Comparative Quantitation of Cytomegalovirus (CMV) DNA in Solid Organ Transplant Recipients with CMV Infection by Using Two High-Throughput Automated Systems

Cytomegalovirus (CMV) DNA quantitation in clinical specimens is progressively becoming a cornerstone in the diagnosis and management of CMV infection in the immunocompromised host. We evaluated two automated and reproducible PCR tests, the LightCycler (Roche Molecular Biochemicals, Indianapolis, Ind.) and the COBAS AMPLICOR CMV Monitor (Roche Diagnostics, Pleasanton, Calif.), for the detection of CMV DNA in blood samples from transplant recipients with CMV infection as determined by shell vial culture. Following a log transformation analysis, the mean CMV DNA in plasma (PL), whole blood (WB), peripheral blood leukocytes (PBL), and peripheral blood mononuclear cells (PBMC) using the LightCycler was 6.79 copies per ml, 7.23 copies per ml, 6.38 copies per 2 x 10(6) cells, and 6.27 copies per 2 x 10(6) cells, respectively. This compares to 7.86 copies per ml, 8.37 copies per ml, 7.59 copies per 2 x 10(6) cells, and 7.44 copies per 2 x 10(6) cells, respectively, using COBAS AMPLICOR CMV Monitor. While higher CMV DNA levels were observed for the various blood compartments analyzed using COBAS AMPLICOR CMV Monitor, a high degree of correlation was evident between the two automated systems (jackknife correlation r = PL 0.77 [95% confidence interval (CI); 0.64, 0.90], WB 0.77 [95% CI; 0.62, 0.92], PBL 0.77 [95% CI; 0.67, 0.88], and PBMC 0.81 [95% CI; 0.72, 0.89], all P < 0.001). Therefore, we conclude that either automated diagnostic system is accurate for CMV DNA quantitation.     

Skin Cancer Risk Associated with Immunosuppressive Therapy in Organ Transplant Recipients

The aim of this review is to summarise the available literature regarding the epidemiology and proposed mechanisms of skin cancer development in organ transplant recipients who are receiving lifelong treatment with immunosuppressive therapy and to review the different strategies for managing complications in this group of patients. Organ transplantation is complicated by an increased incidence of certain cancers, of which non-Hodgkin's lymphoma, Kaposi's sarcoma and squamous cell carcinoma are the most common. The most important risk factor for these cancers is immunosuppressive therapy. The relative importance of different immunosuppressive therapy regimens in relation to the development of skin cancer is still unclear. Immunosuppression per se may play the most important role, but other mechanisms, which are independent of host immunity and which may be different for the various agents used, may also be of importance for the increased risk of cancer. Apart from immunosuppressive therapy, exposure to sunlight and infection with human papillomaviruses are believed to be the most important risk factors for the development of cutaneous squamous cell carcinoma in organ transplant recipients. Human papillomaviruses, no doubt, benefit considerably from immunosuppression, as is indicated by the large number of warts found in these patients, but many questions remain unanswered about their significance in cutaneous oncogenesis. The E6 protein from a range of cutaneous human papillomavirus types effectively inhibits apoptosis in response to ultraviolet light damage. It is, therefore, conceivable that the development of skin cancer in organ transplant recipients is the result of a complex interplay between exposure to ultraviolet radiation, human papillomavirus infection and genetic predisposition. Measures for protection from the sun are important for reducing the risk of skin cancer in organ transplant recipients. Regular surveillance of patients with skin problems and easy access to a dermatologist for these patients is advised. Changing the immunosuppressive regimen from azathioprine to cyclosporin or vice versa does not seem to relieve the skin problems. Tapering the immunosuppressive therapy to the lowest possible dose may be of some advantage. Oral retinoids, e.g. acitretin, have some effect in reducing the number of keratotic skin lesions and in the prevention of skin cancer in organ transplant recipients. Resurfacing the back of the hand can be a successful treatment for patients with multiple skin cancers on the back of the hand and can be used prophylactically in patients with severely actinically damaged skin.     

