[HCV reinfection after liver transplantation for HCV cirrhosis]. / Reinfezione da virus dell'epatite C nei pazienti sottoposti a trapianto epatico per cirrosi da HCV.

Cirrhosis due to hepatitis C virus (HCV) infection is now the most frequent indication for orthotopic liver transplantation (OLT). Recurrence of hepatitis C infection is the major cause of late mortality in patients undergoing OLT for hepatitis C cirrhosis. Recurrent HCV infection develops in 100% of patients HCV + in pre-transplantation time. Histological recurrence occurs in 75-80% of patients after OLT:1/3 of them progress to allograft cirrhosis within 5-7 years. Cholestatic hepatitis C develops in a sub-group of patients who progresses rapidly to graft failure. As a result of this accelerated course of HCV infection, long-term graft and patient survival are significantly reduced in patients undergoing OLT for HCV-related cirrhosis compared with other groups. Moreover, several recurrence's risk factors have been described as predictors of disease severity including those related to the virus, the host, the donor. There are numerous therapeutic strategies to prevent and to treat HCV disease recurrence after OLT. The most common strategy to treat HCV infection post-OLT is based on interferons and ribavirin. Even if clinical trials have shown that the combination of ribavirin with Peg-interferons is more effective than its association with standard interferons, the use of Peg-interferons in transplanted patients is limited by the side-effects of the drug. About treatment of hepatitis C virus infection in the allograft dark and not still cleared points are a lot: the timing and the target of therapy, the dose and duration of pharmacological treatment.

Isolated anti-HBc in chronic hepatitis C predicts a poor response to interferon treatment.

The sustained response to interferon-alpha treatment was evaluated in 147 anti-HCV/HCV-RNA-positive, HBsAg-negative, chronic hepatitis patients, according to HCV genotypes and the presence or absence of anti-HBs and anti-HBc. These patients had been included in a controlled study on the safety, tolerability, and efficacy of three types of interferon-alpha given at a dose of 3 MU three times weekly for 52 weeks. One hundred and two patients had HCV genotype 1, 42 a non-1 HCV genotype and 3 multiple HCV genotypes; 46 were anti-HBs and anti-HBc negative (group A), 50 anti-HBs and anti-HBc positive (group B), and 51 anti-HBs negative and anti-HBc positive ("isolated" anti-HBc, group C). Serum HBV-DNA was detected by polymerase chain reaction in 15 of the 51 (29.4%) patients in group C and in none of those in groups A or B. The Sustained Response rate was higher in patients with a non-1 HCV genotype than those with HCV genotype 1 (31% vs. 17.7%, P > 0.1). Fewer patients in group C showed a sustained response than in group A or group B (7.8% vs. 30.4%, P = 0.009 and 7.8% vs 28%, P = 0.017, respectively). Moreover, the sustained response rate was high in patients with a non-1 genotype, both in group A (42.8%) and in group B (42.8%), intermediate in patients with HCV genotype 1 (23.3% in group A and 22.2% in group B) and low in group C, irrespective of HCV genotype (8.3% for genotype 1 and 7.1% for other genotypes). The data indicate that patients with HCV chronic hepatitis and isolated anti-HBc show a poor response to IFN-alpha, irrespective of the HCV genotype.

HCV genotype and "silent" HBV coinfection: two main risk factors for a more severe liver disease.

