes: nitric-oxide release is expressed in terms of mM/106 cells. Results are presented as mean ± standard deviation of three independent experiments. *Significant difference with respect to CM; #significant difference with respect to SAM; ^significant difference with respect to CQ plus S. aureus-infected macrophages at the P<0.05 level of significance; $significant difference with respect to either ciprofloxacin (CIP) or azithromycin (AZM) plus S. aureus-infected macrophages.Abbreviations: CM, control macrophages; CQ + SAM, chloroquine-pretreated plus S. aureus-infected macrophages; CQ + SAM + AZM, chloroquine-pretreated macrophages infected with S. aureus and then treated with azithromycin; CQ + SAM + CIP, chloroquine-pretreated macrophages infected with S. aureus and then treated with ciprofloxacin; SAM, S. aureus-infected macrophages; SAM + AZM, S. aureus-infected macrophages treated with azithromycin; SAM + CIP, S. aureus-infected macrophages treated with ciprofloxacin.
Mentions: CQ inhibits NO production when added during the stimulation of peritoneal macrophages with S. aureus. A statistically significant decrease in the amount of NO released both in the lysate (Figure 4A) and supernatant (Figure 4B) was observed (P<0.05) in CQ-pretreated macrophages before infection, compared with untreated S. aureus-infected macrophages at 30 and 60 minutes post-incubation. The amount of NO release decreased in the case of macrophages treated with AZM or CIP alone before S. aureus infection compared with untreated S. aureus-infected macrophages (Figure 4A and B). The NO release was significantly lower at 30 and 60 minutes in the supernatant and at 60 minutes in the cell lysate when CQ-pretreated macrophages were incubated with CIP or AZM before S. aureus infection, compared with CIP- or AZM-treated S. aureus-infected macrophages.

Bottom Line: We present here data obtained with a model of S. aureus-infected mouse peritoneal macrophages in which the intracellular growth of the bacteria and the influence of antibiotics was monitored for 30, 60, and 90 minutes in the presence or absence of CQ along with the production of ROS and alteration in levels of antioxidant enzymes and cytokines.It can be suggested that action of AZM in mediating bacterial killing is enhanced by the presence of CQ, indicating enhanced uptake of AZM during early infection that may be essential for bacteria killing by AZM.Based on these observations, one may speculate that in an inflammatory milieu, CQ loaded with AZM elicits a stronger proinflammatory response by increasing the intracellular uptake of AZM or CIP, thus enabling the immune system to mount a more robust and prolonged response against intracellular pathogens.

View Article: PubMed Central - PubMed

Affiliation: Department of Physiology, Immunology laboratory, University of Calcutta, University Colleges of Science and Technology, Calcutta, India.

ABSTRACT
This study aimed to determine any alteration in the killing of Staphylococcus aureus in murine peritoneal macrophages when chloroquine (CQ) is used alone compared with when it is used in combination with ciprofloxacin (CIP) or azithromycin (AZM). The study also aimed to find out the implication of reactive oxygen species (ROS) production and cytokine release in the intracellular killing of S. aureus in macrophages. We present here data obtained with a model of S. aureus-infected mouse peritoneal macrophages in which the intracellular growth of the bacteria and the influence of antibiotics was monitored for 30, 60, and 90 minutes in the presence or absence of CQ along with the production of ROS and alteration in levels of antioxidant enzymes and cytokines. It was observed that S. aureus-triggered cytokine response was regulated when macrophages were co-cultured with CQ and AZM as compared with CQ stimulation only. It can be suggested that action of AZM in mediating bacterial killing is enhanced by the presence of CQ, indicating enhanced uptake of AZM during early infection that may be essential for bacteria killing by AZM. Reduction of oxidative stress burden on the S. aureus-infected macrophages may pave the way for better killing of internalized S. aureus by CQ plus ciprofloxacin (CIP) or CQ plus AZM. Based on these observations, one may speculate that in an inflammatory milieu, CQ loaded with AZM elicits a stronger proinflammatory response by increasing the intracellular uptake of AZM or CIP, thus enabling the immune system to mount a more robust and prolonged response against intracellular pathogens.

No MeSH data available.


Related in: MedlinePlus