Osteoporosis in Transplant Recipients

Osteoporosis is a frequent and severe complication of transplantation that substantially reduces the recipient's quality of life. However, strategies to prevent or to treat osteoporosis have as yet not been established. Determination of bone mass and density is the basis for interventions following transplantation. Therefore, bone mineral density should be measured in all patients awaiting transplantation to assess their individual risk of fracture. Prophylaxis should be considered in patients at high risk in view of the rapid initial bone loss after transplantation. Treatment of acute rejection with corticosteroids induces more bone loss than maintenance therapy with corticosteroids. Once manifest osteoporosis has occurred, treatment with a triple therapy consisting of calcium, vitamin D and either calcitonin or bisphosphonates is possible without negative effects on graft function. Therapy should be initiated according to the general guidelines for osteoporosis therapy, but several aspects specific for this patient group have to be considered. The present recommendations for the management of osteoporosis after transplantation need to be proven by further intervention studies.     

Screening for Hepatitis B Virus DNA in Serum of Organ Donors and Renal Transplant Recipients

In order to determine the impact of screening potential organ donors for hepatitis B virus DNA using a standardized test, the serum of 145 donor candidates was tested. All of the candidates were negative for hepatitis B virus DNA, but the status of one donor was doubtful for hepatitis B virus surface antigen and seven donors tested positive for hepatitis B virus core antibody without hepatitis B virus surface antigen. Nine transplant recipients tested positive for hepatitis B virus surface antibody; they were given kidneys from the donor with a doubtful hepatitis B virus surface antigen result and from four of the seven donors who tested positive for hepatitis B core antibody. Follow-up revealed no case of hepatitis B transmission. In this study, screening for hepatitis B virus DNA was useful and did not lead to donor organ shortage. Patients with hepatitis B virus surface antibodies can safely be given kidneys from donors who are positive for hepatitis B core antibody but negative for hepatitis B virus DNA. Electronic Publication     

Tacrolimus in Heart Transplant Recipients

The development of cyclosporine was pivotal in allowing cardiac transplantation to become an accepted treatment for patients with end-stage heart disease. More recently, tacrolimus has become available as a useful alternative to cyclosporine, and has been successfully combined with either azathioprine or the newer anti-proliferative agents, mycophenolate mofetil or sirolimus. Numerous randomized clinical trials have demonstrated that tacrolimus is comparable to cyclosporine in terms of overall patient survival and at least equally effective in preventing acute rejection. In addition, tacrolimus has been particularly effective as a rescue treatment in cases where recurrent rejection has occurred with cyclosporine. The adverse effects of tacrolimus differ from those of cyclosporine, and the drug particularly shows an improved profile with respect to hypertension, dyslipidemia, and long-term renal function. These data have recently led to the regulatory approval of tacrolimus for primary immunosuppression in patients undergoing cardiac transplantation in the US.     

Study Finds US Organ Transplant Recipients Have High Risk of Developing 32 Types of Cancer

Researchers from the National Cancer Institute (NCI) evaluated medical data from 175,700 transplant recipients and discovered evidence that tied organ transplantation to     

Polymerase Chain Reaction Assays for the Detection of Cytomegalovirus in Organ and Bone Marrow Transplant Recipients

Cytomegalovirus (CMV) infection is ubiquitous and results in a wide spectrum of clinical manifestations ranging from asymptomatic infection to severe life threatening disease. Infection in normal children and adults usually causes no symptoms but in the immuncompromised host, CMV may result in severe opportunistic infections with high morbidity and mortality. Historically, virus detection was dependent on culture of the virus or on a centrifugation culture system referred to as a shell vial assay. The shell vial assay frequently lacked sensitivity and was unable to detect infection in its early phase. Also, as with culture assays, the results were affected by antiviral therapy. The CMV antigenemia assay was developed to provide more rapid results and has gained wide usage. This assay is limited to detection of the virus in white blood cells and is more sensitive than culture or the shell vial assay.

