Early post-transplant neopterin associated with one year survival and bacteremia in liver transplant recipients

Bacterial infections are the most common complications, and the major cause of mortality after liver transplantation (Tx). Neopterin, a marker of immune activation, is produced in monocyte/macrophages in response to inflammation. The aim of our study was to investigate whether early post-operation serum levels of neopterin were associated with post-transplant bacteremia and mortality in liver transplant recipients. We studied 162 of 262 liver Tx patients between January 2008 and February 2011 of whom pre- and early post-Tx sera samples were available.Pre- and early post-operative risk factors of infection and mortality were evaluated in 45 bacteremic patients and 117 non-bacteremic patients. During one-year follow-up, 28 of 262 patients died because of graft failure, septicemia and other diseases.Post-Tx serum neopterin on day 10 (p < 0.001) were significantly higher in bacteriemic patients than in patients without bacteremia. Logistic regression analyses showed that day 10 post-Tx neopterin serum level ⩾40 nmol/l has a predictive value (OR = 6.86: p < 0.001) for bacteremia and mortality (OR = 3.47: p = 0.021).Our results suggest that early post-Tx neopterin serum levels are very sensitive predictive markers of one-year post-Tx bacteremia and mortality in liver Tx recipients.     

Effect of major histocompatibility complex haplotype matching by C4 and MICA genotyping on acute graft versus host disease in unrelated hematopoietic stem cell transplantation

We explored whether matching of human leukocyte antigen (HLA) haplotypes between the recipient and donor of hematopoietic stem cell transplantation (HSCT) predicted by C4 and MICA typing is associated with the incidence of acute graft versus host disease (aGVHD). DNA preparations collected from a total of 81 recipient and donor pairs were used for PCR-based C4 subtyping and/or MICA sequence-based typing. Incidences of aGVHD were compared according to C4 and MICA matching. The six most common MICA alleles were MICA*008:01, *010:01, *002:01, *004, *009:01/049, and *012:01. Among the 59 unrelated pairs, HLA alleles were matched in 34 (57.6%). C4 subtypes were identical between the recipient and donor in 28 (82.4%) HLA-matched unrelated pairs, while MICA genotypes were matched in all HLA-matched unrelated pairs. In the 22 HLA-matched related pairs, all recipients showed identical C4 subtypes with their respective donors. In multivariate analysis, C4 mismatch was a significant risk factor associated with the development of aGVHD in unrelated HSCT (hazard ratio = 3.24, P = 0.006). PCR-based C4 subtyping is a simple method for assessing the genetic identity of the HLA region between a recipient and unrelated donor. This test would be also useful for prediction of aGVHD in HSCT.     

Using EasyMatch® to anticipate the identification of an HLA identical unrelated donor: A validated efficient time and cost saving method

In the absence of an HLA matched familial donor, a search for an unrelated donor or cord blood unit is initiated through worldwide registries. Although a first look-up on available HLA information of donors in the “book” at BMDW (Bone Marrow Donor Worldwide) can provide a good estimation of the number of compatible donors, the variety of resolution typing levels requires confirmatory typing (CT) which are expensive and time consuming. In order to help recipient centers in their work. The French donor registry (France Greffe de Moelle/Agence de la Biomedecine) has recently developed a software program called “EasyMatch®” that uses haplotype frequencies to compute the likelihood of phenotypic match in donors according to various typing resolution levels.The goal of our study is to report a single monocentric user-experience with EasyMatch®, demonstrating that its routine use reduced the cost and the delay of the donor search in our center, allowing the definition of a new strategy to search compatible unrelated donors.The strategy was first established on a retrospective cohort of 217 recipients (185 adults and 32 children = before score) and then validated on a prospective cohort of 171 recipients (160 adults and 11 children = after score).For all patients, we calculated the delay between the registration day and the donor identification day, and the number of CT requested to the donor centre. Considering both groups, we could observe a significant decrease of the number of CT from 8 to 2 (p < 0,001), and a significant decrease of the median delay to identify a suitable donor from 43 to 31 days (p < 0.0001).EasyMatch® estimates the number of potentially identical donors, but doesn’t foresee availability of the donors. It provides us an easy tracking of mismatches, an estimation of the number of potential donors, the selection of population following ethnic origin of patients and a high prediction when probability is high or low. It affords a new approach of donor search in our daily work and improves the efficiency in the great challenge of the compatible donor identification.     

