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Anti-human tissue factor antibody ameliorated intestinal ischemia reperfusion-induced acute lung injury in human tissue factor knock-in mice.

He X, Han B, Mura M, Li L, Cypel M, Soderman A, Picha K, Yang J, Liu M - PLoS ONE (2008)

Bottom Line: Administration of CNTO859 (5 mg/kg, i.v.) attenuated the severity of lung tissue injury, decreased the total cell counts and protein concentration in bronchoalveolar lavage fluid, and reduced Evans blue leakage.In addition, the treatment significantly reduced alveolar fibrin deposition, and decreased tissue factor and plasminogen activator inhibitor-1 activity in the serum.This novel anti-hTF antibody showed beneficial effects on intestinal ischemia-reperfusion induced acute lung injury, which merits further investigation for clinical usage.

View Article: PubMed Central - PubMed

Affiliation: Latner Thoracic Surgery Research Laboratories, Department of Surgery, University Health Network Toronto General Research Institute, University of Toronto, Toronto, Ontario, Canada.

ABSTRACT

Background: Interaction between the coagulation and inflammation systems plays an important role in the development of acute respiratory distress syndrome (ARDS). Anti-coagulation is an attractive option for ARDS treatment, and this has promoted development of new antibodies. However, preclinical trials for these antibodies are often limited by the high cost and availability of non-human primates. In the present study, we developed a novel alternative method to test the role of a humanized anti-tissue factor mAb in acute lung injury with transgenic mice.

Methodology/principal findings: Human tissue factor knock-in (hTF-KI) transgenic mice and a novel humanized anti-human tissue factor mAb (anti-hTF mAb, CNTO859) were developed. The hTF-KI mice showed a normal and functional expression of hTF. The anti-hTF mAb specifically blocked the pro-coagulation activity of brain extracts from the hTF-KI mice and human, but not from wild type mice. An extrapulmonary ARDS model was used by intestinal ischemia-reperfusion. Significant lung tissue damage in hTF-KI mice was observed after 2 h reperfusion. Administration of CNTO859 (5 mg/kg, i.v.) attenuated the severity of lung tissue injury, decreased the total cell counts and protein concentration in bronchoalveolar lavage fluid, and reduced Evans blue leakage. In addition, the treatment significantly reduced alveolar fibrin deposition, and decreased tissue factor and plasminogen activator inhibitor-1 activity in the serum. This treatment also down-regulated cytokine expression and reduced cell death in the lung.

Conclusions: This novel anti-hTF antibody showed beneficial effects on intestinal ischemia-reperfusion induced acute lung injury, which merits further investigation for clinical usage. In addition, the use of knock-in transgenic mice to test the efficacy of antibodies against human-specific proteins is a novel strategy for preclinical studies.

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Related in: MedlinePlus

Anti-hTF mAb treatment reduced cell death.IIR-induced cell death in the lungs was determined by TUNEL staining. CNTO859 effect on the cell death was shown in representative slides (400x) (A, B), and quantified by counting the TUNEL positive cells from 10 randomly chosen fields (C). Caspase 3 activity in the lung tissue was also reduced by the anti-hTF antibody treatment (D). Un-paired t-test was used, 4 animals per group, *: p<0.05.
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pone-0001527-g005: Anti-hTF mAb treatment reduced cell death.IIR-induced cell death in the lungs was determined by TUNEL staining. CNTO859 effect on the cell death was shown in representative slides (400x) (A, B), and quantified by counting the TUNEL positive cells from 10 randomly chosen fields (C). Caspase 3 activity in the lung tissue was also reduced by the anti-hTF antibody treatment (D). Un-paired t-test was used, 4 animals per group, *: p<0.05.

Mentions: In a previous study, we found that IIR induces alveolar epithelial cell death [19]. In the present study, TUNEL positive cells were found in the lung of hTF-KI mice after IIR challenge (Fig. 5A), and they were significantly reduced by CNTO859 treatment (Fig. 5B and 5C). Caspase 3 is a key enzyme for apoptotic cell death. CNTO859 treatment reduced caspase 3 activity in lung tissue homogenates (Fig. 5D).


