Monitoring of Epstein–Barr virus load and antibody in pediatric renal transplant patients

Background: Epstein–Barr virus (EBV) infection can lead to life-threatening post-transplant lymphoproliferative disorder (PTLD). The aim of the present study was to establish EBV monitoring methods to prevent PTLD.
Methods: EBV-DNA load was investigated, using real-time polymerase chain reaction (PCR) and anti-EBV antibody titers, in peripheral blood mononuclear cells of 21 renal transplant patients (seven recipients who were EBV-seronegative, R[−]; 14 who were EBV-seropositive, R[+]) before grafting. The mean age at entry and the mean follow-up period was 7.8 years of age (range, 3.3–12.0 years) and 1.8 years (range, 0.4–4.0 years), respectively, in the R(−) group, and 12.5 years of age (range, 3.9–17.7 years) and 3.8 years (range, 0.8–8.2 years) in the R(+) group, respectively.
Results: The mean maximum load of the EBV genome was 1071 copies/μg DNA (range, 106–20700 copies/μg DNA) in the R(−) group, and 61 copies/μg DNA (range, <50–552 copies/μg DNA) in the R(+) group. During follow up no patient in the R(+) group had any noticeable symptoms that could be related to EBV, but three recipients in the R(−) group developed EBV-related symptoms including adenoid hypertrophy, cervical lymphadenopathy, and PTLD (B cell lymphoma), in one patient each. In the R(−) group the first leukocyte-associated viremia was detected at 30–180 days, and seroconversion at 43–266 days after transplantation.
Conclusions: Viral DNA detection using PCR is a useful tool for EBV surveillance, but the maximum EBV load was not markedly elevated (2474 copies/μg DNA) in a patient with PTLD. Therefore, EBV surveillance using only monitoring of EBV load in peripheral leukocyte may be insufficient. Histology may therefore be necessary to accurately diagnose PTLD.     

Native kidney post-transplant lymphoproliferative disorder in a non-renal transplant patient

Abstract:  PTLD is an important post-transplant complication. Although PTLD affects kidney allografts after renal transplantation, it has not been reported in native kidneys of other solid organ recipients. Herein, we report a child who underwent an orthotropic liver transplant for cryptogenic cholestatic hepatitis and developed fever, generalized lymphadenopathy, chronic EBV viremia, and lymphatic PTLD. Subsequently, she also developed gross hematuria and nephrotic range proteinuria. Kidney histology revealed EBV-positive mononuclear infiltrates within the renal parenchyma consistent with PTLD. Electron microscopy examination demonstrated subepithelial electron-dense deposits consistent with a membranous glomerulopathy pattern. The PTLD was successfully treated with reduced immunosuppression and cyclic cyclophosphamide, rituximab, and prednisone, but the renal disease progressed to end-stage renal failure within two yr. Repeat kidney histology showed chronic nephropathy and membranous glomerulopathy without PTLD infiltrates or detectable EBV staining, although chronic viremia persisted. To our knowledge, this is the first such child to be reported and highlights the importance of remaining vigilant for renal PTLD even in non-kidney organ recipients.     

Various initial presentations of Epstein‐Barr virus infection‐associated post‐transplant lymphoproliferative disorder in pediatric liver transplantation recipients: Case series and literature review

PTLD is a rare but potentially life‐threatening condition, which shows a higher prevalence in children than in adults. From 129 children who underwent LT, we reported 5 cases with biopsy‐proven PTLD at a single teaching hospital. Four patients had shared clinical presentations including fever, lymphadenopathy, and splenomegaly. They were noted to be given a prolonged course of IS due to the management of comorbid complications such as acute cellular rejection or severe food allergy or eosinophilic gastrointestinal disease. The other one patient presented with upper gastrointestinal bleeding from gastric mass during an early post‐transplantation period. Notably, hypoalbuminemia was noted in all reported patients. Similar to previous studies, both EBV serology mismatch between the donor and recipient with high EBV viral load were noted in all except one case, whose EBV serology was unknown before LT. At least one episode of CMV reactivation was also observed in 3 of 5 patients prior to the PTLD diagnosis. The histopathology revealed 1 of 5 early PTLD, 1 of 5 polymorphic PTLD, and 3 of 5 monomorphic PTLD. The treatment included IS withdrawal, chemotherapy, and/or rituximab. One patient died of multiorgan dysfunction, one remains in complete remission, and three patients are either still on treatment or await response evaluation. Even though most of our reported PTLD cases had shared manifestations with fever, lymphadenopathy, splenomegaly, EBV serology mismatch, and high EBV viral load, various initial presentations such as respiratory symptoms, hypoalbuminemia, and prolonged use of IS from other causes such as significant food allergy were noted.     