Clinical polyomavirus BK variants with agnogene deletion are non-functional but rescued by trans-complementation

High-level replication of polyomavirus BK (BKV) in kidney transplant recipients is associated with the emergence of BKV variants with rearranged (rr) non-coding control region (NCCR) increasing viral early gene expression and cytopathology. Cloning and sequencing revealed the presence of a BKV quasispecies which included non-functional variants when assayed in a recombinant virus assay. Here we report that the rr-NCCR of BKV variants RH-3 and RH-12, both bearing a NCCR deletion including the 5′ end of the agnoprotein coding sequence, mediated early and late viral reporter gene expression in kidney cells. However, in a recombinant virus they failed to produce infectious progeny despite large T-antigen and VP1 expression and the formation of nuclear virus-like particles. Infectious progeny was generated when the agnogene was reconstructed in cis or agnoprotein provided in trans from a co-existing BKV rr-NCCR variant. We conclude that complementation can rescue non-functional BKV variants in vitro and possibly in vivo.     

A specific immune tolerance toward offspring cells is to exist after the mother lymphocyte infusion

PurposeTo examine immune tolerance between maternal lymphocytes and offspring tissue after a donor lymphocyte infusion.MethodsMouse models were established by mating female BALB/c mice with male C57BL mice. Splenic lymphocytes from donors of different genetic backgrounds were labeled with carboxyfluorescein succinimidyl ester (CFSE), and 1 × 10⁷ of the labeled cells were intravenously injected into a recipient. At 6 h, 24 h, 72 h and 120 h after the infusion, mononuclear cells in recipient spleen, liver, thymus, lymph nodes, and peripheral blood were collected. CFSE+, CFSE-, CD3+, CD8+, CD4+, CD19+, NK1.1+, CD25+, and CD127+ lymphocytes in those samples were analyzed by flow cytometry. The distribution of donor T cells, B cells, NK cells, helper T cells, cytotoxic T cells, and recipient regulatory T cells in the tissues were then analyzed.ResultsMaternal lymphocytes were more likely to survive in offspring. At 120 h after infusion, the percentages of maternal cells in the offspring were 0.52 ± 0.11% in lymph nodes, 0.97 ± 0.04% in peripheral blood, and 0.97 ± 0.11% in the spleen. Few donor cells, if any, were detected in these tissues at 120 h after aunt to child, father to child, and unrelated allogeneic infusions were performed. The subtype proportion of donor lymphocytes changed significantly in the recipient tissues. Recipient Treg cells increased in the mother to child group, but not in the aunt to child, father to child, and unrelated allogeneic groups, suggesting a decreased cellular immune response to allogeneic cells in the mother to child group. At 120 h after the infusion, no donor cells were detected in the recipient livers and thymuses of all groups, implying that donor cells were barely able to colonize in the liver and thymus.ConclusionSpecific immune tolerance to maternal lymphocytes exists in offspring. An infusion of maternal donor lymphocytes may produce a relatively persistent effect of adoptive immunotherapy with reduced side-effects.     

Human Leukocyte Antigen-G Polymorphisms Association With Cancer Post-Heart Transplantation

BackgroundPost transplantation, a major complication is the development of malignancies. Human Leukocyte Antigen (HLA)-G is a molecule that inhibits the immune system and it is utilized by malignant cells to hide from the immune system. Expression of HLA-G from the donor and recipient cells in transplant patients is regulated by gene variations however, the association between genotype and cancer remains unknown. Our objective was to determine the association between genotype and outcome.MethodsHeart transplant recipients (251) and available corresponding donors (196) samples were genotyped for polymorphisms and the association of polymorphisms to outcome was evaluated with parametric hazard regression models.ResultsRisk of cancer was 22% at 10 years post-transplantation. The mean follow-up was of 4.9 ± 3.6 years. In a multivariable analysis, donor–recipient SNP 3187 matching was identified as a protective factor for cancer (hazard ratio 0.43; 95% confidence interval 0.19–0.93; p = 0.03). While coding region allele (haplotype 6) was identified as an independent risk factor (hazard ratio 3.7; 95% confidence interval 1.36–10.06; p = 0.01).ConclusionIn this investigation, we identified an association between cancer post-transplantation and HLA-G polymorphisms, which may reveal a pathway for potential diagnostic and therapeutic strategies for cancer post-transplantation.     

