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36-P: Phenotypes of acute cellular rejection of lung transplant recipients following induction therapy with T-cell depleting agents
Abstracts
S25
36-P
PHENOTYPES OF ACUTE CELLULAR REJECTION OF LUNG TRANSPLANT RECIPIENTS FOLLOWING INDUCTION THERAPY WITH T-CELL DEPLETING AGENTS. Kathy Spichty, Alin Girnita, Diana Zaldonis, Joseph Pilewski, Kenneth McCurry, Adriana Zeevi. Pathology/Surgery, University of Pittsburgh, Pittsburgh, PA, USA. Aim: Induction therapy with T-cell depleting agents have led to the use of minimal post-transplant immunosuppression (IS). The impact of this protocol on the activation of proinflammatory cytokines/effector molecules that affect cellular rejection is not well known. In this study, we evaluated the relationship between upregulation of T-cell and macrophage-dependent inflammatory cytokines detected by molecular methods and the patient clinical status. Methods: We studied 38 LTx patients who received either Thymo (n⫽8) or Campath-1H (n⫽30) followed by maintenance IS with Tacrolimus and low-dose steroids. We analyzed 197 bronchoalveolar lavage (BAL) mRNA samples (3-8/patient) by Real-Time PCR for IFN-g, GZMB, MCP-1, RANTES, FOXP3, and GAPDH (control). The 7500 Fast System SDS was used and data were analyzed by the dCT and 2-ddCT methods. Results: Early ACR (⬍ 90 days post-LTx) occurring in the Thymo cohort of patients was associated with an increase (8 fold) of macrophage-specific mRNA (MCP-1 and GZMB). In contrast, late ACR (⬎ 90 days post-LTx) occurring in the Campath-1H cohort was associated with a significant upregulation (4-64 fold) of T-cell dependent mRNA (GZMB and IFN-g) in addition to increases of RANTES and MCP-1 mRNA. Also, an upregulation of FOXP3 (⬎ 4 fold) was observed in the Campath-1H patients during episodes of rejection. Conclusions: Early ACR was characterized mainly by macrophage activation while late ACR was associated with cytotoxic T-cell activation. Furthermore, sensitive molecular methods may detect the activation of intra-allograft pro-inflammatory mediators prior to the diagnosis of rejection via transbronchial biopsies and may impact patient management.
37-P
ROLE OF IL-18 AND AIF-1 IN THE DEVELOPMENT OF CORONARY VASCULOPATHY AND CELLULAR REJECTION AFTER CARDIAC TRANSPLANTATION. D. Olga McDaniel,1 Xinchun Zhou,1 Giorgio Aru,1 Tammy Thomas,2 Zhi He,3 Steven Bigler,3 Charles K. Moore.2 1Surgery, University of MS Medical Center, Jackson, MS, USA; 2Medicine, University of MS Medical Center, Jackson, MS, USA; 3Pathology, University of MS Medical Center, Jackson, MS, USA. Aim: Current reliable methods for detection of either CR or CV such as cardiac ventricular biopsy and coronary angiogram are invasive, inconvenient and expensive. Aim of this study was to investigate whether levels of IL-18 or IL-18 receptors and AIF-1 in cardiac biopsy samples could correlate with those observed in PBMCs in association with CV and or CR and might be a reliable method for detecting an early CR or CV. Methods: Surgical biopsy specimens were tested with immunohistochemistry (IHC) stains for IL-18, IL-18 receptors and AIF-1. The biopsy specimens were classified into testing groups based on histopathological assessment for CR and the CV grades were obtained from cardiovascular image database. Results: Only infiltrating monocytes were positive with IL-18 Ab in IHC stain and the density was correlated with the severity of CV and CR. Both monocytes and lymphocytes stained positive with the IL-18 receptor Ab. The distribution of monocytes and lymphocytes stained with IL-18R antibody was higher in biopsy samples with grade 3A rejection than with any grades of CV. The density of AIF-1 was highly correlated with severity of CR and CV. Conclusions: We concluded that IL-18 and its receptor and AIF-1 are important in immunological pathogenesis of CR and CV of cardiac allograft. Local pathological alterations in allograft are proportionally reflected in the PBMCs. This finding provides a possibility to establish a reliable and less invasive method for the detection of cardiac allograft outcomes after transplantation.
