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Synergistic antibacterial and anti-inflammatory activity of temporin A and modified temporin B in vivo.

Capparelli R, Romanelli A, Iannaccone M, Nocerino N, Ripa R, Pensato S, Pedone C, Iannelli D - PLoS ONE (2009)

Bottom Line: Temporins are antimicrobial peptides secreted by the granular glands of the European red frog (Rana temporaria).Administration of 3.4 mg/Kg of temporin A (TA)+1.6 mg/Kg TB-YK, given to individual mice concurrently with a lethal dose of bacteria (gram positive or negative), rescued 100% of the animals.The model offers the possibility to test in a preclinical setting the true potential of TA and TB-YK in combination as antimicrobial and anti-inflammatory agents.

View Article: PubMed Central - PubMed

Affiliation: Department of Soil, Plant, Environment and Animal Production Sciences, University of Naples Federico II, School of Biotechnological Sciences, Napoli, Italy. capparel@unina.it

ABSTRACT
Temporins are antimicrobial peptides secreted by the granular glands of the European red frog (Rana temporaria). They are 10-14 amino acid long polypeptides active prevalently against gram positive bacteria. This study shows that a synthetic temporin B analogue (TB-YK), acquires the capacity to act in synergism with temporin A and to exert antimicrobial and anti-inflammatory activity in vivo against gram positive and gram negative bacteria. Administration of 3.4 mg/Kg of temporin A (TA)+1.6 mg/Kg TB-YK, given to individual mice concurrently with a lethal dose of bacteria (gram positive or negative), rescued 100% of the animals. More importantly, the same doses of temporins, administered one week after experimental infection with a sub lethal dose of bacteria, sterilized 100% of the animals within 3-6 days. Also, it is described an animal model based on the use of sub lethal doses of bacteria, which closely mimics bacterial infection in humans. The model offers the possibility to test in a preclinical setting the true potential of TA and TB-YK in combination as antimicrobial and anti-inflammatory agents.

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Related in: MedlinePlus

Temporins sterilize mice sperimentally infected with S aureus A170 or S.enterica serovar Paratyphi B.The 35/75 pool of temporins, given concurrently with bacteria, sterilize mice infected with a lethal dose (107 CFU/mouse) of S.aureus A170 (A: kidneys from mice infected with S. aureus A170 (closed rhomb) and kidneys from mice infected with S.aureus A170 and immediately after treated with TA plus TB-YK (open box), or a lethal dose (106 CFU/mouse) of S.enterica serovar Paratyphi B (B: gastro instestinal tract from mice infected with S.enterica serovar Paratyphi B (closed rhomb) and gastro intestinal tract from mice infected with S.enterica serovar Paratyphi B and immediately treated with TA plus TB-YK (open box); (C: liver from mice infected with S.enterica serovar Paratyphi B (closed rhomb) and liver from mice infected with S.enterica serovar Paratyphi B and immediately treated with TA plus TB-YK (open box).
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pone-0007191-g006: Temporins sterilize mice sperimentally infected with S aureus A170 or S.enterica serovar Paratyphi B.The 35/75 pool of temporins, given concurrently with bacteria, sterilize mice infected with a lethal dose (107 CFU/mouse) of S.aureus A170 (A: kidneys from mice infected with S. aureus A170 (closed rhomb) and kidneys from mice infected with S.aureus A170 and immediately after treated with TA plus TB-YK (open box), or a lethal dose (106 CFU/mouse) of S.enterica serovar Paratyphi B (B: gastro instestinal tract from mice infected with S.enterica serovar Paratyphi B (closed rhomb) and gastro intestinal tract from mice infected with S.enterica serovar Paratyphi B and immediately treated with TA plus TB-YK (open box); (C: liver from mice infected with S.enterica serovar Paratyphi B (closed rhomb) and liver from mice infected with S.enterica serovar Paratyphi B and immediately treated with TA plus TB-YK (open box).

Mentions: On the basis of the results described above, the therapeutic potential of the TA in combination with TB-YK was tested in a mouse model. The test, if successful, offered the possibility to assess the potential clinical use of the two temporins against septic shock caused by gram positive or gram negative bacteria. Several combinations of the two temporins displayed the capacity to protect mice from lethal doses of bacteria. Among these, the combination of 3.4 mg/Kg TA 0.54 mM in saline +1.6 mg/Kg TB-YK 0.19 mM in saline/mouse (75 µg TA+35 µg TB in 100 µl saline) was chosen for further studies because provided the highest surviving rate of mice infected with gram positive and gram negative bacteria and did not display haemolytic activity. This temporin combination, used in all the experiments to be described, will be referred to as the 75/35 pool. Two groups of mice (24 animals/group; untreated groups) were infected intravenously with S. enterica serovar Paratyphi (106 CFU/mouse) or S. aureus A170 (107 CFU/mouse), respectively. Two more groups of mice (24 animals/group; treated groups) were infected by the same procedure and, immediately after, treated with the 35/75 pool. Untreated mice died all within 4–6 days. The treated mice were all alive 28 days after the temporin treatment. (Figure 5). In an independent experiment, 2 mice infected as described (untreated mice) and 2 mice infected and then treated as above (treated mice) were sacrificed at daily intervals to monitor the bacterial load in the target organs (liver and GI tract of mice infected with S. enterica serovar Paratyphi; the kidneys of mice infected with S. aureus). Treated mice cleared S aureus or S. enterica serovar Paratyphi within 3 (Figure 6A) or 6 Figure 6B–C days from temporin treatment, respectively. Untreated mice, instead, remained heavily infected till the time of death (Figure 6A–C). The results demonstrated the efficacy of the 35/75 pool in vivo. Remarkably, the 35/75 pool was active against the methicillin-resistant S. aureus A170 strain.