Impact of blood donor characteristics on quality of packed red blood cell concentrates

IntroductionImpact of blood donor characteristics on quality of packed red blood cell concentrates.ObjectiveTo determine the impact of donor factors on the quality of packed red blood cell concentrates.Material and methodsThe analytical cross sectional study was conducted on 505 whole blood donors after approval by the Institutional Ethics Committee and written informed consent from blood donors. Two mL EDTA sample was collected for pre-donation haemoglobin estimation and all relevant donor details were recorded. Whole blood was collected in 350 mL double blood bags. PRBCs were prepared as per the departmental SOP. Volume of each PRBC was recorded and sample from each bag was taken for estimation of total haemoglobin content and haematocrit.ResultsOf 505 blood donors, 459 (90.9%) were males and 324 donors (64.2%) were less than 30 years of age. The majority of the donors were repeat donors (61%, n = 308 repeat donors), vegetarians (52.9%, n = 267 vegetarians) and non-smokers (92.7%, n = 468). Mean haemoglobin was found to be significantly higher in males (14.9 vs. 13.3; P ≤ 0.001), donors more than 30 years of age (15 vs. 14.7; P = 0.042), repeat donors (14.9 vs. 14.7), non-vegetarians (15.1 vs. 14.6; P ≤ 0.001) and smokers (15.3 vs. 14.8 g/dL; P = 0.020). PRBC units prepared from male blood donors, repeat donors and non vegetarians had significantly higher mean volume and mean total haemoglobin content. Strong positive correlation was observed between haemoglobin of the blood donor and total haemoglobin content of the PRBC and volume of blood collected.ConclusionsDonor characteristics do have effect on total haemoglobin content of packed red blood cells.     

Prevalence of hepatitis B surface antigen (HBsAg) in blood donor population in Bosnia and Herzegovina: Impact of the pre-donation questionnaire implementation and mandatory hepatitis B virus (HBV) vaccination schedule – 20 years’ experience of the University Clinical Hospital Mostar

ObjectivesCurrently, there are limited data on the prevalence of HBsAg in Bosnia and Herzegovina. This study aimed to evaluate the trend of HBsAg prevalence during a period of 20 years in relation to the implementation of pre-donation questionnaires based on parenteral and sexual risk factors and mandatory HBV vaccination.Material and methodsThis is a retrospective analysis performed on 67,336 blood donors at the University Clinical Hospital Mostar during three distinct periods: before introducing of mandatory HBV vaccination and pre-donation blood donor questionnaires (1998–2002); after introducing of pre-donation blood donor questionnaires, but without mandatory immunized blood donors (2004–2008); and after introducing of mandatory pre-donation blood donor questionnaires and mandatory vaccination (2015–2019).ResultsAccording to implementation of mandatory pre-donation blood donor questionnaires and mandatory HBV vaccination the prevalence of HBsAg significantly decreased among blood donors in all three studied periods of time (0.303% in 1998–2002, 0.236% in 2004–2008, and 0.021% in 2015–2019; P < 0.001). Out of 67,336 tested donors, there were 98 (0.145%) HBsAg-positive donors during analysed periods, with a mean age of 34.76 ± 11.37 years. The mean age of HBsAg-positive donors was 30.72 ± 10.40, 37.56 ± 10.13, and 47.28 ± 12.14) in the 1998–2002, 2004–2008, and 2015–2019 periods, respectively (P < 0.001).ConclusionThis study clearly shows a decreased rate of HBV infections among blood donors with respect to implementation of the HBV vaccination schedule and mandatory pre-donation blood donor questionnaires based on parenteral and sexual risk factors.     

