Treatment Of Transplant Recipients Research Articles

Multidisciplinary Insights Into Optimizing Adherence After Solid Organ Transplantation

Nonadherence to medical treatment in transplant recipients is a major risk factor for graft rejection episodes, and it has significant financial implications. Despite its importance, there is a lack of common understanding across the disciplines involved of the key issues driving nonadherence. A qualitative study, comprising a multidisciplinary workshop, followed by a consultation exercise to validate its outcomes, was initiated to gain further insight into nonadherence behavior and to identify priorities for optimizing adherence to posttransplantation regimens. Eight statements relating to actions necessary to maximize adherence to posttransplantation medication were developed and offered for validation. All but one of these attracted a median score of 9 on an agreement scale of 1 to 10, where 10 was the highest level of agreement. The outcomes generate a structure that will facilitate communication and understanding and informing clinical practice and future research.

Severe Aspergillus endocarditis in a lung transplant recipient with a five-year survival

We report the case of a patient with cystic fibrosis who underwent lung transplant and developed Aspergillus endocarditis and cutaneous relapse. Long-term survival was achieved with surgical and prolonged antifungal treatment. This case report emphasizes the recommendation of life-long antifungal treatment in transplant recipients who survive an episode of fungal endocarditis.

Invasive Aspergillosis in Solid Organ Transplant Recipients

This chapter discusses the current status and evolving trends in the epidemiology, risk factors, diagnostic laboratory assays, and the approach to antifungal prophylaxis and treatment of invasive aspergillosis in Solid Organ Transplant (SOT) recipients. Most invasive fungal infections in these high-risk patients occur within the first month posttransplant; the median time to onset of invasive aspergillosis after renal replacement therapy and retransplantation was 13 and 28 days, respectively, in one study. Other factors associated with invasive aspergillosis in liver transplant recipients include transplantation for fulminate hepatic failure, cytomegalovirus (CMV) infection, and prolonged intensive care unit stay. A retrospective survey documented invasive pulmonary aspergillosis in 7.5% (19/251) of lung transplant recipients, of whom 47% (9/19) had disseminated aspergillosis. In a survey of antifungal prophylactic strategies in lung transplant units in the United States, aerosolized amphotericin B was the most frequently employed antifungal prophylactic agent. In order of preference, the prophylactic agents used in lung transplant centers in this study were inhaled amphotericin B (61%), itraconazole (46%), parenteral amphotericin formulations (25%), and fluconazole (21%); many centers used more than one agent. Preemptive therapy, where antifungal prophylaxis is directed towards lung transplant recipients colonized with Aspergillus spp. immediately before or within 6 to 9 months after transplantation, is another approach to the prevention of invasive aspergillosis in lung transplant recipients.

Transcriptional Profiles in Urine During Acute Rejection, Bacteriuria, CMV Infection and Stable Graft Function After Renal Transplantation

Acute rejection remains an important cause of renal allograft dysfunction and the need for accurate diagnosis is essential to treat transplant recipients successfully. Molecular markers in urine may serve as a diagnostic tool in acute rejection, but controversy still exists regarding the uniqueness of these biomarkers. We measured mRNA of perforin (PRF), granzyme B (GZMB) and granulysin (GNLY) normalized to cyclophilin B in urine specimens from 24 renal allograft recipients with acute rejection, 12 with bacteriuria, 11 with cytomegalovirus (CMV) infections and 17 controls with stable graft function. Measurements were performed using a real-time polymerase chain reaction assay. mRNA levels (means [95% CI]) for all three markers were significantly higher in recipients with acute rejection compared with controls: PRF (0.23 [0.12-0.42] versus 0.04 [0.02-0.07] P<0.001), GZMB (0.14 [0.09-0.23] versus 0.05 [0.03-0.08] P=0.003), GNLY (0.24 [0.14-0.41] versus 0.06 [0.03-0.11] P=0.001). GZMB and GNLY levels during acute rejection were significantly higher when compared with bacteriuria (P=0.011 and P=0.005 respectively), and PRF level during acute rejection was significantly elevated compared with CMV infection (P=0.015). No significant difference was found when comparing marker levels during bacteriuria and CMV infection to controls. Urinary mRNA levels of PRF, GZMB and GNLY are significantly elevated during acute rejection but not during bacteriuria or CMV infections when compared with recipients with stable graft function. The ability to differentiate acute rejection from bacteriuria and CMV infections was only present for some of the markers, that is why careful consideration should be given before applying this technique to clinical practice.

