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Recent evidences have shown that several host genetic factors influence susceptibility or protection to hepatitis C virus (HCV) infection. There are controversial data regarding the associations of human leukocyte antigens (HLA) and the clearance or progression of HCV. The aim of this study was to investigate whether particular HLA molecules were associated with HCV infection in recipients awaiting kidney transplantation considered at high-risk to infection due to protracted hemodialysis treatment. To this purpose, 301 kidney recipients with HCV infection and 1103 uninfected recipients were examined for HLA class I and II molecules. In our case-control study, HLA-A * 26 is positively associated with HCV infection while HLA-A * 29, -B * 40 and -DRB1 * 01 are negatively associated with HCV infection. Multiple logistic regression analysis demonstrated that age (OR = 1.02; 95% CI = 1.01–1.04; p * 26, -A * 29, -B * 40 and -DRB1 * 01 [(OR = 1.54; 95% CI = 1.03–2.30; p = 0.03); (OR = 0.50; 95% CI = 0.26–0.99; p = 0.05); (OR = 0.42; 95% CI = 0.23-0, 7; p = 0.01); (OR = 0.62; 95% CI = 0.41-0, 94; p = 0.03); respectively] are independent predictors of HCV infection. Our results suggest that particular HLA molecules, as host genetic factors, may have a relationship with susceptibility or protection to HCV infection also in recipients awaiting kidney transplantation.

Aim Allogeneic hematopoietic stem cell transplantation (allo-HSCT) is the standard therapy for the treatment of malignant blood diseases or immune system disorders. HLA compatibility influences the transplant outcome and minimizes the risks of rejection or severe graft-versus-host disease. The identification of a compatible unrelated donor depends on the race/ethnicity due to heterogeneity within the various ethnic groups and the high polymorphism of HLA genes that limits the possibility of HSCT from unrelated donors. In Italy, the Italian Bone Marrow Registry (IBMDR) is the main source of unrelated volunteer donors. Here, we report our experience of HLA typing laboratory. Methods The current Italian guidelines include high resolution HLA typing at A, B, C, and DRB1 loci for volunteer unrelated donors in the enrollment time. In 2013, at the Regional Reference Laboratory of Transplant Immunology of Naples (Italy), the donors were typed by sequence specific primers PCR. Results We have identified the HLA-A∗23:18 allele in two different volunteer donors. This is a rare allele in the Italian and Caucasian population (IMGT/HLA AccNo:HLA03170). The allele A∗23:18, for the first time, was identified by SBT in a patient attending HSCT in Italy. To date, this allele is still listed as rare allele in the NMDP rare allele list file version 3.9.0. The HLA typing of these two donors is A∗03:01,∗23:18; B∗07:02,∗14:02; C∗07:02,∗08:02; DRB1∗01:02,∗15:01 and A∗02:05,∗23:18; B∗14:02; C∗08:02; DRB1∗01:02. These donors share the HLA-A∗23:18; B∗14:02; C∗08:02 and DRB1∗01:02 alleles with the patient where this allele was identified for the first time. Both donors are Italians and resident in our region since several generations. Conclusion The high polymorphism and the pattern of HLA alleles and haplotypes distribution in human populations in the world make it difficult to find compatible stem cell donors. However, the role of HLA compatibility at allelic level in allo-HSCT is still debated, especially if mismatches are not localized in exon 2 and 3 of HLA class I genes. Our results highlight that some alleles, considered rare, may be frequent alleles in some regions and therefore it is necessary a more accurate knowledge of allele and haplotype frequencies in order to develop new strategies for the search of HSCT unrelated donors.

