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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

Human tissue factor is functionally expressed in hTF-KI transgenic mice, and effectively and specifically inhibited by the anti-hTF mAb, CNTO859.Pro-coagulant activity of TF in brain extracts from either hTF-KI or wild type (WT) mice, or from human brain tissue was measured with a one-stage clotting assay, and a similar prothrombin time was seen in all brain extracts, indicating a functional replacement of mTF by hTF expressed in the hTF-KI mice (A). Anti-hTF antibody, CNTO859, dose-dependently inhibited TF pro-coagulant activity in the brain extracts from hTF-KI mice and human, but not wild type mice (B). The experiments were repeated three times, and representative data from one experiment are shown.
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pone-0001527-g001: Human tissue factor is functionally expressed in hTF-KI transgenic mice, and effectively and specifically inhibited by the anti-hTF mAb, CNTO859.Pro-coagulant activity of TF in brain extracts from either hTF-KI or wild type (WT) mice, or from human brain tissue was measured with a one-stage clotting assay, and a similar prothrombin time was seen in all brain extracts, indicating a functional replacement of mTF by hTF expressed in the hTF-KI mice (A). Anti-hTF antibody, CNTO859, dose-dependently inhibited TF pro-coagulant activity in the brain extracts from hTF-KI mice and human, but not wild type mice (B). The experiments were repeated three times, and representative data from one experiment are shown.

Mentions: To test anti-TF therapy against hTF in vivo, we used the hTF-KI mice generated by replacing a portion of mTF gene with the open reading frame of hTF. The hTF-KI mice have a similar life span without abnormal pathology in comparison with wild type animals. The tissue distribution and expression levels of hTF in transgenic mice were similar to that of mTF in wild type mice, and no mTF expression was detected in the transgenic mice [16]. The brain extract of hTF-KI mice expressed hTF at 64 ng/mg protein, a level comparable to that from the human brain extract at 80 ng/mg protein. Tested with a one-stage clotting assay, the brain extract from the hTF-KI mice showed a similar pro-coagulation activity as that from wild type mice or human brain tissue (Fig. 1A). These results suggest that hTF functionally substitutes mTF and maintains a normal coagulant activity in the hTF-KI mice.


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)

Human tissue factor is functionally expressed in hTF-KI transgenic mice, and effectively and specifically inhibited by the anti-hTF mAb, CNTO859.Pro-coagulant activity of TF in brain extracts from either hTF-KI or wild type (WT) mice, or from human brain tissue was measured with a one-stage clotting assay, and a similar prothrombin time was seen in all brain extracts, indicating a functional replacement of mTF by hTF expressed in the hTF-KI mice (A). Anti-hTF antibody, CNTO859, dose-dependently inhibited TF pro-coagulant activity in the brain extracts from hTF-KI mice and human, but not wild type mice (B). The experiments were repeated three times, and representative data from one experiment are shown.
© Copyright Policy
Related In: Results  -  Collection

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

pone-0001527-g001: Human tissue factor is functionally expressed in hTF-KI transgenic mice, and effectively and specifically inhibited by the anti-hTF mAb, CNTO859.Pro-coagulant activity of TF in brain extracts from either hTF-KI or wild type (WT) mice, or from human brain tissue was measured with a one-stage clotting assay, and a similar prothrombin time was seen in all brain extracts, indicating a functional replacement of mTF by hTF expressed in the hTF-KI mice (A). Anti-hTF antibody, CNTO859, dose-dependently inhibited TF pro-coagulant activity in the brain extracts from hTF-KI mice and human, but not wild type mice (B). The experiments were repeated three times, and representative data from one experiment are shown.
Mentions: To test anti-TF therapy against hTF in vivo, we used the hTF-KI mice generated by replacing a portion of mTF gene with the open reading frame of hTF. The hTF-KI mice have a similar life span without abnormal pathology in comparison with wild type animals. The tissue distribution and expression levels of hTF in transgenic mice were similar to that of mTF in wild type mice, and no mTF expression was detected in the transgenic mice [16]. The brain extract of hTF-KI mice expressed hTF at 64 ng/mg protein, a level comparable to that from the human brain extract at 80 ng/mg protein. Tested with a one-stage clotting assay, the brain extract from the hTF-KI mice showed a similar pro-coagulation activity as that from wild type mice or human brain tissue (Fig. 1A). These results suggest that hTF functionally substitutes mTF and maintains a normal coagulant activity in the hTF-KI mice.

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