| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
|
|
|
|||||||
|
|||||||||
J Thorac Cardiovasc Surg 2009;137:695-702
© 2009 The American Association for Thoracic Surgery
Cardiothoracic Transplantation |
a Department of Surgery, University of Virginia Health System, Charlottesville, Va
b Department of Medicine, University of Virginia Health System, Charlottesville, Va
Received for publication April 17, 2008; revisions received September 18, 2008; accepted for publication October 13, 2008. * Address for reprints: Victor E. Laubach, PhD, University of Virginia Health System, Department of Surgery, PO Box 801359, Charlottesville, VA 22908. (Email: laubach{at}virginia.edu).
 |
Abstract |
|---|
 Top Abstract IntroductionMaterials and MethodsResultsDiscussionConclusionReferences |
|---|
Methods: An in vivo mouse model of lung ischemia–reperfusion injury was used. C57BL/6 mice were assigned to either the sham group (left thoracotomy) or 7 study groups that underwent 1-hour left hilar occlusion followed by 1 to 24 hours of reperfusion. After in vivo reperfusion, the lungs were perfused ex vivo with buffer whereby pulmonary function was assessed. Lung vascular permeability, edema, neutrophil accumulation, and cytokine/chemokine production (tumor necrosis factor 
, interleukin 17, CCL3, and CXCL1) were assessed based on Evans blue dye leak, wet/dry weight ratio, myeloperoxidase level, and enzyme-linked immunosorbent assay, respectively.
Results: A preliminary study showed that 2 hours of reperfusion resulted in greater pulmonary dysfunction than 1 or 24 hours of reperfusion. The 2-hour reperfusion period was thus used for the remaining experiments. Comparable and significant protection from ischemia–reperfusion injury–induced lung dysfunction and injury occurred after antibody depletion of neutrophils or CD4+ T cells but not CD8+ T cells (P < .05 vs immunoglobulin G control). Lung ischemia–reperfusion injury was proportional to the infiltration of neutrophils but not T cells. Moreover, pulmonary neutrophil infiltration and the production of CXCL1 (KC) were significantly diminished by CD4+ T-cell depletion but not vice versa.
Conclusions: Both CD4+ T lymphocytes and neutrophils accumulate during reperfusion and contribute sequentially to lung ischemia–reperfusion injury. The data suggest that neutrophils mediate ischemia–reperfusion injury; however, CD4+ T cells play a critical role in stimulating chemokine production and neutrophil chemotaxis during ischemia–reperfusion injury.
= interferon ; IgG = immunoglobulin G; IL = interleukin; IRI = ischemia–reperfusion injury; LC = lung compliance; mAb = monoclonal antibody; MPO = myeloperoxidase; TNF- = tumor necrosis factor
|
Introduction |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionConclusionReferences |
|---|
which has been reported to be responsible for up to 30% of patient mortality within 30 days.2 An increasing body of evidence has shown that IRI is associated with enhanced inflammatory responses during reperfusion. Our previous animal experiments have shown that alveolar macrophages and circulating leukocytes contribute importantly to lung IRI, with macrophages serving as triggers and leukocytes (mainly neutrophils) serving as end-effectors.3-6 Furthermore, we recently reported that alveolar epithelial cells, especially type II cells, interact with alveolar macrophages to initiate inflammatory responses during IRI.7 However, the signaling pathways between alveolar macrophages and neutrophils remain to be defined.
There is growing evidence that T cells might also participate in the pathogenesis of lung IRI.8-10 T cells are found to infiltrate the lung and are activated during reperfusion earlier than neutrophils.10 Lymphocyte-deficient rats or mice have decreased IRI.9,10 Cytokines and chemokines that stimulate T-cell chemotaxis and activation, such as interleukin (IL) 8, IL-12, IL-18, CCL5, and CCL2, are increased during lung IRI.7,9,11-13 T cells are known to amplify inflammatory responses through the secretion of lymphokines, including interferon (IFN-), IL-2, IL-4, IL-17, and granulocyte–macrophage colony-stimulating factor.9,14 These stimulate the chemotaxis of neutrophils and monocytes to sites of injury.