Aim Heart transplantation has evolved as the “gold standard” therapy for patients with end-stage heart failure (HF). Although mechanical circulatory support technology is improving, heart transplantation remains the preferred treatment. The mean risk factors for HF include environmental and genetic components. The intricate patterns of gene expression, during normal and disease states, are governed by elaborate signaling pathways that converge on the transcriptional mechanism. The general transcriptional mechanism can be recruited to promoters by Mediator complex (MED). MED is a large multi-subunit complex functioning to integrate diverse cellular signals by multiple mechanisms including recruitment of RNA polymerase II, chromatin modifying proteins and non-coding RNAs to promoters in a context dependent manner. Several MED subunits are involved in many human pathologies, including heart diseases. Particularly, some subunits like MED13 and MED13L are altered in congenital heart diseases. Similarly, MED30 mutation can lead to lethal cardiomyopathy in a mouse model. Methods To gain further insights, we have collected 12 human left atrium samples, which were acquired from hearts of 6 donors and 6 recipients during transplant intervention. Patients were affected by hypertrophic or dilated idiopathic cardiomyopathy. From these samples we have isolated RNA and started to analyze the expression patterns of MED subunits by real-time RT-PCR with specific oligonucleotides. Possible interactions between human leukocyte antigen (HLA) profile and MEDs are under investigation. Furthermore, RNA-seq analysis of samples from both donors and recipients is ongoing in our laboratory and whole transcriptome data will be also shown. Results Our preliminary data confirm that some MED subunits are altered in diseased hearts compared to healthy donor biopsies. Conclusions The identification of such alterations can lead to novel therapeutic targets regulating MED subunits, and HLA in advanced HF.

Aim The interaction of the heart with other organs is well established. In particular, several donor and recipient factors involving the heart are known to be associated with graft loss in kidney transplantation. In this retrospective single-center study, we analyzed the effect of clinical, cardiological and immunological factors on kidney transplantation outcome. The study included 245 transplanted recipients from deceased donors between 2000 and 2006. Methods In donors, age, cause of death, history of hypertension, hypotension or cardiac arrest, length of intensive care unit stay, serum creatinine levels and HLA typing were evaluated; while age, waiting time, HLA typing, antibodies sensitization and allocation were evaluated in recipients. Age donor/recipient matching and HLA mismatches were also considered. Results Cox regression analysis shows that time spent in waiting list increases the risk of restarting dialysis (OR=1.01, 95% CI=1.00-1.03; p Conclusions Our study established that both hypertension and HLA-A mismatch can affect the time of restarting dialysis in patients undergoing kidney transplantation.

Aim The twin epidemics of obesity and Type 2 diabetes continue to increase in industrialized nations. Approximately two thirds of adult Americans and Western Countries are currently overweight or obese and therefore at increased risk for a number of deleterious health conditions including Type 2 diabetes and Cardiovascular Disease (CVD). Consumption of sugar-sweetened beverages may be one of the relevant dietary causes of metabolic disorders. Therefore, substituting sugar with low-calorie sweeteners may be n efficacious weight management strategy. Since Endothelial Progenitor Cells (EPC) are involved in cardiovascular tissue regeneration, we tested the effects of glucose and alternative sweeteners (fructose, aspartame and rebaudioside A) on EPCs. Methods Early-EPCs were isolated from total peripheral blood of healthy human donors and treated with different concentration of D-glucose, fructose, aspartame and rebaudioside A for 4 days. EPC number was evaluated by flow cytometry analysis using CD34 and KDR (CD309/VEGFR-2) monoclonal antibodies. Results As partially expected, high glucose level severely impaired EPC number. Indeed, CD34+/KDR+ dual positive cells were reduced of about 70% at the maximum glucose concentration used. Differently, the other molecules tested were able to reduce the number of total KDR+ cells, but not CD34+/KDR+ dual positive cells. Conclusions Endothelial dysfunction and reduced new blood vessel growth during vascular complication in either Type 1 or Type 2 diabetes, is known to be strictly related to altered number and function of circulating EPCs. The pathogenic role of reduced bioactivity of EPCs is correlated to increased rate of CVD. These preliminary findings suggest that these alternative sweeteners do not alter CD34+/KDR+ dual positive EPC number, although they can alter endothelial cell number through other cellular molecular pathogenic mechanisms.

