Ulrich Binswanger

A blinded, randomized clinical trial of mycophenolate mofetil for the prevention of acute rejection in cadaveric renal transplantation

Mycopehenolate mofetil (MMF) is a powerful immunosuppressant that inhibits the proliferation of T and B lymphocytes by blocking the enzyme inosine monophosphate dehydrogenase. MMF has been shown to prevent acute graft rejection in animal experiments and may have an important role in clinical renal transplantation. We conducted a prospective, double-blind, multi-center trial to compare the efficacy and safety of MMF and azathioprine within standard immunosuppressive regimen for patients receiving a first or second cadaveric renal graft. A total of 503 patients were randomized to groups receiving MMF 3 g (n=164), MMF 2 g (n=173), or azathioprine (AZA) 100-150 mg (n=166) daily. All were treated simultaneously with equivalent doses of cyclosporine and oral corticosteroids and followed for 12 months. The primary endpoint was treatment failure, defined as the occurrence of biopsy-proven graft rejection, graft loss, patient death, or discontinuation of the study drug during the first 6 months after transplantation. Treatment failure occurred in 50.% of patients in the AZA group by 6 months after transplantation, compared with 34.8% in the MMF 3g group (P=0.0045) and 38.2 % in the MMF 2g group (P=0.0287). Biopsy-proven rejection occurred in 15.9% of patients in the MMF 3 g group and 19.7% in the MMF2 g group, compared with 35.5% in the AZA group. Rejection of histologic severity grade II or more developed in 6.1 %, 10.4% and 19.9% of patients in the MMF 3 g, MMF 2 g, and AZA groups, respectively. Patients receiving MMF required less frequent and less intensive treatment for acute rejection: 24.4% of patients on MMF 3 g and 31.0% on MMF 2 g were tested for acute rejection, compared with 47.5% on AZA. Only 4.9% on MMF 3 g and 8.8% on MMF 2 g required antilymphocyte antibodies for treatment of severe or steroid-resistant rejection, compared with 15.4% of the patients on AZA. At 1 year after transplantation, graft survival in the MMF groups was marginally superior to that in the AZA group, although this difference was not statistically significant. Gastrointestinal toxicity and tissue-invasive cytomegalovirus infection were more common in the MMF 3 g group. Noncutaneous malignancies occurred in six patients on MMF 3 g, three patients on MMF 2 g, and four patients on AZA. Lymphoproliferative disorders occurred in two patients per MMF group, compared with one patient receiving AZA. MMF appears to be an important advance in prophylaxis following renal transplantation. It is associated with a significantly lower rate of treatment failure compared with AZA during the first 6 months after renal transplantation and produces a clinically important reduction in the incidence, severity, and treatment of acute graft rejection. These differences persist throughout the first year of follow-up. Clinical benefit was greatest with a dose of MMF 3 g/day, but gastrointestinal effects, invasive cytomegalovirus infection, and malignancies were slightly more common at that dose. The appropriate dose may lie between 2 g and 3 g per day and may require individualization depending on clinical course or other factors.

Topical photodynamic therapy in the treatment of actinic keratoses and Bowen’s disease in transplant recipients

Background. Transplant recipients (TR) have a dramatically increased risk for widespread epithelial neoplasms of the skin. Thus, there is a need to treat initial stages of these neoplasms such as actinic keratoses (AK) and Bowen’s disease (BD) to prevent progression to invasive and potentially fatal squamous cell carcinoma. Topical photodynamic therapy (PDT) has been proven to be an effective treatment for AK and BD in immunocompetent patients, but no prospective trials in immunocompromised TR have been performed so far. Methods. Twenty TR and 20 controls with histologically confirmed AK or BD underwent either a single or two consecutive treatments of topical PDT in an open trial. The application of 20% 5-aminolevulinic acid (ALA) for 5 hours was followed by illumination with 75 J/cm2 of visible light delivered at 80 mW/cm2 by an incoherent light source. Results. The overall complete response rates in TR at 4, 12, and 48 weeks were 0.86, 0.68, and 0.48, respectively. The cure rates in both patient groups were comparable at 4 weeks but were significantly lower in TR than in controls at 12 and 48 weeks (P <0.05). Side effects included erythema, edema, and crust formation after illumination. Cosmetic results were excellent without scar formation or alterations in pigmentation. Conclusions. Topical PDT with 20% 5-ALA is an effective and safe treatment for AK and BD in immunosuppressed TR, with initial response rates comparable with those in immunocompetent patients. It is particularly useful in TR because of the possibility for repeated treatment of large lesional areas.