Whether T cells participate importantly in the inflammatory cascade that results in lung IRI is unclear. In the current study we used an in vivo mouse model of lung IRI to examine the role of T cells in lung IRI. Because neutrophils are end-effectors of lung IRI, we also examined the effect of lymphocyte depletion of neutrophil trafficking into the lung. Monoclonal antibodies (mAbs) were used to render mice deficient in neutrophils, CD4+ T cells, or CD8+ T cells.
|
Materials and Methods |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionConclusionReferences |
|---|
In Vivo Depletion of Neutrophils
Rat anti-mouse Gr-1 mAb was used to deplete circulating neutrophils in mice, as reported by others.15 Briefly, 10 µg of anti-Gr-1 mAb (eBioscience, San Diego, Calif) was injected through the tail vein 24 hours before lung ischemia. Perioperatively, blood (30–40 µL) was obtained by puncturing the left external jugular vein, and leukocyte counts were performed with a HemaVet Hematology System (CDC Technologies, Oxford, Conn).
In Vivo Depletion of CD4+ or CD8+ T Lymphocytes
Depletion of CD4+ or CD8+ T cells was achieved by using selective antibodies, as reported previously.16 Anti–CD4 mAb (GK1.5) or anti–CD8a mAb (53-6.7, eBioscience) was injected intraperitoneally on 2 consecutive days at a dose of 0.2 mg per mouse per day. Two days after the second injection, these animals underwent lung ischemia.
In Vivo Model of Lung Ischemia and Reperfusion
Mice were anesthetized with inhalation of isoflurane, intubated with PE-60 tubing, and connected to a pressure-controlled ventilator (Harvard Apparatus Co, South Natick, Mass). Mechanical ventilation was performed with room air as adjusted to a rate of 150 strokes/min, a stroke volume of 1.0 mL, and a peak inspiratory pressure of less than 20 cm H2O. Heparin (20 U/kg) was administered through external jugular injection. A left thoracotomy was performed by cutting the left fourth rib, and the left hilum was exposed. A 6-0 Prolene suture (Ethicon, Inc, Somerville, NJ) was placed around the hilum facilitated by a tip-curved (22-gauge) gavage needle. Both ends of the suture were then threaded through a 5-mm-long PE-50 tubing. Occlusion was achieved by pulling up on the suture and thus pushing the tube against the hilum to initiate ischemia. A small surgical clip was applied to the suture on top of the tube to maintain tension of the tube against the hilum. The thoracotomy was then closed with sutures, and the mouse was extubated, placed in a cage, and allowed to awaken during the 1-hour hilar occlusion period. Five minutes before reperfusion, the mouse was reanesthetized and reintubated. Reperfusion was achieved by removing the clip and the tube/suture. Again, the chest was closed with sutures. The mouse was extubated and returned to a cage until pulmonary function testing. The temperature was monitored during surgical intervention with an anal probe and maintained between 36.5°C and 37.5°C. Sham animals received only thoracotomy without hilar occlusion. To minimize pain and discomfort, an analgesic (buprenorphine, 0.2 mg/kg) was administered to all animals at the beginning of surgical intervention.
Measurement of Pulmonary Function
At the end of scheduled reperfusion, pulmonary function was evaluated by using an isolated, buffer-perfused mouse lung system (Hugo Sachs Elektronik, March-Huggstetten, Germany), as previously described by our laboratory.6 Briefly, mice were anesthetized with ketamine and xylazine. A tracheostomy was performed, and animals were ventilated with room air at 100 strokes/min and a tidal volume of 7 µL/g body weight with a positive end-expiratory pressure of 2 cm H2O. The animals were exsanguinated by means of inferior caval transection. The pulmonary artery was cannulated through the right ventricle, and the left ventricle was immediately tube vented through a small incision at the apex of the heart. The lungs were then perfused at a constant flow of 60 µL per gram of body weight per minute with Krebs–Henseleit buffer containing 2% albumin, 0.1% glucose, and 0.3% N-2-hydroxyethylpiperazine-N-2-ethanesulfonic acid (335–340 mOsm/kg H2O). The perfusate buffer and isolated lungs were maintained at 37°C throughout the experiment by using a circulating water bath. Once properly perfused and ventilated, the lungs were maintained on the system for a 5-minute equilibration period before data were recorded for an additional 10 minutes. Hemodynamic and pulmonary parameters were recorded during this period by using the PULMODYN data acquisition system (Hugo Sachs Elektronik).
