- Email: [email protected]
Transplantation of “thymoheart” allografts in miniature swine
The Journal of Heart and Lung Transplantation Volume 18, Number 1
I ~~~ i ~,~
at thetlm of recruitmen, L”lEd [IllIn] PAS P-w
I
after ale year L”lDd [ml,
Abstracts
AAS
;I
[W “0, detectable 2St8 73 * 7 5*10 24* 3 76* 6 47ffC W”P c (” -17, 1” =ts, (” =ts, (” =,s, P-values ~ n I. 0,s. 0.000, 0 0004 0 owl T- 0.01c4 lmmunoadsorption led to a significant improvement of cardiac function and dimension. AAB directed against cardiac p ,-receptors seem to be a suitable parameter to assess the success of immunoadsorption. The reason why the cardiac function of the control group also improved may be due to the positive effect of the p- blocker therapy. ww 1 (” -17)
22*3
75f7
&Of 14
38f8
We conclude that, in CHF patients referred for transplantation, the SM fibers mitochondrial properties are correlated with the amount of RV and LV mitochondrial function. This suggests that the SM o&dative abnormalities are linked to ventricular oxidative dysfunction, either as a consequence of LV failure or by circulating ventricular and SM mitochondrial function.
64te
151
149
150
71
TRANSPLANTATION OF “THYMOHEART” ALLOGRAFTS IN MINIATURE SWINE M.T. Menard, K. Mawulawde, K. Yamada, M.L. Schwarze, R.S. Lee, J.K. Slisz, K.S. Allison, D.H. Sachs, J.C. Madsen, Division of Cardiac Surgery and the Transplantation Biology Research Center, Massachusetts General Hospital and Harvard Medical School, Boston, MA 02114 Recent studies from this laboratory have indicated that the thymus plays a critical role in the induction of rapid and stable tolerance in p.artially inbred miniahue swine. Based on these data, we hypothesized that a state of tolerance may be more easily achieved if folly vascuknized and functional donor thymus were transferred to the recipient at the time of heart uansplantation. To this end, we have auccessfolly created the composite “thymoheart” allograft, a heart supporting vascnlarized thymic autografts identical in histologic and flow cytomeuic (PACS) appearance to native thymus. We have now heterotopically transplanted thymohearts into euthymic and thymectomized MHC class I mismatched recipients treated with a short coorse of cyclosporine (lo-13 mgikg/day POD O-l 1). Unmanipulated class I disparate conuol hearts transplanted into euthymic CyA-treated recipients rejected between 33 and SS days. In contrast, class I disparate thymohearts hansplanted into euthymic recipients survived 64 days and >191 days. Survival of donor-matched skin placed on the second animal on POD 159 was prolonged (16 days) compared to skin grafts placed on naive controls (11.7 +/- 2.6 days). Furthermore, when the beating heart was removed from this pig on POD 191, it had nohistologic evidence of interstitiat rejection. Unmanipulated class I disparate control hearts transplanted into thymectomized CyA-treated recipients rejected between 8 and 27 days. However, class I disparate thymohearts tmnsplanted into thymectomized recipimts survived 41.48 and >S6 days. In both experimental groups, the amount of viable thymus present within the thymoheart at the time of autopsy appeared to correlate with allograft survival time. These data indicate that transplwtation of thymoheart allografts has a beneficial effect on cardiac allogaft survival. The strategy of autologous thymic cotransplantation may have particular value in cardiac xenouansplantatim.