Relation of cyclosporine blood levels to adverse effects on lipoproteins.

Hyperlipidemia is common in renal allograft recipients. To elucidate the role of cyclosporine in posttransplant hyperlipidemia, we measured lipids, lipoprotein lipids, and apolipoproteins of thirty-five renal allograft recipients and evaluated their relation to trough cyclosporine blood levels. All patients were on a triple immunosuppressive regimen with equal doses of prednisone and azathioprine, and had stable graft function. Cyclosporine blood levels were significantly correlated to total plasma cholesterol (P = 0.028), low-density lipoprotein cholesterol (P = 0.022), apolipoprotein B (P = 0.017), and the cholesterol/high-density lipoprotein cholesterol ratio (P < 0.002), but not to plasma triglycerides. Significant inverse correlations were found between cyclosporine blood levels and high-density lipoprotein cholesterol (P = 0.034), high-density lipoprotein3 cholesterol (P = 0.025), and apolipoprotein A-1 (P = 0.047), but not high-density lipoprotein2 cholesterol. The independent relation of cyclosporine blood levels to each of the measured lipid parameters was investigated by a stepwise regression model including age, body mass index, interval from transplantation, diabetes mellitus, plasma creatinine, and intake of diuretics and beta-blockers. After correction for these 7 variables, cyclosporine blood levels remained significantly associated with high-density lipoprotein cholesterol, high-density lipoprotein3 cholesterol, apolipoprotein A-1, apolipoprotein B, low-density lipoprotein cholesterol, and the cholesterol/high-density lipoprotein cholesterol ratio. These data suggest that cyclosporine causes atherogenic dyslipidemia.

Disseminated varicella infection in adult renal allograft recipients: four cases and a review of the literature.

Disseminated varicella-zoster (VZV) infection is a rare complication after renal allotransplantation in adults. We report four patients, among them one with combined VZV and cytomegalovirus infection. The main complications were hepatitis, pneumonitis, and disseminated intravascular coagulation. A review of the literature from 1981 to 2000 revealed 34 additional cases of disseminated varicella infection in adult renal allograft recipients with an overall mortality of 34%. Among these patients 82% suffered from primary varicella, 18% had a reactivation. High-dose acyclovir therapy combined with reduction of immunosuppression lead to reduction of mortality from 53% before 1990 to 22% after 1990. No immunosuppressive drug is significantly associated with a higher risk of disseminated VZV infection. Immunization against VZV in adults is still a matter of controversy. Whereas passive immunization is performed only for prophylactic but not therapeutic purpose, active immunization is routinely performed in children and may also be recommended for adults before renal transplantation.

Human Parathyroid Hormone IMMUNOLOGICAL CHARACTERIZATION OF ANTIBODIES AGAINST A GLANDULAR EXTRACT AND THE SYNTHETIC AMINO-TERMINAL FRAGMENTS 1-12 AND 1-34 AND THEIR USE IN THE DETERMINATION OF IMMUNOREACTIVE HORMONE IN HUMAN SERA

Antibodies to a urea-trichloroacetic acid extract [hPTH-(TCA)] of human parathyroid tumors and to the synthetic NH(2)-terminal fragments of human parathyroid hormone hPTH-(1-12) and -(1-34) were developed in goats to characterize immunochemically various PTH preparations and to estimate immunoreactive PTH (iPTH) in human sera. They were quantitated on the basis of their capacity to bind [(131)I]-hPTH-(1-12), [(131)I]hPTH-(1-34) or [(131)I]bovine PTH (bPTH-(1-84)). The quality of the antibodies was assessed by reference to inhibition of their interaction with labeled peptides by synthetic hPTH comprising 34 NH(2)-terminal amino acid residues or fragments thereof [hPTH-(1-12), -(13-34), -(18-34), -(25-34), -(18-24)] or by the Sephadex G-100-purified full-length peptide hPTH-(1-84) [hPTH-(1-84)G-100]. The synthetic peptides used in this work correspond in their structure to the NH(2)-terminal amino acid sequence 1-34, as elucidated by Brewer and collaborators (1972. Proc. Natl. Acad. Sci. U. S. A.69: 3583-3588). Inhibition studies were also carried out with bPTH-(1-34) and bPTH-(1-84). Anti-hPTH-(TCA) exhibited specificities directed to determinants in the COOH-terminal and NH(2)-terminal part of hPTH-(1-84) and exhibited cross-reactivity with bPTH-(1-84). Anti-hPTH-(1-34), on the other hand, showed immunological specificities mainly directed to antigenic determinants located in the COOH-terminal half of hPTH-(1-34). In addition, some reactivity with the NH(2)-terminal hPTH-(1-12) and with the extractive full-length peptides of human and bovine origin was observed. Antibodies to hPTH-(1-12) cross-reacted with hPTH-(1-34) and -(1-84)G-100.IPTH WAS RADIOIMMUNOLOGICALLY DETERMINED IN HUMAN SERA BY THE FOLLOWING SYSTEMS: (a) [(131)I]bPTH-(1-84), anti-hPTH-(TCA) and hPTH-(1-84)G-100 as standard; (b) [(131)I]hPTH-(1-34), anti-hPTH-(1-34) and hPTH-(1-34) as standard. With system (a), COOH-terminal fragments of hPTH-(1-84) having a molecular weight of approximately 7,000 were detected, and there was an almost total discrimination of serum iPTH levels in normal and in hyperparathyroid subjects. With system (b), on the other hand, several molecular species of iPTH were detected, including a component larger than hPTH-(1-84) and others similar to hPTH-(1-84) and to a fragment co-eluting with the NH(2)-terminal fragment hPTH-(1-34). When serum iPTH was assayed in system (b), there was a large overlap of iPTH levels in control subjects and in patients with primary hyperparathyroidism.

