Limits...
Depolymerase improves gentamicin efficacy during Klebsiella pneumoniae induced murine infection.

Bansal S, Harjai K, Chhibber S - BMC Infect. Dis. (2014)

Bottom Line: Highly significant reduction in neutrophil infiltration and levels of pro-inflammatory and anti-inflammatory cytokines was also observed.In addition, as confirmed through flow cytometry, the hyperimmune sera raised against the enzyme did not neutralize its activity.Moreover, since this decapsulating protein does not possess a lytic activity therefore there would be no chances of development of bacterial resistance against it.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology, Panjab University, Sector-14, Chandigarh 160014, India. sanjaychhibber8@gmail.com.

ABSTRACT

Background: Presence of capsule enhances the virulence of bacteria that cause pneumonia, meningitis, cystic fibrosis, dental caries, periodontitis. Capsule is an important virulence factor for Klebsiella pneumoniae and infections due to this pathogen have been associated with high mortality rates. In the present study, use of an Aeromonas punctata derived capsule depolymerase against K. pneumoniae, to reinstate the efficacy of gentamicin during pneumonia and septicemia was investigated.

Methods: Depolymerase was administered in mice intraperitoneally (50 μg) alone as well in combination with gentamicin (1.5 mg/kg), 24 h post infection during acute lung infection and 6 h later during septicemia. Bacterial load, neutrophil infiltration and cytokine levels were estimated. The immunogenicity of protein was also studied.

Results: In comparison to groups treated with gentamicin alone, combination treatment with depolymerase and gentamicin significantly reduced (P < 0.01) bacterial titer in the lungs, liver, kidney, spleen and blood of experimental animals. Highly significant reduction in neutrophil infiltration and levels of pro-inflammatory and anti-inflammatory cytokines was also observed. This indicated an efficient capsule removal by the enzyme, that improved gentamicin efficacy in vivo. Although the enzyme was found to be immunogenic, but no significant reduction in treatment efficacy was observed in the preimmunized as well as naïve mice. In addition, as confirmed through flow cytometry, the hyperimmune sera raised against the enzyme did not neutralize its activity.

Conclusion: The results confirm that administration of enzyme 'depolymerase' along with gentamicin not only checked the virulence of K. pneumoniae in vivo but it also increased its susceptibility to gentamicin at a lower concentration. Such a strategy would help to avoid exposure to higher concentration of gentamicin. Moreover, since this decapsulating protein does not possess a lytic activity therefore there would be no chances of development of bacterial resistance against it. Therefore, it should be studied further for its successful inclusion in our prophylactic/therapeutic regimes.

Show MeSH

Related in: MedlinePlus

Cytokine levels during septicemia: a) IL 1ß b) IL 10 c) TNFalevels (pg/ml) 24 h post infection in various organs and blood of mice infected via intra peritoneal route withK. pneumoniaeB5055 and treated with gentamicin (1.5 mg/kg) and/or depolymerase (50 μg). G1, G2, G3, G4 represent Groups 1, 2, 3, 4. Arrows indicate significant reduction in cytokine levels in all organs and blood from mice belonging to different groups. Error bars represent standard deviation (S.D) from four independent values.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
getmorefigures.php?uid=PMC4150946&req=5

Fig4: Cytokine levels during septicemia: a) IL 1ß b) IL 10 c) TNFalevels (pg/ml) 24 h post infection in various organs and blood of mice infected via intra peritoneal route withK. pneumoniaeB5055 and treated with gentamicin (1.5 mg/kg) and/or depolymerase (50 μg). G1, G2, G3, G4 represent Groups 1, 2, 3, 4. Arrows indicate significant reduction in cytokine levels in all organs and blood from mice belonging to different groups. Error bars represent standard deviation (S.D) from four independent values.

Mentions: In animals receiving infection intraperitoneally, systemic spread of infection resulted in marked upregulation in pro-inflammatory as well as anti-inflammatory cytokine levels within 24 h. A modest downregulation in levels of IL-1ß [~1.56 fold (lungs), ~1.37 fold (liver), ~1.52 fold (kidney), ~1.65 fold (spleen)], TNFa [~1.72 fold (lungs), ~1.56 fold (liver), ~1.75 fold (kidney), ~1.79 fold (spleen)] and IL-10 [~1.46 fold (lungs), ~1.86 fold (liver), ~1.92 fold (kidney), ~1.75 fold (spleen)] was observed in the infected, enzyme alone treated animals (Figure 4a, b, c). In contrast, cytokine levels in the blood serum samples were significantly reduced (P < 0.01) by ~2.1 fold (IL-1ß), ~2.43 fold (TNFa and ~3.26 fold (IL-10) in this group. When depolymerase was used along with gentamicin, negligible levels of all cytokines (P < 0.01) were observed in lungs, liver, kidney, spleen and blood (Figure 4a, b, c).Figure 4


Depolymerase improves gentamicin efficacy during Klebsiella pneumoniae induced murine infection.