Risk factors for cytomegalovirus DNAemia following haploidentical stem cell transplantation and its association with host hepatitis B virus serostatus

BackgroundThe risk factors associated with CMV DNAemia are not well known after haploidentical stem cell transplantation (SCT).ObjectivesThis study investigated the risk factors and prognosis for CMV DNAemia among CMV seromatched donors and recipients (D+/R+).Study designA retrospective study of patients undergoing haploidentical stem cell transplantation (SCT) between January 2010 and January 2012 was conducted. Cox regression analysis was performed to identify the risk factors for CMV DNAemia. These possible factors included recipient/donor age, recipient/donor gender, gender disparity, recipient HBsAg serostatus, diagnosis, risk stratification, anti-thymocyte globulin (ATG) dose (6 mg/kg,10 mg/kg), early neutrophil engraftment (≤12 days, >12 days), absolute lymphocyte count on day 30 (ALC30) and the occurrence of acute GVHD before CMV DNAemia.ResultsThe total number of patients was 248 with median age of 31 years (range, 14–56). The cumulative incidence of CMV DNAemia (146/248) was 59.5%. CMV DNAemia was first detected after a median of +35 days (range,12–82). Seventeen patients (17/146, 11.6%) developed CMV disease. Multivariate analysis identified HBsAg seropositivity (P = 0.002, hazard ratio (HR) = 1.833; 95%CI = 1.257–2.673) and the occurrence of acute GVHD before CMV DNAemia (P = 0.014; HR = 1.520; 95%CI = 1.088–2.124) as risk factors for CMV DNAemia. CMV DNAemia was associated with subsequent II-IV acute graft-versus-host disease (GVHD) (P = 0.014), III-IV aGVHD (P = 0.013) and chronic GVHD (P = 0.008). Totally, CMV DNAemia was found to be a poor prognostic factor in terms of non-relapse mortality (NRM) (P = 0.003, HR = 2.730; 95%CI = 1.406–5.197), and overall survival (OS) (P = 0.045, HR = 1.654; 95%CI = 1.012–2.701).ConclusionsOur data showed HBsAg seropositivity was associated with an increased risk of cytomegalovirus DNAemia. Detection of CMV DNAemia proved to be a poor prognostic factor for haploidentical patients.     

Reporting of delayed adverse donor reactions in whole blood donors: Just the tip of an iceberg!

Background and objectivesThe study was planned to determine the incidence and analyze how various epidemiological factors tend to be associated with delayed adverse donor reactions (ADR).Material and methodsThe prospective observational study was conducted in Department of Transfusion Medicine of tertiary care hospital from January to December 2019. Eligible blood donors were observed for any adverse reactions after 15 minutes of removal of phlebotomy needle. Further, telephonic calls were made to each enrolled blood donor on day-2 and day-7 of the whole blood donation. For each day, two calls were made at an interval of 4 hours before declaring the participant to be non-responder.ResultsA total of 1540 (84.1%) blood donors responded on day-2 and 1610 (87.9%) responded on day-7 of follow-up. Total 180 (11.2%) blood donors experienced delayed ADRs. Two donors (1.1%) experienced on-site while 178 (98.89%) reported off-site delayed ADRs when followed-up telephonically. The commonest delayed ADRs reported were bruise (n = 72; 30.9%), arm-pain (n = 61; 26.2%) and generalised weakness (n = 44; 18.9%). Female donors (27.3% vs. 11.2%; P = 0.004), first time donors (15.2 vs. 9.9%; P = 0.002), donors with low body-weight (range of 45–60 kg; 15.9% vs. 11.5% vs. 6.1%; P = 0.011) and body mass index < 18.5 (24% vs. 12.5% vs. 9.7% vs. 11.3%; P = 0.028) experienced more delayed ADRs.ConclusionBlood donors do experience delayed ADRs but these are not reported to the blood centers as these are usually mild. However, it is important to capture these delayed adverse donor reactions and report it to National Hemovigilance Program so that strategies can be formulated to prevent their occurrence and recurrence.     

Genetic analysis of JC virus and BK virus from a patient with progressive multifocal leukoencephalopathy with hyper IgM syndrome

A case of acute progressive multifocal leukoencephalopathy (PML) with hyper IgM syndrome 1 is reported. Viral DNA and VP1 protein of JC virus (JCV) and BK virus (BKV) were detected by immunohistochemistry, in situ hybridization, semi-nested polymerase chain (PCR) and PCR-restriction enzyme analysis. JCV DNA and VP1 protein were found in the nuclei of oligodendrocytes. The non-coding control region (NCCR) and VP1 region of the JCV genome were sequenced; this revealed a novel rearrangement pattern of the NCCR in the brain tissue. The VP1 regions of brain and urine JCV were identical and of genotype type 2A. The BKV in the urine sample was genotype I. No BKV genome was found in the brain. The novel genomic rearrangement of the JCV NCCR in the brain tissue may have altered JCV pathogenesis to induce PML; the impaired immunity from hyper IgM syndrome 1 may have enabled the rearrangement. The JCV NCCR rearrangement in the brain may have originated from the archetypal form in the urine through deletion and duplication.     

