Pier Luigi Calvo

Comparative Analysis of rs12979860 SNP of the IFNL3 Gene in Children with Hepatitis C and Ethnic Matched Controls Using 1000 Genomes Project Data

The rs12979860 single nucleotide polymorphism located on chromosome 19q13.13 near the interferon L3 gene (formerly and commonly known as interleukin 28B gene) has been associated in adults with both spontaneous and treatment induced clearance of hepatitis C virus. Although the exact mechanism of these associations remains unclear, it suggests that variation in genes involved in the immune response against the virus favours viral clearance. Limited and preliminary data are available on this issue in children. The aim of the present study was to evaluate, in a representative cohort of children with perinatal infection, the potential association between rs12979860 single nucleotide polymorphism and the outcome of hepatitis C virus infection. Alleles and genotypes frequencies were evaluated in 30 children who spontaneously cleared the virus and in 147 children with persistent infection and were compared with a population sample of ethnically matched controls with unknown hepatitis C status obtained using the 1000 Genomes Project data. The C allele and the C/C genotype showed greater frequencies in the clearance group (76.7% and 56.7%, respectively) when compared with both children with viral persistence (C allele 56.5%, p = 0.004; C/C genotype 32.7%, p = 0.02) and with the ethnically matched individuals (C allele 59.7%, p = 0.02; C/C genotype 34.7%, p = 0.03). Children with the C/C genotype were 2 times more likely to clear hepatitis C virus relative to children with the C/T and T/T genotypes combined (odds ratio: 2.7; 90% confidence intervals: 1.3–5.8). The present study provides the evidence that the rs12979860 single nucleotide polymorphism influences the natural history of hepatitis C virus in children.

Interleukin 28B rs12979860 single-nucleotide polymorphism predicts spontaneous clearance of hepatitis C virus in children.

Objective: Recent genome-wide association studies performed in adults correlated single-nucleotide polymorphisms (SNPs rs12979860 and rs8099917) located on chromosome 19, upstream of the interleukin 28B gene, with spontaneous clearance of hepatitis C virus and with response to treatment with paginated interferon and ribavirin. The aim of the present collaborative study was to evaluate the rs12979860 SNP in a large cohort of Italian children with perinatal acquisition of hepatitis C. Methods: Children were prospectively enrolled in 2 Italian centers. The interleukin 28B rs12979860 SNP was studied according to the diagnosis of chronic infection or spontaneous clearance. Results: One hundred thirty children (86.7%) with chronic infection and 23 (13.3%) with spontaneous clearance of the virus were enrolled. Overall, the interleukin 28B C/C and C/T-T/T genotypes were found in 57 (37.3%) and 96 (62.7%) children, respectively. The proportion of C/C genotype was higher among children who cleared infection (14/23; 60.9%) compared with children with chronic infection (43/130; 33.1%; P = 0.01; odds ratio 3.15; 90% confidence intervals 1.34–7.53). Conclusions: The present study showed that, as already demonstrated in adults, children with the rs12979860 C/C SNP of the interleukin 28B gene have a higher probability of spontaneous clearance of hepatitis C virus.

EBV-encoded microRNAs profile evaluation in pediatric liver transplant recipients

BACKGROUND Epstein-Barr virus (EBV) infects 90% of the world population, commonly causing self-limiting infectious mononucleosis or rarely inciting a range of malignancies. EBV microRNAs (miRNAs) were discovered by sequencing libraries of small RNAs generated from several EBV-positive cell lines. Little is known about their roles, but their high stability and easy quantification make these molecules potential biomarkers. OBJECTIVES In this study a stem-loop MGB real-time RT-PCR has been used to detect and quantify miR-BART2-5p, miR-BART15 and miR-BART22 EBV miRNAs levels. STUDY DESIGNS The profiles of EBV miRNAs levels were evaluated in 51 serum samples of 37 pediatric liver transplant patients subdivided in 3 study groups: EBV seronegative, EBV seropositive and PCR negative, EBV seropositive and PCR positive. RESULTS miR-BART22 serum levels in patients with positive EBV PCR were significantly higher than those in patients with negative EBV PCR (p=0.0005). On the contrary, miR-BART2-5p and miR-BART15 did not exhibit significant difference in positive and negative EBV PCR patients (p=0.5511 and p=0.3523, respectively). CONCLUSION This study described a method for quantitative detection of miR-BART 22, miR-BART2-5p and miR-BART15 EBV miRNAs in liver transplanted patients, and suggests the use of miR-BART22 as a potential biomarker for EBV reactivation.