Bansal S, Harjai K, Chhibber S - BMC Infect. Dis. (2014)

Cytokine levels during septicemia: a) IL 1ß b) IL 10 c) TNFalevels (pg/ml) 24 h post infection in various organs and blood of mice infected via intra peritoneal route withK. pneumoniaeB5055 and treated with gentamicin (1.5 mg/kg) and/or depolymerase (50 μg). G1, G2, G3, G4 represent Groups 1, 2, 3, 4. Arrows indicate significant reduction in cytokine levels in all organs and blood from mice belonging to different groups. Error bars represent standard deviation (S.D) from four independent values.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4150946&req=5

Fig4: Cytokine levels during septicemia: a) IL 1ß b) IL 10 c) TNFalevels (pg/ml) 24 h post infection in various organs and blood of mice infected via intra peritoneal route withK. pneumoniaeB5055 and treated with gentamicin (1.5 mg/kg) and/or depolymerase (50 μg). G1, G2, G3, G4 represent Groups 1, 2, 3, 4. Arrows indicate significant reduction in cytokine levels in all organs and blood from mice belonging to different groups. Error bars represent standard deviation (S.D) from four independent values.
Mentions: In animals receiving infection intraperitoneally, systemic spread of infection resulted in marked upregulation in pro-inflammatory as well as anti-inflammatory cytokine levels within 24 h. A modest downregulation in levels of IL-1ß [~1.56 fold (lungs), ~1.37 fold (liver), ~1.52 fold (kidney), ~1.65 fold (spleen)], TNFa [~1.72 fold (lungs), ~1.56 fold (liver), ~1.75 fold (kidney), ~1.79 fold (spleen)] and IL-10 [~1.46 fold (lungs), ~1.86 fold (liver), ~1.92 fold (kidney), ~1.75 fold (spleen)] was observed in the infected, enzyme alone treated animals (Figure 4a, b, c). In contrast, cytokine levels in the blood serum samples were significantly reduced (P < 0.01) by ~2.1 fold (IL-1ß), ~2.43 fold (TNFa and ~3.26 fold (IL-10) in this group. When depolymerase was used along with gentamicin, negligible levels of all cytokines (P < 0.01) were observed in lungs, liver, kidney, spleen and blood (Figure 4a, b, c).Figure 4

Bottom Line: Highly significant reduction in neutrophil infiltration and levels of pro-inflammatory and anti-inflammatory cytokines was also observed.In addition, as confirmed through flow cytometry, the hyperimmune sera raised against the enzyme did not neutralize its activity.Moreover, since this decapsulating protein does not possess a lytic activity therefore there would be no chances of development of bacterial resistance against it.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology, Panjab University, Sector-14, Chandigarh 160014, India. sanjaychhibber8@gmail.com.

ABSTRACT

Background: Presence of capsule enhances the virulence of bacteria that cause pneumonia, meningitis, cystic fibrosis, dental caries, periodontitis. Capsule is an important virulence factor for Klebsiella pneumoniae and infections due to this pathogen have been associated with high mortality rates. In the present study, use of an Aeromonas punctata derived capsule depolymerase against K. pneumoniae, to reinstate the efficacy of gentamicin during pneumonia and septicemia was investigated.

Methods: Depolymerase was administered in mice intraperitoneally (50 μg) alone as well in combination with gentamicin (1.5 mg/kg), 24 h post infection during acute lung infection and 6 h later during septicemia. Bacterial load, neutrophil infiltration and cytokine levels were estimated. The immunogenicity of protein was also studied.

Results: In comparison to groups treated with gentamicin alone, combination treatment with depolymerase and gentamicin significantly reduced (P < 0.01) bacterial titer in the lungs, liver, kidney, spleen and blood of experimental animals. Highly significant reduction in neutrophil infiltration and levels of pro-inflammatory and anti-inflammatory cytokines was also observed. This indicated an efficient capsule removal by the enzyme, that improved gentamicin efficacy in vivo. Although the enzyme was found to be immunogenic, but no significant reduction in treatment efficacy was observed in the preimmunized as well as naïve mice. In addition, as confirmed through flow cytometry, the hyperimmune sera raised against the enzyme did not neutralize its activity.

Conclusion: The results confirm that administration of enzyme 'depolymerase' along with gentamicin not only checked the virulence of K. pneumoniae in vivo but it also increased its susceptibility to gentamicin at a lower concentration. Such a strategy would help to avoid exposure to higher concentration of gentamicin. Moreover, since this decapsulating protein does not possess a lytic activity therefore there would be no chances of development of bacterial resistance against it. Therefore, it should be studied further for its successful inclusion in our prophylactic/therapeutic regimes.

Show MeSH
Related in: MedlinePlus