CHARACTERIZATION OF TEE SKELETAL MUSCLE (SM) MITOCHONDRIAL. RESPIRATION IN SEVERE CHRONIC HEART FAILURE (CHF) AND ITS RFL4TIONS TO THE VENTRICULAR OXIDATIVE CAPACITY, J Zoll, B Mettaoer, H Sanchez, E Epailly, E Lonsdorfer, E Lampert, AX Bigard, V Veksler, R Ventura-Clapier. Project PROGRES-INSERM, Physiology dept. Faculty of Medicine, Suasbourg France, INSERM U 446, Faculty of Pharmacy, ChatenayMalabry, France and CRSSA Grenoble - la Tronche, France. SM oxidative impairement is a constant featore of CHF. It has been argued that the SM abnormalities may develop at least partially independently t?om the decrease in let? ventricular (LV) function and thus be one of the main contributor of the decrease in peak O2 consumption (mVOJ in these patients. Since heart transplantation is a unique opportunity to obtain &sh severely failing ventricular tissue this study was aimed at comparing the SM and right ventricular (RV) or LV maximal mitochondrial in-situ respiration. RV and LV fibers samples from the explanted failing heart were obtained simultaneously with SM tissue from the vastus lateralis at the time of transplantation in 7 severe CHF patients (age 539k8.3 yrs) with a mVOl of 14.6&l .9 ml/tin/kg. AtIer saponin sarcolemmal skinning, the fibers 02 consumption (VO1) was measured at saturating substrate and O2 concentrations in oxygrapbic Strathkelvin respiration chambers to obtain the ADP-free (VO) and the maximal ADP-stimulated (Vmax) V02 @moles O&in/g dry weight). The acceptor control ratio (ACR) was defmed as VmaxNO. Although neither SM fibers Vmax or ACR were related to the severity of LV failure as defined by the LV ejection fraction, hemodynamics, or circulating neuro-hormone levels, the SM ACR was correlated fibers ACR (r = 0.88, p = 0.009).
STAT4-MEDIATED THI RESPONSES OFGRAFT ARTERIOSCLEROSIS
,- D.BOP . o.ooe ‘*
‘.aL”f!h”‘J ACR‘O”
factors that simultaneously
PROMOTE
“’ alter
THE DEVELOPMENT
J6rg Koglin, Silpa Gadiraju, Mary E. Russell, Cardiovascular Biology Laboratory, Harvard School of Public Health, Boston, Massachusetts As a way of modifying Thl and Th2 responses. we used Stat4- and Stat6-knockout mice as recipients in our mouse cardiac transplant model of chronic rejection (CBA into C57BL6x129, anti-CD418 for 28 days). At day 55 after transplantation, cardiac grafts placed into Stat4 -/- (n=lO) had significantly reduced frequency (24f2%) and severity (9i4%) of vascular occlusion when compared to wildtype controls (n=7, frequency 7Oil2% [p
152
FIVE-YEAR FOLLOW-UP OF A RANDOMIZED TRIAL OF PRAVASTATIN IN HEART TRANSPLANT PATIENTS. 3.A. Kobashigawa, J.D. Moriguchi, H. Laks, T.K. Ro, M.A. Hamilton, A. Hage, L. Wener, N. Kawata. University of California, Los Angeles, Calif. The original publication of this trial’s first-year data was the first to suggest significant benefit in outcome in heart transplant patients randomized to pravastatin (PVS), an HMG-CoA reductase inhibitor, or no PVS at the time of transplant surgery. We now report the 5-year follow-up of this study. 97 heart transplant patients given triple drug immunosuppression, between 7/92 and 2/94, were randomized to PVS or no PVS within 2 weeks after transplant. After the fmt year of the smdy, there was a considerable number of control patients (71%) who crossed over to take PVS due to the first year study results demonstrating benefit of PVS. Five-year survival continues to be superior in the PVS group with greater divergence in survival compared to the first year results (see Table). Major causes of death in both groups were rejection and transplant coronary artery disease (TCAD). In the PVS group vs. the control group, there were 5 vs. 10 patients who died due to rejection and 2 vs. 5 patients who died due to TCAD. Intravascular ultrasound (IVUS) of 24 patients with paired stodies (10 control and 14 PVS patients) from baseline (6 weeks after transplant) to 5 years after transplant revealed a significant reduction in the change in intimal area (IA) in the PVS group vs. the control group. Although not significant, there was a numerically lower change in intimal index (II) and lumen area (LA) and more patients free from TCAD (angiography) and/or death in the PVS group vs. control group. 5-VCU AIWS lbaseline to 5 vrs) Freedom from IA II LA Survival TCAD and/or death (mm2) (mm*) Prnva Group 83% * 64% 0.42’ .06 -3.14 62% 55% 1.36 .ll -3.49 Control Group * p < 0.05
Conclusion: The use of pravastatin early after heart transplantation appears to have continued beneficial effects in survival, rejection, and the development of TCAD at 5 years after Uansplant. A large crossover to PVS from the control group suggests ihat the early use of PVS appears important for good outcome.
