A recombinant soluble chimeric complement inhibitor composed of human CD46 and CD55 reduces acute cardiac tissue injury in models of pig-to-human heart transplantation.

BACKGROUND: Inasmuch as complement plays a critical role in many pathological processes and in xenograft rejection, efficient complement inhibitors are of great interest. Because the membrane-associated complement inhibitors are very effective, recombinant soluble molecules have been generated. METHODS: We tested the efficacy of complement activation blocker-2 (CAB-2), a recombinant soluble chimeric protein derived from human decay accelerating factor (DAF, CD55) and membrane cofactor protein (MCP, CD46), in two models of pig-to-human xenotransplantation in which tissue injury is complement mediated. The in vitro model consisted of porcine aortic endothelial cells and human serum, and the ex vivo model consisted of a porcine heart perfused with human blood. RESULTS: In vitro, addition of CAB-2 to serum inhibited cytotoxicity and the deposition of C4b and iC3b on the endothelial cells. Ex vivo, addition of CAB-2 to human blood prolonged organ survival from 17.3 +/- 6.4 min in controls to 108 +/- 55.6 min with 910 nM (100 microg/ml) CAB-2 and 219.8 +/- 62.7 min with 1820 nM (200 microg/ml) CAB-2. CAB-2 also retarded the onset of increased coronary vascular resistance. The complement activity of the perfusate was reduced by CAB-2, as was the generation of C3a and SC5b-9. The myocardial tissues had similar deposition of IgG, IgM, and Clq; however, CAB-2 reduced the deposition of C3, C4, and C9. Hearts surviving >240 min demonstrated trace to no deposition of C9 and normal histologic architecture. CONCLUSION: These results indicate that CAB-2 can function as an inhibitor of complement activation and markedly reduce tissue injury in models of pig-to-human xenotransplantation and thus may represent a useful therapeutic agent for xenotransplantation and other complement-mediated conditions.

Potential of anticomplement therapy in clinical transplantation.

The activation of complement contributes to tissue damage in many ways. Prevention of complement activation in organ transplantation has largely centred on studies in xenotransplantation, where complement plays a key role in the pathogenesis of hyperacute rejection. Transgenic porcine organs expressing human regulators of complement activation in combination with soluble inhibitors as well as appropriate immunosuppression may be sufficient to alleviate the major immunological barriers that currently prevent the use of xenogeneic organs for human transplantation.

Risk factors for the development of bronchiolitis obliterans syndrome after lung transplantation.

OBJECTIVE: This study identifies specific clinical and immunologic factors in lung transplant recipients that influence the subsequent development of chronic allograft dysfunction. METHODS: The study group consisted of 132 consecutive patients who received lung allografts (76 single, 25 bilateral single, and 31 heart-lung) and survived at least 90 days. One hundred twenty-one patients were used in the analysis that modeled time to development of histologic obliterative bronchiolitis or bronchiolitis obliterans syndrome. RESULTS: Variables noted to have an effect on the time to development of bronchiolitis obliterans syndrome included cytomegalovirus pneumonitis (RR = 3.2, p = 0.001), late acute rejection (RR = 1.3, p = 0.02), human leukocyte antigen mismatches at the A loci (RR = 1.8, p = 0.02), total human leukocyte antigen mismatches (RR = 1.4, p = 0.04), and absence of donor antigen-specific hyporeactivity (52% vs 100% survival free from bronchiolitis obliterans syndrome at 2 years; p = 0.005). Cytomegalovirus pneumonitis had a significant effect on time to obliterative bronchiolitis (RR = 3.6, p = 0.0005), as did donor antigen-specific hyporeactivity (52% vs 100% survival free from obliterative bronchiolitis at 2 years; p = 0.01). In multivariate analysis, cytomegalovirus pneumonitis (RR = 3.2, p = 0.02), human leukocyte antigen mismatches at the A loci (RR = 2.4, p = 0.006), and late acute rejection (RR = 1.3, p = 0.02) were identified as predictors of bronchiolitis obliterans syndrome. Cytomegalovirus pneumonitis was associated with time to development of histologic obliterative bronchiolitis (RR = 2.3, p = 0.02). CONCLUSIONS: Several risk factors were associated with the development of chronic allograft dysfunction, which, in turn, had a significant impact on long-term survival. Early identification of lung allograft recipients with risk factors for the development of bronchiolitis obliterans syndrome may allow modification in immunosuppression and antiviral therapy to potentially decrease the prevalence of this disorder.

Early and late airway complications after lung transplantation: incidence and management.

