- Email: [email protected]
Making a Case for EPLET MatchingEplet Matching Is Associated with Improved Baseline Spirometry and Reduced Risk of CLAD Following Lung Transplantation
S160
The Journal of Heart and Lung Transplantation, Vol 36, No 4S, April 2017
Methods: Plasma samples from LTx recipients (LTxRs) were analyzed by high-density protein microarrays containing 9000 unique full-length human proteins in native conformation. We pooled 12 LTxRs that developed CLAD and compared that with 11 LTxRs that remained CLAD-free. Longitudinal samples from LTxRs with CLAD were collected and stratified as pre-DSA, peak-DSA and DSA-resolved, and plasma samples from stable LTxRs that did not develop CLAD were stratified into 3, 9 and 15 month post-LTx periods. The samples were analyzed by Protoarray microchips and results were determined using Protoarray Prospector. The antigenic targets were analyzed by Ingenuity Pathway Analysis and DAVID Bioinformatics Resources. Results: Ab mapping by protein microarray revealed a unique repertoire in LTxRs that developed CLAD compared with stable LTxRs. Overall, CLAD patients demonstrated a > 10 fold higher Ab signal for 1756 proteins of which 35 proteins had > 50 fold higher signal intensities compared to that in stable LTxRs. Furthermore, peak-DSA plasma exhibited an expanded Ab repertoire compared to that of pre-DSA, which later contracted when DSA was resolved with reductions in both number of target antigens and signal intensities. In addition, de novo DSA was positively associated with the spread of nonHLA Ab repertoire. Conclusion: We demonstrate a vast repertoire of antigens involved in the development of immune responses following human LTx. Further, de novo DSA was strongly associated with development of immune responses to non-HLA SAgs. We conclude that de novo DSA is a major trigger for the development of immune responses to SAgs involved in the immunopathogenesis of CLAD. 4( 08) Early Elevation of Anti-HLA Immunoglobulin M Level Is Associated with Subsequent Lung Transplant Rejection and Worse Outcomes K. Miyahara , K. Miyoshi, T. Kurosaki, S. Otani, S. Sugimoto, M. Yamane, T. Oto. Department of Thoracic Surgery, Organ Transplant Center, Okayama University Hospital, Okayama-shi,Okayama prefecture, Japan. Purpose: Posttransplant immunoreactivity against allografts potentially differs widely in each case of transplantation. Early optimization of immunosuppressive intensity is crucial for later favorable outcome. However no useful noninvasive method exists for monitoring immunoreactivity. Immunoglobulin (Ig) M production stimulated by alloantigen is substantially sensitive to immunoreaction since primitive alloresponses can easily allow a mild B-cell response to produce IgM. The aim of this study is to examine relation between serum level of anti-human leukocyte antigen (HLA) IgM early after lung transplant and subsequent posttransplant outcomes. Methods: Consecutive 20 patients who underwent deceased lung transplantation in Okayama University Hospital from July 2013 to April 2015 were included. Immunoreactivity against HLA was retrospectively analyzed by detecting anti-HLA IgM of the serum sampled at day2, 4, 7, 10, and 14 after transplant. Flow cytometry crossmatching technique with Flow PRA HLA class I beads was used. The ratio of the level at each day to day2 (anti-classI IgM level) was obtained. The patients were divided into positive and negative groups according to the maximal IgM level during the first 14 days after lung transplantation. Results: Four patients exhibited significant elevation (max IgM level > 2.0, positive group) and 16 stayed negative throughout the monitoring period (negative group). In the positive group, the incidence of acute rejection during the first 30 days after transplant was significantly higher than negative group (p= 0.02). Two of four patients in positive group died of chronic lung allograft dysfunction (CLAD) at 9 and 13 months after lung transplantation respectively, while no patients in the negative group died of CLAD for 20 months after transplantation. IgM levels are independent of inflammatory event representing C-reactive protein value. Conclusion: Elevation of anti-HLA IgM levels during the first 14 days after lung transplantation may correlate to the higher incidence of later rejection and worse outcomes. Monitoring the level of anti-HLA IgM early after transplant can contribute to subsequent optimal immunosuppressive adjustments. 4( 09) Making a Case for EPLET Matching...Eplet Matching Is Associated with Improved Baseline Spirometry and Reduced Risk of CLAD Following Lung Transplantation