Screening for PTLD in lung and heart-lung transplant recipients by measuring EBV DNA load in bronchoalveolar lavage fluid using real time PCR

Abstract:  Pediatric L-HLTx recipients are at risk for developing PTLD with the lung being a primary site of disease. We hypothesized that BALF is a better sample than peripheral blood for measuring EBV DNA load in this high-risk population. Archived BALF specimens from pediatric L-HLTx recipients with and without PTLD were assayed for EBV DNA load using a quantitative real time TaqMan PCR assay. These values were compared with values determined in peripheral blood by a competitive PCR assay. Fifty-five BALF specimens from 16 L-HLTx patients were evaluated. Three patients with PTLD had mean BALF EBV DNA load values almost 50-fold higher than subjects without PTLD (4.6 × 10⁵ copies/mL vs. 1.0 × 10⁴ copies/mL). Patients who were EBV seronegative pretransplantation (i.e., high risk for PTLD) had elevated EBV DNA load values vs. patients who were EBV seropositive pretransplantation, regardless of the diagnosis of PTLD (mean values of 3.2 × 10⁵ copies/mL vs. 1.1 × 10⁴ copies/mL). Lastly, BALF analysis identified all subjects with PTLD, whereas peripheral blood analysis identified only one of these cases. Therefore, it can be concluded that monitoring EBV DNA load in BALF following L-HLTx facilitates detection of PTLD in high-risk patients and may be superior to peripheral blood assays.     

Four-country surveillance of intestinal intussusception and diarrhoea in children

Aim:   Establishment of baseline epidemiology of intussusception in developing countries has become a necessity with the possibility of reintroduction of rotavirus vaccine. The current study assessed the seasonal trend in cases admitted with intussusceptions and dehydrating acute watery diarrhoea in children aged 2 months to 10 years.
Methods:   In a prospective surveillance study, teaching and research hospital sites in India (Lucknow and Nagpur), Brazil (Fortazela), Egypt (Ismailia) and Kenya (Nairobi) established a surveillance where a network of hospitals with surgical facilities catered to a reference population of about 1–2 million for reporting of intussusception. One large hospital per site also recruited admitted cases of acute watery diarrhoea.
Results:   From April 2004 to March 2006, 173 and 2346 cases of intussusception and diarrhoea, respectively, were recruited. Cases of intussusception had no apparent seasonality. Most cases of intussusception (61.3%) (107/173) were in the ≤1 year age group, with males comprising 68.8% (119/173) of all cases. Hospital mortality of intussusception was 4.2% (4/96). Cases of diarrhoea peaked in March, with 56.6% (1328/2346) of admitted cases being males. Majority (83.1%) of cases of diarrhoea had received antibiotics, and the hospital mortality was 0.8% (18/2280).
Conclusion:   Intussusception in the four participating countries exhibited no seasonal trend. We found that it is feasible to establish a surveillance network for intussusception in developing countries. Future efforts must define population base before the introduction of rotavirus vaccine and continue for some years thereafter.     

Perforated appendicitis and appendicolith in a child presenting as intussusception: a case report

Simultaneous diagnoses of intussusception and appendicitis in the same patient have rarely been described in the pediatric literature. A case of a 30-month-old boy is presented with an initial diagnosis of intussusception that was successfully reduced by air contrast enema. When the patient's condition deteriorated, a diagnostic search surprisingly revealed a missed perforated appendicitis with an appendicolith that had been part of the intussusception. The patient's hospitalization and surgical course is described along with a discussion of the intermingling of intussusception and appendicitis in a young child. This case illustrates the need to consider alternative diagnoses when a patient's course takes an unexpected and confusing turn.     

