Diabetes mellitus in transplantation: 2002 consensus guidelines.

Diabetes mellitus is a serious complication following organ transplantation that is underdiagnosed, possibly due to the inadequate definitions used in published literature and the lack of standardized screening. Diabetes in transplantation amplifies the already increased risk of cardiovascular disease among transplant patients, and increases the risk of graft loss and death. Patients at risk of developing diabetes in transplantation should therefore be prospectively identified and given individualized immunosuppressive therapy to minimize the risk of developing this disease. These guidelines are intended to: (1) help identify patients at risk for diabetes after transplantation; (2) set down a standard definition of posttransplant diabetes mellitus (PTDM); (3) create a standard monitoring protocol for the diagnosis of PTDM; and (4) optimize the management of patients at risk of developing or who develop diabetes after transplantation. With improved diagnosis, individualization of therapy, and proper early management, the incidence of diabetes in transplantation, and the accompanying additional burden of illness the disease carries, may be diminished. In turn, this will help achieve the therapeutic goals of reducing the risk of graft complications, improving quality of life, and reducing postoperative morbidity and mortality in transplant patients.

The failure of Daudi cells to express the cellular prion protein is caused by a lack of glycosyl-phosphatidylinositol anchor formation.

The cellular prion protein (PrPc) is a glycosyl-phosphatidylinositol (GPI)-linked cell surface protein, which is expressed at high density on nervous tissues and at lower levels on most other solid-organ tissues. It is also expressed on peripheral blood mononuclear cells (PBMC) of all lineages. In lymphocytes, its level of expression is dependent upon the state of cell activation, and polyclonal anti-PrP antisera partially block lectin-induced T-cell activation, suggesting a functional role of the protein in this process. Using the monoclonal antibody (mAb) 3F4 we examined PrPc surface immunoreactivity on leukaemic cell lines of T- and B-cell origin, and unexpectedly observed a complete lack of PrPc cell-surface expression in Daudi cells, while all other cell lines displayed discernible reactivity. We demonstrated the intracellular presence of PrP-specific mRNA and PrP protein. The lack of surface PrPc is unrelated to the well-known defect of beta2-microglobulin (beta2m) expression in Daudi cells as other beta2m-deficient cells, such as the melanoma cell line F0-1 and spleen cells from beta2m gene-deleted mice, were not deficient in cell-surface PrPc. Daudi cells failed to bind antibodies directed against all GPI-linked cell surface proteins. In somatic hybridization experiments using murine spleen cells as partners, we observed de novo expression of human PrPc, CD55 and CD59, thus demonstrating in Daudi cells the availability of these gene products for GPI linkage and cell-surface expression.

Anti-glomerular basement membrane glomerulonephritis after extracorporeal shock wave lithotripsy.

Extracorporeal shock wave lithotripsy (ESWL) is a common noninvasive procedure for removal of upper urinary tract stones. We present a case of a man who developed anti-glomerular basement membrane (anti-GBM) disease after ESWL and review the two other cases described in the medical literature. In all cases, the affected individuals expressed the HLA DR2/HLA DR15 major histocompatibility antigen and developed a rapidly progressive anti-GBM-induced glomerulonephritis 3 to 7 months after ESWL. Anti-GBM disease may be a rare complication of ESWL in susceptible individuals and should be considered in patients who develop acute renal failure after lithotripsy.

Two-hour cyclosporine level determination is the appropriate tool to monitor Neoral therapy.

