Conversion from cyclosporin A to tacrolimus is safe and decreases blood pressure, cholesterol levels and TGF-beta 1 type I receptor expression.

To determine whether conversion from cyclosporin A (CsA) to tacrolimus (TAC)-based immunosuppressive therapy is safe and might lead to improvement in the clinical side effect profile we studied 55 cardiac allograft recipients. Ten stable patients were electively converted (0.2-1.5 yr after transplantation; group I) and 45 patients were converted on indication (0.5-14 yr after transplantation; group II). We studied blood pressure, cholesterol level and renal function in all patients. To unravel the mechanisms by which CsA may exert its toxic effects and to evaluate whether conversion is associated with immune activation, we analyzed the transforming growth factor (TGF)-beta 1 system and intragraft interleukin (IL)-2 and IL-15 mRNA expression by real-time reverse transcription-polymerase chain reaction (RT-PCR) and quantitative flow cytometry in the selectively converted patients (group I). Conversion did not result in immune activation as no clinical, histological or molecular signs of immune activation (increased intragraft IL-2 and IL-15 messenger RNA (mRNA) expression) leading to rejection were found. It did not improve renal function neither in patient group I nor in patient group II. However, after conversion the blood pressure decreased (group I: systolic 154+/-16 vs 143+/-21 mmHg, p=0.03, diastolic: 99+/-11 vs 90+/-11, p=0.02 and group II: systolic 155+/-17 vs 142+/-14, p<0.001, diastolic: 99+/-11 vs 91+/-8 mmHg, p<0.001). Likewise, the cholesterol levels improved (group I: 6.6+/-0.5 vs 5.7+/-0.3 mmol/L, p=0.001 and group II: 7.1+/-1.7 vs 6.1+/-1.7 mmol/L, p=0.001). When patients were treated with TAC the ongoing rejections (n=4) resolved and gum hyperplasia disappeared (n=5). Conversion was associated with a two-fold lower TGF-beta 1 type I receptor expression on peripheral lymphocytes and monocytes (p=0.02 and p=0.002, respectively). Conversion from CsA to TAC results in improvement of blood pressure and cholesterol levels and does not induce immune activation. These beneficial effects were accompanied with lower TGF-beta 1 type I receptor expression.

Impact of heart transplantation on the safety and feasibility of the dobutamine stress test.

BACKGROUND: Dobutamine myocardial perfusion imaging is a useful method for evaluation of coronary artery disease. However, this technique does not allow for ischemia monitoring, which may have an impact on the safety of the test in heart transplant recipients due to cardiac sensory denervation. The aim of this study was to assess the impact of heart transplantation on the feasibility and complications of the dobutamine stress test. METHODS: We studied 225 heart transplant recipients (mean age 57 +/- 7 years) and a control group of 225 patients without previous transplant matched for age and gender by dobutamine (up to 40 microg/kg per minute) stress myocardial perfusion imaging. RESULTS: During the test, transplant recipients had a lower prevalence of premature ventricular contractions (23% vs. 37%, p < 0.001) and ventricular tachycardia (0.04% vs 7.5%, p < 0.0001) compared with control patients. By multivariate analysis, heart transplantation was a powerful independent variable associated with a reduced risk of ventricular arrhythmias (chi(2) = 20.8, p < 0.0001) and minor side effects (nausea, dizziness, anxiety, flushing, chills) (chi(2) = 20, p < 0.0001) during dobutamine stress. The target heart rate was reached in 82% of transplant recipients and in 77% of the control group. Overall feasibility (achievement of the target heart rate and/or an ischemic end-point) was 87% in the transplant and 86% in the control group. CONCLUSIONS: Dobutamine stress myocardial perfusion imaging is a safe and feasible method for evaluation of coronary artery disease in heart transplant recipients. The prevalence of arrhythmias and minor complications using the dobutamine stress test is lower in heart transplant recipients compared with control patients. The independent association between heart transplantation and reduced risk of arrhythmias and minor side effects of the dobutamine stress test indicates that cardiac sensory and autonomic nerve function plays a major role in the induction of these complications during the test.

Renal failure after clinical heart transplantation is associated with the TGF-beta 1 codon 10 gene polymorphism.