I ~~~ i ~,~
at thetlm of recruitmen, L”lEd [IllIn] PAS P-w
I
after ale year L”lDd [ml,
Abstracts
AAS
;I
[W “0, detectable 2St8 73 * 7 5*10 24* 3 76* 6 47ffC W”P c (” -17, 1” =ts, (” =ts, (” =,s, P-values ~ n I. 0,s. 0.000, 0 0004 0 owl T- 0.01c4 lmmunoadsorption led to a significant improvement of cardiac function and dimension. AAB directed against cardiac p ,-receptors seem to be a suitable parameter to assess the success of immunoadsorption. The reason why the cardiac function of the control group also improved may be due to the positive effect of the p- blocker therapy. ww 1 (” -17)
22*3
75f7
&Of 14
38f8
We conclude that, in CHF patients referred for transplantation, the SM fibers mitochondrial properties are correlated with the amount of RV and LV mitochondrial function. This suggests that the SM o&dative abnormalities are linked to ventricular oxidative dysfunction, either as a consequence of LV failure or by circulating ventricular and SM mitochondrial function.
64te
151
149
150
71
TRANSPLANTATION OF “THYMOHEART” ALLOGRAFTS IN MINIATURE SWINE M.T. Menard, K. Mawulawde, K. Yamada, M.L. Schwarze, R.S. Lee, J.K. Slisz, K.S. Allison, D.H. Sachs, J.C. Madsen, Division of Cardiac Surgery and the Transplantation Biology Research Center, Massachusetts General Hospital and Harvard Medical School, Boston, MA 02114 Recent studies from this laboratory have indicated that the thymus plays a critical role in the induction of rapid and stable tolerance in p.artially inbred miniahue swine. Based on these data, we hypothesized that a state of tolerance may be more easily achieved if folly vascuknized and functional donor thymus were transferred to the recipient at the time of heart uansplantation. To this end, we have auccessfolly created the composite “thymoheart” allograft, a heart supporting vascnlarized thymic autografts identical in histologic and flow cytomeuic (PACS) appearance to native thymus. We have now heterotopically transplanted thymohearts into euthymic and thymectomized MHC class I mismatched recipients treated with a short coorse of cyclosporine (lo-13 mgikg/day POD O-l 1). Unmanipulated class I disparate conuol hearts transplanted into euthymic CyA-treated recipients rejected between 33 and SS days. In contrast, class I disparate thymohearts hansplanted into euthymic recipients survived 64 days and >191 days. Survival of donor-matched skin placed on the second animal on POD 159 was prolonged (16 days) compared to skin grafts placed on naive controls (11.7 +/- 2.6 days). Furthermore, when the beating heart was removed from this pig on POD 191, it had nohistologic evidence of interstitiat rejection. Unmanipulated class I disparate control hearts transplanted into thymectomized CyA-treated recipients rejected between 8 and 27 days. However, class I disparate thymohearts tmnsplanted into thymectomized recipimts survived 41.48 and >S6 days. In both experimental groups, the amount of viable thymus present within the thymoheart at the time of autopsy appeared to correlate with allograft survival time. These data indicate that transplwtation of thymoheart allografts has a beneficial effect on cardiac allogaft survival. The strategy of autologous thymic cotransplantation may have particular value in cardiac xenouansplantatim.