To assess the safety profile of Neoral dose adjustment using cyclosporine (CsA) trough levels (C0) compared with levels obtained 2 h after the morning dose (C2), 30 stable adult heart transplant patients 1 yr or more after surgery were converted from Sandimmune to Neoral. After a baseline visit (before conversion), initial follow-up included two visits (2 and 4-6 wk after conversion). After the first visit, patients were randomized to Group I (C0: 100-200 ng/ml) or Group II (C2: 200-400 ng/ml). Abbreviated pharmacokinetics were obtained for the estimation of the AUC0-4 h. Renal function was assessed by serum creatinine and the cimetidine-modified creatinine clearance. C2 correlated better than C0 with the AUC0-4 h (r = 0.91 vs. 0.63). Initial Neoral dose (mg/kg/d) was similar in both groups (2.8 +/- 0.5 and 2.8 +/- 0.8), and was lower in Group II at the second visit (2.0 +/- 0.7 vs. 3.0 +/- 0.6, p = 0.0001). C2 levels decreased in Group II from 912 +/- 438 to 555 +/- 271 ng/ml (p = 0.01), without evidence of acute rejection on endomyocardial biopsies. After the second visit,-both groups were monitored with C2, and the range was increased to 300-600 ng/ml. At the last visit (additional follow-up of 5 +/- 1 months), Neoral dose (mg/kg/d) was reduced to 2.0 +/- 0.3 in Group I (p < 0.001) and 1.8 +/- 0.4 in Group II. Serum creatinine was lower in Group II at the second visit (138 +/- 59 vs. 168 +/- 37 mumol/L, p = 0.01) and was similar in both groups at the last visit. Neoral dose reduction based on C2 levels was not associated with acute rejection. The better correlation with the AUC0-4 h suggests that C2 may be more reliable than C0 for Neoral dose adjustment.

Safety and tolerability of cyclosporine and cyclosporine microemulsion during 18 months of follow-up in stable renal transplant recipients: a report of the Canadian Neoral Renal Study Group.

BACKGROUND: There has been concern that the increased drug exposure associated with treatment with cyclosporine microemulsion (CsA-ME) would lead to an increase in adverse events. METHODS: The long-term safety and tolerability of conventional cyclosporine (CsA) and CsA-ME were compared in a randomized, multicenter, pharmacoepidemiologic study involving 1097 stable renal transplant patients after 18 months of follow-up. RESULTS: No significant difference was seen in change in serum creatinine or calculated creatinine clearance between the two groups. Episodes of deterioration in renal function (change in serum creatinine > or = 20%) were categorized with the following results for CsA-ME versus CsA, respectively: acute rejection, 4.5% vs. 4.5%; chronic rejection, 8% vs. 11%; CsA nephrotoxicity, 12% vs. 7% (P=0.008); transient changes, 17% vs. 12%; other causes, 4% vs. 6%. During the first 6 months of the study, a transient increase in the incidence of gastrointestinal and neurological adverse events was seen in the CsA-ME group compared with the CsA group. Up to 18 months, patients in the CsA group reported significantly fewer hearing and vestibular disorders, but more cardiovascular problems than those in the CsA-ME group (P=0.035). CONCLUSIONS: Tolerance to CsA and CsA-ME was similar. Renal function over 18 months was not adversely affected by the increased drug exposure with CsA-ME, although there was a transient increase in nephrotoxicity. The frequency of acute and chronic rejection did not change.

Treatment of steroid-resistant and recurrent acute cardiac transplant rejection with a short course of antibody therapy.

The purpose of this study was to assess the efficacy of short courses of OKT3 and ATG, respectively, for steroid resistant or recurrent acute allograft cardiac rejection (AR). Between June 1988 and March 1994, 101 heart transplant patients were treated with a quadruple sequential immunosuppression protocol (ATG, azathioprine, CsA, and prednisone). AR was diagnosed by endomyocardial biopsy (EMB), and patients with scores > 2 (ISHLT) received pulse methylprednisolone, 500 mg i.v. on 3 consecutive days. In cases of steroid-resistant or recurrent AR, OKT3 (5 mg/d) or ATG (1.5-2.5 mg/kg/d), was administered for 5-7 d instead of the usual 10-14 d course. OKT3 (17 courses; 10 steroid resistant, 7 recurrent AR; 5.3 +/- 0.7 doses) was given to 16 patients (4F/12M, 45 +/- 11 yr), 29-269 d after transplantation. ATG (8 courses; 5 steroid resistant, 3 recurrent AR; 4.9 +/- 0.6 doses) was given to 8 patients (1F/7M, 53 +/- 7 yr), 23-503 d after transplantation. Successful treatment of AR with a score < 2 at the first and second EMB after treatment was 88% and 88% with OKT3, and 87.5% and 100% with ATG, respectively. Throughout follow-up (50 +/- 22 months after OKT3; 49 +/- 28 months after ATG), there was a trend towards lower incidence of subsequent AR after ATG (25% vs. 69%, P = 0.09), and similar incidence of infections, graft atherosclerosis and mortality. No cases of lymphoproliferative disorder were observed. We conclude that short courses of OKT3 or ATG are safe and effective for the treatment of steroid resistant or recurrent AR, with a similar incidence of complications. These results may have cost-effectiveness implications and need to be confirmed in a randomized study.

