Alessandra Coiana

Treatment of Chronic Hepatitis D with Interferon Alfa-2a

BACKGROUND AND METHODS Chronic hepatitis D is a severe and rapidly progressive liver disease for which no therapy has been proved effective. To evaluate the efficacy of treatment with interferon, we studied 42 patients with chronic hepatitis D who were randomly assigned to receive either 9 million or 3 million units of recombinant interferon alfa-2a (three times a week for 48 weeks) or no treatment. RESULTS By the end of the treatment period, serum alanine aminotransferase values had become normal in 10 of 14 patients receiving 9 million units (71 percent), as compared with 4 of 14 treated with 3 million units (29 percent, P = 0.029) and 1 of 13 untreated controls (8 percent, P = 0.001). Seven patients treated with the higher dose of interferon (50 percent) had a complete response (normal levels of alanine aminotransferase and no detectable serum hepatitis delta virus [HDV] RNA), as compared with three of those who received the lower dose (21 percent, P = 0.118), and none of the controls (P = 0.004). Treatment with 9 million units of interferon was associated with a marked improvement in the histologic findings (reduced periportal necrosis and portal and lobular inflammation), whereas in the untreated controls there was considerable histologic deterioration. In 5 of the 10 patients treated with 9 million units of interferon whose alanine aminotransferase values became normal, the biochemical responses persisted for up to 4 years (mean, 39 months), but the effects of treatment on viral replication were not sustained. In contrast, none of those who received 3 million units and none of the untreated controls had a sustained biochemical or virologic response. CONCLUSIONS In about half the patients with chronic hepatitis D treated with high doses of interferon alfa-2a (9 million units three times a week for 48 weeks), the serum alanine aminotransferase level becomes normal, HDV RNA becomes undetectable in serum, and there is histologic improvement. However, a relapse is common after treatment has been stopped.

Early changes in hepatitis C viral quasispecies during interferon therapy predict the therapeutic outcome

Despite recent treatment advances, the majority of patients with chronic hepatitis C fail to respond to antiviral therapy. Although the genetic basis for this resistance is unknown, accumulated evidence suggests that changes in the heterogeneous viral population (quasispecies) may be an important determinant of viral persistence and response to therapy. Sequences within hepatitis C virus (HCV) envelope 1 and envelope 2 genes, inclusive of the hypervariable region 1, were analyzed in parallel with the level of viral replication in serial serum samples obtained from 23 patients who exhibited different patterns of response to therapy and from untreated controls. Our study provides evidence that although the viral diversity before treatment does not predict the response to treatment, the early emergence and dominance of a single viral variant distinguishes patients who will have a sustained therapeutic response from those who subsequently will experience a breakthrough or relapse. A dramatic reduction in genetic diversity leading to an increasingly homogeneous viral population was a consistent feature associated with viral clearance in sustained responders and was independent of HCV genotype. The persistence of variants present before treatment in patients who fail to respond or who experience a breakthrough during therapy strongly suggests the preexistence of viral strains with inherent resistance to IFN. Thus, the study of the evolution of the HCV quasispecies provides prognostic information as early as the first 2 weeks after starting therapy and opens perspectives for elucidating the mechanisms of treatment failure in chronic hepatitis C.

Experimental Transmission of Hepatitis C Virus-Associated Fulminant Hepatitis to a Chimpanzee

Hepatitis C virus (HCV) was transmitted from a patient with fulminant hepatitis C to a chimpanzee. The patient had developed two episodes of fulminant hepatitis C, each occurring after a separate liver transplantation. Serial serum and liver samples from the patient and the chimpanzee were analyzed for HCV replication, genotype, quasispecies heterogeneity, and antibodies. In the patient, the levels of HCV replication in serum and liver correlated with the degree of hepatocellular necrosis and the clinical expression of fulminant hepatitis. The same HCV strain, genotype 1a, was recovered from both episodes of fulminant hepatitis. An unusually severe acute hepatitis was also observed in the chimpanzee. The viruses recovered from the patient and the chimpanzee were almost identical and displayed relatively little quasispecies heterogeneity. Thus, the same HCV strain induced two episodes of fulminant hepatitis in a single patient and severe hepatitis in a chimpanzee, suggesting that the pathogenicity or virulence of a specific HCV strain may be important in the pathogenesis of fulminant hepatitis C.

Peripheral blood lymphocyte response to exogenous interleukin 2 by PHA-prestimulated and non-PHA-prestimulated cells in patients with cancer.

The study aims were 1) to assess the response of peripheral blood lymphocytes (PBL) of cancer patients to exogenous Interleukin 2 (IL 2) either by PHA-prestimulated or non PHA-prestimulated PBL, and 2) to carry out preliminary experiments for a direct quantitative evaluation of endogenous IL 2 production by PBL cultures of cancer patients in order to define the actual role of IL 2 in the disease. Analysis of PBL subsets was also carried out with monoclonal antibodies in a selected group of patients. A total of 134 patients entered the study. Cancer sites were: larynx 32, breast 36, lung (NSC) 24, colorectal 17 and gynecologic 25. In the former 3 cancer sites staging showed localized or only locally advanced disease, and in the last 2 sites disseminated disease. Our results provided evidence that cancer patients exhibit a T-cell functional immunodepression, which progresses during tumor growth, so that the localized disease shows a low-grade defect, and advanced disease a high-grade defect. Our data also clearly suggested that the factor involved with a primary role in this functional immune impairment is the IL 2 deficiency. A perspective may be drawn on the therapeutic administration in vivo of IL 2 and IL 2-activated lymphokine-activated killer cells in controlled clinical trials of selected groups of cancer patients.

