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1034-LBO
498
Abstracts / Human Immunology 75 (2014) 479–501
1034-LBO DIFFERENTIAL MICRORNA EXPRESSION PROFILING IN LUNG ALLOGRAFT RECIPIENTS WHO DEVELOP DONOR SPECIFIC ANTIBODY AND BRONCHIOLITIS OBLITERANS SYNDROME. Zhongping Xu 1, Sabarinathan Ramachandran 1, Nayan Sarma 1,
Venkataswarup Tiriveedhi 1, Baskaran Gautam 3, Aviva Aloush 1, Nancy Steward 1, Ramsey Hachem 3, Elbert Trulock 3, G. Alexander Patterson 1, Thalachallour Mohanakumar 1,2. 1 Surgery, Washington Univ Sch Med, St. Louis, United States; 2 3
Pathology & Immunology, Washington Univ Sch Med, St. Louis, United States; Medicine, Washington Univ Sch Med, St. Louis, United States.
Aim: Bronchiolitis Obliterans Syndrome (BOS), clinically diagnosed as chronic rejection, has remained a major setback following lung transplantation (LT). However, its pathogenesis is poorly understood and no predictive biomarkers have been identified. Expression of microRNA (miRNA) in blood is deregulated in physiopathological conditions including immunological disorders. Methods: In this study, we hypothesized that miRNA profiles from peripheral blood mononuclear cells (PBMCs) in LT recipients (LTxR) could identify patients at risk of BOS. MiRNA expression profiling was performed using TaqMan Human MicroRNA Array in PBMCs collected from three groups of LTxR: 10 stable, 10 who developed DSA and 10 who developed BOS. Results: There were 98 miRNAs upregulated in LTxR with BOS compared to that of stable transplants (relative fold>2.5); and 32 miRNAs downregulated with BOS (relative fold>2.5). In addition, we found that there were 16 miRNAs upregulated (including miR-99a, miR-147, and miR-124⁄) with BOS compared to that of DSA; and 5 miRNAs downregulated (including miR-34b, miR-363⁄, and miR-155⁄) in BOS compared to DSA, indicating that these miRNAs may play a role in BOS with DSA. We validated those differential miRNAs in independent PBMCs and broncheoalveolar lavage cells using Taqman real-time PCR. In addition we focused on one miRNA, miR-99a, which caused upregulation of 2.8-fold in LTxR with BOS compared to that of DSA. Bioinformatics prediction indicated that miR-99a can mediate gene-expression regulation of TNIK (TRAF2 and NCK interacting kinase) and TRAF4 (TNF receptor-associated factor 4), which were involved in regulating NF-jB signaling pathway. Conclusions: In conclusion our results demonstrate that differential miRNA expression profile has the potential to predict BOS and development of DSA following LT.
1035-LBP DEVELOPMENT OF ANTIBODIES TO HLA IN PATIENTS ON LEFT VENTRICULAR ASSISTING DEVICE (LVAD) LEADS TO INCREASED PROINFLAMMATORY CYTOKINES AND IMMUNE RESPONSES TO CARDIAC SELF-ANTIGENS, MYOSIN AND VIMENTIN. Babak Banan 1,
Venkataswarup Tiriveedhi 1, Donna Phelan 2, Greg Ewald 3, Thalachallour Mohanakumar 1,4. 1 Surgery, Washington Univ Sch Med, St. Louis, MO, United States; 2
HLA Laboratory, Barnes-Jewish Hospital, St. Louis, MO, United States; 3 Medicine, Washington Univ Sch Med, St. Louis, MO, United States; 4 Pathology & Immunology, Washington Univ Sch Med, St. Louis, MO, United States.