Plateletpheresis donation through donor awareness,motivation, and recruitment drives: A unique concept and experience from a tertiary care oncology centre in India

ObjectivesThe majority of blood donor motivational and awareness activities are directed toward whole blood donation and not much emphasis is given to the plateletpheresis. The study was designed to analyze the effectiveness of the unique concept of platelet drives (PD) to increase voluntary plateletpheresis donations.Materials and methodsThe study was a retrospective study conducted at a tertiary care oncology hospital-based blood transfusion services (BTS).ResultsA total of 13 PDs were conducted from January 2016 to December 2020. A total of 559 potential donors came for the donor registration and 125 donors got deferred on medical history. A total of 434 donors gave their samples for the testing of plateletpheresis. The median age of potential male and female donors was 32 and 30 years respectively. A total of 58 males and two females have donated single donor platelets (14.3% vs. 7.4%, P = 0.319). The median age of male and female donors was 36 and 42 years, respectively. In male donors, 48 had donated once, seven donated twice, two donated thrice and one donor donated four times. Out of two female donors, one donor donated twice and the other donor donated four times. The female donors were more committed to repeat donations (P = 0.004). Since the start of PDs, the number of voluntary donations has increased considerably over the years.ConclusionPDs have helped in increasing the number of voluntary plateletpheresis donors. All the BTS must have the standard operating procedures in place for these kinds of awareness and motivational drives.     

A 3-month course of ciprofloxacin does not prevent BK virus replication in heavily immunosuppressed kidney-transplant patients

BackgroundIn vitro and retrospective studies of kidney-transplant patients have shown that quinolones can efficiently prevent BK virus (BKV) replication. However, in a prospective study, a 3 month-course of levofloxacin did not decrease the rate of BK viruria in kidney-transplant patients treated with standard immunosuppression.ObjectivesThe aim of this study was to assess the effect of a 3-month course of ciprofloxacin prophylaxis on BKV replication in kidney-transplant patients that had received heavy immunosuppression (plasma exchange or immunoadsorption and rituximab) to achieve desensitization before undergoing HLA- and/or ABO-incompatible (ABOi) transplantation.Study designTwenty-nine patients were given ciprofloxacin (500 mg/d) for 3 months, starting immediately after transplantation. The results were compared with results from a previous study where patients had received a similar immunosuppression regimen without ciprofloxacin prophylaxis (n = 43). Around 60% of patients had undergone a retransplantation. After transplantation, all patients were given induction therapy, tacrolimus, mycophenolic acid and steroids. BK viruria and viremia were monitored at months 1, 3, 6 and 12 post-transplantation.ResultsThe rates of BK viruria, BK viremia, and BKV-associated nephropathy did not differ between patients who were given or not given ciprofloxacin prophylaxis. These rates were also identical when patients received quinolones at any time within the first year after transplantation compared to those that had not. The rate of bacterial infection was also similar in patients who had or had not received ciprofloxacin.ConclusionThe use of quinolones seemed to not have any beneficial effect in preventing BKV replication in kidney-transplant patients receiving heavy immunosuppression.     

Is high hemoglobin a hindrance factor for blood donation? A pilot observational study from the coastal region of India

BackgroundBlood donors with high Hb are often deferred for the presumed risk of polycythemia vera (PV). However, adequate data to substantiate or refute this hypothesis is lacking.MethodologyWe conducted an observational study on blood donors found to have high hemoglobin (Hb ≥ 18 g/dL) during the pre-donation screening process using a portable hemoglobinometer at our blood center for four months. We adopted a cost effective methodology wherein a questionnaire was used to elicit the secondary causative factors of high hemoglobin and a complete blood count test to observe the blood cell parameters and JAK2V617F mutation test was performed in a subset of donors lacking secondary erythrocytosis (SE) history.ResultsOf the total 7076 donors enrolled, 112 male donors (1.58%) had high hemoglobin. The majority (70.4%) were repeat donors with mean age of 31.4 years. About 61% of the donors had attributable factors for SE like smoking, occupational exposure to carbon monoxide. The mean hemoglobin value of capillary and venous hemoglobin demonstrated a statistically significant difference (P < 0.05) where 2.7% of donors had venous Hb < 18 g/dL. The hematological profile of all the donors showed increased RBC but normal platelet and WBC count. Of 24 donors included for the JAK2V617F test, none had a positive report.ConclusionThis study suggests high hemoglobin in blood donors is less likely due to PV. Hence, re-considering their deferral may help alleviate donor anxiety and allow donor return. However, multi-centric studies are required to develop consensus statements on PV risk status and blood donation eligibility.     