S25
36-P
PHENOTYPES OF ACUTE CELLULAR REJECTION OF LUNG TRANSPLANT RECIPIENTS FOLLOWING INDUCTION THERAPY WITH T-CELL DEPLETING AGENTS. Kathy Spichty, Alin Girnita, Diana Zaldonis, Joseph Pilewski, Kenneth McCurry, Adriana Zeevi. Pathology/Surgery, University of Pittsburgh, Pittsburgh, PA, USA. Aim: Induction therapy with T-cell depleting agents have led to the use of minimal post-transplant immunosuppression (IS). The impact of this protocol on the activation of proinflammatory cytokines/effector molecules that affect cellular rejection is not well known. In this study, we evaluated the relationship between upregulation of T-cell and macrophage-dependent inflammatory cytokines detected by molecular methods and the patient clinical status. Methods: We studied 38 LTx patients who received either Thymo (n⫽8) or Campath-1H (n⫽30) followed by maintenance IS with Tacrolimus and low-dose steroids. We analyzed 197 bronchoalveolar lavage (BAL) mRNA samples (3-8/patient) by Real-Time PCR for IFN-g, GZMB, MCP-1, RANTES, FOXP3, and GAPDH (control). The 7500 Fast System SDS was used and data were analyzed by the dCT and 2-ddCT methods. Results: Early ACR (⬍ 90 days post-LTx) occurring in the Thymo cohort of patients was associated with an increase (8 fold) of macrophage-specific mRNA (MCP-1 and GZMB). In contrast, late ACR (⬎ 90 days post-LTx) occurring in the Campath-1H cohort was associated with a significant upregulation (4-64 fold) of T-cell dependent mRNA (GZMB and IFN-g) in addition to increases of RANTES and MCP-1 mRNA. Also, an upregulation of FOXP3 (⬎ 4 fold) was observed in the Campath-1H patients during episodes of rejection. Conclusions: Early ACR was characterized mainly by macrophage activation while late ACR was associated with cytotoxic T-cell activation. Furthermore, sensitive molecular methods may detect the activation of intra-allograft pro-inflammatory mediators prior to the diagnosis of rejection via transbronchial biopsies and may impact patient management.
37-P
ROLE OF IL-18 AND AIF-1 IN THE DEVELOPMENT OF CORONARY VASCULOPATHY AND CELLULAR REJECTION AFTER CARDIAC TRANSPLANTATION. D. Olga McDaniel,1 Xinchun Zhou,1 Giorgio Aru,1 Tammy Thomas,2 Zhi He,3 Steven Bigler,3 Charles K. Moore.2 1Surgery, University of MS Medical Center, Jackson, MS, USA; 2Medicine, University of MS Medical Center, Jackson, MS, USA; 3Pathology, University of MS Medical Center, Jackson, MS, USA. Aim: Current reliable methods for detection of either CR or CV such as cardiac ventricular biopsy and coronary angiogram are invasive, inconvenient and expensive. Aim of this study was to investigate whether levels of IL-18 or IL-18 receptors and AIF-1 in cardiac biopsy samples could correlate with those observed in PBMCs in association with CV and or CR and might be a reliable method for detecting an early CR or CV. Methods: Surgical biopsy specimens were tested with immunohistochemistry (IHC) stains for IL-18, IL-18 receptors and AIF-1. The biopsy specimens were classified into testing groups based on histopathological assessment for CR and the CV grades were obtained from cardiovascular image database. Results: Only infiltrating monocytes were positive with IL-18 Ab in IHC stain and the density was correlated with the severity of CV and CR. Both monocytes and lymphocytes stained positive with the IL-18 receptor Ab. The distribution of monocytes and lymphocytes stained with IL-18R antibody was higher in biopsy samples with grade 3A rejection than with any grades of CV. The density of AIF-1 was highly correlated with severity of CR and CV. Conclusions: We concluded that IL-18 and its receptor and AIF-1 are important in immunological pathogenesis of CR and CV of cardiac allograft. Local pathological alterations in allograft are proportionally reflected in the PBMCs. This finding provides a possibility to establish a reliable and less invasive method for the detection of cardiac allograft outcomes after transplantation.