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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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Administration of anti-hTF mAb ameliorated intestinal ischemia-reperfusion (IIR)-induced acute lung injury in hTF-KI mice.IIR challenged hTF-KI mice were treated with CNTO859 (5 mg/kg, i.v.) or saline. The representative histology (H&E, x400) from the intestine (A, B) and lung (C, D) was shown. The lung injury was scored by a pathologist in a blind fashion (E) (▪ inflammatory cells infiltration; ▪ alveolar wall edema; ▪ hemorrhage; atelectasis). □ Lung injury scores of 4 categories were analyzed with Kruskall-Wallis test, n = 4 animals/group, *: p<0.05. The pulmonary permeability was determined by Evans Blue dye assay (F, G, H). Administration of CNTO859 also reduced the wet/dry lung weight ratio (I), albumin concentration (J), and total cell counts (K) in the BAL fluid. Panels I-K: n = 4 animals/group,*: p<0.05, un-paired t-test.
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pone-0001527-g002: Administration of anti-hTF mAb ameliorated intestinal ischemia-reperfusion (IIR)-induced acute lung injury in hTF-KI mice.IIR challenged hTF-KI mice were treated with CNTO859 (5 mg/kg, i.v.) or saline. The representative histology (H&E, x400) from the intestine (A, B) and lung (C, D) was shown. The lung injury was scored by a pathologist in a blind fashion (E) (▪ inflammatory cells infiltration; ▪ alveolar wall edema; ▪ hemorrhage; atelectasis). □ Lung injury scores of 4 categories were analyzed with Kruskall-Wallis test, n = 4 animals/group, *: p<0.05. The pulmonary permeability was determined by Evans Blue dye assay (F, G, H). Administration of CNTO859 also reduced the wet/dry lung weight ratio (I), albumin concentration (J), and total cell counts (K) in the BAL fluid. Panels I-K: n = 4 animals/group,*: p<0.05, un-paired t-test.

Mentions: A clinically relevant model of extrapulmonary ARDS has been used to induce acute lung injury by intestinal ischemia-reperfusion in C57BL/6 mice, and a high mortality rate was observed in these studies during 4 h of reperfusion period when animals were ventilated with oxygen [19], [20]. It is known that C57BL/6 mice are sensitive to hyperoxia [27]. In the present study, we used hTF-KI mice, which are of a hybrid strain of 129SvBrd (∼20%) and C57BL/6 (∼80%). We first conducted a pilot study with a modified protocol, in which animals were not subject to mechanical ventilation and pure oxygen during the reperfusion period. There was no mortality during the first 24 h of reperfusion (data not shown). The IIR challenge induced a significant intestinal (Fig. 2A) and lung injury (Fig. 2C) in hTF-KI mice, which are very similar to that in wild type mice (data not shown) and as observed in our previous study [19], [20]. These results suggest that hTF not only substituted mTF for coagulation, but also may play a similar role in acute inflammatory response related to acute lung injury.


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)

Administration of anti-hTF mAb ameliorated intestinal ischemia-reperfusion (IIR)-induced acute lung injury in hTF-KI mice.IIR challenged hTF-KI mice were treated with CNTO859 (5 mg/kg, i.v.) or saline. The representative histology (H&E, x400) from the intestine (A, B) and lung (C, D) was shown. The lung injury was scored by a pathologist in a blind fashion (E) (▪ inflammatory cells infiltration; ▪ alveolar wall edema; ▪ hemorrhage; atelectasis). □ Lung injury scores of 4 categories were analyzed with Kruskall-Wallis test, n = 4 animals/group, *: p<0.05. The pulmonary permeability was determined by Evans Blue dye assay (F, G, H). Administration of CNTO859 also reduced the wet/dry lung weight ratio (I), albumin concentration (J), and total cell counts (K) in the BAL fluid. Panels I-K: n = 4 animals/group,*: p<0.05, un-paired t-test.
© Copyright Policy
Related In: Results  -  Collection

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

pone-0001527-g002: Administration of anti-hTF mAb ameliorated intestinal ischemia-reperfusion (IIR)-induced acute lung injury in hTF-KI mice.IIR challenged hTF-KI mice were treated with CNTO859 (5 mg/kg, i.v.) or saline. The representative histology (H&E, x400) from the intestine (A, B) and lung (C, D) was shown. The lung injury was scored by a pathologist in a blind fashion (E) (▪ inflammatory cells infiltration; ▪ alveolar wall edema; ▪ hemorrhage; atelectasis). □ Lung injury scores of 4 categories were analyzed with Kruskall-Wallis test, n = 4 animals/group, *: p<0.05. The pulmonary permeability was determined by Evans Blue dye assay (F, G, H). Administration of CNTO859 also reduced the wet/dry lung weight ratio (I), albumin concentration (J), and total cell counts (K) in the BAL fluid. Panels I-K: n = 4 animals/group,*: p<0.05, un-paired t-test.
Mentions: A clinically relevant model of extrapulmonary ARDS has been used to induce acute lung injury by intestinal ischemia-reperfusion in C57BL/6 mice, and a high mortality rate was observed in these studies during 4 h of reperfusion period when animals were ventilated with oxygen [19], [20]. It is known that C57BL/6 mice are sensitive to hyperoxia [27]. In the present study, we used hTF-KI mice, which are of a hybrid strain of 129SvBrd (∼20%) and C57BL/6 (∼80%). We first conducted a pilot study with a modified protocol, in which animals were not subject to mechanical ventilation and pure oxygen during the reperfusion period. There was no mortality during the first 24 h of reperfusion (data not shown). The IIR challenge induced a significant intestinal (Fig. 2A) and lung injury (Fig. 2C) in hTF-KI mice, which are very similar to that in wild type mice (data not shown) and as observed in our previous study [19], [20]. These results suggest that hTF not only substituted mTF for coagulation, but also may play a similar role in acute inflammatory response related to acute lung injury.

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