Elisabetta Tanzi

Interferon Alfa-2a Therapy in Cryoglobulinemia Associated with Hepatitis C Virus

BACKGROUND Essential mixed cryoglobulinemia is frequently associated with hepatitis C virus (HCV) infection. A beneficial effect of interferon alfa therapy has been reported, but we do not know whether the antiviral activity of the drug affects the clinical and biochemical manifestations of disease. METHODS In a prospective randomized, controlled trial, we studied 53 patients with HCV-associated type II cryoglobulinemia. A group of 27 patients received recombinant interferon alfa-2a thrice weekly at a dose of 1.5 million units for a week and then 3 million units thrice weekly for the following 23 weeks. The 26 control patients did not receive anything apart from previously prescribed treatments. All patients were then followed for an additional 24 to 48 weeks. RESULTS Interferon was usually well tolerated, but it was permanently discontinued in two patients because of atrial fibrillation and depression. Two of the 26 patients in the control group were lost to follow-up. After the treatment period, serum HCV RNA was undetectable in 15 of the remaining 25 patients who received interferon alfa-2a, but in none of the controls. In comparison with the control group, the 15 patients with undetectable levels of HCV RNA in serum had significant improvement in cutaneous vasculitis (P = 0.04) and significant decreases in serum levels of anti-HCV-antibody activity (P = 0.007), cryoglobulins (P = 0.002), IgM (P = 0.002), rheumatoid factor (P = 0.001), and creatinine (P = 0.006). After treatment with interferon alfa-2a was discontinued, viremia and cryoglobulinemia recurred in all 15 HCV RNA-negative patients. On resumption of treatment, three of four patients had a virologic, clinical, and biochemical response. CONCLUSIONS The therapeutic efficacy of interferon alfa-2a in HCV-associated cryoglobulinemia is closely related to its antiviral activity, thus supporting the idea that HCV infection may be a cause of this disease.

Hepatitis C virus infection in patients with essential mixed cryoglobulinemia

OBJECTIVE To study the association between hepatitis C virus (HCV) infection and essential mixed cryoglobulinemia. SETTING Wards and clinics of the Ospedali Riuniti di Bergamo and Ospedale di Treviglio e Caravaggio, Italy. PATIENTS Fifty-one patients with essential mixed cryoglobulinemia associated with glomerulonephritis and 45 controls with noncryoglobulinemic glomerulopathies. MEASUREMENTS Antibodies to hepatitis C virus (anti-HCV) in sera from patients with essential mixed cryoglobulinemia and from controls, using two enzyme-linked immunosorbent assays (c100 ELISA and c22/c200 ELISA) and a recombinant immunoblot assay (4-RIBA); cryoprecipitate anti-HCV before and after use of dithiothreitol, a substance able to destroy IgM antibodies with rheumatoid factor activity, in patients with essential mixed cryoglobulinemia; serum HCV RNA by polymerase chain reaction in patients with essential mixed cryoglobulinemia. RESULTS In patients with essential mixed cryoglobulinemia, the c22/c200 ELISA detected anti-HCV in 98% of serum samples (95% CI, 90% to 100%), whereas the rate of reactivity remained at 2% (CI, 0% to 12%) in the control group (P less than 0.0001). These results were confirmed by the 4-RIBA in 66% of patients with essential mixed cryoglobulinemia. The study of cryoprecipitate by c100 ELISA showed anti-HCV in 41% (Cl, 28% to 56%) of patients. After dithiothreitol, the rate of reactivity increased to 94% (CI, 84% to 99%; P less than 0.0001 by the McNemar paired chi-square test), suggesting that the elimination of rheumatoid factor leads to unmasking of anti-HCV in cryoprecipitate. Polymerase chain reaction detected HCV RNA in 13 of 16 sera from patients with essential mixed cryoglobulinemia. CONCLUSIONS The extremely high prevalence of anti-HCV in serum and cryoprecipitate along with the frequently associated serum HCV RNA suggests a close relation between essential mixed cryoglobulinemia and chronic HCV infection.

