- Email: [email protected]
C4d in antibody-mediated rejection-how sensitive is it?
The Journal of Heart and Lung Transplantation Volume 24, Number 2S
graded semiquantitatively 1–5 where 1 was negative and 5 was obvious and generalized presence of the finding. Results: 768/3169 biopsies had IF findings diagnostic of AMR (occurring alone or as mixed AMR and cellular rejection). We performed a Receiver Operator Characteristic (ROC) curve to assess the sensitivity and specificity of VASC and ENDO parameters (used in combination) to predict AMR diagnosed by IF (See ROC curve and enclosed table). Conclusion: We conclude that histologic features which are proposed as screening tools to detect AMR on endomyocardial biopsies will not be sufficiently sensitive. Immunohistochemical testing remains necessary in the majority of cases to identify AMR.
Abstracts
S145
Conclusion: 1) C4d has outstanding specificity but limited sensitivity in the diagnosis of AMR. In view of the limited sensitivity, it does not appear feasible to use this marker alone to diagnose AMR. 2) While the deleterious impact of positive IF has been well documented, the implications of IF positive/C4d negative status remain to be determined.
C4D and IF in Antibody-Mediated Rejection N
IF positive
IF negative
C4D positive C4D negative
219 143
138 1465
317 THE CELLULAR INFILTRATE IN THE HUMAN ARTERY WALL WITH TRANSPLANT CORONARY ARTERY DISEASE: A T-HELPER 1 MEMORY RESPONSE J. van Kuik,1 J. van Loosdregt,1 D.F. van Wichen,1 M.F.M. van Oosterhout,1 A.H. Bruggink,1 C.C. Baan,2 N. de Jonge,1 F.H.J. Gmelig-Meyling,1 R.A. de Weger,1 1Pathology, University Medical Centre Utrecht, Utrecht, Netherlands; 2Internal Medicine, Erasmus Medical Center, Rotterdam, Netherlands
316 C4d IN ANTIBODY-MEDIATED REJECTION–HOW SENSITIVE IS IT? J. Stehlik,1 A.G. Kfoury,1 D.G. Renlund,1 E.M. Gilbert,1 J.C. Stringham,1 B. Dabbas,1 E.H. Hammond,1 1U.T.A.H. Cardiac Transplant Program, Salt Lake City, UT Background: Diagnostic criteria for antibody-mediated rejection (AMR) are evolving. We have previously described a method of histologic examination and routine immunofluorescence (IF) with classification of results as cellular, antibody-mediated, or mixed rejection. The aim of this study was to assess performance of C4d in diagnosing AMR. Methods: All endomyocardial biopsies from the first 6 post-transplant weeks in patients transplanted between 1999 and 2003, as well as selected biopsies done at other intervals, were tested for C4d on frozen sections simultaneously with IF, in addition to routine light microscopy. Cellular, AMR, or mixed rejection pattern was established based on IF positivity independent of the C4d result. Results: There were 1965 biopsies done in 213 patients. Twenty percent of the biopsies showed cellular rejection, 10% AMR, and 8% mixed cellular and AMR. C4d was detected in 219 of 357 (61%) IF positive biopsies (see Table). C4d was also detected in 138 of 1465 (9.4%) IF negative biopsies. Most of the C4d positive tests in IF negative samples occurred in patients with a concurrent cellular rejection episode; a third had simultaneous expression of C3d without immunoglobulin, suggesting complement activation without antibody deposition. We were unable to identify predictors of C4d negativity in IF positive patients. Considering our current practice as institutional standard, the sensitivity of C4d alone as a diagnostic test for antibody-mediated rejection was 60%, the specificity was 91%.