Aim: Recent studies have demonstrated that patients with left ventricular assist devices (LVAD) are at increased risk of development of antibody mediated rejection (AMR) and cardiac allograft vasculopathy (CAV) post heart transplantation (HTx). However, the role of pre-existing antibodies (Abs) to HLA in inducing immune responses to cardiac self-antigens (CSAgs) is yet to be defined. In this study, we determined the kinetics of development of Abs to HLA and immune responses to CSAgs during the pre-transplant period in patients on LVAD. Methods: Sera from 70 HTx (40 LVAD, 30 Non-LVAD) were enrolled and serially analyzed for Abs to HLA using Luminex and for CSAgs by ELISA. Serum cytokines (IFNc, IL-17, IL-1b and IL-10) were analyzed by LUMINEX. Sera from 32 age and gender matched normal subjects were the controls. Results: Among LVAD patients, 22/40 (55% vs. 3.1%, p < 0.01) had anti-HLA prior to HTx. LVAD with antiHLA 19/22 (86.4% vs. 13.6%, p < 0.01) had Abs to CSAgs. 23/40 (57.5% vs. normal 6.3%, p < 0.01) developed MYO-Abs, 25/40 (62.5% vs. normal 3.1%, p < 0.01) developed VIM-Abs. In contrast, Non-LVAD group only 1/ 30 (3.3% vs. 3.1%, p > 0.08) had anti-HLA prior to HTx. In this group, 5/30 (16.7% vs. normal 6.3%, p < 0.05) developed MYO-Abs, 6/30 (20% vs. normal 3.1%, p < 0.05) developed VIM-Abs. No significant difference in Abs to collagen-II (control) were noticed among the three cohorts namely, LVAD vs non-LVAD vs control. Further, patients with Abs to HLA and CSAgs have increased circulating levels of pro-inflammatory cytokines, IFNc (2.4 fold), IL-17 (1.9 fold), and IL-1b (2.7 fold) along with decrease in IL-10 (3.1 fold). Conclusions: LVAD patients have increased risk for development of anti-HLA prior to transplantation and anti-HLA positive patients further develop immune responses to CSAgs. LVAD patients with anti-HLA and anti-CSAgs have increased circulating pro-inflammatory cytokines prior to transplant which we hypothesize to play a significant role in the development of AMR and CAV following HTx.
Abstracts / Human Immunology 75 (2014) 479–501
1034-LBO DIFFERENTIAL MICRORNA EXPRESSION PROFILING IN LUNG ALLOGRAFT RECIPIENTS WHO DEVELOP DONOR SPECIFIC ANTIBODY AND BRONCHIOLITIS OBLITERANS SYNDROME. Zhongping Xu 1, Sabarinathan Ramachandran 1, Nayan Sarma 1,
Venkataswarup Tiriveedhi 1, Baskaran Gautam 3, Aviva Aloush 1, Nancy Steward 1, Ramsey Hachem 3, Elbert Trulock 3, G. Alexander Patterson 1, Thalachallour Mohanakumar 1,2. 1 Surgery, Washington Univ Sch Med, St. Louis, United States; 2 3
Pathology & Immunology, Washington Univ Sch Med, St. Louis, United States; Medicine, Washington Univ Sch Med, St. Louis, United States.
Aim: Bronchiolitis Obliterans Syndrome (BOS), clinically diagnosed as chronic rejection, has remained a major setback following lung transplantation (LT). However, its pathogenesis is poorly understood and no predictive biomarkers have been identified. Expression of microRNA (miRNA) in blood is deregulated in physiopathological conditions including immunological disorders. Methods: In this study, we hypothesized that miRNA profiles from peripheral blood mononuclear cells (PBMCs) in LT recipients (LTxR) could identify patients at risk of BOS. MiRNA expression profiling was performed using TaqMan Human MicroRNA Array in PBMCs collected from three groups of LTxR: 10 stable, 10 who developed DSA and 10 who developed BOS. Results: There were 98 miRNAs upregulated in LTxR with BOS compared to that of stable transplants (relative fold>2.5); and 32 miRNAs downregulated with BOS (relative fold>2.5). In addition, we found that there were 16 miRNAs upregulated (including miR-99a, miR-147, and miR-124⁄) with BOS compared to that of DSA; and 5 miRNAs downregulated (including miR-34b, miR-363⁄, and miR-155⁄) in BOS compared to DSA, indicating that these miRNAs may play a role in BOS with DSA. We validated those differential miRNAs in independent PBMCs and broncheoalveolar lavage cells using Taqman real-time PCR. In addition we focused on one miRNA, miR-99a, which caused upregulation of 2.8-fold in LTxR with BOS compared to that of DSA. Bioinformatics prediction indicated that miR-99a can mediate gene-expression regulation of TNIK (TRAF2 and NCK interacting kinase) and TRAF4 (TNF receptor-associated factor 4), which were involved in regulating NF-jB signaling pathway. Conclusions: In conclusion our results demonstrate that differential miRNA expression profile has the potential to predict BOS and development of DSA following LT.