New mutations in DYNC2H1 and WDR60 genes revealed by whole-exome sequencing in two unrelated Sardinian families with Jeune asphyxiating thoracic dystrophy

Jeune asphyxiating thoracic dystrophy (JATD; Jeune syndrome, MIM 208500) is a rare autosomal recessive chondrodysplasia, phenotypically overlapping with short-rib polydactyly syndromes (SRPS). JATD typical hallmarks include skeletal abnormalities such as narrow chest, shortened ribs, limbs shortened bones, extra fingers and toes (polydactyly), as well as extraskeletal manifestations (renal, liver and retinal disease). To date, disease-causing mutations have been found in several genes, highlighting a marked genetic heterogeneity that prevents a molecular diagnosis of the disease in most families. Here, we report the results of whole-exome sequencing (WES) carried out in four JATD cases, belonging to three unrelated families of Sardinian origin. The exome analysis allowed to identify mutations not previously reported in the DYNC2H1 (MIM 603297) and WDR60 (MIM 615462) genes, both codifying for ciliary intraflagellar transport components whose mutations are known to cause Jeune syndrome.

Hodgkin's disease presenting in 1 of 4 siblings affected by hereditary spinocerebellar ataxia: clinical, immunological and genetic study

One of 4 siblings affected by hereditary spinocerebellar ataxia (HSCA) of Maries type developed Hodgkins disease (HD): the stage was IV B, the patient was submitted to conventional chemo- and radiotherapy and achieved complete remission. An accurate clinical, genetic and immunological study was carried out on all his family, including a complete HLA typing, a chromosome study, the immunophenotyping of peripheral blood mononuclear cells (PBMC), the PBMC response to polyclonal mitogens, to interleukin 2 (IL-2), to the association of PHA + IL-2 and the evaluation of the IL-2 receptor expression. No association was clearly demonstrable between an HLA haplotype and HSCA, while the patient with HSCA and HD was HLA-B18- and DQw3-positive (the last at homozygous level), two antigens known to be strongly associated with HD, mainly among the Sardinian ethnic group. The mode of inheritance of HD susceptibility is however completely different from that of Maries HSCA. The chromosome study did not show any characteristic pattern of the karyotype, neither of the HSCA affected nor of the unaffected members. The immunological investigations did not elucidate any characteristic behavior of the family members, apart from the typical findings of HD seen on patients with HSCA and HD. Our study could not demonstrate any genetic and/or immunologic common background shared by the two diseases, HSCA and HD. Their coexistence in our patient, although the statistic probability is very low, seems to be a fortuitous coincidence more than the result of a common genetic and pathogenetic mechanism.

Preconceptional identification of cystic fibrosis carriers in the Sardinian population: A pilot screening program

BACKGROUND In Sardinia the mutational spectrum of CFTR gene is well defined. A mutation detection rate of 94% can be achieved by screening for 15 CFTR mutations with a frequency higher than 0.5%. The efficiency of this molecular test suggests that Sardinians may represent a suitable population for a preconceptional screening. METHODS Five hundred couples of Sardinia descent were screened for 38 mutations using a semi-automated reverse-dot blot and PCR-gel electrophoresis assays. This mutation panel included the 15 most frequent CF alleles in Sardinia. RESULTS We identified 38 CF carriers, revealing an overall frequency of 1/25 (4%). The most common CF allele was the p.Thr338Ile (T338I) (65%), followed by the p.Phe508del (F508del) (22.5%). We also identified one couple at risk and an asymptomatic female homozygote for the p.Thr338Ile allele. CONCLUSIONS In spite of the low number of the couples tested, the results herein reported demonstrate the efficacy and efficiency of the preconceptional screening program and the high participation rate of the Sardinian population (99%).

A Novel pattern of uridine diphosphate glucuronosyltransferase polymorphisms associated with hyperbilirubinemia during nilotinib treatment

[Letter to the Editor] Here we report on a patient who developed hyperbilirubinemia during nilotinib treatment, showing a pattern of UGT1A1 polymorphisms so far undescribed in this clinical setting.

Transfusion-associated Hepatitis (TAH) in Polytransfused Thalassemic Children Following the Introduction of Anti-HCV Donor Screening in Sardinia

Until recently, polytransfused thalassemic children were at high risk of developing non-A, non-B hepatitis. The purpose of our study was to investigate the incidence of transfusion-associated hepatitis (TAH) in these patients, following the introduction of anti-HCV donor screening. Forty thalassemic children were studied who had received hepatitis B virus (HBV) vaccine, experienced anti-HBs seroconversion, had no evidence of preexisting liver disease, and were negative for antibodies to hepatitis C virus (anti-HCV). All children were monitored for serum alanine aminotransferase (ALT) at 3-week intervals and an aliquot of serum was stored at −30°C for virological testing. A total of 2954 blood units, prescreened for anti-HCV, were given to the children (median 80 units, range 9–116). Among them, 14 (35%) developed TAH, which was due to HCV in only one case. The etiologic agent was undefined in 9 cases.

Epidemiology of infection with HIV-1 in Sardinia: A multicentre prospective study

To study the spread of human immunodeficiency virus type 1 (HIV-1) in Sardinia, we conducted a multicentre prospective study of the prevalence of antibody to HIV-1 (anti-HIV-1) in various populations during 1985–1989. The highest anti-HIV-1 prevalence (61.4%) was found in intravenous drug users. Anti-HIV-1 was found in 32% of haemophiliacs, 4.2% of thalassemics and less than 1% in the other groups. We conclude that control ofHIV infection in Sardinia will require a major expansion of prevention and treatment programs for drug addiction.