Situación actual de las infecciones fúngicas invasivas. Las nuevas técnicas diagnósticas y los nuevos antifúngicos

In the last few years, major advances in the treatment of transplant recipients, with hemato-oncological diseases or admitted to the intensive care unit, has been accompanied by an increase in classical fungal infections and by the emergence of uncommon fungal infections. Despite the development of new diagnostic techniques such as galactomannan detection and the availability of new antifungal agents, these opportunistic infections continue to pose a diagnostic challenge, prolong length of hospital stay, and increase costs. In addition, mortality from these infections is high. The present chapter provides a brief review of the epidemiology of these infections, diagnostic advances, and the new antifungal agents that have been developed in the last few years.

Update on the epidemiology and diagnosis of invasive fungal infection

In recent years, the most important advances in the treatment of transplant recipients, patients with haematological neoplasm and critically ill patients have been accompanied by an increase in the incidence of common fungal diseases and the emergence of some less common ones. Although new techniques (e.g. galactomannan detection) and new antifungals have appeared, these opportunistic infections remain difficult to diagnose and have a high mortality. New diagnostic techniques could improve this outlook, although they are far from becoming available in daily practice.

MRD Kinetics in Adult Transplantation Recipients with Relapsed Acute Lymphoblastic Leukemia Depend on Initial Molecular Response to Front-Line Chemotherapy.

Background: Adult patients with relapsed acute lymphoblastic leukemia (ALL) are candidates for allogeneic stem cell transplantation (SCT), if feasible. Despite the intensity of SCT treatment at least 40% of patients relapse. Survival is additionally negatively influenced by a high transplant related mortality. Therefore, markers that distinguish patients who will benefit from SCT from those who might profit from different therapeutic modalities or modification of SCT are highly warranted. Kinetics of minimal residual disease (MRD) can serve as molecular parameter of chemoresistance of the leukemia. The purpose of the current study was to investigate whether molecular resistance to front-line therapy can be overcome by allo-SCT following myeloablative conditioning.Methods: MRD was analyzed before and after myeloablative SCT in adult patients with Philadelphia negative relapsed ALL. MRD kinetics were compared to previous molecular response to first-line treatment. For this purpose, cases with persistent detectable disease >1×E–04 at day+71/112 of first-line therapy were classified as primarily molecularly chemoresistant (PMR), cases with MRD values below 1×E–04 around day+71/112 of first-line treatment were categorized as primarily molecularly chemosensitive (PMS). Relapsed ALL patients were included if they had been treated according to the GMALL trials 06/99 or 07/03 with prospective MRD monitoring, and if they had bone marrow samples taken after relapse until day+100 after SCT. Real-time quantitative (RQ)-PCR analysis of patient specific immunoglobulin and T-cell receptor gene rearrangements were used as targets for quantification of MRD.Results: 25 patients were eligible (15 T- and 10 B-lineage ALL). Median age was 22 (16–45) years. Median disease-free survival after front-line therapy was 17 (3 to 52) months. Eleven cases were classified as molecularly chemoresistant to front-line therapy, 14 ALL cases were categorized as primarily molecularly chemosensitive. In keeping with molecular response to initial treatment, PMR cases showed only a modest reduction of MRD in response to salvage chemotherapy with a median MRD value of 3×E–01 (range 2×E–03 to 1×E+00) before SCT. Median MRD levels within the first 100 days after myeloablative SCT decreased to 4×E–04 with a range between MRD negativity and 1.0×E+00. PMS cases showed a significantly better response to salvage chemotherapy: In contrast to PMR cases who were MRD positive prior to SCT in all analysed cases, four out of 7 PMS cases were MRD negative pre-SCT (range: MRD negativity to 4.2×E–03). Also within 100 days after SCT 13/16 analysed samples were MRD negative (compared to only 4/11 analysed PMR samples, p=0.02, see Figure). Taken together, these results show for relapsed ALL that molecular chemoresistance to front-line therapy correlates with a poor molecular response to second-line treatment in transplant recipients. [Display omitted]