Mother-to-infant transmission of hepatitis C virus

To assess the risk of mother-to-infant transmission of hepatitis C virus (HCV), we followed up 116 babies of anti-HCV positive mothers, of whom 22 were coinfected with HIV and 94 had HCV alone. None of the babies whose mothers had HCV alone acquired HCV, while 8 babies (36%; p < 0.001) of mothers co-infected with HIV acquired HCV (5 babies) or HCV and HIV (3). There was no association between any specific maternal HCV genotype and enhanced risk of neonatal infection. HCV-RNA levels were significantly higher (p < 0.05) in mothers with HIV coinfection than in those with HCV alone. These data indicate that maternal HIV status correlates with enhanced level of viraemia which favours neonatal infection.

Identification of a novel variant of hepatitis E virus in Italy

Hepatitis E infection is typically associated with areas in which hepatitis E virus (HEV) is endemic. Except for a few cases in Europe and in the United States, acute hepatitis E is usually associated with travel to endemic areas. We set out to determine the etiologic role of HEV in acute non‐A‐C hepatitis in Italy. The presence of HEV‐RNA and antibody was determined in 218 patients diagnosed with acute viral non‐A‐C hepatitis. Acute hepatitis E infection was defined by the presence of HEV‐RNA in sera and positivity for IgM anti‐HEV and seroconversion to IgG anti‐HEV. Acute hepatitis E was found in 10.1% of the patients with acute non‐A‐C, with 95.5% exhibiting a benign course. A more severe course was observed in a patient co‐infected with HAV and HEV. Most cases were travelers to endemic areas, although 18.2% reported no travel. One patient was from a household with an infected patient. Sequence analyses of the polymerase chain reaction (PCR) product derived from a patient who never visited endemic areas, identified an isolate that is divergent significantly from all reported isolates of HEV (79.5–85.8% nucleotide identity). Evidence from this study suggests that HEV accounts for approximately 10% of acute non‐A‐C viral hepatitis in Italy, diagnosed generally in travelers returning from endemic areas. However, the identification of a new HEV variant in an individual who never indicated travel or contact with individuals associated with endemic areas, suggests that this virus may be native to Italy. J. Med. Virol. 57:356–360, 1999.

Perinatal transmission and manifestation of hepatitis C virus infection in a high risk population.

We studied the perinatal transmission of hepatitis C virus (HCV) in 70 high risk mother/infant pairs. Seventy-six percent of the mothers (53 of 70) were coinfected with human immunodeficiency virus (HIV) and 79% (55 of 70) had a history of drug addiction. During the follow-up HCV RNA was detected in 14 of 70 (20%) infants: 12% (2 of 17) in infants born to HIV-negative mothers; and 23% (12 of 53) in infants to HIV-positive mothers. The rate of vertical transmission was significantly higher in vaginally delivered infants than in those delivered by cesarean section (32% vs. 6%; P < 0.05). All 56 uninfected infants lost passively acquired anti-HCV by age 9 \pm 4 months and only 2 of 56 infants (4%) had evidence of HIV infection. Four of 14 HCV RNA-positive infants (29%) had evidence of HTV coinfection. We observed 3 clinical patterns of HCV infection: a transient viremia in 2 infants; an acute pattern in 2 infants; and a chronic pattern in 10 infants. All 4 HIV-coinfected infants had chronic HCV infection. All infants with a chronic pattern, had increased alanine aminotransferase values for more than 6 months and 5 had a liver biopsy that showed signs of chronic persistent hepatitis. HCV perinatal transmission was more frequent in infants born to HIV-coinfected mothers than in infants born to HIV-noninfected women, particularly when delivered vaginally.

A Prospective Study on Mother-to-Infant Transmission of Hepatitis C Virus

To assess the rate of mother-to-infant transmission of hepatitis C virus (HCV) and to identify potential risk factors for transmission, we followed up (mean 22.4 months, range 1–7.5 years) a cohort of 291 babies born to anti-HCV-positive mothers, 40 of whom were also HIV coinfected. Seventeen (5.8%) babies acquired HCV infection, but none became icteric. All babies developed chronic HCV infection with 16 babies showing elevated levels of ALT. The rate of transmission was higher in babies born to mothers coinfected with HIV than in those born to mothers with HCV alone (22.5 vs. 3.2%, p < 0.0001). No association was seen between a specific maternal HCV genotype and an increased risk of neonatal infection. The median level of HCV-RNA was higher in mothers who transmitted infection than in those who did not, although the ranges overlapped. In this study, maternal history of chronic liver disease, mode of delivery and type of feeding were not predictive of HCV infection.