M. Paraskeva ,1 D.C. Walton,2 S.J. Hiho,2 J. Fuller,3 L.S. Cantwell,2 G.P. Westall,1 G.I. Snell.1 1Lung Transplant Service, Alfred Hospital, Melbourne, Australia; 2Victorian Transplantation and Immunogenetics Service, Australian Red Cross Blood Service, Melbourne, Australia; 3Monash University, Melbourne, Australia. Purpose: Increasingly HLA compatibility has been defined in terms of epitopes as compared to HLA antigen matching. We investigated the impact of eplet matching on the attainment of baseline or “best” spirometry following lung transplant and whether eplet matching predicted long term maintenance of baseline. Methods: All adult lung transplant recipients undergoing first bilateral lung transplantation between August 2008 and July 2012 with information available to allow eplet matching were included. Baseline FEV1 and FVC were defined as per ISHLT criteria. Normal spirometry was defined as FEV1 > 80% plus FVC> 80% plus FER> 70%. Results: 145 recipients were included. Those with < 40 eplet mismatches were more likely to have normal spirometry at baseline and took longer to achieve baseline FEV1 and FVC. Additionally they were less likely to develop CLAD.Conversely, those with ≥ 40 eplet mismatches were more likely to have abnormal spirometry at baseline and reach their peak FEV1 and FVC early.
FEV1 Best FVC Best HLA-DR/DQ Normal Abnormal Time (Mean Time (Mean Eplet MM Spirometry Spirometry days) days) CLAD
No CLAD
< 40 N= 65 77%(50) ≥ 40 N= 80 56%(45)
83%(54) 53%(42)
23%(15) 44%(35)
456 371
649 567
17%(11) 48%(38)
Conclusion: Eplet matching influences the attainment of baseline ("best") spirometry. Lung transplant recipients with fewer DR/DQ-eplet mismatches are more likely to normalise their spirometry and demonstrate long periods of good function with reduced incidence of CLAD.
4( 10) The Histomorphological Spectrum of Restrictive Chronic Lung Allograft Dysfunction (rCLAD): Implications for Its Etiopathogenesis and Evidence for Prognostic Patterns of Fibrosis J.H. von der Thüsen ,1 E. Vandermeulen,2 R. Vos,2 B. Weynand,3 E. Verbeken,3 S.E. Verleden.2 1Department of Pathology, Erasmus MC, Rotterdam, Netherlands; 2Lung Transplant Unit, Department of Clinical and Experimental Medicine, KU Leuven, Leuven, Belgium; 3Department of Pathology, UZ Gasthuisberg, Leuven, Belgium. Purpose: Chronic lung allograft dysfunction (CLAD) continues to be the main contributor to poor long-term allograft survival after lung transplantation. The restrictive phenotype of CLAD (rCLAD) carries a particularly poor prognosis. Little is known about the pathogenetic mechanisms involved in rCLAD. In this study, we undertook detailed histomorphological and immunohistochemical analysis of the lungs of patients with rCLAD. Methods: Material was obtained after death/redo transplantation or VATS biopsy from 21 rCLAD patients. Histopathologic patterns of rejection, fibrosis and vascular changes were scored after routine histochemical stains and additional immunohistochemistry for endothelial markers and C4d. Results: 75% of cases showed evidence of acute cellular rejection, ranging from minimal (Grade A1) to moderate (Grade A3). Lymphocytic bronchiolitis was absent in most lungs (B1R was found in only 21%), while in 55% there was obliterative bronchiolitis. Almost half of the cases showed a pattern of intra-alveolar fibrosis consistent with pleuroparenchymal fibroelastosis (PPFE) (n= 10, 47%), a minor subset showed non-specific interstitial pneumonia (NSIP) (n= 5) or bland fibrosis with emphysema (n= 5). Fibrinous exudates were frequently seen in association with fibrosis (n= 6), but no diffuse alveolar damage (DAD) was found. Evidence of microvascular damage was present in most cases. An emphysematous pattern of fibrosis was associated with a better survival (p= 0.0011), while fibrinous exudates were associated with a worse survival (p= 0.0007).