Application of the polymerase chain reaction (PCR) to these problems has resulted in the development of assays for CMV which are more sensitive than previously available methods. This method employs liquid hybridization with ³²P labeled probes and gel retardation analysis for detection of amplified DNA specific for each virus.

A comparison of the detection of CMV by an antigenemia assay or the PCR method in the leukocytes of renal transplant patients revealed that the PCR assay detects cytomegalovirus earlier and more consistently than the antigenemia assay.

Finally, the application of a fluorescent dye detection system and image analysis of the acrylamide gel with a laser scanner provides additional sensitivity to the detection of cytomegalovirus, as well as avoiding the use of radioactivity, making the assay more adaptable to the clinical laboratory.     

Cross-Infection of Solid Organ Transplant Recipients by a Multidrug-Resistant Klebsiella pneumoniae Isolate Producing the OXA-48 Carbapenemase,Likely Derived from a Multiorgan Donor

We describe two cases of bacteremic infections caused by a multidrug-resistant Klebsiella pneumoniae isolate producing the OXA-48 carbapenemase that occurred in two solid organ transplant (liver and kidney) recipients, which was apparently transmitted with the allografts. This finding underscores the risk of donor-derived infections by multidrug-resistant Gram-negative pathogens in solid organ transplant recipients and emphasizes the need for rapid screening of organ donors for carriage of similar pathogens.     

Ultrastructural Findings in Cardiac Transplant Recipients

Ultrastructural findings in 350 endomyocardial biopsy specimens and 59 autopsy or explanted hearts from cardiac transplant recipients are reviewed. Myocyte degeneration can be readily distinguished from necrosis by the technique described. Vascular changes of endothelial activation, endothelial cell damage, and basement membrane reduplication can be readily identified. In addition, the myofilament composition of ischemic hearts in patients with allograft coronary artery disease is distinctive: There is a disproportionate loss of actin over myosin, giving a coarse appearance to the myofilaments. These changes are useful in further defining the morphologic features associated with rejection and ischemia in cardiac transplant recipients.     

Psychophysiological Reactivity in Cardiac Transplant Recipients

To assess the contribution of the heart's autonomic innervation to reactivity to psychological stressors, hemodynamic responsiveness of the denervated human heart was examined in two studies. In Study 1, cardiac output measured by thermodilution. heart rate, and systolic and diastolic blood pressure responses to a 4-min mental arithmetic task were studied in 7 cardiac transplant patients during routine post-transplant cardiac catheterization. In Study II, 6 cardiac transplant patients, 5 normal controls, and 5 renal transplant patients participated in a 78-min psychophysiological stress protocol during which heart rate, systolic and diastolic pressure, and cardiac output (measured noninvasively by impedance cardiography) as well as serum epinephrine and norepinephrine were measured at baseline and while subjects performed mental arithmetic and reaction time tasks. In Study I, transplant patients showed significant increases, relative to baseline, in heart rate, systolic blood pressure, and cardiac output in response to mental arithmetic. The diastolic blood pressure response was marginally significant. In Study II, mental arithmetic produced significant reactivity in systolic blood pressure and marginally significant increases in heart rate and diastolic blood pressure in cardiac transplant patients. Reaction time produced only marginally significant diastolic blood pressure reactivity. Hemodynamic reactivity of the cardiac transplant group generally was lower than that of the two innervated groups, which generally were similar to each other. Although the small number of subjects makes conclusions tentative, these data suggest that: 1) Cardiac transplant patients are capable of significant reactivity to psychological stressors despite the absence of innervation of the heart, and 2) reactivity to these stressors is diminished relative to innervated control subjects. In the absence of cardiac innervation, reactivity is due to the vascular system and cardiac effects mediated by humoral factors.     