Synergistic antibacterial and anti-inflammatory activity of temporin A and modified temporin B in vivo.

Capparelli R, Romanelli A, Iannaccone M, Nocerino N, Ripa R, Pensato S, Pedone C, Iannelli D - PLoS ONE (2009)

Temporins sterilize mice sperimentally infected with S aureus A170 or S.enterica serovar Paratyphi B.The 35/75 pool of temporins, given concurrently with bacteria, sterilize mice infected with a lethal dose (107 CFU/mouse) of S.aureus A170 (A: kidneys from mice infected with S. aureus A170 (closed rhomb) and kidneys from mice infected with S.aureus A170 and immediately after treated with TA plus TB-YK (open box), or a lethal dose (106 CFU/mouse) of S.enterica serovar Paratyphi B (B: gastro instestinal tract from mice infected with S.enterica serovar Paratyphi B (closed rhomb) and gastro intestinal tract from mice infected with S.enterica serovar Paratyphi B and immediately treated with TA plus TB-YK (open box); (C: liver from mice infected with S.enterica serovar Paratyphi B (closed rhomb) and liver from mice infected with S.enterica serovar Paratyphi B and immediately treated with TA plus TB-YK (open box).
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2747021&req=5

pone-0007191-g006: Temporins sterilize mice sperimentally infected with S aureus A170 or S.enterica serovar Paratyphi B.The 35/75 pool of temporins, given concurrently with bacteria, sterilize mice infected with a lethal dose (107 CFU/mouse) of S.aureus A170 (A: kidneys from mice infected with S. aureus A170 (closed rhomb) and kidneys from mice infected with S.aureus A170 and immediately after treated with TA plus TB-YK (open box), or a lethal dose (106 CFU/mouse) of S.enterica serovar Paratyphi B (B: gastro instestinal tract from mice infected with S.enterica serovar Paratyphi B (closed rhomb) and gastro intestinal tract from mice infected with S.enterica serovar Paratyphi B and immediately treated with TA plus TB-YK (open box); (C: liver from mice infected with S.enterica serovar Paratyphi B (closed rhomb) and liver from mice infected with S.enterica serovar Paratyphi B and immediately treated with TA plus TB-YK (open box).
Mentions: On the basis of the results described above, the therapeutic potential of the TA in combination with TB-YK was tested in a mouse model. The test, if successful, offered the possibility to assess the potential clinical use of the two temporins against septic shock caused by gram positive or gram negative bacteria. Several combinations of the two temporins displayed the capacity to protect mice from lethal doses of bacteria. Among these, the combination of 3.4 mg/Kg TA 0.54 mM in saline +1.6 mg/Kg TB-YK 0.19 mM in saline/mouse (75 µg TA+35 µg TB in 100 µl saline) was chosen for further studies because provided the highest surviving rate of mice infected with gram positive and gram negative bacteria and did not display haemolytic activity. This temporin combination, used in all the experiments to be described, will be referred to as the 75/35 pool. Two groups of mice (24 animals/group; untreated groups) were infected intravenously with S. enterica serovar Paratyphi (106 CFU/mouse) or S. aureus A170 (107 CFU/mouse), respectively. Two more groups of mice (24 animals/group; treated groups) were infected by the same procedure and, immediately after, treated with the 35/75 pool. Untreated mice died all within 4–6 days. The treated mice were all alive 28 days after the temporin treatment. (Figure 5). In an independent experiment, 2 mice infected as described (untreated mice) and 2 mice infected and then treated as above (treated mice) were sacrificed at daily intervals to monitor the bacterial load in the target organs (liver and GI tract of mice infected with S. enterica serovar Paratyphi; the kidneys of mice infected with S. aureus). Treated mice cleared S aureus or S. enterica serovar Paratyphi within 3 (Figure 6A) or 6 Figure 6B–C days from temporin treatment, respectively. Untreated mice, instead, remained heavily infected till the time of death (Figure 6A–C). The results demonstrated the efficacy of the 35/75 pool in vivo. Remarkably, the 35/75 pool was active against the methicillin-resistant S. aureus A170 strain.

Bottom Line: Temporins are antimicrobial peptides secreted by the granular glands of the European red frog (Rana temporaria).Administration of 3.4 mg/Kg of temporin A (TA)+1.6 mg/Kg TB-YK, given to individual mice concurrently with a lethal dose of bacteria (gram positive or negative), rescued 100% of the animals.The model offers the possibility to test in a preclinical setting the true potential of TA and TB-YK in combination as antimicrobial and anti-inflammatory agents.

View Article: PubMed Central - PubMed

Affiliation: Department of Soil, Plant, Environment and Animal Production Sciences, University of Naples Federico II, School of Biotechnological Sciences, Napoli, Italy. capparel@unina.it

ABSTRACT
Temporins are antimicrobial peptides secreted by the granular glands of the European red frog (Rana temporaria). They are 10-14 amino acid long polypeptides active prevalently against gram positive bacteria. This study shows that a synthetic temporin B analogue (TB-YK), acquires the capacity to act in synergism with temporin A and to exert antimicrobial and anti-inflammatory activity in vivo against gram positive and gram negative bacteria. Administration of 3.4 mg/Kg of temporin A (TA)+1.6 mg/Kg TB-YK, given to individual mice concurrently with a lethal dose of bacteria (gram positive or negative), rescued 100% of the animals. More importantly, the same doses of temporins, administered one week after experimental infection with a sub lethal dose of bacteria, sterilized 100% of the animals within 3-6 days. Also, it is described an animal model based on the use of sub lethal doses of bacteria, which closely mimics bacterial infection in humans. The model offers the possibility to test in a preclinical setting the true potential of TA and TB-YK in combination as antimicrobial and anti-inflammatory agents.

Show MeSH
Related in: MedlinePlus