Metabolic aspects of phosphate replacement therapy for hypophosphatemia after renal transplantation: Impact on muscular phosphate content, mineral metabolism, and acid/base homeostasis

Hypophosphatemia caused by renal phosphate loss occurs frequently after kidney transplantation. In assumption of systemic phosphorus depletion, the presumed deficit commonly is replaced by oral phosphate supplements. However, such treatment is debatable, because intracellular phosphorus stores have not been assessed in this setting and may not be accurately reflected by serum phosphate concentrations. Moreover, disturbances in mineral metabolism from chronic renal failure, such as hypocalcemia and hyperparathyroidism, may be prolonged with oral phosphate supplements. Conversely, a neutral phosphate salt might improve renal acid excretion and systemic acid/base homeostasis for its properties as a urinary buffer and a poorly reabsorbable anion. Twenty-eight patients with mild early posttransplantation hypophosphatemia (0.3-0.75 mmol/L) were randomly assigned to receive either neutral sodium phosphate (Na(2)HPO(4)) or sodium chloride (NaCl) for 12 weeks and examined with regard to (1) correction of serum phosphate concentration and urinary phosphate handling; (2) muscular phosphate content; (3) serum calcium and parathyroid hormone (PTH); and, (4) renal acid handling and systemic acid/base homeostasis. Mean serum phosphate concentrations were similar and normal in both groups after 12 weeks of treatment; however, more patients in the NaCl group remained hypophosphatemic (93% versus 67%). Total muscular phosphorus content did not correlate with serum phosphate concentrations and was 25% below normophosphatemic controls but was completely restored after 12 weeks with and without phosphate supplementation. However, the percentage of the energy-rich phosphorus compound adenosine triphosphate (ATP) was significantly higher in the Na(2)HPO(4) group, as was the relative content of phosphodiesters. Also, compensated metabolic acidosis (hypobicarbonatemia with respiratory stimulation) was detected in most patients, which was significantly improved by neutral phosphate supplements through increased urinary titratable acidity. These benefits of added phosphate intake were not associated with any adverse effects on serum calcium and PTH concentrations. In conclusion, oral supplementation with a neutral phosphate salt effectively corrects posttransplantation hypophosphatemia, increases muscular ATP and phosphodiester content without affecting mineral metabolism, and improves renal acid excretion and systemic acid/base status.

The acute parathyroid hormone response to changes in ionized calcium during phosphate infusions in the cow.

Abstract. The concentration of plasma immunoreactive parathyroid hormone ([IPTH]) increased within one minute after the plasma ionized calcium concentration ([Ca++]) had been lowered by phosphate infusions in 8 cows. The decrease in [Ca++] could be accounted for by a rise in nonultrafiltrable calcium. The plasma total calcium concentration ([CaTot]) remained unchanged during the first 4 minutes of the phosphate infusion. Until this time [IPTH] was inversely related to [Ca++] and directly related to plasma phosphorus concentrations, but not to [CaTot]. Peak levels of [IPTH] were attained at 4 minutes, before the nadir of [Ca++] was reached and prior to a significant fall in [CaTot]. The data suggest that initial decreases in ionized but not in total calcium stimulate parathyroid hormone secretion. They provide evidence for a model of parathyroid hormone secretion which includes a small storage pool available for immediate release in response to a lowering of the [Ca++]. Between 4 and 12 minutes [IPTH] remained approximately constant in association with a continued fall in [Ca^], whereas between 13 and 16 minutes (the end of the phosphate infusions) [IPTH] was decreasing in association with still falling [Ca++]. It can be speculated that the synthesis of PTH is insufficient to account for a sustained increase in [IPTH], or that abrupt decreases of [Ca++] inside the parathyroid cells inhibit the secretion coupling mechanisms. Finally after 16 minutes [IPTH] continued decreasing in relation to the rising [Ca++].