1035-LBP DEVELOPMENT OF ANTIBODIES TO HLA IN PATIENTS ON LEFT VENTRICULAR ASSISTING DEVICE (LVAD) LEADS TO INCREASED PROINFLAMMATORY CYTOKINES AND IMMUNE RESPONSES TO CARDIAC SELF-ANTIGENS, MYOSIN AND VIMENTIN. Babak Banan 1,
Venkataswarup Tiriveedhi 1, Donna Phelan 2, Greg Ewald 3, Thalachallour Mohanakumar 1,4. 1 Surgery, Washington Univ Sch Med, St. Louis, MO, United States; 2
HLA Laboratory, Barnes-Jewish Hospital, St. Louis, MO, United States; 3 Medicine, Washington Univ Sch Med, St. Louis, MO, United States; 4 Pathology & Immunology, Washington Univ Sch Med, St. Louis, MO, United States.
Aim: Recent studies have demonstrated that patients with left ventricular assist devices (LVAD) are at increased risk of development of antibody mediated rejection (AMR) and cardiac allograft vasculopathy (CAV) post heart transplantation (HTx). However, the role of pre-existing antibodies (Abs) to HLA in inducing immune responses to cardiac self-antigens (CSAgs) is yet to be defined. In this study, we determined the kinetics of development of Abs to HLA and immune responses to CSAgs during the pre-transplant period in patients on LVAD. Methods: Sera from 70 HTx (40 LVAD, 30 Non-LVAD) were enrolled and serially analyzed for Abs to HLA using Luminex and for CSAgs by ELISA. Serum cytokines (IFNc, IL-17, IL-1b and IL-10) were analyzed by LUMINEX. Sera from 32 age and gender matched normal subjects were the controls. Results: Among LVAD patients, 22/40 (55% vs. 3.1%, p < 0.01) had anti-HLA prior to HTx. LVAD with antiHLA 19/22 (86.4% vs. 13.6%, p < 0.01) had Abs to CSAgs. 23/40 (57.5% vs. normal 6.3%, p < 0.01) developed MYO-Abs, 25/40 (62.5% vs. normal 3.1%, p < 0.01) developed VIM-Abs. In contrast, Non-LVAD group only 1/ 30 (3.3% vs. 3.1%, p > 0.08) had anti-HLA prior to HTx. In this group, 5/30 (16.7% vs. normal 6.3%, p < 0.05) developed MYO-Abs, 6/30 (20% vs. normal 3.1%, p < 0.05) developed VIM-Abs. No significant difference in Abs to collagen-II (control) were noticed among the three cohorts namely, LVAD vs non-LVAD vs control. Further, patients with Abs to HLA and CSAgs have increased circulating levels of pro-inflammatory cytokines, IFNc (2.4 fold), IL-17 (1.9 fold), and IL-1b (2.7 fold) along with decrease in IL-10 (3.1 fold). Conclusions: LVAD patients have increased risk for development of anti-HLA prior to transplantation and anti-HLA positive patients further develop immune responses to CSAgs. LVAD patients with anti-HLA and anti-CSAgs have increased circulating pro-inflammatory cytokines prior to transplant which we hypothesize to play a significant role in the development of AMR and CAV following HTx.