Clinical Profiles and Outcomes of Cardiac Transplant Recipients Using Allomap and Cylex Immuknow Assays

Abstracts S295 on early- and long-term survival were evaluated with the use of multivariable analysis. Results: PGD-RV was diagnosed in 3 patients (0.8%) and PGD-LV occurred in 38 patients (9.8%): mild (n= 2, 0.5%), moderate (n= 26, 6.7%), and severe (n= 10, 2.6%). On multivariable analysis, pre-operative extracorporeal membrane oxygenation (ECMO) support (relative risk, 4.05, 95% confidence interval [CI], 1.31-12.58; P= 0.015) and longer total ischemic time (>  3 hours) (relative risk, 2.60, 95% CI, 1.29-5.25; P= 0.008) emerged as significant and independent predictors for moderate-to-severe PGD-LV. During a median follow-up period of 57.3 months (interquartile range, 26.2-106.6), 79 patients (20.4%) died. Patients sustaining moderate-to-severe PGD-LV had a higher risk of early mortality as well as overall mortality during the whole study period: the risk of 30-day mortality of 21.88 (95% CI, 3.86124.08; P< 0.001) and the risk of overall mortality of 2.57 (95% CI, 1.315.04; P= 0.006). Conclusion: Moderate-to-severe PGD-LV defined by the new criteria is a good predictor for early- as well as long-term mortality. A refined preoperative management of the recipient in critical status and the better donor heart preservation may reduce the incidence of PGD-LV and improve the overall survival. 8( 09) Clinical Profiles and Outcomes of Cardiac Transplant Recipients Using Allomap and Cylex Immuknow Assays S.R. Krim, P.T. Campbell , S. Desai, S. Mandras, A. Bansal, H. Patel, C. Eiswirth, H.O. Ventura.  Cardiology, Ochsner Heart and Vascular Institute, New Orleans, LA. Purpose: This study aimed to examine how clinical profiles based on Allomap scores and Cylex can predict 1-year clinical outcomes (allograft rejection and infection) in patients with heart transplantation. Methods: Allomap scores and Cylex measured at 6 months post cardiac transplantation from 80 consecutive patients from a single heart transplant center. Clinical profiles were defined based on Allomap scores and Cylex; Profile 1 (Allomap< 34/Cylex:226-524); Profile 2 (Allomap< 34/Cylex< 225); Profile 3 (Allomap< 34/Cylex> 525); Profile 4 (Allomap > 34/Cylex:226-524); Profile 5(Allomap> 34/Cylex< 225); Profile 6 (Allomap> 34/Cylex> 525). Clinical outcomes included composite of acute cellular or antibody mediated rejection and infection at 1 year post-transplant. Results: In the total cohort, mean age was 52±12 years, 61% were males. The following clinical profiles were identified: Profile 1 (46%); Profile 2 (25 %); Profile 3 (9%); Profile 4 (10%); Profile 5 (9%) and Profile 6 (1 %). Patients with profile 5 (Allomap> 34/Cylex< 225) were more likely to reach the clinical end point of composite of acute cellular or antibody mediated rejection and infection at 1 year post-transplant when compared to patients with profile 4 (Allomap > 34/Cylex:226-524) (57.1% vs. 12.5% respectively). Allograft rejection and infection were more common in cardiac transplant recipients with Profiles 2 and 5. Interestingly, cardiac transplant recipients with Profile 4 (Allomap > 34/Cylex: 226-524) had the lowest rates of rejection and infection (0% and 12.5% respectively). Conclusion: The utilization of clinical profiles defined by Allomap and Cylex scores may be useful in predicting the presence clinical outcomes of allograft rejection, infection at 1 year post-transplant. 