Mycophenolate Mofetil in Lupus Glomerulonephritis

Mycophenolate mofetil (MMF) is an immunosuppressive agent initially used in the treatment of transplant recipients. MMF has been used in renal, heart, and liver transplantation, where it seems more effective than other immunosuppressive regimens in reducing the incidence of acute rejection episodes. MMF has a variety of immunosuppressive effects, including selective suppression of T and B lymphocyte proliferation, and has been more recently used in many autoimmune inflammatory conditions. Systemic lupus erythematosus (SLE) is an autoimmune disease that can potentially involve any organ or system of the human body. Glomerulonephritis (GLN) has been recognized as the most frequent severe manifestation of SLE, leading to poor long-term prognosis. In the treatment of lupus GLN, several therapeutic approaches, all including immunosuppressive drugs, such as cyclophosphamide, azathioprine, or cyclosporine A, have been used. The short- and long-term toxicity of these drugs limits their use in a substantial number of patients. Over the last few years, MMF has emerged as an alternative therapeutic regimen in lupus GLN, mainly for patients refractory to other therapies. These studies have shown that it is highly effective and generally well tolerated.

Costimulation Blockade-Induced Cardiac Allograft Tolerance: Inhibition of T Cell Expansion and Accumulation of Intragraft cD4+Foxp3+ T Cells

Previous studies have demonstrated that anti-CD40L or anti-B7 requires the presence of CD4(+)CD25(+) regulatory T cells (Treg) to induce antigen specific hyporesponsiveness. Other tolerance strategies involving Treg have shown a dependency on interleukin (IL)-10. The objective of this study was to investigate the role of CD4(+)CD25(+) Treg and IL-10 when treating transplant recipients with cytotoxic T lymphocyte-associated antigen (CTLA)-4 immunoglobulin (Ig), anti-CD40L, and anti-lymphocyte function-associated antigen (LFA)-1. Recombinase activating gene-deficient (Rag1(-/-) mice were transplanted with BALB/c hearts and adoptively transferred with IL-10(-/-) CD4(+) T cells, CD4(+)CD25(-) T cells or CD4(+)CD25(-)CD103(-) T cells and treated with costimulation blockade. Intragraft T cells from C57BL/6 recipients were analyzed for the expression of the Foxp3 protein after tolerance induction. Mice reconstituted with IL-10(-/-) CD4(+) T cells, CD4(+)CD25(-) T cells or CD4(+)CD25(-) CD103(-) T cells and treated with costimulation blockade accepted allografts permanently. Analysis of cells from recipient mice adoptively transferred with CD4(+)CD25(-) T cells contained a population of CD4(low)CD25(+) T cells 100 days after transplantation. Costimulation blockade partially prevented the homeostatic proliferation of CD4(+)CD25(-)CD103(-) T cells in Rag-1(-/-) recipients. Accepted allografts contained an elevated number of CD4(+)Foxp3(+) T cells. These results indicate that T-cell derived IL-10 is not essential for induction of graft acceptance in mice treated with costimulation blockade, but that treatment limits T-cell expansion in the recipients. The results further indicate that tolerance is maintained by intragraft CD4(+)Foxp3(+) T cells.

No Improvement of Patient or Graft Survival in Transplant Recipients Treated with Angiotensin-Converting Enzyme Inhibitors or Angiotensin II Type 1 Receptor Blockers

It was reported recently that treatment of kidney transplant recipients with angiotensin-converting enzyme inhibitors (ACEI) or angiotensin II type 1 receptor blockers (ARB) is associated with strikingly improved long-term graft and patient survival. This finding has important implications for future posttransplantation therapy recommendations. In an analysis of 17,209 kidney and 1744 heart transplant recipients, an association of treatment with ACEI/ARB with improved transplant outcome could not be confirmed. It is concluded that recommendations for a widespread use of ACEI/ARB treatment in transplant recipients are unwarranted.