Hodgkin's lymphoma after post-transplant lymphoproliferative disease in a renal transplant recipient

Lymphoid malignancies such as post-transplant lymphoproliferative disease (PTLD) are a major complication of solid organ transplantation. Hodgkin's lymphoma (HL) is not part of the typical spectrum of PTLD, but has rarely been reported as a separate complication. We report a case of HL occurring after previous PTLD in a renal transplant recipient. A 9-yr-old girl with end-stage autosomal recessive polycystic kidney disease received a cadaveric renal transplant at 1 yr of age. She developed polymorphic PTLD localized to the bone marrow at 6 yr post-transplant. She was treated with reduction of immunosuppression and alpha-interferon. No chemotherapy or anti-B cell antibody was administered. The PTLD resolved and kidney graft function remained stable. At 9 yr post-transplant, she presented again with fever of 2 wk duration, associated with enlarged lymph nodes at multiple sites. A lymph node biopsy revealed the presence of classic Reed Sternberg cells positive for CD15, CD30 and EB RNA. She was treated with standard combination chemotherapy for HL with COPP/ABV. All immunosuppressive agents were discontinued except for low dose prednisone. The patient had an excellent response, with resolution of her lymphadenopathy and maintenance of stable graft function. RS like cells have been reported in the setting of PTLD, but these cells possess an activated B cell phenotype, are EBV negative and CD15 negative. True HL following PTLD has been reported in only three previous cases, with good response to standard chemotherapy in each.     

Cat scratch disease and acute rejection after pediatric renal transplantation

Cat scratch disease (CSD) can lead to unexplained fever, generalized lymphadenopathy and organomegaly in immunocompetent individuals. CSD has rarely been reported in immunocompromised transplant recipients, where its clinical features would mimic the more common post-transplant lymphoproliferative disease (PTLD). We report three cases of CSD seen recently in children who had received prior kidney transplants. The three children were between 7 and 9 yr old, and had received kidney transplants 2-4 yr prior, with stable renal function. In each case, there was unexplained fever with either lymphadenopathy or organomegaly. The diagnosis of CSD was suggested by a history of new cats being introduced into each household and confirmed in all cases by the serological presence of a significant titer (> 1 : 64) of IgM antibodies to Bartonella henselae. Tests for other bacterial infections, cytomegalovirus and Epstein-Barr virus infections were negative. All the patients showed a clinical improvement with anti-microbial therapy. In patients A and B, the CSD was associated with an acute rejection episode shortly after diagnosis. The rejection episodes were reversed by intravenous steroid pulse therapy. Only four cases of CSD have been previously reported following solid organ transplantation. Acute rejection following CSD has not been previously reported. CSD should be included in the differential diagnosis of fever in the post-transplant setting, especially where PTLD is suspected.     

Sinusoidal CD30+ diffuse large B‐cell lymphoma can masquerade as anaplastic large cell lymphoma in pediatric posttransplant lymphoproliferative disorders

Posttransplant lymphoproliferative disorder (PTLD) is a known complication of solid organ transplantation. Diffuse large B‐cell lymphoma (DLBCL) is frequently seen in this setting. However, CD30+ DLBCL with sinusoidal pattern of involvement has not been reported in pediatric PTLD. We are reporting a 9‐year‐old female child presented with diffuse lymphadenopathy postheart transplantation. The pattern of involvement was suggestive of anaplastic large cell lymphoma, but the malignant cells were positive for B‐cell markers and negative for anaplastic lymphoma kinase. The patient was treated aggressively with multiagent chemotherapy and rituximab. Accurate diagnosis in PTLD is paramount in making management decisions.     

Choledochal cyst in two pediatric heart transplant patients

Abstract:  Choledochal cyst is a relatively uncommon entity in Western countries. No reports of choledochal cyst in heart transplant patients have been reported to date. We report two cases of choledochal cyst in pediatric heart transplant recipients, one with post-transplant lymphoproliferative disorder (PTLD) within the cyst. The first patient had abdominal pain, increased liver enzymes and was seropositve for Epstein-Barr virus. A choledochal cyst with PTLD was removed 4 years after heart transplantation. The second patient presented 14 years after heart transplantation with a choledochal cyst that was excised for severe abdominal pain. This previously unreported association between choledochal cysts in conjunction with PTLD and heart transplantation is interesting and a possible common pathogenesis is proposed. The management and alternative treatments were briefly noted. We recommend an aggressive treatment for patients with suspected choledochal cyst after heart transplantation because of the increased potential for malignant transformation.     