The Journal of Heart and Lung Transplantation, Vol 36, No 4S, April 2017
Methods: Plasma samples from LTx recipients (LTxRs) were analyzed by high-density protein microarrays containing 9000 unique full-length human proteins in native conformation. We pooled 12 LTxRs that developed CLAD and compared that with 11 LTxRs that remained CLAD-free. Longitudinal samples from LTxRs with CLAD were collected and stratified as pre-DSA, peak-DSA and DSA-resolved, and plasma samples from stable LTxRs that did not develop CLAD were stratified into 3, 9 and 15 month post-LTx periods. The samples were analyzed by Protoarray microchips and results were determined using Protoarray Prospector. The antigenic targets were analyzed by Ingenuity Pathway Analysis and DAVID Bioinformatics Resources. Results: Ab mapping by protein microarray revealed a unique repertoire in LTxRs that developed CLAD compared with stable LTxRs. Overall, CLAD patients demonstrated a > 10 fold higher Ab signal for 1756 proteins of which 35 proteins had > 50 fold higher signal intensities compared to that in stable LTxRs. Furthermore, peak-DSA plasma exhibited an expanded Ab repertoire compared to that of pre-DSA, which later contracted when DSA was resolved with reductions in both number of target antigens and signal intensities. In addition, de novo DSA was positively associated with the spread of nonHLA Ab repertoire. Conclusion: We demonstrate a vast repertoire of antigens involved in the development of immune responses following human LTx. Further, de novo DSA was strongly associated with development of immune responses to non-HLA SAgs. We conclude that de novo DSA is a major trigger for the development of immune responses to SAgs involved in the immunopathogenesis of CLAD. 4( 08) Early Elevation of Anti-HLA Immunoglobulin M Level Is Associated with Subsequent Lung Transplant Rejection and Worse Outcomes K. Miyahara , K. Miyoshi, T. Kurosaki, S. Otani, S. Sugimoto, M. Yamane, T. Oto. Department of Thoracic Surgery, Organ Transplant Center, Okayama University Hospital, Okayama-shi,Okayama prefecture, Japan. Purpose: Posttransplant immunoreactivity against allografts potentially differs widely in each case of transplantation. Early optimization of immunosuppressive intensity is crucial for later favorable outcome. However no useful noninvasive method exists for monitoring immunoreactivity. Immunoglobulin (Ig) M production stimulated by alloantigen is substantially sensitive to immunoreaction since primitive alloresponses can easily allow a mild B-cell response to produce IgM. The aim of this study is to examine relation between serum level of anti-human leukocyte antigen (HLA) IgM early after lung transplant and subsequent posttransplant outcomes. Methods: Consecutive 20 patients who underwent deceased lung transplantation in Okayama University Hospital from July 2013 to April 2015 were included. Immunoreactivity against HLA was retrospectively analyzed by detecting anti-HLA IgM of the serum sampled at day2, 4, 7, 10, and 14 after transplant. Flow cytometry crossmatching technique with Flow PRA HLA class I beads was used. The ratio of the level at each day to day2 (anti-classI IgM level) was obtained. The patients were divided into positive and negative groups according to the maximal IgM level during the first 14 days after lung transplantation. Results: Four patients exhibited significant elevation (max IgM level > 2.0, positive group) and 16 stayed negative throughout the monitoring period (negative group). In the positive group, the incidence of acute rejection during the first 30 days after transplant was significantly higher than negative group (p= 0.02). Two of four patients in positive group died of chronic lung allograft dysfunction (CLAD) at 9 and 13 months after lung transplantation respectively, while no patients in the negative group died of CLAD for 20 months after transplantation. IgM levels are independent of inflammatory event representing C-reactive protein value. Conclusion: Elevation of anti-HLA IgM levels during the first 14 days after lung transplantation may correlate to the higher incidence of later rejection and worse outcomes. Monitoring the level of anti-HLA IgM early after transplant can contribute to subsequent optimal immunosuppressive adjustments. 4( 09) Making a Case for EPLET Matching...Eplet Matching Is Associated with Improved Baseline Spirometry and Reduced Risk of CLAD Following Lung Transplantation