Beneficial Effects of Simultaneous Treatment With 15-deoxyspergualin and Monoclonal Antibodies to CD45RB and CD154 on Murine Islet Transplantation Recipients

Treatment of transplant recipients with either 15-deoxyspergualin (DSG) or monoclonal antibodies (mAbs) to T-cell proteins CD45RB and CD154 (a two-signal blockade) has been shown to prolong islet graft survival. Therefore, we investigated the combined effect of DSG, anti-CD45RB, and anti-CD154 in murine islet model. Chemically induced diabetic C57BL/6 mice underwent allografting with islets from BALB/c mice or xenografting with rat islets. After transplantation, they were treated with either DSG, the two-signal blockade, or both (the triple treatment). The tolerogenic effects of the posttransplant treatments were measured with an intraperitoneal glucose tolerance test (IPGTT), immunohistology, enzyme-linked immunosorbent assays, and flow cytometry. Blood glucose profiles measured after glucose challenges were improved in all islet recipients. Enhancement of xenograft survival in triple-treated groups was not statistically significant (P = 0.08), compared to graft survival in group received only the two-signal blockade. However, 15 days after transplantation, xenografts in the triple-treated group showed a significant decrease in the proportion of CD4, CD8, and CD4CD45RB T-cells, and in the expression of interleukin-10 and interferon-gamma, relative to grafts in the other treatment groups. In addition, reduced infiltration of the xenografts by CD3 T-cells was observed in groups that had received either the two-signal blockade or the triple treatment. With long-term (>248 days) xenografts, only those in the triple-treated group were free of inflammatory infiltrates. These grafts also exhibited larger islet clusters and contained more insulin- and glucagon-positive cells, relative to grafts in the other treatment groups. Triple treatment has a beneficial effect in murine islet xenotransplantation.

Immunosuppression, Skin Cancer, and Ultraviolet A Radiation

Azathioprine is an immunosuppressant commonly used to treat transplant recipients. A recent study suggests that it renders the DNA of skin cells more sensitive to ultraviolet A radiation.

Prolonged administration of FTY720 does not cause renal toxicity in mice

Prevention of allograft rejection without renal toxicity caused by calcineurin-inhibitor immunosuppression is a major goal for transplantation. FTY720 is a synthetic drug that modulates immune responses of transplantation in many animal models. FTY720 modulating mechanisms relate to lymphocyte migration to peripheral lymph nodes instead of inflammatory sites. The drug has no effect on T-cell proliferation or cytokine production, and therefore prevents generalized immunosuppression. However, it is still to be confirmed that FTY720 has no nephrotoxic effects when administered continuously. In the present study FTY720 was administered orally for 21 days to C57BL/6 mice that underwent weekly evaluations. FTY720 (1 mg/kg per day) impaired body weight gain, but had no significant effect on either renal function or structure. Our findings suggest that FTY720 may provide a reasonable add-on therapy in calcineurin-inhibitor–treated transplant recipients.

Evaluation of an immunosuppressant side effect instrument

Clinicians continue to be compelled to evaluate the impact of immunosuppressive medication side effects on the quality of life of transplant recipients. We Were asked to develop an instrument to measure side effects in immunosuppressed transplant recipients. To construct an instrument that measures the impact and severity of side effects of immunosuppressive medications used in transplantation and to assess the reliability and validity of the newly developed instrument called the Memphis Survey. The instrument was constructed by a panel of physicians, nurses, and pharmacists with experience in treating transplant recipients. A small group of kidney transplant recipients (n= 13) provided pilot data for refining and testing the instrument. A national sample of kidney, liver, and heart transplant recipients (n = 505) provided data that were used to further develop the instrument. Factor analysis was used to determine the psychological dimensions underlying the instrument and to guide the construction of scales from the survey items. The instrument scales were then computed from the dataset of 505 transplant recipients to quantify the impact of immunosuppressant side effects on the quality of life of transplant recipients. Analyses showed the final instrument scales to be valid and reliable. Exploratory analysis suggests the need for further testing of the instrument to determine gender differences.