BACKGROUND: To determine whether genetic factors are involved in the development of renal dysfunction due to cyclosporine nephrotoxicity, we analyzed 2 polymorphisms in the signal sequence of the transforming growth factor (TGF)-beta 1 gene; codon 10 (Leu(10) --> Pro) and codon 25 (Arg(25) --> Pro). METHOD: Using sequence specific oligonucleotide probing, we analyzed both TGF-beta1 gene polymorphisms in cardiac allograft recipients (n = 168) who survived at least 1 year with minimal follow-up of 7 years. Patients received cyclosporine and steroids as maintenance immunosuppressive therapy. Renal dysfunction was defined as a serum creatinine > or = 250 micromol/liter. RESULTS: Renal dysfunction was observed in 2% (3/168) of the patients at 1 year, in 7% (11/160) at 3 years, in 12% (18/152) at 5 years, and in 20% (26/131) at 7 years post-transplantation. The genotypic distributions for TGF-beta1 codon 10 were 7% Pro/Pro, 61% Pro/Leu, and 32% Leu/Leu, and for codon 25 these percentages were 1% Pro/Pro, 12% Pro/Arg, and 87% Arg/Arg. We found an association between the TGF-beta 1 genotype encoding proline at codon 10 and renal dysfunction. At 7 years post-transplantation, 26% (23/89) of the patients with the heterozygous Pro/Leu or homozygous Pro/Pro genotype had renal dysfunction vs only 7% (3/42) of the patients with the homozygous Leu/Leu genotype (p = 0.017). For the TGF-beta1 codon 25 genotypes, we found no association between TGF-beta 1 genotypes and renal dysfunction. CONCLUSION: Our data support the hypothesis that TGF-beta 1 is involved in the process leading to renal insufficiency in cyclosporine-treated cardiac allograft recipients. In these patients the presence of TGF-beta 1 Pro(10) might be a risk factor.

Accuracy of dobutamine tetrofosmin myocardial perfusion imaging for the noninvasive diagnosis of transplant coronary artery stenosis.

BACKGROUND: Exercise stress myocardial perfusion scintigraphy has been used for the diagnosis of transplant coronary artery stenosis (TCAS) in cardiac allograft recipients. However, the role of pharmacologic stress myocardial perfusion imaging has not been evaluated. Aim of the study is to assess the accuracy of dobutamine stress 99m technetium tetrofosmin myocardial perfusion imaging for the diagnosis of TCAS in heart transplant recipients. PATIENTS AND METHODS: We studied 50 patients (age 56 +/- 8 year, 45 men) at a mean of 6.4 +/- 2.8 years after cardiac transplant with dobutamine (up to 40 ìg/kg/min) stress 99m technetium tetrofosmin SPECT. Resting images were acquired 24 hours after the stress study. Significant TCAS was defined as > or =50% luminal diameter stenosis by coronary angiography. RESULTS: Significant TCAS was detected in 30 patients (60%). Myocardial perfusion abnormalities (reversible and/or fixed defects) were detected in 27 of the 30 patients with and in 9 of the 20 patients without significant TCAS (sensitivity = 90%, CI 82-98, specificity = 55% CI 41-69, positive predictive value = 75%, CI 63-87, negative predictive value = 79%, CI 67-90 and accuracy = 76%, CI 64-88). Patients with multivessel TCAS had a larger stress perfusion defect score (5.6 +/- 3.1 vs 3.2 +/- 2.4, p < 0.05) compared to patients with single vessel TCAS. Among patients with abnormal perfusion who had no significant TCAS, 2 had lesions <50%, 2 had luminal irregularities and 5 had no abnormalities at angiography. Therefore specificity was 62% (8/13) in patients without any detectable angiographic abnormalities. CONCLUSIONS: Dobutamine stress tetrofosmin myocardial perfusion imaging is a highly sensitive method for the detection of TCAS in recipients of cardiac allografts. The high negative predictive value of the test indicates that patients who demonstrate normal perfusion by this method may be excluded from further invasive studies.

Functional responses of T cells blocked by anti-CD25 antibody therapy during cardiac rejection.