Catheter-Related Complications During Continuous Ambulatory Peritoneal Dialysis (CAPD): A Retrospective Study on Sixty-Two Double-Cuff Tenckhoff Catheters

From May 1980 to April 1985, a total of 62 double-cuff Tenckhoff catheters were surgically implanted in 54 patients through a low medial laparotomy. The follow-up on continuous ambulatory peritoneal dialysis accounted for 1,029 patient-monthly. The patients who used a detachable double-bag system developed a total of four exit-site infections, one tunnel infection, three outer cuff erosions, four catheter dislocations, and two dialysate leaks. In five patients, the catheter had to be removed (one exit-site infection, two catheter breaks, and two catheter dislocations). Cumulative catheter survival using the method of life-time analysis was 81% and 70% after 1 and 2 years, respectively, considering all catheters implanted, and 92% and 92%, respectively, when peritonitis-related removal was excluded.

Stimulation of Na+/H+ exchange activity by endothelin in opossum kidney cells

Endothelin-1 (ET-1) controls multiple aspects of kidney function. In this study we have analysed the effects of ET-1 on apical Na+/H+ exchange activity in opossum kidney (OK) cells. ET-1 (at 10−10 M and 10−8 M) activated Na+/H+ exchange activity within 5 min of exposure. ET-1 (10−8 M) prevented PTH-induced (parathyroid hormone; 10−8 M) inhibition of Na+/H+ exchange activity; it also abolished transport inhibition in response to 10−3 M IBMX (isobutylmethylxanthine) and 3×10−7 M TPA (phorbol 12-myristate 13-acetate), but had no effect on the 8-bromo-cAMP-induced (10−4 M) decrease of transport rate. Basal cAMP content, IBMX- and PTH-stimulated cAMP production were unaffected by ET-1 (10−8 M). The stimulatory action of ET-1 (10−8 M) on Na+/H+ exchange activity was prevented by calphostin C (10−8 M). These data document that OK cells might serve as a useful in vitro model for analysis of cellular mechanisms involved in endothelin action; proteine kinase C activation seems to participate in the observed endothelin effects.

1,25-Dihydroxyvitamin D3 and the Synthetic Vitamin D Analogue, KH 1060, Modulate the Growth of Mouse Proximal Tubular Cells

Apart from its classical role in bone and mineral metabolism, 1,25-dihydroxyvitamin D3 (1,25-vitamin D3) has recently been shown to affect cell growth and differentiation from a variety of tissues primarily not involved in mineral metabolism and to provide immunomodulatory functions. Findings in human mesangial cells in vitro as well as in rat model of compensatory renal growth in vivo suggest that renal cells are a target for 1,25-vitamin D3. In the present study the effects of 1,25-vitamin D3 and a synthetic vitamin D analogue, KH 1060, on proliferation and protein synthesis were investigated in a mouse proximal tubular cell line (MCT). 1,25-vitamin D3 and KH 1060 inhibited cell proliferation dose dependently (10(-7) to 10(-12) M) and specifically as assessed by 3H-thymidine incorporation (DNA synthesis) and cell counting. The cellular protein concentration, protein synthesis (3H-methionine incorporation) and protein/DNA ratio were not influenced by 1,25-vitamin D3 and KH 1060. By analyzing c-fos and c-myc expression using semiquantitative RT-PCR, a constant basal expression of both protooncogenes, even under serum-free conditions, was found in MCT cells. Phorbol 12-myristate 13-acetate (TPA) further stimulated c-fos and c-myc expression, whereas treatment with 1,25-vitamin D3 (10(-7) M) it had no effect either in unstimulated or in TPA-stimulated cells. In conclusion, 1,25-vitamin D3 and a synthetic vitamin D analogue, KH 1060, provide growth-regulating effects on renal proximal tubular cells in vitro. These effects are not mediated by regulation of c-fos and c-myc. Further studies will have to clarify whether 1,25-vitamin D3 plays a physiologic role in renal development and growth in vivo.