Evaluation of an Immunosuppressant Side Effect Instrument

Clinicians continue to be compelled to evaluate the impact of immunosuppressive medication side effects on the quality of life of transplant recipients. We Were asked to develop an instrument to measure side effects in immunosuppressed transplant recipients. To construct an instrument that measures the impact and severity of side effects of immunosuppressive medications used in transplantation and to assess the reliability and validity of the newly developed instrument called the Memphis Survey. The instrument was constructed by a panel of physicians, nurses, and pharmacists with experience in treating transplant recipients. A small group of kidney transplant recipients (n= 13) provided pilot data for refining and testing the instrument. A national sample of kidney, liver, and heart transplant recipients (n = 505) provided data that were used to further develop the instrument. Factor analysis was used to determine the psychological dimensions underlying the instrument and to guide the construction of scales from the survey items. The instrument scales were then computed from the dataset of 505 transplant recipients to quantify the impact of immunosuppressant side effects on the quality of life of transplant recipients. Analyses showed the final instrument scales to be valid and reliable. Exploratory analysis suggests the need for further testing of the instrument to determine gender differences.

Simultaneous determination of four immunosuppressants by means of high speed and robust on-line solid phase extraction–high performance liquid chromatography–tandem mass spectrometry

In this study immunosuppressants, i.e. cyclosporin A (CyA), tacrolimus (TRL), sirolimus (SRL) and everolimus (RAD) were quantified in whole blood samples from immunosuppressant treated transplant recipients by an integrated on-line solid phase extraction–high performance liquid chromatography–tandem mass spectrometry (SPE–HPLC–MS/MS) system. This method has been developed to improve the following characteristics: speed, robust analysis, simultaneous determination and low cost. This can be achieved by the use of a perfusion column as an extraction cartridge in combination with a short HPLC column and highly selective and sensitive atmospheric pressure ionisation tandem mass spectrometry (API–MS/MS) in the multiple reaction monitoring (MRM) detection mode. This high throughput technique is perfectly appropriate for routine therapeutic drug monitoring (TDM) of organ transplanted patients.

Effect of mycophenolate mofetil regimen on peripheral blood lymphocyte subsets in kidney transplant recipients

Background and aimsThere is growing evidence of the effects of immunosuppressive agents on “immune targets” in renal transplantation. Immunological monitoring could indirectly measure the suppressive effect of these drugs and guide early preventive interventions in transplant recipients. Due to the selective antiproliferative effect of mycophenolate mofetil (MMF) on lymphocytes, our goal was to determine whether MMF modulates peripheral blood lymphocyte subsets (PBLS) in kidney allograft patients. MethodsWe assessed absolute CD3+, CD3+CD4+, CD3+CD8+, CD19+, CD16+CD3− PBLS counts and CD4/CD8 ratios for 12 months in three groups of kidney allograft patients stratified according to maintenance immunosuppressive regimen: group A (n = 31), which started MMF with prednisone (P) + cyclosporine A (CyA), and two control groups, B (n = 19) and C (n = 15) on P + CyA + azathioprine (Aza) and P + CyA regimens, respectively. We compared intra- and intergroup lymphocyte counts and ratios. ResultsIntergroup comparisons showed a significant reduction in all PBLS in group A (CD19+ from 3 months and other subsets from 6 months), whereas there were no significant changes in PBLS in the other group analyses or comparisons. ConclusionsOur findings suggest that (1) MMF modulates all PBLS in kidney allograft patients, causing a progressive reduction occurring earlier in CD19+, and (2) we can rule out that these changes were caused by the “natural immunological evolution” of the transplantation. These results could offer a new method for immunological monitoring of transplant patients.

Anti-CD20 monoclonal antibody treatment of Epstein-Barr virus-induced intrahepatic lymphoproliferative disorder following liver transplantation