Introduction: There is circumstantial evidence from animal studies that chemokines play an important role in the regulation of the inflammatory response that causes Transplant Coronary Artery Disease (TCAD). Research on chemokines and chemokine receptors (C&CR) in coronary arteries (CA) of human heart transplantation patients (HTx) is limited. Therefore, we characterized the infiltrating cells and their C&CR expression in the CA of these patients. Methods: CA were obtained from HTx patients and compared to control CA of non-transplanted patients with and without arteriosclerosis. Tissue sections of complete or laser microdissected CA were analyzed by immunohistochemistry (IHC) for a variety of cell markers (HLA-DR, CD3, CD4, CD8, CD20, CD68 and CD79a), chemokines (MIG, IP-10, ITAC, RANTES and fractalkine), and their receptors (CXCR3, CCR5, CCR4, and CX3CR1). Q-PCR was used to study the mRNA expression of these C&CR and of the cytokines IL-4, IL-10, and ␥-IFN. Results: In normal CA almost no infiltrating cells were found. After transplantation only in CA with TCAD (TCAD⫹) a large number of infiltrating cells was observed. In these infiltrates activated (HLA-DR ⫹) helper T-cells prevailed (CD4:CD8 ⫽ 2) and only a few B-cells and macrophages were detected. IHC analysis of TCAD⫹ CA showed a strong expression of all C&CR. Q-PCR of the TCAD⫹ CA showed a stronger expression of all C&CR (except CCR4) and ␥-IFN, as compared to the CA without TCAD. IL-4 and IL-10 expression was not significantly different in both groups, and in the controls. Conclusion: The difference in CA of HTx patients with or without TCAD indicated that TCAD is induced and mediated by activated (HLA-DR⫹), helper-1 (␥-IFN⫹), memory (ITAC, CXCR3 and CX3CR1) T-cells and not T helper 2 cells (IL-4, IL-10 and CCR4). 318 ABDOMINAL ONLY APPROACH FOR HEARTMATE LVAD PUMP REPLACEMENT A.H. Healy,1 C.R. Wecker,1 K.E. Nelson,1 R. Khodaverdian,1 J.W. Long,1 1Utah Artificial Heart Program, LDS Hospital, Salt Lake City, UT Background: A shortage of donor hearts and an increase in left ventricular assist device (LVAD) destination therapy (DT) for endstage heart failure patients has led to an increase in the duration of support for LVAD patients. End of life pump failure will eventually
graded semiquantitatively 1–5 where 1 was negative and 5 was obvious and generalized presence of the finding. Results: 768/3169 biopsies had IF findings diagnostic of AMR (occurring alone or as mixed AMR and cellular rejection). We performed a Receiver Operator Characteristic (ROC) curve to assess the sensitivity and specificity of VASC and ENDO parameters (used in combination) to predict AMR diagnosed by IF (See ROC curve and enclosed table). Conclusion: We conclude that histologic features which are proposed as screening tools to detect AMR on endomyocardial biopsies will not be sufficiently sensitive. Immunohistochemical testing remains necessary in the majority of cases to identify AMR.
Abstracts
S145
Conclusion: 1) C4d has outstanding specificity but limited sensitivity in the diagnosis of AMR. In view of the limited sensitivity, it does not appear feasible to use this marker alone to diagnose AMR. 2) While the deleterious impact of positive IF has been well documented, the implications of IF positive/C4d negative status remain to be determined.