Bronchoalveolar Lavage
After pulmonary function measurements, the left lungs were lavaged with 0.4 mL of normal saline. A microclamp was used to occlude the right hilum before lavage. The bronchoalveolar lavage (BAL) fluid was immediately centrifuged at 4°C (500g for 5 minutes), and the supernatant was stored at –80°C until further analysis.
Lung Wet/Dry Weight Ratio
In separate groups the left lung was harvested, weighed, and then placed in a vacuum oven (at 58°C) until a stable dry weight was achieved. The ratio of lung wet weight to dry weight was then calculated.
Pulmonary Microvascular Permeability
IRI-induced microvascular permeability in the lungs was determined by using the Evans blue dye extravasation technique.17 Evans blue (20 mg/kg; Sigma–Aldrich, St Louis, Mo) was injected intravenously 30 minutes before death. The pulmonary vasculature was then perfused for 15 minutes by using the isolated, buffer-perfused lung system to remove intravascular dye. Lungs were then homogenized in phosphate-buffered saline to extract the Evans blue and centrifuged. The absorption of Evans blue was measured in the supernatant at 620 nm and corrected for the presence of heme pigments as follows: 
.
Immunohistochemistry
A standard immunohistochemistry protocol for paraformaldehyde-fixed tissue was used, as detailed previously.16 Briefly, the left lung was harvested, cut into 4 short-axis slices, and immediately fixed in 1% paraformaldehyde in phosphate-buffered saline (pH 7.4) for paraffin embedding. Immunostaining was performed with rat anti-mouse neutrophil antibody (Serotec, Inc, Oxford, United Kingdom), anti-CD3 antibody (Santa Cruz Biotechnology, Santa Cruz, Calif), or anti-Mac2 antibody (Accurate Chemical & Scientific Corp, Westbury, NY). Three lung tissue slides (1 slide per mouse) from each group were used for semiquantitative cell counts in peripheral lung tissue. These cell counts did not distinguish among cells in various components of the lung (eg, airspace, interstitial, or marginated) but included all cells in peripheral (alveolar) lung tissue. On each slide, the lung tissue was divided into 4 to 6 parallel zones, and 1 photo was taken at each zone where the target cells were found in the highest numbers at 100x magnification.
Measurement of Myeloperoxidase
Myeloperoxidase (MPO) was measured in BAL fluid by using a mouse MPO enzyme-linked immunosorbent assay kit (Cell Sciences, Canton, Mass).
Measurement of Cytokines/Chemokines
Cytokines/chemokines in BAL fluid were quantified by using the Bioplex Bead Array technique with a multiplex cytokine panel assay (Bio-Rad Laboratories, Hercules, Calif), as previously done by our laboratory.7 The samples were analyzed as instructed with the Bioplex array reader, which is a fluorescent-based flow cytometer using a bead-based multiplex technology, each of which is conjugated with a reactant specific for a different target molecule.
Statistical Analysis
All data are presented as the mean ± standard error of the mean. Data were compared with 1-way analysis of variance, followed by the Student's t test for unpaired data with the Bonferroni correction. Square roots of tissue cell counts were compared by using 1-way analysis of variance.
|
Results |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionConclusionReferences |
|---|
Changes in Circulating Leukocyte Numbers
Whole blood samples were collected from antibody-treated mice before ischemia and after 2 hours of reperfusion, and blood cells were counted with a HemaVet Hematology System (Figure 1
). In neutrophil-depleted mice there was a greater than 80% reduction in the number of neutrophils (P < .05); however, there was also a significant reduction in monocytes (80%) compared with that seen in immunoglobulin G (IgG) isotype control mice (Figure 1, A). After reperfusion, the total number of circulating white blood cells and lymphocytes was significantly reduced in both IgG control and neutrophil-depleted mice compared with that seen before ischemia (Figure 1, B). Monocytes were further significantly reduced in IgG control mice after reperfusion but not in neutrophil-depleted mice. On the contrary, neutrophils were significantly increased by 2-fold after reperfusion in IgG control mice (compared with before ischemia) and not significantly increased in neutrophil-depleted mice (Figure 1, B).