Economic analysis of Neoral in de novo renal transplant patients in Canada.

This pilot economic evaluation was performed as part of the Canadian arm of an international randomized, controlled, double-blind safety and tolerability trial (OLM-105/NOF-2). The clinical study compared the safety and tolerability of a new microemulsion oral formulation of cyclosporine A (Neoral) with the oral cyclosporine. A preparation currently in use (Sandimmune SGC)/(SGC). To assess the economic impact of Neoral in newly grafted renal transplant patients, primary cost data were collected at the five participating Canadian centers and evaluated from the Ministry of Health (MOH) and hospital perspectives. The results of this cost analysis are presented in this paper. Since the new formulation has shown more consistent absorption and a more predictable pharmacokinetic profile, medical resource utilization and, consequently, cost of treatment could be expected to be lower for those renal transplant recipients treated with Neoral than for those receiving standard SGC. The findings of this study support this hypothesis. Robustness of the conclusion was confirmed with sensitivity analyses. Reduced health care costs for patients treated with Neoral were primarily a result of fewer hospitalization days and lower physician costs for inpatient and outpatient procedures.

A randomized, prospective multicenter pharmacoepidemiologic study of cyclosporine microemulsion in stable renal graft recipients. Report of the Canadian Neoral Renal Transplantation Study Group.

BACKGROUND: The safety, tolerability, and pharmacokinetics of conventional cyclosporine (ConCsA) and cyclosporine microemulsion (MeCsA) were compared under conditions of normal clinical practice in a prospective, randomized, concentration-controlled, pharmacoepidemiologic study. METHODS: Between September 1994 and March 1995, 1097 stable renal transplant recipients in 14 Canadian centers were randomized 2:1 to treatment with MeCsA or ConCsA. Patients were commenced on each study drug at a dose equal to their previous therapy with ConCsA, and the dose was adjusted to maintain predose whole blood cyclosporine concentrations within the therapeutic range established for each center. Prednisone and azathioprine were continued unless dose adjustment was required for clinical reasons. RESULTS: The mean cyclosporine concentration was comparable in both treatment groups at all time points throughout the 6 months of follow-up. The mean dose of cyclosporine was 3.6 mg/kg/day in both treatment groups at entry to the study, and declined by 0.3% and by 2.8% in patients receiving ConCsA and MeCsA, respectively. The nature and severity of adverse events were similar in both treatment groups, but there was a transient increase in neurological and gastrointestinal complications in the group receiving MeCsA within the first month after conversion (P<0.05). Serum creatinine and creatinine clearance did not change in either treatment group throughout the study. Biopsy-proven acute rejection occurred in three patients (0.8%) receiving ConCsA and in seven patients (0.9%) receiving MeCsA, with non-histologically proven acute rejection in an additional three patients (0.8%) receiving ConCsA and five patients (0.6%) receiving MeCsA (P=NS). Serum creatinine rose transiently in 35 patients (9.8%) receiving ConCsA and 138 patients (18.7%) receiving MeCsA (P<0.05) and resolved either spontaneously or after a reduction in the cyclosporine dose. One graft was lost in the MeCsA group due to irreversible rejection, and seven patients died, three in the group receiving ConCsA and four of those receiving MeCsA (P=NS). Absorption of cyclosporine was more rapid and complete from MeCsA than from ConCsA during the first 4 hr of the dosing interval, resulting in almost 40% greater exposure to the drug (P<0.001). There was close correlation between area under the time-concentration curve (AUC) over the first 4 hr of the 12-hr dosage interval and AUC over the entire 12-hr dosage interval for both formulations, making AUC over the first 4 hr a good predictor of total cyclosporine exposure. Using this parameter, patients with low absorption randomized to receive MeCsA showed a marked increase in drug exposure by months 3 and 6, whereas there was no change in those who continued on ConCsA. A limited sampling strategy utilizing samples at the predose and postdose trough levels provided an excellent correlation with drug exposure, particularly for patients receiving MeCsA (r2=0.94 MeCsA vs. r2=0.89 ConCsA). CONCLUSIONS: MeCsA appears to be a safe and effective therapy in stable renal transplant patients and provides superior and more consistent absorption of cyclosporine when compared with ConCsA. Transient toxicity after conversion to MeCsA occurs in some patients, and may reflect the increased exposure to cyclosporine. Use of a limited sampling approach combining trough and 2-hr postdose concentrations may provide an effective way to monitor this exposure.

