Limits...
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.

Show MeSH

Related in: MedlinePlus

Anti-hTF mAb treatment attenuated IIR induced coagulapathy and protected pulmonary endothelium.Plasma TF and PAI-1 activities were determined as described in Methods. Administration of CNTO859 significantly inhibited both TF (A) and PAI-1 (B) activities in comparison with the saline treated control (*: p<0.05, n = 4 animals/group, un-paired t-test). The fibrin staining showed that CNTO859 ameliorated IIR-induced fibrin deposition (pink) in the alveoli (C, D). The lung tissues were stained for vWF, a specific marker for endothelial integrity. Weaker staining of vWF (pink) of endothelium in the pulmonary vessels was noted in the saline control group (E). In CNTO859 treated animals stronger vWF staining was observed in the endothelium of pulmonary vessels (F). The pulmonary endothelial injury was further examined with electron microscopy. IIR challenge led to significant endothelial cell swelling (G), which was protected by CNTO859 treatment (H).
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC2211395&req=5

pone-0001527-g003: Anti-hTF mAb treatment attenuated IIR induced coagulapathy and protected pulmonary endothelium.Plasma TF and PAI-1 activities were determined as described in Methods. Administration of CNTO859 significantly inhibited both TF (A) and PAI-1 (B) activities in comparison with the saline treated control (*: p<0.05, n = 4 animals/group, un-paired t-test). The fibrin staining showed that CNTO859 ameliorated IIR-induced fibrin deposition (pink) in the alveoli (C, D). The lung tissues were stained for vWF, a specific marker for endothelial integrity. Weaker staining of vWF (pink) of endothelium in the pulmonary vessels was noted in the saline control group (E). In CNTO859 treated animals stronger vWF staining was observed in the endothelium of pulmonary vessels (F). The pulmonary endothelial injury was further examined with electron microscopy. IIR challenge led to significant endothelial cell swelling (G), which was protected by CNTO859 treatment (H).

Mentions: It is known that activation of TF can trigger a pro-coagulation status, and lead to fibrin deposition in the lung. Administration of CNTO859 significantly reduced both TF and PAI-1 activities in the plasma (Fig. 3A and 3B), and dramatically attenuated the IIR-induced alveolar fibrin deposition (Fig. 3C and 3D).


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 attenuated IIR induced coagulapathy and protected pulmonary endothelium.Plasma TF and PAI-1 activities were determined as described in Methods. Administration of CNTO859 significantly inhibited both TF (A) and PAI-1 (B) activities in comparison with the saline treated control (*: p<0.05, n = 4 animals/group, un-paired t-test). The fibrin staining showed that CNTO859 ameliorated IIR-induced fibrin deposition (pink) in the alveoli (C, D). The lung tissues were stained for vWF, a specific marker for endothelial integrity. Weaker staining of vWF (pink) of endothelium in the pulmonary vessels was noted in the saline control group (E). In CNTO859 treated animals stronger vWF staining was observed in the endothelium of pulmonary vessels (F). The pulmonary endothelial injury was further examined with electron microscopy. IIR challenge led to significant endothelial cell swelling (G), which was protected by CNTO859 treatment (H).
© Copyright Policy
Related In: Results  -  Collection

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

pone-0001527-g003: Anti-hTF mAb treatment attenuated IIR induced coagulapathy and protected pulmonary endothelium.Plasma TF and PAI-1 activities were determined as described in Methods. Administration of CNTO859 significantly inhibited both TF (A) and PAI-1 (B) activities in comparison with the saline treated control (*: p<0.05, n = 4 animals/group, un-paired t-test). The fibrin staining showed that CNTO859 ameliorated IIR-induced fibrin deposition (pink) in the alveoli (C, D). The lung tissues were stained for vWF, a specific marker for endothelial integrity. Weaker staining of vWF (pink) of endothelium in the pulmonary vessels was noted in the saline control group (E). In CNTO859 treated animals stronger vWF staining was observed in the endothelium of pulmonary vessels (F). The pulmonary endothelial injury was further examined with electron microscopy. IIR challenge led to significant endothelial cell swelling (G), which was protected by CNTO859 treatment (H).
Mentions: It is known that activation of TF can trigger a pro-coagulation status, and lead to fibrin deposition in the lung. Administration of CNTO859 significantly reduced both TF and PAI-1 activities in the plasma (Fig. 3A and 3B), and dramatically attenuated the IIR-induced alveolar fibrin deposition (Fig. 3C and 3D).

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