C4D and IF in Antibody-Mediated Rejection N
IF positive
IF negative
C4D positive C4D negative
219 143
138 1465
317 THE CELLULAR INFILTRATE IN THE HUMAN ARTERY WALL WITH TRANSPLANT CORONARY ARTERY DISEASE: A T-HELPER 1 MEMORY RESPONSE J. van Kuik,1 J. van Loosdregt,1 D.F. van Wichen,1 M.F.M. van Oosterhout,1 A.H. Bruggink,1 C.C. Baan,2 N. de Jonge,1 F.H.J. Gmelig-Meyling,1 R.A. de Weger,1 1Pathology, University Medical Centre Utrecht, Utrecht, Netherlands; 2Internal Medicine, Erasmus Medical Center, Rotterdam, Netherlands
316 C4d IN ANTIBODY-MEDIATED REJECTION–HOW SENSITIVE IS IT? J. Stehlik,1 A.G. Kfoury,1 D.G. Renlund,1 E.M. Gilbert,1 J.C. Stringham,1 B. Dabbas,1 E.H. Hammond,1 1U.T.A.H. Cardiac Transplant Program, Salt Lake City, UT Background: Diagnostic criteria for antibody-mediated rejection (AMR) are evolving. We have previously described a method of histologic examination and routine immunofluorescence (IF) with classification of results as cellular, antibody-mediated, or mixed rejection. The aim of this study was to assess performance of C4d in diagnosing AMR. Methods: All endomyocardial biopsies from the first 6 post-transplant weeks in patients transplanted between 1999 and 2003, as well as selected biopsies done at other intervals, were tested for C4d on frozen sections simultaneously with IF, in addition to routine light microscopy. Cellular, AMR, or mixed rejection pattern was established based on IF positivity independent of the C4d result. Results: There were 1965 biopsies done in 213 patients. Twenty percent of the biopsies showed cellular rejection, 10% AMR, and 8% mixed cellular and AMR. C4d was detected in 219 of 357 (61%) IF positive biopsies (see Table). C4d was also detected in 138 of 1465 (9.4%) IF negative biopsies. Most of the C4d positive tests in IF negative samples occurred in patients with a concurrent cellular rejection episode; a third had simultaneous expression of C3d without immunoglobulin, suggesting complement activation without antibody deposition. We were unable to identify predictors of C4d negativity in IF positive patients. Considering our current practice as institutional standard, the sensitivity of C4d alone as a diagnostic test for antibody-mediated rejection was 60%, the specificity was 91%.
Introduction: There is circumstantial evidence from animal studies that chemokines play an important role in the regulation of the inflammatory response that causes Transplant Coronary Artery Disease (TCAD). Research on chemokines and chemokine receptors (C&CR) in coronary arteries (CA) of human heart transplantation patients (HTx) is limited. Therefore, we characterized the infiltrating cells and their C&CR expression in the CA of these patients. Methods: CA were obtained from HTx patients and compared to control CA of non-transplanted patients with and without arteriosclerosis. Tissue sections of complete or laser microdissected CA were analyzed by immunohistochemistry (IHC) for a variety of cell markers (HLA-DR, CD3, CD4, CD8, CD20, CD68 and CD79a), chemokines (MIG, IP-10, ITAC, RANTES and fractalkine), and their receptors (CXCR3, CCR5, CCR4, and CX3CR1). Q-PCR was used to study the mRNA expression of these C&CR and of the cytokines IL-4, IL-10, and ␥-IFN. Results: In normal CA almost no infiltrating cells were found. After transplantation only in CA with TCAD (TCAD⫹) a large number of infiltrating cells was observed. In these infiltrates activated (HLA-DR ⫹) helper T-cells prevailed (CD4:CD8 ⫽ 2) and only a few B-cells and macrophages were detected. IHC analysis of TCAD⫹ CA showed a strong expression of all C&CR. Q-PCR of the TCAD⫹ CA showed a stronger expression of all C&CR (except CCR4) and ␥-IFN, as compared to the CA without TCAD. IL-4 and IL-10 expression was not significantly different in both groups, and in the controls. Conclusion: The difference in CA of HTx patients with or without TCAD indicated that TCAD is induced and mediated by activated (HLA-DR⫹), helper-1 (␥-IFN⫹), memory (ITAC, CXCR3 and CX3CR1) T-cells and not T helper 2 cells (IL-4, IL-10 and CCR4). 318 ABDOMINAL ONLY APPROACH FOR HEARTMATE LVAD PUMP REPLACEMENT A.H. Healy,1 C.R. Wecker,1 K.E. Nelson,1 R. Khodaverdian,1 J.W. Long,1 1Utah Artificial Heart Program, LDS Hospital, Salt Lake City, UT Background: A shortage of donor hearts and an increase in left ventricular assist device (LVAD) destination therapy (DT) for endstage heart failure patients has led to an increase in the duration of support for LVAD patients. End of life pump failure will eventually