CHARACTERIZATION OF TEE SKELETAL MUSCLE (SM) MITOCHONDRIAL. RESPIRATION IN SEVERE CHRONIC HEART FAILURE (CHF) AND ITS RFL4TIONS TO THE VENTRICULAR OXIDATIVE CAPACITY, J Zoll, B Mettaoer, H Sanchez, E Epailly, E Lonsdorfer, E Lampert, AX Bigard, V Veksler, R Ventura-Clapier. Project PROGRES-INSERM, Physiology dept. Faculty of Medicine, Suasbourg France, INSERM U 446, Faculty of Pharmacy, ChatenayMalabry, France and CRSSA Grenoble - la Tronche, France. SM oxidative impairement is a constant featore of CHF. It has been argued that the SM abnormalities may develop at least partially independently t?om the decrease in let? ventricular (LV) function and thus be one of the main contributor of the decrease in peak O2 consumption (mVOJ in these patients. Since heart transplantation is a unique opportunity to obtain &sh severely failing ventricular tissue this study was aimed at comparing the SM and right ventricular (RV) or LV maximal mitochondrial in-situ respiration. RV and LV fibers samples from the explanted failing heart were obtained simultaneously with SM tissue from the vastus lateralis at the time of transplantation in 7 severe CHF patients (age 539k8.3 yrs) with a mVOl of 14.6&l .9 ml/tin/kg. AtIer saponin sarcolemmal skinning, the fibers 02 consumption (VO1) was measured at saturating substrate and O2 concentrations in oxygrapbic Strathkelvin respiration chambers to obtain the ADP-free (VO) and the maximal ADP-stimulated (Vmax) V02 @moles O&in/g dry weight). The acceptor control ratio (ACR) was defmed as VmaxNO. Although neither SM fibers Vmax or ACR were related to the severity of LV failure as defined by the LV ejection fraction, hemodynamics, or circulating neuro-hormone levels, the SM ACR was correlated fibers ACR (r = 0.88, p = 0.009).
STAT4-MEDIATED THI RESPONSES OFGRAFT ARTERIOSCLEROSIS
,- D.BOP . o.ooe ‘*
‘.aL”f!h”‘J ACR‘O”
factors that simultaneously
PROMOTE
“’ alter
THE DEVELOPMENT
J6rg Koglin, Silpa Gadiraju, Mary E. Russell, Cardiovascular Biology Laboratory, Harvard School of Public Health, Boston, Massachusetts As a way of modifying Thl and Th2 responses. we used Stat4- and Stat6-knockout mice as recipients in our mouse cardiac transplant model of chronic rejection (CBA into C57BL6x129, anti-CD418 for 28 days). At day 55 after transplantation, cardiac grafts placed into Stat4 -/- (n=lO) had significantly reduced frequency (24f2%) and severity (9i4%) of vascular occlusion when compared to wildtype controls (n=7, frequency 7Oil2% [p
152
FIVE-YEAR FOLLOW-UP OF A RANDOMIZED TRIAL OF PRAVASTATIN IN HEART TRANSPLANT PATIENTS. 3.A. Kobashigawa, J.D. Moriguchi, H. Laks, T.K. Ro, M.A. Hamilton, A. Hage, L. Wener, N. Kawata. University of California, Los Angeles, Calif. The original publication of this trial’s first-year data was the first to suggest significant benefit in outcome in heart transplant patients randomized to pravastatin (PVS), an HMG-CoA reductase inhibitor, or no PVS at the time of transplant surgery. We now report the 5-year follow-up of this study. 97 heart transplant patients given triple drug immunosuppression, between 7/92 and 2/94, were randomized to PVS or no PVS within 2 weeks after transplant. After the fmt year of the smdy, there was a considerable number of control patients (71%) who crossed over to take PVS due to the first year study results demonstrating benefit of PVS. Five-year survival continues to be superior in the PVS group with greater divergence in survival compared to the first year results (see Table). Major causes of death in both groups were rejection and transplant coronary artery disease (TCAD). In the PVS group vs. the control group, there were 5 vs. 10 patients who died due to rejection and 2 vs. 5 patients who died due to TCAD. Intravascular ultrasound (IVUS) of 24 patients with paired stodies (10 control and 14 PVS patients) from baseline (6 weeks after transplant) to 5 years after transplant revealed a significant reduction in the change in intimal area (IA) in the PVS group vs. the control group. Although not significant, there was a numerically lower change in intimal index (II) and lumen area (LA) and more patients free from TCAD (angiography) and/or death in the PVS group vs. control group. 5-VCU AIWS lbaseline to 5 vrs) Freedom from IA II LA Survival TCAD and/or death (mm2) (mm*) Prnva Group 83% * 64% 0.42’ .06 -3.14 62% 55% 1.36 .ll -3.49 Control Group * p < 0.05
Conclusion: The use of pravastatin early after heart transplantation appears to have continued beneficial effects in survival, rejection, and the development of TCAD at 5 years after Uansplant. A large crossover to PVS from the control group suggests ihat the early use of PVS appears important for good outcome.