Anti-human tissue factor antibody ameliorated intestinal ischemia reperfusion-induced acute lung injury in human tissue factor knock-in mice.

He X, Han B, Mura M, Li L, Cypel M, Soderman A, Picha K, Yang J, Liu M - PLoS ONE (2008)

Anti-hTF mAb treatment reduced cell death.IIR-induced cell death in the lungs was determined by TUNEL staining. CNTO859 effect on the cell death was shown in representative slides (400x) (A, B), and quantified by counting the TUNEL positive cells from 10 randomly chosen fields (C). Caspase 3 activity in the lung tissue was also reduced by the anti-hTF antibody treatment (D). Un-paired t-test was used, 4 animals per group, *: p<0.05.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2211395&req=5

pone-0001527-g005: Anti-hTF mAb treatment reduced cell death.IIR-induced cell death in the lungs was determined by TUNEL staining. CNTO859 effect on the cell death was shown in representative slides (400x) (A, B), and quantified by counting the TUNEL positive cells from 10 randomly chosen fields (C). Caspase 3 activity in the lung tissue was also reduced by the anti-hTF antibody treatment (D). Un-paired t-test was used, 4 animals per group, *: p<0.05.
Mentions: In a previous study, we found that IIR induces alveolar epithelial cell death [19]. In the present study, TUNEL positive cells were found in the lung of hTF-KI mice after IIR challenge (Fig. 5A), and they were significantly reduced by CNTO859 treatment (Fig. 5B and 5C). Caspase 3 is a key enzyme for apoptotic cell death. CNTO859 treatment reduced caspase 3 activity in lung tissue homogenates (Fig. 5D).

Bottom Line: Administration of CNTO859 (5 mg/kg, i.v.) attenuated the severity of lung tissue injury, decreased the total cell counts and protein concentration in bronchoalveolar lavage fluid, and reduced Evans blue leakage.In addition, the treatment significantly reduced alveolar fibrin deposition, and decreased tissue factor and plasminogen activator inhibitor-1 activity in the serum.This novel anti-hTF antibody showed beneficial effects on intestinal ischemia-reperfusion induced acute lung injury, which merits further investigation for clinical usage.

View Article: PubMed Central - PubMed

Affiliation: Latner Thoracic Surgery Research Laboratories, Department of Surgery, University Health Network Toronto General Research Institute, University of Toronto, Toronto, Ontario, Canada.

ABSTRACT

Background: Interaction between the coagulation and inflammation systems plays an important role in the development of acute respiratory distress syndrome (ARDS). Anti-coagulation is an attractive option for ARDS treatment, and this has promoted development of new antibodies. However, preclinical trials for these antibodies are often limited by the high cost and availability of non-human primates. In the present study, we developed a novel alternative method to test the role of a humanized anti-tissue factor mAb in acute lung injury with transgenic mice.

Methodology/principal findings: Human tissue factor knock-in (hTF-KI) transgenic mice and a novel humanized anti-human tissue factor mAb (anti-hTF mAb, CNTO859) were developed. The hTF-KI mice showed a normal and functional expression of hTF. The anti-hTF mAb specifically blocked the pro-coagulation activity of brain extracts from the hTF-KI mice and human, but not from wild type mice. An extrapulmonary ARDS model was used by intestinal ischemia-reperfusion. Significant lung tissue damage in hTF-KI mice was observed after 2 h reperfusion. Administration of CNTO859 (5 mg/kg, i.v.) attenuated the severity of lung tissue injury, decreased the total cell counts and protein concentration in bronchoalveolar lavage fluid, and reduced Evans blue leakage. In addition, the treatment significantly reduced alveolar fibrin deposition, and decreased tissue factor and plasminogen activator inhibitor-1 activity in the serum. This treatment also down-regulated cytokine expression and reduced cell death in the lung.

Conclusions: This novel anti-hTF antibody showed beneficial effects on intestinal ischemia-reperfusion induced acute lung injury, which merits further investigation for clinical usage. In addition, the use of knock-in transgenic mice to test the efficacy of antibodies against human-specific proteins is a novel strategy for preclinical studies.

Show MeSH
Related in: MedlinePlus