Seroprevalency of transfusion-transmitted infections in first-time volunteer and replacement donors in Tunisia

BackgroundReplacement donors are considered as having a major risk of transmission of infections to recipients mainly by the World Health Organisation.Study design and methodsSeroprevalency of HBV, HCV, HIV and syphilis were determined in 19,783 whole blood donations collected in the Tunisian National Blood Transfusion Centre during the year 2010 (12,968 [65.55%] replacement donations and 6815 [34.44%] voluntary blood donations). For HBV, HCV and syphilis, we performed a univariate analysis to determine whether age, sex and type of donation were risk factors, then multivariate logistic regression, to see if these factors were independent.ResultsMean age of donors was 30.1 years (replacement donors 34.5 years, first time non-remunerated donors 34.5 years, P < 0.001). The predominant age group was 30–39 years (35.51%) in replacement donors and 20–29 years (54.15%) in first time non-remunerated donors. Male gender was significantly predominant (73.00% men vs 27.00% women, P < 10^?6). There were significantly more men among replacement donors (82.27% vs 55.38%, P < 10^?3). There were more women in the age groups 18–19 and 20–29 years. Only one HIV seropositive donation was noted in a male first time non-remunerated donor aged 18. Replacement type of donation, male sex and age were three independent risk factors for the HBs Ag carriage. For anti-HCV antibodies and TPHA, only replacement type of donation and age were found out to be risk factors and only age was independent.ConclusionIn Tunisia, replacement blood donors were at higher risk of infection transmission, but only for hepatitis B.     

Alteration in biochemical parameters during plateletpheresis in healthy donors: A compendious analysis

ObjectivesDuring plateletpheresis, citrate induces hypocalcemia and hypomagnesemia, which are usually transient and self-limiting, but they can lead to significant donor discomfort. The aim of study was to determine the effect of citrate infusion on a multitude of biochemical parameters during plateletpheresis in healthy donors and to correlate changes with adverse donor reactions.MethodsThe study was conducted on 60 healthy plateletpheresis donors. Blood samples were drawn on three occasions, a baseline pre-donation sample, 30 min at start of procedure and 30 min post procedure. Heparinized samples were taken to measure ionized calcium and plain samples to measure serum calcium, serum magnesium, parathyroid hormone, total protein and serum albumin.ResultsThere was statistically significant decline in mean total calcium (9.27 ± 0.66 mg/dl to 8.72 ± 0.87 mg/dl) and ionized calcium (3.8 ± 0.51 mg/dl to 2.9 ± 0.67 mg/dl) from baseline until 30 min after the start of procedure respectively. A significant fall in serum magnesium, total protein and serum albumin was observed. The mean parathyroid hormone showed significant increase from baseline levels till at the completion of procedure (19.94 ± 12.1 pg/ml to 92.08 ± 36.78 pg/ml). If the yield was set constant, there was negative correlation between ACD used and pre-donation platelet count. Majority of adverse donor reactions were hypocalcemic reactions, which were more with Amicus double yield plateletpheresis and were managed with calcium supplementation.ConclusionPlateletpheresis induces marked reduction in serum calcium and magnesium levels. Moreover, increase in parathyroid hormone levels was significant. In addition, decline in total protein and serum albumin may be a concern in donors also participating in plasmapheresis.     