Effect of cyclosporin weaning on glomerular filtration rate in renal transplantation.

The objective of this study was to determine the effect of weaning cyclosporin on glomerular filtration rate in 55 renal transplant patients from the Renal Transplant Unit at the Royal Victoria Hospital. Men or women, older than 20 years of age, who received cadaveric or living-related renal transplant and who were treated with the complete cyclosporin protocol, were included. Weaning of cyclosporin was started 15 days after the prednisone dose was reduced to 15 mg/day and this weaning process was completed at day 105 posttransplant. The daily cyclosporin dose was decreased by 25 mg once every 2 weeks, and completely withdrawn after treatment when 50 mg/day was tolerated for 2 weeks. The duration of the weaning process varied because of fixed dose reduction. Renal graft function was assessed by plasma creatinine concentrations. We used the time point of cyclosporin discontinuation as time zero. Serum creatinine decreased over the time from 150 +/- 61 mmol/l at time zero to 100 +/- 18 mmol/l in the last determination, while GFR had a significant increment from 66 +/- 18 to 77 +/- 20. A total of 4 rejection episodes were observed 6 months after cyclosporin discontinuation. There were no graft failures and deaths. To determine predictors of improvement, we carried out multiple regression analysis and we found that prophylactic antilymphocyte globulin and the onset of graft function after transplantation were predictors.

Restricted TCR V beta repertoire in the human mixed lymphocyte reaction is determined by the TCR on the responder cell and not by the stimulating antigen on the stimulator cell.

The use of a restricted TCR repertoire has been reported in antigen-stimulated T cells. We have examined by flow cytometry and polymerase chain reaction the changes in the TCR variable beta (V beta) repertoire as they occur in the mixed leukocyte reaction. The HLA class I-negative lymphoblastoid B cell line, Daudi, an HLA class I-positive variant of the Daudi cell line, and the homozygous typing cell line HTC 9062 were used to stimulate HLA-mismatched responder cells from 8 individuals. The magnitude of HLA incompatibility in various combinations ranged from single DR10 and DR13 differences to full 6-antigen differences at the HLA-A, -B, and -DR loci. Using a panel of 19 V beta-specific oligonucleotide primers, changes in the level of TCR V beta mRNA were assessed. The observed V beta repertoire was not specific for the stimulator cell antigens used. We did not find a correlation between the number of mismatched HLA antigens and the number of V beta elements involved. It rather appeared that individual responder cells displayed a uniform V beta pattern following exposure to single and multiple HLA antigens. We conclude that the V beta repertoire was largely determined by factors associated with the TCR on the responder cells, and not as obviously influenced by the HLA antigen(s) on the stimulator cells.

A solitary plasmacytoma of donor origin arising 14 years after kidney allotransplantation.