M. Paraskeva ,1 D.C. Walton,2 S.J. Hiho,2 J. Fuller,3 L.S. Cantwell,2 G.P. Westall,1 G.I. Snell.1 1Lung Transplant Service, Alfred Hospital, Melbourne, Australia; 2Victorian Transplantation and Immunogenetics Service, Australian Red Cross Blood Service, Melbourne, Australia; 3Monash University, Melbourne, Australia. Purpose: Increasingly HLA compatibility has been defined in terms of epitopes as compared to HLA antigen matching. We investigated the impact of eplet matching on the attainment of baseline or “best” spirometry following lung transplant and whether eplet matching predicted long term maintenance of baseline. Methods: All adult lung transplant recipients undergoing first bilateral lung transplantation between August 2008 and July 2012 with information available to allow eplet matching were included. Baseline FEV1 and FVC were defined as per ISHLT criteria. Normal spirometry was defined as FEV1 > 80% plus FVC> 80% plus FER> 70%. Results: 145 recipients were included. Those with < 40 eplet mismatches were more likely to have normal spirometry at baseline and took longer to achieve baseline FEV1 and FVC. Additionally they were less likely to develop CLAD.Conversely, those with ≥ 40 eplet mismatches were more likely to have abnormal spirometry at baseline and reach their peak FEV1 and FVC early.
FEV1 Best FVC Best HLA-DR/DQ Normal Abnormal Time (Mean Time (Mean Eplet MM Spirometry Spirometry days) days) CLAD
No CLAD
< 40 N= 65 77%(50) ≥ 40 N= 80 56%(45)
83%(54) 53%(42)
23%(15) 44%(35)
456 371
649 567
17%(11) 48%(38)
Conclusion: Eplet matching influences the attainment of baseline ("best") spirometry. Lung transplant recipients with fewer DR/DQ-eplet mismatches are more likely to normalise their spirometry and demonstrate long periods of good function with reduced incidence of CLAD.
4( 10) The Histomorphological Spectrum of Restrictive Chronic Lung Allograft Dysfunction (rCLAD): Implications for Its Etiopathogenesis and Evidence for Prognostic Patterns of Fibrosis J.H. von der Thüsen ,1 E. Vandermeulen,2 R. Vos,2 B. Weynand,3 E. Verbeken,3 S.E. Verleden.2 1Department of Pathology, Erasmus MC, Rotterdam, Netherlands; 2Lung Transplant Unit, Department of Clinical and Experimental Medicine, KU Leuven, Leuven, Belgium; 3Department of Pathology, UZ Gasthuisberg, Leuven, Belgium. Purpose: Chronic lung allograft dysfunction (CLAD) continues to be the main contributor to poor long-term allograft survival after lung transplantation. The restrictive phenotype of CLAD (rCLAD) carries a particularly poor prognosis. Little is known about the pathogenetic mechanisms involved in rCLAD. In this study, we undertook detailed histomorphological and immunohistochemical analysis of the lungs of patients with rCLAD. Methods: Material was obtained after death/redo transplantation or VATS biopsy from 21 rCLAD patients. Histopathologic patterns of rejection, fibrosis and vascular changes were scored after routine histochemical stains and additional immunohistochemistry for endothelial markers and C4d. Results: 75% of cases showed evidence of acute cellular rejection, ranging from minimal (Grade A1) to moderate (Grade A3). Lymphocytic bronchiolitis was absent in most lungs (B1R was found in only 21%), while in 55% there was obliterative bronchiolitis. Almost half of the cases showed a pattern of intra-alveolar fibrosis consistent with pleuroparenchymal fibroelastosis (PPFE) (n= 10, 47%), a minor subset showed non-specific interstitial pneumonia (NSIP) (n= 5) or bland fibrosis with emphysema (n= 5). Fibrinous exudates were frequently seen in association with fibrosis (n= 6), but no diffuse alveolar damage (DAD) was found. Evidence of microvascular damage was present in most cases. An emphysematous pattern of fibrosis was associated with a better survival (p= 0.0011), while fibrinous exudates were associated with a worse survival (p= 0.0007).