CO4 THERAPEUTICAL IMPLICATIONS IN THE STUDY OF MUTATION OF THE CYP450 3 A5*3 IN THE METABOLISM OF TACROLIMUS IN PAEDIATRIC LIVER TRANSPLANTATION: THE EXPERIENCE OF A SINGLE MEDICAL CENTRE

liver abnormalities. Anti-PG-PS IgA antibodies were elevated in spite of normal H2BTs. A multivariate analysis of studied parameters showed that the significant decrease of ALT (average variation vs. placebo, p 0.03) and of anti-PG-PS IgA antibodies (average variation vs. placebo, p 0.03) values after probiotic treatment was independent from changes of BMI z score and visceral fat. TNF-α, and US bright liver parameters remained fairly stable. The baseline t test (T0 placebo vs T0 probiotic) was not significant for all variables evaluated (ALT, BMI z score, US visceral fat, TNF-α, US ROI ratio, anti-PG-PS IgA antibodies). Conclusion: Results of the present pilot study confirm that lactobacillus rhamnosus strain GG deserves consideration as a therapeutic tool for improving hypertransaminasemia but not bright liver in hepatopatic obese children who are unable to follow slimming diets and/or to change lifestyle. Further studies on a larger scale are therefore warranted.

PP47 CLINICAL SIGNIFICANCE OF HERPESVIRUS 6 (HHV6) IN PEDIATRIC TRANSPLANTATION

Case report: We report the case of an 80-day-old infant referred to our department with refractory severe dermatitis, resembling Acrodermatitis Enteropathica (AE), associated with moderate failure to thrive. She had been hospitalized in a different clinic when she was 50 days old. During that time she was treated with oral steroids for suspected atopic dermatitis with no lasting benefits. The baby had been exclusively breast fed. At admission, the patient was well below the 3rd percentile for weight and at the 25th percentile for length, while the respective percentiles at birth were both at the 50th. Dermatological examination revealed erythematous, scaly plaques with crusts and ulcerations, predominantly in the diaper area, perioral area and extremities. Mucous membranes and nails were not involved. Laboratory tests revealed anemia, hypoalbuminemia and zinc deficiency. Laboratory abnormalities included the following: albumin minimum values of 16.2 and 19.2 g/dl (38–54 g/dl), haemoglobin minimum values of 5.5 and 6.7 g/dl (10–13 g/dl). Two albumin infusions and two blood transfusions were required. A sweat test was done but the amount of sweat collected was insufficient for an accurate result, influenced by the skin condition. However, stools were positive for fat analysis; steatocrit was 28% ( 5 U/g). With the suspect of CF pancreatic enzyme supplementation was started and was associated with clinical and laboratory findings improvement. The dermatitis showed a gradual resolution, preceded by an extensive desquamation. Finally, mutation analysis for CF showed homozygous mutation 508. After three months of therapy, the patient’s weight increased from the 3rd to the 25th percentile, the dermatitis had completely resolved and laboratory findings had improved. Conclusions: Skin manifestations of CF are under-estimated; however, dermatitis can be the presenting sign of the disease, prior to pulmonary and gastrointestinal symptoms. AE-like dermatitis may concur and allows a correct and early diagnosis in atypical presentations of CF, particularly in association with other symptoms such as failure to thrive or laboratory findings such as anemia and hypoalbuminemia. Even in cases of negative neonatal screening, further investigations such as the sweat test and/or genetic analysis may be required if there are symptoms suggestive of CF. It is important to offer genetic counselling to parents with a family history of CF so that they are aware of the possibility of performing CF carrier screening and prenatal diagnosis.