BACKGROUND: Airway anastomosis complications continue to be a source of morbidity for lung transplant recipients. METHODS: This study analyzes incidence, treatment, and follow-up of airway anastomotic complications occurring in 127 consecutive lung transplant airway anastomoses (77 single lung and 25 bilateral sequential lung). Complications were categorized as stenosis (11), granulation tissue (8), infection (7), bronchomalacia (5), or dehiscence (3). Follow-up after treatment ranged from 6 months to 4 years. RESULTS: Nineteen airway anastomosis complications (15.0%) occurred in 18 patients. Telescoping the airway anastomosis reduced the complication rate to 12 of 97 (12.4%), compared with 7 of 30 (23.3%) for omental wrapping, (p = 0.15). Complications developed in 13 of 77 single-lung airway anastomoses (16.9%) versus 6 of 50 bilateral sequential lung recipients (12.0%). Treatment consisted of stenting (9 airway anastomoses), bronchodilation (8), laser debridement (4), rigid bronchoscopic debridement (2), operative revision (2), and growth factor application (2). There was no difference in actuarial survival between patients with or without airway anastomosis complications (p = 1.0). CONCLUSIONS: Airway anastomosis complications can be successfully managed in the immediate or late postoperative period with good outcome up to 4 years after intervention.

Heart transplantation for patients over age 60.

As the proportion of people over age 60 in our society continues to rise, so does the number of potential heart transplant candidates. Advanced recipient age, however, has long been used as an exclusion criterion for transplantation, though the upper age limit remains poorly defined. The purpose of our study was to analyze the outcomes of 31 heart transplant recipients over age 60 at our institution. They were followed with regard to early and late morbidity and mortality. Mean follow-up time was 50 months. The 1- and 5-yr survival rates were 90% and 85%. These rates were not significantly different compared with younger (age 18 to 59) recipients transplanted in the same time period. The older recipients more frequently developed osteoporotic changes as well as cutaneous and visceral malignancies, but had infrequent rejection episodes. Overall, heart transplantation is safe and effective for patients over age 60 with end-stage congestive heart failure. Both 1- and 5-yr survival rates are well within the acceptable range and do not differ significantly from younger recipients. Individualized immunosuppression might help reduce the incidence of malignancy in older recipients, but further studies are needed. Matching donor and recipient age would make the best use of available organs.

Coronary sinus injury during retrograde cardioplegia: a report of three cases.

Coronary sinus injuries related to the use of retrograde cardioplegia are uncommon. In most cases injuries are encountered with overinflation of the coronary sinus catheter balloon or traumatic catheter insertion. This article describes three cases of coronary sinus injury during retrograde cardioplegia administration in patients with ventricular hypertrophy, while the heart was manually retracted to expose the posterior myocardium. We propose that the risk of coronary sinus injury during retrograde cardioplegia, in patients with left ventricular hypertrophy, can be minimized by avoiding excessive retraction of the heart, deflation of the retrograde catheter during retraction, and the use of a left ventricular vent.

Primary pulmonary hypertension as a risk factor for the development of obliterative bronchiolitis in lung allograft recipients.

STUDY OBJECTIVES: Obliterative bronchiolitis (OB) is a major factor limiting long-term survival after lung transplantation. The etiology of this disease process remains incompletely understood. Several risk factors have been identified previously, including acute rejection and cytomegalovirus pneumonitis. The purpose of this study was to evaluate primary pulmonary hypertension (PPH) as a potential risk factor for the development of OB after lung transplantation. DESIGN AND PATIENTS: We retrospectively analyzed 107 lung allograft recipients (28 heart-lung, 18 bilateral sequential single-lung, 61 single-lung) who underwent transplantation between May 1, 1986, and April 30, 1994, and survived at least 3 months posttransplant. Mean follow-up posttransplant was 28.6 months (range, 3.5 to 99 months). Actuarial survival was estimated for patients with or without PPH and for those who did or did not develop OB. RESULTS: In all, 25 patients (23.4%) developed OB, diagnosed by strict histologic criteria. Of 23 patients with PPH, 9 (39.1%) developed OB, compared with 16 (19.0%) of 84 patients without PPH (p = 0.044). Actuarial survival, sex, time on waiting list, and follow-up posttransplant were not significantly different between groups. PPH was the major determinant for the development of OB (p = 0.0468) when evaluating PPH and cytomegalovirus pneumonitis together as risk factors. Patients with PPH also developed OB significantly earlier posttransplant, compared with patients with other primary disease (p = 0.05). CONCLUSIONS: Patients with PPH who undergo lung transplantation are at increased risk for the development of OB, which also occurs at a shorter time interval posttransplant. This subgroup needs aggressive monitoring for diagnosis and treatment of OB.