Safety and immunogenicity of influenza vaccination in individuals infected with HIV

Influenza can cause severe complications in HIV infected individuals leading to increases in hospitalisation and mortality. Vaccination is recommended for such individuals, but some studies reported that immunisation against influenza may stimulate an increase of HIV viral load and decrease of CD4+ cells count. A review of published studies, including our study carried out in HIV former drug addicts, indicates that vaccination against influenza is well tolerated in both children and adult individuals with HIV, but response to vaccination is lower than that observed in immunocompetent individuals. Most studies, including our own, show that vaccination does not induce significant changes in viral load and CD4+ cell counts. In studies reporting modifications of such parameters there is a general agreement that the increased viral replication is usually transient and unable to determine a clear, measurable progression of the underlying HIV disease. Therefore, vaccination against influenza can be safely administered to HIV infected people.

Multicenter trial on mother-to-infant transmission of GBV-C virus

Evidence indicates that the GBV‐C or hepatitis G virus can cause persistent infection in humans, but little is known on the importance of vertical transmission. To assess the risk of mother‐to‐infant transmission and the clinical outcome of infected babies, we investigated 175 anti‐HCV positive mothers and followed‐up their children for 3–33 months. GBV‐C RNA was detected by RT‐PCR and anti‐E2 antibody was assayed by EIA. Thirty‐four (19.4%) women were GBV‐C RNA positive and transmission occurred to 21 (61.8%) babies; 20 (95.2%) acquired GBV‐C alone, and one (4.8%) GBV‐C and HCV. Maternal factors such as intravenous drug use, HIV coinfection, HCV‐RNA positivity, and type of feeding were not correlated with GBV‐C transmission. GBV‐C RNA remained persistently positive in all infected babies but one baby who seroconverted to anti‐E2. Seven (35%) babies with GBV‐C alone developed marginally elevated ALT; the baby with HCV and GBV‐C co‐infection had the highest ALT peak value (664 IU/l). Seven of the 141 (5%) babies born to the GBV‐C RNA negative mothers acquired HCV and six (85.7%) had abnormal ALT. The mean ALT peak value was significantly higher (P < 0.05) for babies with HCV than for those with GBV‐ C. None of the children with GBV‐C or with HCV became icteric. GBV‐C is frequently present in anti‐HCV positive women. The infection is transmitted efficiently from mother to baby and rate of transmission is much higher than that for HCV. GBV‐C can cause persistent infection in babies but usually without clear evidence of liver disease. J. Med. Virol. 54: 107–112, 1998.

Dominant Role of Host Selective Pressure in Driving Hepatitis C Virus Evolution in Perinatal Infection

ABSTRACT The dynamics of the genetic diversification of hepatitis C virus (HCV) populations was addressed in perinatal infection. Clonal sequences of hypervariable region 1 of the putative E2 envelope protein of HCV were obtained from four HCV-infected newborns (sequential samples spanning a period of 6 to 13 months after birth) and from their mothers (all samples collected at delivery). The data show that the variants detected between birth and the third month of life in samples from the four newborns were present in the HCV populations of their mothers at delivery. In the newborns, a unique viral variant (or a small group of closely related variants) remained stable for weeks despite active viral replication. Diversification of the intrahost HCV population was observed 6 to 13 months after birth and was substantially higher in two of the four subjects, as documented by the intersample genetic distance (GD) (P = 0.007). Importantly, a significant correlation between increasing GD and high values for the intersampleKa/Ks ratio (the ratio between antonymous and synonymous substitutions; an index of the action of selective forces) was observed, as documented by the increase of both parameters over time (P = 0.01). These data argue for a dominant role of positive selection for amino acid changes in driving the pattern of genetic diversification of HCV populations, indicate that the intrahost evolution of HCV populations is compatible with a Darwinian model system, and may have implications in the designing of future antiviral strategies.

Vaccination against hepatitis B: the Italian strategy

Viral hepatitis B, which is a major health problem worldwide, is endemic in Italy. In response to this, vaccination became compulsory in 1991. Vaccine is administered to neonates and 12-year-old adolescents; in 12 years time, all Italians under the age of 24 will be immune to HBV.