8( 10) Inverse Monocytic Subset Profile in Blood and Tissue During Human Heart Transplant Rejection With a Simultaneous Predominance of M2 Macrophages at the Tissue Level T.P. van den Bosch ,1 M.D. Kraaij,1 K. Caliskan,2 A.A. Constantinescu,2 O.C. Manintveld,1 P.J. Leenen,3 C.C. Baan,1 M.C. van Groningen,4 A.T. Rowshani.1  1Internal Medicine, Erasmus Medical Center, Rotterdam, Netherlands; 2Cardiology, Erasmus Medical Center, Rotterdam, Netherlands; 3Immunology, Erasmus Medical Center, Rotterdam, Netherlands; 4Pathology, Erasmus Medical Center, Rotterdam, Netherlands. Purpose: Increased monocytic infiltration of heart transplant tissue is documented during acute rejection. It is still unclear how changes in circulating monocyte pool relate to counterpart alterations of monocyte-macrophage infiltrates on tissue level. We aimed to establish blood and tissue monocyte/ macrophage profiles using serial blood samples and endomyocardial biopsies in 10 heart transplant recipients experiencing rejection in comparison to time points prior to rejection intra-individually. Blood profiles of heart transplant recipients were compared with 33 healthy individuals using a cross-sectional approach. Methods: Flow cytometric expression of co-stimulatory and migrationrelated molecules; HLA-DR, CD40, CD80, and CD54 were studied on different monocyte subsets. TNF-α , IFN-γ , IL-1β , IL-6, IL-10, IL-12 production capacity of monocytes were measured after LPS stimulation. Using immunohistochemistry and immunofluorescence we studied the expression of CD14, CD16, CD56, CD68, CD80 and CD163 in endomyocardial biopsies. Digital imaging and confocal laser scan microscopy were used to quantify. Results: Increased percentages of classical CD14++CD16- monocytes and simultaneously decreased fractions of intermediate CD14++CD16+ and non-classical CD14+CD16++ monocytes signifies the subset composition of circulating monocytes in heart transplant recipients as compared to healthy individuals with no differences between rejection and non-rejection. However, rejection was reflected by significantly increased expression of CD54 and HLA-DR within the CD16+ monocyte pool indicative of a higher antigen presentation potential and migration capacity. Cytokine production potential was consistently high and independent of rejection. Significantly more CD16+ monocytes were found in rejecting endomyocardial biopsies compared to non-rejection. Significantly more CD68+ CD163+ M2 macrophages were documented during rejection parallel to this increase in intra-graft CD16+ monocyte infiltration. In contrary, CD68+CD80+ M1 macrophages did not differ between rejection and non- rejection. Conclusion: Our data show inverse monocytic subset profile in blood and tissue during human heart transplant rejection with a simultaneous predominance of M2 anti-inflammatory macrophages. 8( 11) Pathogenicity of Pre-Transplant Donor-Specific Anti-HLA Antibodies in Heart Transplant Recipients M. Kubanek ,1 T. Gazdic,2 E. Svobodova,3 A. Slavcev,3 I. Netuka,2 J. Pirk,2 I. Malek.1  1Cardiology, Institute for Clinical and Experimental Medicine, Prague, Czech Republic; 2Cardiac Surgery, Institute for Clinical and Experimental Medicine, Prague, Czech Republic; 3Immunogenetics, Institute for Clinical and Experimental Medicine, Prague, Czech Republic. Purpose: Presence of preformed donor-specific anti-HLA antibodies (DSA) has been associated with antibody mediated rejection (AMR), cardiac allograft vasculopathy (CAV) and unfavourable prognosis in heart transplant (HTx) recipients. We assessed several characteristics of pre-transplant DSA using a solid phase assay (Luminex®) and C1q assay (C1q Screen™) to predict their pathogenicity after heart transplantation (HTx). Methods: The study group included 264 HTx recipients [211 males (80%), age 50±12 years, pre-transplant VAD 68 (26%)], who had a negative prospective CDC crossmatch in case of panel reactive antibodies ≥ 10%.They received induction with antithymocyte globulin followed by standard immunosuppression (tacrolimus/cyclosporine + mycophenolate mophetil + prednisone). We analysed pre-transplant sera using a solid phase assay Luminex® and C1q binding capacity of DSA by C1qScreen™. Occurrence of pathological AMR, CAV and all-cause mortality were analysed during a median follow-up of 39 months (19-66).