Continuous infusion of thymoglobulin for induction therapy in pediatric heart transplant recipients; experience and outcomes with a novel strategy for administration

Abstract:  Minimal data exist on the perioperative use of TG for induction in pediatric HTx recipients. We report our experience using continuous infusion of TG on (i) perioperative adverse events, (ii) rejection, (iii) CAV, and (iv) PTLD. TG was infused via peripheral intravenous intra- and perioperatively as a continuous infusion (24 h/day). Starting dose was 1.5 mg/kg/day titrated to achieve target lymphocyte count of 0.1–0.3 × 10⁹/L. Fifty-five patients received TG; mean age at HTx was 4.4 yr (1 day–17.8 yr). The mean duration of TG was three and a half days (2–7 days). Median platelet count during TG infusion was 95 × 10⁹/L (28–228). Five patients had TG stopped for low platelets (at 4–6 days post-HTx) – all started maintenance immunosuppression. There was no perioperative mortality due to infection. Mean follow-up of 46 survivors was 2.3 yr (0.6–5.8 yr). Fifty-one percent had ≥ ISHLT 2R rejection at a median time of 33 days post-HTx (7 days–2 yr). One patient developed PTLD 1.4 yr post-HTx; three patients developed mild-moderate CAV. TG as a continuous infusion appears to have a good safety profile. Though mild thrombocytopenia was prevalent, there was no bleeding attributable solely to TG. Whether early depletion of T-cell function will translate into long-term benefits remains to be determined.     

Multifocal cutaneous and systemic plasmablastic lymphoma in an infant with combined living donor small bowel and liver transplant

Abstract:  Small bowel allograft recipients have a relatively high risk (approximately 20%) of developing PTLD. Onset of PTLD is usually soon after transplant (median of eight months). Children are at a higher risk than adults. Although PBL was originally described in 1997 by Delecluse et al. as a human immunodeficiency virus-associated neoplasm typically presenting in the oral cavity, it is now recognized as a PTLD. We describe an unusual and interesting case and to our knowledge the first case of an infant who developed diffuse multifocal cutaneous and systemic PBL shortly after small bowel and liver transplant. We report a case of a 14-month-old female child who received a small bowel and liver transplant from her father. She had excellent graft function with no rejection episodes. Five months post-transplant she developed a sudden gastrointestinal bleed and was noted to have a constantly rising EBV titer despite ongoing maximal antiviral therapy. A patchy erythematous rash was noted on her abdomen that was diagnosed as PBL–PTLD. By the time of this diagnosis, she had developed multiorgan failure unresponsive to therapy.     

Recombinant growth hormone use pretransplant and risk for post-transplant lymphoproliferative disease--a report of the NAPRTCS

Abstract:  rhGH, widely used to optimize linear growth in children with ESRD, also modulates B-cell precursor development and may be associated with malignancy development. To determine if rhGH use in children was associated with higher risk of PTLD, we analyzed retrospectively collected data on children with CRI, on dialysis or with renal transplants in a large multi-center registry of children with ESRD. Of the 194 LPD patients currently listed in the registry, 41 were previously enrolled in the CRI registry and 18/41 (43.9%) used rhGH during their period with CRI. Among CRI patients who later received a transplant, rates of PTLD post-transplant were significantly higher among rhGH users (18/407 or 4.4%) compared to patients who never used rhGH during their CRI follow-up and received a transplant (23/1240 or 1.9%, p = 0.009). After adjusting for the confounders of recipient age (at CRI and at transplant) and transplant era, the use of rhGH pretransplant was associated with a borderline higher risk for PTLD (odds ratio 1.88, 95% CI = 1.00–3.55, p = 0.05). In contrast, use of rhGH during dialysis or post-transplant only was not associated with a higher risk for PTLD. Continued monitoring is recommended.     

Chronic high Epstein-Barr viral load carriage in pediatric liver transplant recipients

Abstract:  Development of EBV disease and PTLD is usually accompanied by the detection of a high EBV load in peripheral blood. However, many children undergoing primary EBV infection following LTx will maintain chronically elevated EBV loads in the absence of clinical symptoms. To better understand this phenomenon, we retrospectively reviewed the records of children undergoing LTx at our center from 1997 to 2007 to identify chronic high EBV load carriers in this population. A CHL state was defined by the presence of a high load for >50% of samples for greater than or equal to six months following either asymptomatic or complete clinical resolution of EBV disease/PTLD. A total of 35 CHL carriers were identified. Pretransplant serologies were available for 29 of the 35; 22/29 (76%) were EBV negative prior to LTx; eight of these 22 developed their CHL state at the time of their primary EBV infection. Fourteen of the 35 had EBV disease (n = 7) or PTLD (n = 7) prior to development of the CHL state. Only one of 35 CHL carriers developed PTLD or lymphoma while they were a high load carrier. In all, 23/35 resolved their CHL state without apparent sequelae while 11 children continue to be asymptomatic high load carriers. These data provide important information about the outcome of chronic EBV high load carriage in pediatric liver transplant recipients.     