Allotransplantation of solid organs transfers passenger leucocytes which may give rise to a state of persistent microchimaerism. In this report we describe the case of a patient who developed a solitary plasmacytoma in a transplanted kidney more than 10 years after allografting. The diagnosis was established on the basis of the presence of a monoclonal IgG kappa peak in the serum, and light chain proteinuria, the plasmacytoid features of tumour cells including cell surface expression of IgG, kappa light chains, CD20, CD38 and CD56, the absence of lytic bone lesions and a normal bone marrow biopsy, and the disappearance of the monoclonal IgG peak after graft nephrectomy. A donor origin of the tumour was established by HLA DNA typing of tumour, tumour-free kidney tissue, and peripheral blood leucocytes, respectively.

Expression of early and late activation markers on peripheral blood T lymphocytes does not reliably reflect immune events in transplanted hearts.

Monoclonal antibodies directed against early (receptors for interleukin-2 and transferrin [IL-2R, TfR]) and late (PTA1, alpha 1 integrin VLA-1) activation antigens were used as probes to monitor cardiac transplant patients for episodes of acute graft rejection. Age- and sex-matched patient control groups consisting of 11 patients awaiting cardiac transplantation and 13 kidney transplant recipients with long-term grafts, respectively, were used to define an upper limit for normal activation antigen expression (mean + 3 SD) in patients. Expression of all cell markers was significantly higher in both patient control groups than in healthy control individuals. Therefore, the level of activation marker expression in heart patients awaiting transplantation was used as comparison for the patient population under study. Sequential monitoring of 24 heart transplant recipients failed to demonstrate a significant correlation of increased activation marker expression with clinical events of immune activation. Subsequently 62 consecutive endomyocardial biopsy scores in 36 patients were compared with the expression of IL-2R, TfR and VLA-1 on peripheral blood T cells. Neither increased cellular infiltration of the endocardium, nor of the myocardium, was associated with increasing proportions of IL-2R, TfR, or VLA-1 positive T cells. Elevated T-cell expression of the three markers combined indicated acute graft rejection with a sensitivity, specificity, and overall accuracy of 38%, 52%, and 43%, respectively. Acute graft rejection in biopsies with associated myofiber damage (biopsy rejection scores 2 and 3A,B) was not associated with a change in the proportion of activated T cells in circulation within the first 6 months after transplantation.(ABSTRACT TRUNCATED AT 250 WORDS)

Prolonged survival after heart-lung transplantation in systemic lupus erythematosus.

Patients with multisystem involvement of connective tissue disorders are generally excluded from consideration for heart-lung and lung transplantation because of profound donor organ shortages. A 23-year-old woman with systemic lupus erythematosus (SLE) was referred for evaluation of severe, progressive pulmonary hypertension. She underwent an uneventful heart-lung transplant and received cyclosporine A, azathioprine, and prednisone on a long-term basis. Bronchiolitis obliterans resulted in the development of moderate airflow obstruction 18 months after transplantation, but the process was stabilized with augmented immunosuppression consisting of high-dose parenterally administered corticosteroids, and subsequently a course of antithymocyte globulin. Four years after transplant, despite the persistence of reduced complement levels, the patient remains functionally well without clinical manifestations of SLE. This patient's long-term successful outcome indicates that connective tissue disorders such as SLE do not necessarily represent absolute contraindications to heart-lung and lung transplantation.

Cyclosporine-induced elevation in circulating endothelin-1 in patients with solid-organ transplants.