|
Changes in Leukocyte Numbers and BAL MPO Levels
In sham and antibody-treated mice leukocytes in peripheral (alveolar) lung tissue were semiquantitatively evaluated in the left lung by means of immunohistochemistry. There were no significant differences in macrophage numbers between antibody-treated (after IRI) and sham mice (Table 1
and Figure 2, B
, bottom row). Both CD4+ and CD8+ T cells express CD3 antigen, and thus immunohistochemistry with anti-CD3 antibody was used to assess combined CD4+ and CD8+ T cells. CD3+ T cells were significantly increased after IRI in IgG control and neutrophil-depleted mice compared with those seen in sham mice (Table 1). CD3+ T-cell counts were not altered in CD8-depleted mice but were significantly reduced in CD4-depleted mice (Table 1 and Figure 2, B, middle row). Neutrophil numbers were significantly increased in IgG control and CD8- and CD4-depleted mice but not in neutrophil-depleted mice compared with those seen in sham mice. However, neutrophil numbers were significantly lower in CD4-depleted mice compared with those seen in IgG control and CD8-depleted mice (Table 1 and Figure 2, B, top row). MPO levels in BAL fluid, an indicator of neutrophil infiltration into alveolar airspace, showed no difference between sham, neutrophil-depleted, and CD4-depleted mice but were significantly increased in IgG control and CD8-depleted mice (Figure 2, A).
|
|
|
|
Changes in BAL Fluid Cytokine/Chemokine Expression
CCL3 (macrophage inflammatory protein 1), tumor necrosis factor (TNF-), IL-17, and CXCL1 (KC) levels in BAL fluid were significantly increased after IRI in IgG control mice and neutrophil- and CD8-depleted mice (Figure 5
). No significant induction of IFN- or IL-12 was observed after IRI (data not shown). Neutrophil depletion significantly reduced expression of IL-17. Depletion of CD4+ T cells resulted in significantly reduced CCL3, TNF-, IL-17, and CXCL1 levels versus those seen in IgG control mice. Depletion of CD8+ T cells did not significantly alter cytokine/chemokine expression versus that seen in IgG control mice.
|
|
Discussion |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionConclusionReferences |
|---|
, IL-17, and CXCL1 (KC) production after reperfusion, which were all significantly reduced by depletion of CD4+ T cells. Taken together, these results indicate that both neutrophils and CD4+ T cells contribute importantly to acute lung IRI.
Mouse Model of Lung IRI
Inflammatory responses during lung IRI play a critical role in early graft failure after lung transplantation.1,2,4,9,11,14,18 Most in vivo lung IRI models entail ligation of the left hilum for 60 to 90 minutes, followed by release of the ligature. Here the mice are typically maintained on ventilation until some point during reperfusion, and thus ventilator-induced lung injury could have significantly contributed to these models.19 The current study used a model with similar surgical intervention; however, the mechanical ventilation time was shortened to less than 20 minutes to minimize the potential of ventilator-induced lung injury. The absence of ventilator-induced injury is reflected in the current study by the sham mice, which display minimal lung injury and well-preserved lung function compared with that seen in the IRI group. In addition, lung function is relatively stable throughout 2 hours of continuous ventilation and perfusion in sham mice (data not shown), which would be expected to increase significantly over time if ventilator-induced injury was a significant component of this model.
Neutrophils Are End-effectors During Lung IRI
Studies have suggested that neutrophil accumulation appears to be the prime cellular mediator of pulmonary tissue destruction during IRI.1-4 Lung IRI induces all the characteristics of an acute inflammatory response, such as oxidative stress; activation of complement, macrophages, and mast cells; elaboration of cytokines and chemokines; release of chemotactic factors; expression of cell adhesion molecules; neutrophil infiltration; and pulmonary necrosis.1,3,4,6,10,20,21 Compelling evidence from a variety of animal models and clinical studies indicates that neutrophils are the principle end-effectors of IRI.3,20,21 Significant numbers of activated neutrophils accumulate in the lung after reperfusion, as determined by means of tissue immunostaining10 or increased activity of MPO,20 which is found almost exclusively within neutrophils.22 Neutrophils are generally believed to exacerbate tissue injury through the release of a variety of cytotoxic mediators, such as reactive oxygen species and proteases.23,24 The current study is entirely consistent with a critical role of neutrophils in causing lung IRI. More than 80% of neutrophils were successfully depleted with anti-Gr-1 antibody. However, in contrast to other reports,15 a significant number of circulating monocytes (80%) was also lost (Figure 1, A). The reduction in monocytes might have been due to the use of intravenous injection rather than intraperitoneal injection, as used previously.15 There was no significant reduction in lymphocytes. In the neutrophil-depleted mice lung IRI was significantly reduced, despite significant infiltration of CD3+ T cells. Furthermore, we found that protection from lung IRI correlated with a significant reduction in neutrophil infiltration but not other subtypes of leukocytes. These results demonstrate that activated neutrophils are end-effectors, which directly cause lung IRI. Although T lymphocytes might have some direct toxic effects on pulmonary tissues, they predominantly appear to amplify an inflammatory response that is chemotactic to neutrophils.