Unusual case of Epstein-Barr virus DNA tissue positive: blood negative in a patient with post-transplant lymphoproliferative disorder

Abstract:  Detection of PTLD uses PCR to detect circulating EBV DNA in the blood or in situ hybridization to identify EBV DNA in tissue biopsies. EBV DNA was detected in the tissue section using both real-time PCR and in situ hybridization. We report an unusual presentation of PTLD with no detectable EBV DNA in the blood using EBER-1 and EBNA-1 PCR assays. This report suggests that the use of EBV-PCR for the early detection of PTLD in blood samples may not be 100% effective in detecting disease.     

Unmanipulated donor lymphocytes for EBV-related PTLD after T-cell depleted HLA-haploidentical transplantation

Epstein-Barr virus (EBV)-related post-transplantation lymphoproliferative disorder (PTLD) is a life-threatening complication in patients given T-cell-depleted hematopoietic stem cell transplantation from an HLA-haploidentical relative (haplo-HSCT). We report the case of a child who developed severe EBV-related PTLD after haplo-HSCT from his mother. Despite receiving the anti-CD20 monoclonal antibody, the patient presented with intestinal obstruction due to huge abdominal lymphadenopathy, hematemesis, and nodulary pulmonary lesions. Histology showed that the lesions were due to CD20-/CD19+ large neoplastic B cells. The patient underwent double intestinal resection with partial abdominal lymphadenectomy and then received 3 monthly doses of donor-derived unmanipulated mononuclear cells. The initial dose of CD3+ cells was 3?10(5)/kg recipient body weight. The 2 additional doses consisted of 5?10(5) CD3+ cells/kg. No sign or symptom attributable to graft-versus-host disease was observed, and the patient completely cleared EBV-related lesions. The child was disease-free for 13 months after the first lymphocyte infusion. This case demonstrates that repeated infusions of controlled numbers of donor CD3+ cells cure EBV-related PTLD in haplo-HSCT without inducing graft-versus-host disease.     

T-cell PTLD presenting as acalculous cholecystitis

Abstract:  We report a five-yr-old child, presenting three yr after heart transplant with acalculous cholecystitis. Histology revealed EBV negative T-cell PTLD. The disease involved the gallbladder, liver, lungs, and mesenteric lymph nodes. He was treated with chemotherapy, went into remission, but relapsed after 11 months and died.     

Neurological symptoms in children with intussusception

Aim:  The classical combination of abdominal pain, vomiting, rectal blood loss and a palpable abdominal mass is only present in a minority of children with intussusception. Neurological signs and symptoms have been described, but are not a well understood phenomenon. We performed a retrospective study to ascertain the frequency and nature of these symptoms and to describe the characteristics of the patients presenting in this atypical way.
Methods:  The records of 58 children presenting with intussusception from 2003 to 2008 were reviewed for abdominal and neurological signs and symptoms, duration of symptoms and effectiveness of treatment.
Results:  In 10 out of 58 patients (17%), one or more neurological symptoms were recorded at presentation, with lethargy being the most frequent, followed by hypotonia and fluctuating consciousness. The patients with neurological abnormalities were significantly younger and presented with a shorter duration of symptoms. Therapy was more invasive, although not statistically significant, in this patient category.
Conclusion:  Intussusception should be considered in the differential diagnosis in young children presenting with lethargy, hypotonia and/or sudden alterations of consciousness even in the absence of the classical symptoms of intussusception.     

超声诊断和水压灌肠治疗小儿肠套叠

目的：　评价超声诊断和超声监视下水压灌肠治疗小儿急性肠套叠的方法和疗效。方法：　对173例可疑肠套叠患儿进行超声检查,发现有“同心圆”征的同时在超声监视下进行水压灌肠复位治疗。结果：　超声确诊肠套叠32例,诊断准确率100%,其中30例水压灌肠复位成功,复位成功率93.8%。3例回回结型中1例水压灌肠成功,2例失败改手术治疗。结论：　超声诊断小儿肠套叠准确率高,水压灌肠复位成功率高。对回回结型超声可以确诊,但复位成功率低