Unilateral volume reduction after single-lung transplantation for emphysema.

BACKGROUND: Single-lung transplantation has become accepted therapy for patients with end-stage emphysema. Hyperinflation of the native lung can occur after single-lung transplantation with mediastinal shifting and compression of the transplanted lung. A volume reduction operation (pneumectomy) [corrected] may relieve symptoms of dyspnea and improve exercise tolerance. METHODS: Three of 66 patients who underwent single-lung transplantation for emphysema had development of native lung hyperexpansion and mediastinal shifting causing compression of the transplanted contralateral lung at 12, 17, and 42 months after transplantation. There were 2 men and 1 woman. Unilateral volume reduction was performed without complication in all 3 patients. RESULTS: All patients were noted to have marked improvement in chest radiographs after volume reduction, substantial relief of dyspnea, and improvement in exercise tolerance. An improvement in pulmonary function test results was noted in 1 patient, but tests were not done for the other 2 patients. CONCLUSIONS: Patients with chronic obstructive pulmonary disease who undergo single-lung transplantation may have symptomatic hyperexpansion of the native lung requiring volume reduction months to years after transplantation. Unilateral volume reduction can be safely performed in the posttransplantation period.

Selective IgM depletion prolongs organ survival in an ex vivo model of pig-to-human xenotransplantation.

In the pig-to-primate model, xenograft hyperacute rejection (HAR) is mediated by antibody and complement. Previous studies have implicated xenoreactive IgM natural antibody (nAb) as the predominant immunoglobulin involved in HAR. To further evaluate the role of IgM, we selectively reduced IgM levels in human blood, without changing IgG and IgA levels, and then used this blood to perfuse porcine hearts ex vivo. Specific IgM depletion was accomplished with an immunoabsorption column containing sheep anti-human IgM (mu-chain specific) conjugated to Sepharose beads. Human blood was separated into plasma and cellular components. For control experiments, those components were unmodified and recombined in the perfusion system. For experiments with IgM reduced blood, the plasma was passed through the IgM column. Immunoabsorption resulted in approximately 90% reduction in xenoreactive IgM levels, as measured by ELISA. Porcine hearts perfused with unmodified human blood survived 25 +/- 5.6 min (n=5). Porcine hearts perfused with human blood containing reduced levels of IgM survived 229 +/- 45.2 min (n=4; P<0.01). Organ survival was negatively associated with xenoreactive IgM nAb levels measured immediately before perfusion (r=-0.83; P=0.01), and not with IgG nAb levels (r=-0.21; P=0.62). The ability of plasma from IgM-depleted blood to elicit complement activation, measured by iC3b binding to porcine aortic endothelial cells in vitro, was also strongly associated with IgM xenoreactive nAb levels (r=0.92; P<0.0001). Control hearts perfused with unmodified human blood showed typical widespread histologic features of HAR, while porcine hearts perfused with IgM-reduced blood demonstrated milder and less uniform changes. Immunopathological analysis of heart tissues obtained at the completion of each study showed similar deposition of IgG between groups but markedly less IgM, C3, C4, and C9 in the IgM reduction group. These results suggest that selective IgM reduction delays HAR with prolongation of survival and that xenoreactive IgM may be the predominant immunoglobulin involved in HAR in the pig-to-human combination.

Expression of human CD59 in transgenic pig organs enhances organ survival in an ex vivo xenogeneic perfusion model.

The serious shortage of available donor organs for patients with end stage organ failure who are in need of solid organ transplantation has led to a heightened interest in xenotransplantation. The major barrier to successful discordant xenotransplantation is hyperacute rejection. Hyperacute rejection results from the deposition of preformed antibodies that activate complement on the luminal surface of the vascular endothelium, leading to vessel occlusion and graft failure within minutes to hours. Endogenous membrane-associated complement inhibitors normally protect endothelial cells from autologous complement -- however, these molecules are species-restricted and therefore are ineffective at inhibiting activated xenogeneic complement. To address the pathogenesis of hyperacute rejection in the pig-to-human combination, F1 offspring were generated from a transgenic founder animal that was engineered to express the human terminal complement inhibitor hCD59. High-level cell surface expression of hCD59 was detected in the hearts and kidneys of these transgenic F1 animals, similar to expression levels in human kidney tissue. The hCD59 was expressed on both large vessel and capillary endothelium. Ex vivo perfusion experiments, using human blood as the perfusate, were performed with transgenic porcine hearts and kidneys to evaluate the ability of hCD59 to inhibit hyperacute rejection. These experiments demonstrated that transgenic organs expressing hCD69 resisted hyperacute rejection, as measured by increased organ function for both the hearts and the kidneys, as compared with control pig organs. Hearts from hCD59-expressing animals demonstrated a five-fold prolongation in function compared with controls, 109.8 +/- 20.7 min versus 21.2 +/- 2.9 min (P = 0.164). The hCD59-expressing kidneys also demonstrated significantly prolonged function at 157.8 +/- 27.0 min compared with 60.0 +/- 6.1 min for controls (P = 0.0174). Deposition of C9 neoantigen In the vasculature of porcine organs perfused with human blood was markedly reduced in organs expressing hCD59. These studies demonstrate that C5b-9 plays an important role in hyperacute rejection of a porcine organ perfused with human blood and suggest that donor pigs transgenic for hCD59 may be an integral component of successful clinical xenotransplantation.