CD4+ T Lymphocytes Mediate Inflammatory Responses During Reperfusion
An increasing body of evidence has shown that T cells contribute importantly to lung IRI. T cells are activated and infiltrate into the lung during reperfusion earlier than neutrophils.10 Inhibition of T cells before reperfusion has been shown to attenuate inflammation and decrease lung IRI.20,25 These data are consistent with the current results, which suggest that CD4+ T cells, or a subset of these cells, are activated during lung IRI.
To identify the role of CD4+ T cells in lung IRI, we used mice with antibody-induced depletion of either CD4+ or CD8+ T cells. After reperfusion, significantly less infiltration of CD3+ T cells was found in CD4- and CD8-depleted mice than in IgG control and neutrophil-depleted mice (Table 1). In sham mice there were higher numbers of peripheral lung T cells and macrophages than neutrophils. There was no significant change in the number of alveolar macrophages after 2 hours of reperfusion in all IRI groups; however, in the IgG control group there was a further increase in the number of infiltrating neutrophils and T cells, with the change of neutrophils being greater (Table 1). This increase in T-cell numbers might represent chemotaxis of circulating lymphocytes because circulating lymphocytes were significantly reduced during reperfusion. Although significant numbers of neutrophils are trapped (marginated or infiltrated) in the lungs, circulating neutrophil numbers were increased after reperfusion (statistical significance was reached only in IgG control mice). The reason for the increase in circulating neutrophil numbers is not clear but possibly reflects mobilization from bone marrow in response to granulocyte–macrophage colony-stimulating factor released by activated CD4+ T cells. Compared with CD8+ T cell–depleted mice, CD4+ T cell–depleted mice had significantly less infiltration of CD3+ T cells and neutrophils with correspondingly less lung IRI. However, neutrophil-depleted mice still exhibited increased numbers of CD3+ T cells, as seen in IgG control mice. Thus neutrophils have no effect on T-cell activation during lung IRI. Taken together, these results demonstrate that CD4+ T cells and not CD8+ T cells are activated during lung IRI and play a critical role in amplifying an inflammatory response, which culminates in the activation of neutrophils.
In lung IRI CD4+ T cells could become activated through either antigen-independent or antigen-dependent pathways. Antigen-independent mechanisms for T-cell activation have been described, which involve IL-12, TNF-, CCL2, CCL3, CCL5, and IFN-–inducible protein 10.12,13,26 We found that CCL3, TNF-, IL-17, and CXCL1 were significantly increased after IRI in all antibody-treated mice except CD4-depleted mice. In CD4-depleted mice production of IL-17 and CXCL1 were significantly lower than seen in other antibody-treated mice after IRI (except for IL-17 in neutrophil-depleted mice), indicating that CD4+ T-cell activation precedes CXCL1 induction. These results are consistent with an antigen-independent pathway leading to CD4+ T-cell activation during lung IRI. IL-17 activates alveolar macrophages and epithelial type II cells to induce CXCL1, which is a strong chemotactic mediator for neutrophil infiltration.7,27 Another possibility is that CD4+ T cells are activated directly by factors produced during reperfusion to release cytokines/chemokines that transactivate other leukocytes and epithelial cells.