BK polyomavirus-associated hemorrhagic cystitis among pediatric allogeneic bone marrow transplant recipients: Treatment response and evidence for nosocomial transmission

BackgroundBK polyomavirus-associated hemorrhagic cystitis (BK-PyVHC) is a significant complication of allogenic hematopoietic stem cell transplantation (HSCT), but risk factors and treatment are currently unresolved. BK-PyVHC typically presents with clinical cystitis, macrohematuria, and increasing urine and blood BKV loads.ObjectivesCharacterization of children undergoing allogeneic HSCT with BK-PyVHC and their clinical and antibody response to cidofovir treatment.Study designBy prospective screening of urine and plasma in 50 pediatric allogenic HSCT performed between 2008 and 2010, we identified 6 (12%) children with BK-PyVHC. Cidofovir was administered intravenously to 5 patients and intravesically to 4 patients (3 double treatments).ResultsDecreasing BKV viremia of > 2 log₁₀ copies/mL and clinical resolution was seen in 4 patients over 5–12 weeks. Responses occurred only in patients mounting BKV-specific IgM and IgG responses. Epidemic curve plots, BKV genotyping and contact tracing provided evidence of transmission between 2 BKV-seronegative patients, but ruled out transmission among the remaining four patientsConclusionsThe data suggest that BK-PyVHC may be the result of nosocomial transmission in children with low/undetectable BKV antibodies and raises urgent questions about appropriate infection control measures and the role of cidofovir.     

Association of cytokine gene polymorphisms with the complications of allogeneic haematopoietic stem cell transplantation

The purpose of our study was to confirm the prevalence of the association between single nucleotide polymorphisms present in genes encoding cytokines and the complications occurring after haematopoietic stem cell transplantation (HSCT). 108 recipients and 81 donors were typed for TNF-α ( ?308), TGF-β1 (codon 10, 25), IL-10 (?1082, ?819, ?592), IL-6 (?174) and INF-γ (+874). Our studies have shown a tendency toward association between the occurrence of acute form of graft versus host disease (aGVHD) and IL-6 genotype. Homozygote C/C was less likely to develop aGVHD (p = 0,09). Genotype GCC/ATA in IL-10 recipient gene alone had protective effect against the occurrence of aGVHD (p = 0,01). Furthermore, GCC/ATA protected the host against developing the disease in the clinically relevant grades (II-IV) (p = 0,03). In addition, the recipient’s T/T G/G genotype (TGF-β1) predisposed to the development of both acute (p = 0,06 – trend) and chronic (p = 0,04) GVHD and also severe aGVHD (p = 0,004). We also observed a statistically significant association between the genotype of recipient and the risk of infection – the protective function of the G/C IL-6 in the bloodstream infections (p = 0,001). Our results suggest that IL-6, IL-10 and TGF-β1 genotypes of recipient are the most associated with the risk of complications after HSCT.     

Comparison of three real-time PCR for the quantification of polyomavirus BK

BackgroundReactivation of latent polyomavirus BK is associated with nephropathy (PVAN) after renal transplantation. BK viral load determinations are a highly sensitive and specific method for predicting risk for PVAN.Objectives and study designThe performance of three real-time PCR for BKV DNA quantification (MultiCode^®-RTx BK virus ASR [MC-RTx], MGB-Alert BKV ASR [MGB] and a laboratory developed assay [LDA]) were evaluated against a conventional PCR (test of record, TOR) in terms of linearity, dynamic range, and accuracy.ResultsThe LOD (log₁₀ copies/ml) were 2.0, 2.0 and 3.0 for MC-RTx, MGB and LDA, respectively with a commercial plasma panel and 2.0, 2.6 and 3.5 with a urine panel. These assays demonstrated excellent linearity (r² = 1.0) and reproducibility (CV range = 0.7–20.4%, 0.9–13.2%, and 0.5–13%, respectively). In an analysis of 100 clinical specimens, all 76 samples defined as true positive for BKV DNA (positive by two or more methods or a recent history of positivity) were detected with MC-RTx, while only 64 were detected with MGB and 55 were detected with LDA. BKV DNA was not detected by any method in the true negative specimens. Based on these results, the sensitivities were 100% for MC-RTx, 84% for MGB and 72% for LDA. The greatest linear correlation with the mean concentration was observed with MC-RTx (r² = 0.96) with two samples (3%) with greater than 0.5 log₁₀ variance in quantification versus seven (11%) with MGB and ten (18%) with LDA.ConclusionsThese real-time assays for BKV load demonstrated excellent performance characteristics, with the MC-RTx demonstrating the greatest sensitivity.