Does donor organ ischemia over four hours affect long-term survival after lung transplantation?

BACKGROUND: Lung transplantation is currently limited by a donor shortage and the need for a short organ ischemic time. The purpose of this analysis was to evaluate prolonged donor organ ischemia and its effect on overall survival. METHODS: We conducted a retrospective analysis of 83 patients undergoing single (n = 62) or bilateral sequential lung transplantation (n = 21) from June 1, 1989, through July 31, 1994. All allografts were flushed with modified EuroCollins solution at 4 degrees C and stored in cold saline solution. Ischemic time was measured from aortic crossclamping at organ procurement to reperfusion. Ischemic times were divided into three groups: group I < 240 minutes (n = 39), group II 240 to 360 minutes (n = 36), and group III > 360 minutes (n = 8). Ischemic times ranged from 97 to 708 minutes (median, 245 minutes; mean, 252 minutes). Bilateral sequential and single lung transplantations were considered together. RESULTS: Actuarial survival was not significantly different among groups (p = 0.09). We found no significant difference in time spent in the intensive care unit (p = 0.27) or in total hospital stay (p = 0.57) after transplantation, in forced expiratory volume in 1 second at 1 month after transplantation (p = 0.74), or in the number of acute rejection episodes (p = 0.65). In addition, length of follow-up was similar among groups (p = 0.24). CONCLUSIONS: Prolonged donor allograft ischemic times were not associated with an adverse effect on survival. The use of allografts with ischemic times through 6 hours achieved acceptable 2-year survival rates after transplantation. The use of donor organs with prolonged ischemic times should prompt the United Network for Organ Sharing to move toward better allocation of donor organs.

Complement inhibition with an anti-C5 monoclonal antibody prevents acute cardiac tissue injury in an ex vivo model of pig-to-human xenotransplantation.

Prevention of hyperacute xenograft rejection in the pig-to-primate combination has been accomplished by removal of natural antibodies, complement depletion with cobra venom factor, or prevention of C3 activation with the soluble complement inhibitor sCR1. Although these strategies effectively prevent hyperacute rejection, they do not address the relative contribution of early (C3a, C3b) versus late (C5a, C5b-9) activated complement components to xenogeneic organ damage. To better understand the role of the terminal complement components (C5a, C5b-9) in hyperacute rejection, an anti-human C5 mAb was developed and tested in an ex vivo model of cardiac xenograft rejection. In vitro studies demonstrated that the anti-C5 mAb effectively blocked C5 cleavage in a dose-dependent manner that resulted in complete inhibition of both C5a and C5b-9 generation. Addition of anti-C5 mAb to human blood used to perfuse a porcine heart prolonged normal sinus cardiac rhythm from a mean time of 25.2 min in hearts perfused with unmodified blood to 79,296, or > 360 min when anti-C5 mAb was added to the blood at 50 micrograms/ml, 100 micrograms/ml, or 200 micrograms/ml, respectively. In these experiments, activation of the classical complement pathway was completely inhibited. Hearts perfused with blood containing the highest concentration of anti-C5 mAb had no histologic evidence of hyperacute rejection and no deposition of C5b-9. These experiments suggest that the activated terminal complement components C5a and C5b-9, but not C3a or C3b, play a major role in tissue damage in this porcine-to-human model of hyperacute rejection. They also suggest that targeted inhibition of terminal complement activation by anti-C5 mAbs may be useful in clinical xenotransplantation.

Deep venous thrombosis and pulmonary embolism after lung transplantation.