The long-term use of cyclosporine is associated with significant complications, including hypertension and renal failure. Recent data from animal experiments suggest that alterations in renal function induced by high-dose CsA may be mediated by endothelin-1 (ET-1), a potent endothelium-derived vasoconstrictor and mitogenic peptide. The aim of the present study was to determine the effect of oral CsA on circulating levels of ET-1 in patients receiving standard immunosuppressive therapy following solid-organ transplantation (13 renal, 7 heart, 1 heart-lung, 1 liver). Plasma levels of ET-1 were measured by radioimmunoassay over a 24-hr period beginning with the oral administration of a single daily dose of CsA in 18 patients (5.6 +/- 0.5 mg/kg), or similar immunosuppressive therapy without CsA in 4 patients. Blood levels of CsA (parent compound and metabolites) were measured in 10 of the patients by RIA. Predose levels of ET-1 were similar in the two groups (1.73 +/- 0.32 and 1.29 +/- 0.9 pg/ml, respectively). Patients not receiving CsA showed no change in plasma ET-1 over the 24-hr period. In contrast, in the CsA-treated group there was a significant increase in plasma ET-1, reaching a peak at 6 hr (2.45 +/- 0.56 pg/ml, P < 0.03) that followed the peak increase in CsA parent compound and preceded the peak increase in metabolites. No significant differences were found between peak and trough levels of ET-1 in patients with moderate renal dysfunction (creatinine (Cr) > or = 150 mumol/L) compared with those with near-normal renal function, or patients receiving renal compared with nonrenal grafts. However, patients with long-term grafts (> 60 days) showed an exaggerated response to CsA, with a fractional increase in plasma ET-1 of 3.67 +/- 0.52 (n = 8) compared with 2.16 +/- 0.28 (n = 10) for patients with more recent grafts (P < 0.05). Therefore, oral administration of CsA causes an increase in circulating ET-1 in patients with solid-organ transplants that might contribute to CsA-associated nephrotoxicity and hypertension, particularly during long-term immunosuppressive therapy.

The utility of cyclosporine weaning in renal transplantation.

Abrupt conversion of cyclosporine immunosuppression to conventional treatment with azathioprine and prednisone avoids long-term cyclosporine nephrotoxicity, albeit at the cost of a 20% to 40% rejection rate. The authors investigated the benefits and risks of a cyclosporine weaning protocol in 24 cadaveric and 9 live donor kidney recipients treated with a sequential quadruple immunosuppressive protocol. In cadaver kidney recipients, slow tapering of cyclosporine resulted in a 19% (p less than 0.001) improvement in the glomerular filtration rate, as estimated by the inverse ratio of the plasma creatinine concentration. Cadaver kidney recipients were stratified according to graft function (GFR ratio greater than 0.76, less than 0.76) at the of cyclosporine discontinuation. In 12 patients with well-functioning grafts, a 24% improvement was observed, whereas in 12 patients with poor graft function, the gain was limited to 13%. Patients with limited graft function tended to have more acute rejection episodes before cyclosporine weaning (0.92 +/- 0.64 versus 0.42 +/- 0.64, not significant). When the 24 cadaver kidney recipients were stratified according to onset of graft function after transplantation (days to plasma creatinine of 250 mumol/L), need for dialysis, panel reactive antibodies (PRA), and duration of cyclosporine treatment, no significant differences in graft function were observed at the onset or end of cyclosporine weaning. Acute graft rejection before cyclosporine weaning was the only variable associated with a significantly lower estimated glomerular filtration rate ratio at the end of cyclosporine treatment (0.83 +/- 0.11 versus 0.67 +/- 0.16, p less than 0.01). Weaning of cyclosporine was associated with a minimal risk of acute graft rejection. A single patient with stable graft function at the onset of the weaning process experienced an acute but reversible rejection episode 2 months after cyclosporine was discontinued. In summary, gradual weaning of cyclosporine improves graft function, and eliminates the excessive risk of acute graft rejection without the need for additional corticosteroid treatment.

Cellular isoform of the scrapie agent protein participates in lymphocyte activation.

The scrapie agent protein (Sp33-37 or PrPSc) is the disease-associated isoform of a normal cellular membrane protein (Cp33-37 or PrPC) of unknown function. We report that normal human lymphocytes and lymphoid cell lines, but not erythrocytes or granulocytes, express PrPC mRNA and protein. PrPC is detectable on the surface of lymphocytes; the surface immunoreactivity is sensitive to phosphatidylinositol-specific phospholipase C, indicating glycosyl-phosphatidylinositol membrane anchorage. Lymphocyte PrPC surface abundance is increased by cell activation, and polyclonal antibodies to PrPC suppress mitogen-induced activation. We conclude that PrPC is a lymphocyte surface molecule that may participate in cell activation.