Purpose: Normothermic ex vivo lung perfusion (EVLP) of high risk donor lungs prior to transplantation results in outcomes similar to that of con- temporaneous lung transplantation. The underlying mechanism for this effect remains to be identified. We performed a time-course microarray analysis on rejected donor lungs subjected to 12 h of EVLP to further investigate the mechanism. Methods and Materials: Rejected human lungs were subjected to 12 h of EVLP(n6). Tissue biopsies taken at time zero and every three hours hence were snap-frozen and stored at 80C, resulting in a total of 30 samples. RNA was extracted from all samples (RIN 7) and utilized for microarray (Affy U133plus2) analysis. Preprocessing was performed using the PLIER method, followed by time-course analysis and clustering using the STEM method. Gene ontology was utilized on the resulting gene- clusters to identify the enriched functional modules and pathways. Results: About 6% of all genes showed at least 2-fold variability across the time-course (2013 genes). Of these, about two-thirds (1356/2013) were assigned by STEM to a statistically significant cluster, indicating a coher- ent temporal trend. More genes showed decreasing expression (58.1%; 788 genes) than increasing expression (33.8%; 458 genes). A small fraction showed transient changes (8.1%; 110 genes). Down-regulated genes were enriched for genes involved in immune response (p0.001), for oxi- doreductases(p0.002), and for genes involved in the defense respon- se(p0.016) and include genes such as CCL5, CCR2, CCR5, CCL17, TLR7, and TLR5. In contrast, genes which increased in expression during EVLP were enriched for those involved in signal transduction (p0.001) and include genes such as SMAD3, HMGA1, and FOXE3. Conclusions: During EVLP, genes involved in the immune and inflam- matory response become downregulated over time. In contrast, genes involved in the regulation of cellular signaling become upregulated during EVLP. These changes may pre-condition the lungs to better tolerate rep- erfusion post-transplantation. 297 Non-HLA Antibody Screening after Heart Transplantation Identifies High Risk for Cardiac Allograft Vasculopathy M.J. Barten, 1 D. Dragun, 2 M.-T. Dieterlen, 1 S. von Salisch, 1 J. Garbade, 1 S. Klein, 1 S. Dhein, 1 F.W. Mohr, 1 H.B. Bittner. 11 Clinic for Cardiac Surgery, Heart Center Leipzig, Leipzig, Germany; 2 Clinic for Nephrology and Intensive Care Medicine, Charite Campus Virchow Clinic, Berlin, Germany. Purpose: Cardiac allograft vasculopathy (CAV) after heart transplantation (HTx) is a major therapeutic challenge, occuring in over 50% of HTx recipients in the first years after transplantation. Antibodies against human leukocyte antigens (HLA) or non-HLA antigens like major histocompati- bility complex class I-related chain A (MICA), angiotensin type 1 receptor (AT1R) or endothelin receptor A (ETAR) gain in importance as modula- tors of allograft function and survival. Methods and Materials: Sera of 114 HTx recipients were screened post- transplantation by Luminex-technology for HLA and MICA antibodies and by ELISA for AT1R and ETAR antibodies. For statistical analysis gender, age, status of CAV, PRA level before HTx and the number of blood transfusions was documented. Results: CAV was detected in n43 recipients. There was no significant difference in gender and number of blood transfusions between recipients with or without antibodies. HTx recipients developed antibodies against HLA class I or class II (12.9%) to a lower extend than against non-HLA antigens, especially against AT1R (35.3%) and ETAR (47.4%). CAV appeared in 27.1% of recipients with non-HLA antibodies, whereas 5.8% of the recipients with HLA antibodies developed CAV. Significant more recipients with CAV were positive for AT1R and ETAR antibodies, 30.2% and 37.2%, compared to CAV-positive recipients which were positive for HLA class I, HLA class II and MICA antibodies, 2.3%, 9.3% and 13.9% respectively (p0.05). Furthermore, recipients with non-HLA antibodies developed CAV earlier (69.1mo) than recipients without these antibodies (80.1mo). Conclusions: Non-HLA antibodies are linked to earlier and higher inci- dence of CAV after HTx. The screening of HLA and especially of non- HLA antibodies after HTx is recommended to identify patients with an increased risk for CAV. 298 Pre-Transplant Non-Cytotoxic Anti-HLA Antibodies Predict Multiple Adverse Events after Heart Transplantation L. Potena, 1 A. Bontadini, 2 F. Fruet, 2 S. Iannelli, 2 F. Barberini, 1 M. Nardozza, 1 V. Manfredini, 1 M. Masetti, 1 G. Magnani, 1 F. Grigioni, 1 A. Branzi. 