Hematopoietic Stem Cell Transplantation in Solid Organ Recipients with Emphasis on Transplant Complications: A Nationwide Retrospective Survey on Behalf of the Japan Society for Hematopoietic Stem Cell Transplantation Transplant Complications Working Group

Little is known about stem cell transplantation in solid organ transplantation (SOT) recipients. We conducted a nationwide retrospective survey of Japan Society for Hematopoietic Stem Cell Transplantation centers. A total of 19 patients who underwent 22 hematopoietic stem cell transplantations (HSCTs) after SOT were identified: 5 autologous HSCTs and 17 allogeneic HSCTs were performed. Patients who underwent autologous HSCT received a liver (n = 4) or kidney (n = 1) transplant. All 5 patients achieved neutrophil engraftment, and 2 of 3 patients with hepatoblastoma were alive at 1 year after HSCT. Allogeneic HSCT was performed in 16 patients (7 liver transplant recipients and 9 kidney transplant recipients). Among these, 2 donors were identical for both transplantations. All but 1 patient achieved neutrophil engraftment. The 5-year overall survival rate was 41.7%, but that in patients with malignant disease (n = 13) was much lower than the overall rate (23.1%). Only 1 patient with malignant disease underwent allogeneic HSCT in nonremission. In allogeneic HSCT after kidney transplantation, post-transplantation (1 year) kidney function in 5 evaluable patients was significantly lower than that before allogeneic HSCT, and 3 patients experienced renal rejection. However, no severe hepatic rejection was noted. In SOT recipients, HSCT is a potentially curable treatment for hematologic disorders, but it must be performed with caution, especially in patients with malignancy.     

CMV Infection in Bone Marrow and Solid Organ Transplant Patients in the Era of Antiviral Prophylaxis

Recent developments in the diagnosis and therapy of cytomegalovirus (CMV) infection have helped to reduce CMV-associated morbidity and mortality following allogeneic bone marrow and solid organ transplantation. The clinical symptoms of active CMV infection and the prevalence of life-threatening CMV disease vary widely between different patient populations according to the type of transplant and the intensity of immunosuppression employed. Antiviral prophylaxis with aciclovir, valaciclovir and ganciclovir has been shown to reduce CMV infection and disease following organ transplantation. Antiviral drugs, in particular ganciclovir and foscarnet, have varying sideeffects, however, and antiviral resistance due to prolonged administration of ganciclovir and foscarnet has been reported recently. Short courses of pre-emptive antiviral therapy for documented CMV infection help to reduce the duration and sideeffects of therapy, offering an alternative strategy to antiviral prophylaxis. Studies are required to compare the efficacy and costs of antiviral prophylaxis with pre-emptive therapy.     

Management of Medication Noncompliance in Solid-Organ Transplant Recipients

Solid-organ transplantation has emerged as one of the most significant medical advances in the management of end-stage organ disease to date. However, a long term successful outcome after transplantation relies heavily upon the extended, if not lifelong, intake of immunosuppressive medication. Noncompliance with the medication regimen may have devastating effects on the graft and the patient. Furthermore, the effects of noncompliance place an additional burden on the medical resources available and the already scarce organ supply. The magnitude of post-transplant noncompliance and factors associated with noncompliance with various immunosuppressant drugs are reviewed. Patient, physician, social and family relationships interact in a complex manner in the post-transplant scenario and problems here could underlie noncompliance. The paper also includes a review of the methods of evaluating and monitoring noncompliance. Preventive and remedial measures that may help the transplant team to effectively manage this problem are suggested. The multidisciplinary nature of post-transplant patient management and the need for a cohesive approach toward the patient is emphasised. With the identification of patients at higher risk for noncompliance, close monitoring and early intervention, it may be possible to effectively control the effects of noncompliance until newer strategies are developed which permit immunosuppression-free transplantation.     

Nocardia bacteraemia in an HIV positive patient - a case report

Nocardiosis has been recognized in recent times as an unusual opportunistic infection associated with HIV. Bacteraemia due to this pathogen is even rarer and only few cases have been reported in the literature. We report here a case of pulmonary nocardiosis with bacteraemia, which was initially diagnosed as pulmonary tuberculosis. A high index of suspicion is required to diagnose this infection as the clinical presentation and radiographic features mimic pulmonary tuberculosis.