Aim Previous clinical studies indicate that implantation of Bone Marrow Cells (BMCs) into ischemic limbs may improve peripheral ischemia. We previously demonstrated that intraarterial administration of autologous BMC and antioxidants and L-arginine therapy is safe and effective in patients with advanced atherosclerotic peripheral artery disease (PAD) with positive effects until 18 months. Here, we assessed the long-term follow-up of patients until 36 months. Methods In the original study protocol (NCT00306085), we studies 18 patients with PAD (advanced III/IV Fontaine stage) and an additional group of 18 patients taking maximal drug therapy that refused BMC therapy as control. Therapeutic neo-angiogenesis was estimate by clinical assessment, angiography and laser Doppler/capillaroscopy. Results At 18 months of follow-up, among conservative control patients, 10 underwent amputation in comparison with two BMC-treated patients (55.6 vs. 13.3%; p=0.014). Ischemic ulcers improved in 13 patients (after 6-12 months). The follow-up at 36 months of remaining patients confirmed a favorable trends in patients treated with BMCs in comparison to controls (p Conclusions This follow-up study confirmed safety and feasibility of intraarterial autologous BMCs in patients with critical ischemia and advanced atherosclerotic PAD.

Aim Recent evidences have shown that several host genetic factors influence susceptibility or protection to hepatitis C virus (HCV) infection. There are controversial data regarding the associations of human leukocyte antigens (HLA) and clearance or progression of HCV. The aim of this study was to investigate whether particular HLA molecules were associated with HCV infection in recipients awaiting kidney transplantation, considered at high-risk to infection due to protracted hemodialysis treatment. Methods This case-control study included 1404 of 3000 total chronic kidney diseases patients on dialysis, in waiting list for transplantation, enrolled at the Laboratory of Transplantation Immunology of Second University of Naples, Italy. Particularly, we consecutively considered 301 patients with HCV infection and we randomly selected 1103 HCV-negative as control group. None of the patients carried hepatitis B surface antigen or had human immunodeficiency virus antibody to exclude additional viral factors that could affect our results. All patients were tested for HCV antibody detection by serological Chemiluminescent Microparticle Immunoassay (CMIA, Abbott, Italy). All kidney recipients were tested for HLA class I (HLA-A and -B) and class II (-DR) antigens by serological and molecular HLA typing. Results In our case-control study, HLA-A26 is positively associated with HCV infection, while HLA-A29, -B40 and -DR1 are negatively associated with HCV infection. Multiple logistic regression analysis demonstrated that age (OR=1.02; 95% CI=1.01-1.04; p Conclusions Our results suggest that particular HLA molecules, as host genetic factors, may have a relationship with susceptibility or protection to HCV infection also in recipients awaiting kidney transplantation.

Aim Mediator (MED) complex functions as a pivotal adaptor between transcription factors bounded at gene regulatory elements, RNA polymerase II and general transcription factors. Different ancestral humanMED complexes including at least 30 distinct MED subunits (MEDs) have been isolated. Because of the importance of ancestral MED role in the transcription of the eukaryotic genes, disruption of MED function may have relevant pathophysiological consequence also in the cardiovascular system and transplantation. Here, we have analyzed the expression data relative to MEDs in human endothelial progenitor cells (EPCs) obtained by RNA-Seq on a next generation sequencing platform. The introduction of next generation sequencing (NGS) technologies has revealed the complexity of mammalian transcriptomes, enabling to effectively explore, with an unprecedented throughput capacity, simple and complex genomes and even their differences in health and disease conditions. Methods RNA was isolated from early human EPCs and after ribodepletion it was used for the RNA-Seq through SOLID System. Results By analysis of RNA-Seq data and RT-PCR validation we have identified novel transcripts in several MED genes (including MED1, MED 15, MED 17 and MED23). Some of these transcripts are different in their 5’ and 3’ UTRs. Other transcripts are differently spliced thus excluding or including known/new exons. Conclusions This findings contribute to the characterization of novel MED transcripts in EPCs that could participate to the regulation of genes involved in different cell states. Our findings could have relevant implications in the regenerative action of EPCs in the cardiovascular system immune response after cardiac transplantation.