Abstract Epstein-Barr virus (EBV)-associated post-transplant lymphoproliferative disorders (PTLDs) are a common cause of death in transplant patients. Their incidence following liver transplantation is reported to be between 0.5% and 4%. Despite various therapeutic approaches, there is still no consensus on a treatment strategy. The treatment of transplant recipients with monoclonal antibodies directed against B-cell antigens is a new, therapeutic approach with which, however, little clinical experience has so far been gained. Two patients developed intrahepatic PTLD 7 and 15 months, respectively, after transplantation. In one case, this was diagnosed as polymorphic PTLD, in the other as monomorphic, monoclonal PTLD. After having their immunosuppression terminated, 4 weeks after establishment of the diagnosis, both patients were treated with anti-CD20 antibodies (rituximab) at a dose of 375 mg/m2 on days 1, 8, 15 and 22. Treatment with rituximab was tolerated well by both patients. One of the patients in whom cholestasis parameters remained high underwent re-transplantation. In one of the cases, the histological work-up confirmed necrosis of 90% of the tumour cells, and complete remission in the other. Both patients died of secondary complications 10 weeks and 10 months, respectively, after the diagnosis of PTLD. We can conclude that treatment of PTLD with Rituximab led to remission in both of our patients. Nevertheless, progression of cholestasis persisted, and both patients ultimately died of complications unrelated to PTLD.

Anti-CD20 monoclonal antibody treatment of Epstein-Barr virus-induced intrahepatic lymphoproliferative disorder following liver transplantation.

Epstein-Barr virus (EBV)-associated post-transplant lymphoproliferative disorders (PTLDs) are a common cause of death in transplant patients. Their incidence following liver transplantation is reported to be between 0.5% and 4%. Despite various therapeutic approaches, there is still no consensus on a treatment strategy. The treatment of transplant recipients with monoclonal antibodies directed against B-cell antigens is a new, therapeutic approach with which, however, little clinical experience has so far been gained. Two patients developed intrahepatic PTLD 7 and 15 months, respectively, after transplantation. In one case, this was diagnosed as polymorphic PTLD, in the other as monomorphic, monoclonal PTLD. After having their immunosuppression terminated, 4 weeks after establishment of the diagnosis, both patients were treated with anti-CD20 antibodies (rituximab) at a dose of 375 mg/m(2) on days 1, 8, 15 and 22. Treatment with rituximab was tolerated well by both patients. One of the patients in whom cholestasis parameters remained high underwent re-transplantation. In one of the cases, the histological work-up confirmed necrosis of 90% of the tumour cells, and complete remission in the other. Both patients died of secondary complications 10 weeks and 10 months, respectively, after the diagnosis of PTLD. We can conclude that treatment of PTLD with Rituximab led to remission in both of our patients. Nevertheless, progression of cholestasis persisted, and both patients ultimately died of complications unrelated to PTLD.

Measurement of human cytomegalovirus loads by quantitative real-time PCR for monitoring clinical intervention in transplant recipients.

Quantitative monitoring of human cytomegalovirus (HCMV) infection is helpful in determining appropriate antiviral management of transplant recipients. Quantitative PCR technologies have demonstrated accuracy in measuring systemic HCMV loads. A total of 298 consecutive whole-blood specimens submitted to the Clinical Virology Laboratory at Vanderbilt University Medical Center from 15 February to 31 October 1999 were included in the study. In addition to a qualitative colorimetric microtiter plate PCR assay (MTP-PCR) and a semiquantitative pp65 antigenemia assay, each specimen was measured for HCMV loads by a quantitative PCR assay performed on an ABI PRISM 7700 Sequence Detection System (TaqMan). Compared to results of the MTP-PCR, the sensitivity, specificity, positive predictive value, and negative predictive value were 70.5, 97.5, 87.8, and 92.8% for the antigenemia assay and were 96.7, 92.0, 75.6, and 99.1% for the TaqMan assay, respectively. There was a high correlation between antigenemia values and HCMV loads as determined by the TaqMan (r = 0.989; P < 0.001). Antigenemia values of 0, 1 to 10, 11 to 100, 101 to 1,000, and over 1,000 positive cells per 2 x 10(5) leukocytes corresponded to median HCMV loads measured by TaqMan of 125, 1,593, 5,713, 16,825, and 5,425,000 copies/ml, respectively. Corresponding to antigenemia values of 1 to 2, 10, and 50 positive cells per 2 x 10(5) leukocytes, HCMV viral loads of 1,000, 4,000, and 10,000 copies/ml are proposed as cutoff points for initiating antiviral therapy in patient groups with high, intermediate, and low risk of CMV diseases.