The incidence of deep venous thrombosis or pulmonary embolism after lung or heart-lung transplantation has not been well defined. Pulmonary embolism may be of particular concern in the postoperative period owing to an inadequately developed or absent collateral bronchial circulation and potential risk of pulmonary infarction. Fourteen (12.1%) of 116 patients undergoing either lung (n = 87) or heart-lung (n = 29) transplantation developed thromboembolic complications 10 days to 36 months after operation. Deep vein thrombosis developed in nine patients, including three with upper body thrombosis related to indwelling central venous catheters. Seven patients (6%) had pulmonary embolism, and three of them died. Resolution of pulmonary embolism was successfully accomplished by selective pulmonary artery infusion of urokinase in three patients without complications. Our experience indicates that deep vein thrombosis and pulmonary embolism are significant problems after lung transplantation. Mortality is high in those patients in whom pulmonary embolism develops. Therefore, a comprehensive prevention protocol is warranted.

Antibody removal by column immunoabsorption prevents tissue injury in an ex vivo model of pig-to-human xenograft hyperacute rejection.

Hyperacute rejection of a pig-to-primate organ xenograft is triggered by binding of anti-pig endothelial cell antibodies to the vascular endothelium of the xenograft and complement activation. Xenograft survival can be prolonged by pretransplant depletion of antibody with plasmapheresis or organ perfusion. However, these techniques have disadvantages for use immediately pretransplant or in the post-transplant period, including a marked reduction in coagulation proteins. To remove IgM and IgG from human plasma we employed a reusable Ig-binding column containing polyclonal anti-human IgG (heavy chain- and light chain-specific) conjugated to Sepharose beads (Therasorb, Baxter Corp.). Human blood was separated into plasma and cell fractions. Column absorption of plasma followed by recombination of plasma and cell fractions in the perfusion system resulted in 90.5 and 86.0% reduction in total IgG and IgM, respectively, and in a 47.0 and 69.4% reduction in IgG and IgM anti-pig endothelial cell antibodies, respectively. When the cellular fraction was recombined with untreated plasma and used to perfuse pig hearts in an ex vivo perfusion system, there was rapid cessation of normal cardiac rhythm (25.2 +/- 5.6 min) and intense deposition of Igs, complement proteins, and fibrin in the tissues. In contrast, perfusion with blood containing column-absorbed plasma was able to sustain cardiac function, with normal sinus rhythm maintained for 258 +/- 48.1 min, without tissue deposition of IgM or complement proteins and minimal deposition of IgG. We conclude that column absorption can be used effectively to deplete plasma of anti-pig endothelial cell antibodies.(ABSTRACT TRUNCATED AT 250 WORDS)

A model of unilateral pulmonary lobar transplantation.

Unilateral lung transplantation has become an accepted treatment for patients with end-stage pulmonary disease. Donor shortage, however, is a major limitation, with up to 87% of patients dying of their pulmonary disease while awaiting transplantation. This is especially true in neonatal and pediatric patient populations. The use of organ segments from cadaveric or living donors may provide a solution. The purpose of this study, therefore, was to evaluate the function and hemodynamic response to pulmonary lobar transplantation using a swine model. Five transplants were performed for acute study, while 10 were performed for 6-week survival. The left lower lobe was harvested from a 70- to 75-kg donor animal. The lobe was then transplanted into a 20 to 25-kg recipient following left pneumonectomy. Graft function was determined by pulmonary arterial and venous blood gas analysis. Cardiac output, pulmonary pressure, and pulmonary vascular resistance were measured under two experimental conditions: (1) baseline and (2) with the right pulmonary artery occluded, forcing the entire cardiac output through the lobar graft. All grafts showed excellent acute and long-term function with regard to gas exchange. The lobar grafts, however, were characterized by high pulmonary vascular resistance both acutely and 6 weeks post-transplant. Contralateral pulmonary artery occlusion resulted in hemodynamic instability and right heart failure. No animal was able to be solely supported by the lobar transplant for more than one hour. These results have prompted a bilateral lobar transplant model and current studies are in progress.

Suicide right ventricle after lung transplantation for Eisenmenger syndrome.

Right ventricular outflow tract obstruction, or "suicide right ventricle," rarely has been observed after single or bilateral single-lung transplantation for the treatment of Eisenmenger syndrome. We describe our experience in 2 patients with Eisenmenger syndrome in whom right ventricular outflow tract obstruction developed, in 1 after single-lung transplantation and ventricular septal defect repair and in the other after bilateral single-lung transplantation. Both patients suffered progressive deterioration and hemodynamic instability that was unresponsive to aggressive medical therapy. Diagnosis was confirmed in both patients by transesophageal echocardiography. Operative intervention was undertaken 72 and 24 hours after transplantation, and consisted of myectomy and outflow tract patching. One patient survived; the other died intraoperatively. The index of suspicion for this problem should be high during the intraoperative performance of transesophageal echocardiography, as well as during direct gradient measurement, with consideration of immediate management of severe right ventricular outflow tract obstruction at the time of transplantation.