BACKGROUND: Despite anti-CD25 (interleukin [IL]-2 receptor alpha chain) monoclonal antibody (mAb) therapy, rejection can still occur. T-cell activation through the IL-2 receptor beta and gamma chains by IL-2 or other growth factors may contribute to this rejection. Recently, we have demonstrated that the T-cell growth factor IL-15 was abundantly present in rejecting cardiac grafts during anti-CD25 mAb treatment. METHODS: To test whether IL-2- and IL-15-responsive T cells play an active role in rejection during anti-CD25 mAb therapy, we measured the frequency of IL-2- and IL-15-proliferative T cells in peripheral blood from treated patients during rejection (n=12). Measurements were made by limiting dilution analysis in the absence and presence of extra in vitro-added mouse anti-human CD25 mAb. RESULTS: In the absence of anti-CD25 mAb, the frequencies of peripheral T cells responding to recombinant human (rh)IL-2 and rhIL-15 from patients were lower than those measured in samples of healthy controls (n=7): median of IL-2-responding T cells 78 per 10(6) (range 31-210 per 10(6)) vs. 154 per 10(6) (122-484 per 10(6), P=0.008) and median of IL-15-responding T cells 62 per 10(6) (range 19-207 per 10(6)) vs. 129 per 10(6) (range 79-192 per 10(6), P=0.02), respectively. In the presence of extra in vitro-added anti-CD25 mAb, frequencies of IL-2-responding T cells from patients significantly decreased, although a considerable number of T cells still proliferated on rhIL-2 (median 85%, range 46-100%). In contrast, the frequencies of IL-15 T cells still responding remained stable (median 2%, range 0-50%, P<0.001). CONCLUSIONS: Treatment with anti-CD25 mAbs cannot provide complete suppression of T-cell function because significant numbers of IL-2- and IL-15-responsive T cells remain present in the peripheral blood of allograft recipients during anti-CD25 mAb treatment.

Anti-CD25 therapy reveals the redundancy of the intragraft cytokine network after clinical heart transplantation.

BACKGROUND: Despite blockade of the interleukin-2/interleukin 2 receptor (IL-2/IL-2R) pathway by the murine anti-CD25 (i.e., IL-2R alpha chain) monoclonal antibody BT563, cardiac rejection can still occur. In these cases, growth factors other than IL-2 may contribute to allograft rejection. We studied the expression of IL-15, a macrophage-derived cytokine associated with T-cell activation, which interacts with the beta and gamma chains of the IL-2R during rejection episodes under anti-CD25 therapy. METHODS: We measured intragraft IL-15 mRNA expression and the number of IL-15- and CD68-positive cells in posttransplantation endomyocardial biopsies (EMBs; n=45) and in nontransplanted, donor-heart specimens (n=11) by competitive template reverse transcription-polymerase chain reaction and immunohistochemistry, respectively. RESULTS: IL-15 mRNA expression was present in the majority of posttransplantation EMB specimens (91%, 41/45) and in nontransplanted donor-heart specimens (91%, 10/11). Relative IL-15 mRNA levels were neither associated with transplantation nor with rejection status. After transplantation, the number of IL-15- and CD68-positive cells significantly increased (P<0.001), but IL-15-positive cell counts did not reflect the histological rejection grade. Anti-CD25 treatment, in contrast to its effects on the IL-2/IL-2R complex, had no influence on intragraft IL-15 mRNA and protein production. In rejection EMB specimens, during (n=5) and after (n=8) anti-CD25 therapy, no differences in relative IL-15 mRNA levels, or in IL-15- and CD68-positive cell counts, were measured. CONCLUSIONS: After heart transplantation, high numbers of IL-15- and CD68-positive cells infiltrate the graft. This phenomenon is independent of the rejection status. IL-15 remains present during blockade of the IL-2/IL-2R pathway by anti-CD25 monoclonal antibodies, and it may participate in T cell-dependent donor-directed immune responses, thereby explaining the occurrence of rejection in the absence of IL-2.

Spark erosion myectomy in hypertrophic obstructive cardiomyopathy.

The design features of the cutting electrode and the electrical characteristics of a monopolar electrosurgical device were specially adapted for performing a septal myectomy in patients with hypertrophic obstructive cardiomyopathy. Both the cutting behavior and electrode design were found to facilitate myectomy.