11 Cardiovascular Department, University of Bologna, Bologna, Italy; 2 Immunogenetics Unit, Academic Hospital S. Orsola- Malpighi, Bologna, Italy. Purpose: Detection of positive panel reactive antibodies (PRA) in trans- plant candidates has been associated with poor outcome after transplant. High sensitive techniques now allow detecting low titer of anti HLA antibodies (HLA-ab) but their clinical impact on post-heart transplant (HT) follow-up is controversial. Methods and Materials: Aiming to clarify this issue, by using luminex screening, we assayed for HLA-ab pre-transplant sera from patients with negative cytotoxic PRA who received a HT in our center between 2000 and 2005, before luminex was available, and after we started to routinely use thymoglobuline induction. Of note, these patients received HT blinded for positive non-cytotoxic HLA-ab. Results: Out of the 174 consecutive patients tested (aged 5213y; 16% females; 47% with ischemic etiology), 33 (19%) tested positive for HLA- ab: 12 (7%) for both class I and II, 17 (10%) for class I only, and 4 (2%) for class II only. HLA-ab were much more common in female recipients (46% vs. 13%; P0.01). Survival rate at 5-y was significantly lower in patients with any HLA-ab as compared with negative patients (658 vs. 853%; P0.03), accounting for a doubled mortality risk, even after adjusting for female sex and other baseline characteristics (OR: 2.21; P0.04). In addition, anti-HLA ab detection was associated with increased prevalence of early graft failure (35 vs 15%; P0.05) and cellular rejection occurring later than year 1 after HT (29 vs. 11%; P0.03). Only a subgroup of 37 patients was tested for pathological signs of AMR during the follow-up. The 9 patients with pAMR grade 2 were more likely to have positive anti HLA ab before HT (P0.06). Conclusions: By retesting sera of historical patients who received HT, we found that detection of non-cytotoxic anti HLA antibody in HT candidates predicts several negative outcomes after transplant, including early graft failure, late cellular rejection, AMR, and 5 years mortality. Careful allo- cation methods and desensitization protocols need to be developed and prospectively tested in these patients. 299 Cardiac Allograft Remodeling Is Associated with Increased Inflammatory Burden of Coronary Atherosclerotic Plaque E. Raichlin, Y. Matsuo, S.S. Kushwaha, A.L. Clavell, R.P. Frantz, R.J. Rodeheffer, B.S. Edwards, B.A. Boilson, J.A. Schirger, A. Lerman, N.L. Pereira. William J. von Liebig Transplant Center, Mayo Clinic, Rochester, MN; Division of Cardiovascular Diseases, Center for Coronary Physiology and Imaging, Mayo Clinic, Rochester, MN; Division of Cardiovascular Diseases, Univarsity of Nebraska Medical Center, Omaha, NE. Purpose: Cardiac allograft remodeling (CAR) is associated with increased mortality after heart transplantation (HTx). The association between CAR and cardiac allograft vasculopathy (CAV) as assessed with 3D- VH (Vir- tual Histology) intravascular ultrasound (IVUS) has not been described. Methods and Materials: A total of 83 HTx recipients (mean age 53.515.7 years) underwent 3D VH-IVUS examination of the left anterior descending artery 4.12.9 years following HTx. Coronary plaques were defined as inflammatory(VH-IP: necrotic core and dense calcium 30%) and noninflammatory (VH-NIP: necrotic core and dense calcium 30%). Echocardiographic studies were performed at the time of IVUS. Cohort was divided into 2 groups based on determination of left ventricle mass and relative wall thickness: 1. NG - normal geometry (n15) and 2. CAR - cardiac allograft remodeling includes recipients with concentric remodel- ing (n40) and hypertrophy (n18). Results: Coronary vessel, lumen and plaque volumes were significantly greater in the CAR group as compared to the NG group but no difference in plaque index was observed. The percentage of necrotic core and the presence of VH-IP were significantly higher in the CAR group. S107 Abstracts