To evaluate whether HCV genotype and a "silent" HBV infection may be related to a more severe clinical presentation of liver disease, 205 anti-HCV/HCV-RNA positive, HBsAg/anti-HBs negative patients with chronic hepatitis (113 males and 92 females; median age 55 years, range 18-77), were studied on presentation at the Liver Unit from January 1993 to December 1997. Presence of serum anti-HBc, in the absence of HBsAg and anti-HBs, was considered a marker of "silent" HBV infection. Of the 205 patients, 134 had undergone percutaneous liver biopsy. Two main diagnosis groups were established: the mild liver disease group (76 patients), and the severe liver disease group (109 patients); 20 patients who had refused to undergo liver biopsy were not included in the clinical and virological evaluation because the diagnosis was uncertain. The prevalence of severe liver disease was similar in the genotype 1 and non-1 groups (61.3% of 98 patients with genotype 1 and 52.9% of 70 patients with a non-1 genotype). Instead, the 88 patients with "silent" HBV infection showed a higher percentage of severe liver disease than the 97 anti-HBc negative patients (72.7% vs. 46.4%, respectively: P < 0.0005). Of the 88 anti-HBc positive patients, the prevalence of those with severe liver disease was similar in the 32 cases with serum HBV-DNA as detected by PCR and in the 56 HBV-DNA negative (81.2% vs. 67.8%, P = 0.4). The relation between "silent" HBV infection and severe liver disease was observed both in genotype 1 and non-1 infected patients. Nevertheless, the anti-HBc negative patients infected by genotype 1 showed a severe liver disease more frequently than those infected by a non-1 genotype, with a difference that is significant to the statistical analysis (P < 0.05). The findings suggest that "silent" HBV infection in anti-HCV positive chronic hepatitis enhances the severity of the liver disease. Evidence was also found that in patients without "silent" HBV infection there is a correlation between the presence of HCV genotype 1 and the severity of liver disease.

[Anti-HAV screening patients with chronic hepatitis]. / Screening per anti-HAV in pazienti con epatite cronica.

Following the decrease in HAV endemic levels in the last decades, nowadays in Italy most people aged less than 30 years are susceptible to the infection. We have tested for serum anti HAV IgG 194 chronic hepatitis patients from Naples to evaluate the level of protection against HAV in a category of patients in whom an acute necrosis due to HAV might induce liver failure. The study shows that 90.7% of Neapolitan chronic hepatitis patients has serologic evidence of prior HAV infection. Specifically, 98.1% of the patients over 40 years of age, but only 54.6% of those under 40 tested positive for anti HAV IgG. 102 patients had cirrhosis and only one tested negative. The data suggest that HAV vaccination in susceptible chronic hepatitis patients should be judged by the physicians in care on the basis of the severity of disease, patient's age, real risk of exposure to the infection, and level of education.

Immortalization of a cell line showing some characteristics of the oligodendrocyte phenotype.

We have used the polyoma middle T oncogene to immortalize cells from rat embryo encephalon. Immunostaining experiments with monoclonal antibodies demonstrated that the cells of one of the obtained lines, named CEINGE CL3, are stained by anti-vimentin and anti-S100 antibodies, are not stained by anti-neurofilaments (NF) or anti-glial fibrillary acidic-protein (GFAP) antibodies. Only a subset of the CEINGE CL3 cells (20-30%) is stained by an anti-galactocerebroside antibody. Northern blot analysis demonstrated that these cells express low levels of proteolipid protein mRNA, whereas polymerase chain reaction (PCR) amplification failed to evidentiate the presence of both NF and GFAP mRNAs. Either retinoic acid or forskolin treatments or a combination of them are able to induce morphological changes that are accompanied by a complete growth arrest.

A rat brain mRNA encoding a transcriptional activator homologous to the DNA binding domain of retroviral integrases.

We have isolated a rat cDNA, named FE65, hybridizing to an mRNA of about 2,300 nucleotides present in rat brain, undetectable in rat liver and very poorly represented in other tissues. An mRNA of the same size is present in human neuroblastoma cells and is absent from other human cell lines. The FE65 cDNA contains an open reading frame (ORF) coding for a polypeptide of 499 amino acids in which 143 residues can be aligned with the DNA binding domain of the integrases encoded by mammalian immunodeficiency viruses. The remaining part of the FE65 ORF is not homologous with the correspondent regions of the integrases; the first 206 residues of the FE65 ORF show numerous negative charges and a short sequence not dispensable for the function of the transactivating acidic domain of the jun family transcriptional factors. A plasmid which expresses FE65 amino acids 1-232 fused to the yeast GAL4 DNA binding domain was co-transfected with a plasmid containing five GAL4 binding sites upstream of a minimal Adenovirus promoter controlling the expression of the CAT gene. This experiment showed that the fused protein GAL4-FE65 is able to obtain a 30-40 fold increase of the CAT gene expression compared to the expression observed in the presence of the GAL4 DNA binding domain alone. Two types of FE65 mRNA are present in rat brain, differing only for six nucleotides. We demonstrate that this is the consequence of a neuron-specific alternative splicing of a six-nucleotide miniexon, which is also present in the human genome, in an intron/exon context very similar to that of the rat FE65 gene.