Clinical Effect
Identification of the initiatory signaling cascade through CD4+ T cells and macrophages will likely lead to specific pharmacologic interventional targets for the amelioration of lung IRI. For example, recent studies by our group have identified one such potential therapeutic agent to be the A2A adenosine receptor, which resides on leukocytes, including CD4+ T cells and neutrophils. Here we have shown that agonists that specifically activate A2A adenosine receptors significantly reduce neutrophil infiltration and attenuate lung IRI after transplantation.4,28 The possible protective role of A2A adenosine receptors on CD4+ T cells in the setting of lung IRI and the underlying molecular mechanisms are yet to be elucidated and are currently under investigation.
|
Conclusion |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionConclusionReferences |
|---|
|
Footnotes |
|---|
|
References |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionConclusionReferences |
|---|
This article has been cited by other articles:
|
|
 |
|
  D. P. Mulloy, A. K. Sharma, L. G. Fernandez, Y. Zhao, C. L. Lau, I. L. Kron, and V. E. Laubach Adenosine A3 Receptor Activation Attenuates Lung Ischemia-Reperfusion Injury Ann. Thorac. Surg., May 1, 2013; 95(5): 1762 - 1767. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 K. Tsutahara, M. Okumi, Y. Kakuta, T. Abe, K. Yazawa, S. Miyagawa, K. Matsunami, H. Otsuka, J. Kaimori, S. Takahara, et al. The blocking of CXCR3 and CCR5 suppresses the infiltration of T lymphocytes in rat renal ischemia reperfusion Nephrol. Dial. Transplant., October 1, 2012; 27(10): 3799 - 3806. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 D. J. LaPar, V. E. Laubach, A. Emaminia, I. K. Crosby, V. A. Hajzus, A. K. Sharma, H. M. Sumner, D. V. Webb, C. L. Lau, and I. L. Kron Pretreatment strategy with adenosine A2A receptor agonist attenuates reperfusion injury in a preclinical porcine lung transplantation model J. Thorac. Cardiovasc. Surg., October 1, 2011; 142(4): 887 - 894. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. K. Sharma, D. J. LaPar, Y. Zhao, L. Li, C. L. Lau, I. L. Kron, Y. Iwakura, M. D. Okusa, and V. E. Laubach Natural Killer T Cell-derived IL-17 Mediates Lung Ischemia-Reperfusion Injury , June 1, 2011; 183(11): 1539 - 1549. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. Pelletier, A. Micheletti, and M. A. Cassatella Modulation of human neutrophil survival and antigen expression by activated CD4+ and CD8+ T cells J. Leukoc. Biol., December 1, 2010; 88(6): 1163 - 1170. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
F. Anvari, A. K. Sharma, L. G. Fernandez, T. Hranjec, K. Ravid, I. L. Kron, and V. E. Laubach Tissue-derived proinflammatory effect of adenosine A2B receptor in lung ischemia-reperfusion injury J. Thorac. Cardiovasc. Surg., October 1, 2010; 140(4): 871 - 877. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 R. K. Tadagavadi, W. Wang, and G. Ramesh Netrin-1 Regulates Th1/Th2/Th17 Cytokine Production and Inflammation through UNC5B Receptor and Protects Kidney against Ischemia-Reperfusion Injury J. Immunol., September 15, 2010; 185(6): 3750 - 3758. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. F. Rodrigues and D. N. Granger Role of blood cells in ischaemia-reperfusion induced endothelial barrier failure Cardiovasc Res, July 15, 2010; 87(2): 291 - 299. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
H. Sahara, A. Shimizu, K. Setoyama, M. Okumi, M. Oku, E. Samelson-Jones, and K. Yamada Carbon monoxide reduces pulmonary ischemia-reperfusion injury in miniature swine J. Thorac. Cardiovasc. Surg., June 1, 2010; 139(6): 1594 - 1601. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. K. Sharma, V. E. Laubach, S. I. Ramos, Y. Zhao, G. Stukenborg, J. Linden, I. L. Kron, and Z. Yang Adenosine A2A receptor activation on CD4+ T lymphocytes and neutrophils attenuates lung ischemia-reperfusion injury J. Thorac. Cardiovasc. Surg., February 1, 2010; 139(2): 474 - 482. [Abstract] [Full Text] [PDF]
|
|
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
| ANN THORAC SURG | ASIAN CARDIOVASC THORAC ANN | EUR J CARDIOTHORAC SURG |
| J THORAC CARDIOVASC SURG | ICVTS | ALL CTSNet JOURNALS |
P < .05 versus neutrophil-depleted mice.