Analysis of full-length hepatitis A virus genome in sera from patients with fulminant and self-limited acute type A hepatitis

Fujiwara, Keiichi; Yokosuka, Osamu; Fukai, Kenichi; Imazeki, Fumio; Saisho, Hiromitsu; Omata, Masao

doi:10.1016/s0168-8278(01)00074-5

Cited by 89 publications

(69 citation statements)

References 23 publications

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“…The alignment of the 24 clinical and reference sequences with the genotype I consensus showed 137 variable positions over the 675 studied nucleotides. Among those present in both reference and clinical sequences, we found G324A and C372G/T reported by Fujiwara et al as potentially associated with benign hepatitis (14). These mutations were observed in clinical sequences from both benign (G1B5 and G1B6) and severe (G1S1 and G1F2) hepatitis, and also in the three genotype IB reference sequences, including HM-175.…”

Section: Resultssupporting

confidence: 54%

“…The finding that these mutations were associated with viral attenuation in vivo supports the hypothesis of viral determinants of virulence in the 5Ј UTR (15). Among the few clinical studies which have addressed this question, Fuji-wara et al, by comparing full-length HAV genomes obtained from Japanese patients with benign or fulminant hepatitis, found less nucleotide variation in the 5Ј UTRs from patients with fulminant hepatitis (12,13) and suggested that two IRES mutations (G324A and C372G/T) might influence the course of HAV infection (14).…”

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confidence: 99%

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Nucleotide Variability and Translation Efficiency of the 5′ Untranslated Region of Hepatitis A Virus: Update from Clinical Isolates Associated with Mild and Severe Hepatitis

Mackiewicz

Cammas

Desbois

et al. 2010

J Virol

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Mutations in the internal ribosome entry site (IRES) of hepatitis A virus (HAV) have been associated with enhanced in vitro replication and viral attenuation in animal models. To address the possible role of IRES variability in clinical presentation, IRES sequences were obtained from HAV isolates associated with benign (n ‫؍‬ 8) or severe (n ‫؍‬ 4) hepatitis. IRES activity was assessed using a bicistronic dual-luciferase expression system in adenocarcinoma (HeLa) and hepatoma (HuH7) cell lines. Activity was higher in HuH7 than in HeLa cells, except for an infrequently isolated genotype IIA strain. Though globally low, significant variation in IRES-dependent translation efficiency was observed between field isolates, reflecting the low but significant genetic variability of this region (94.2% ؎ 0.5% nucleotide identity). No mutation was exclusive of benign or severe hepatitis, and variations in IRES activity were not associated with a clinical phenotype, indirectly supporting the preponderance of host factors in determining the clinical presentation.Hepatitis A virus (HAV) is a nonenveloped RNA virus of the Picornaviridae family. The viral genome consists of an approximately 7,500-nucleotide (nt)-long, positive-stranded RNA divided in three parts: a 5Ј untranslated region (5Ј UTR), a single open reading frame that encodes both structural and nonstructural proteins, and a 3Ј UTR with a short poly(A) tail. By sequencing of the VP1-2A junction and the VP1 gene, 3 genotypes (I, II, and III) divided into A and B subtypes have been described in humans (7,27). HAV is the main cause of acute viral hepatitis worldwide. The majority of cases follow a benign course, but some may be present with fulminant forms, characterized by acute liver failure (factor V levels of Ͻ50% and encephalopathy). HAV-induced liver disease appears to result primarily from immunologic mechanisms, chiefly on the basis of in vitro studies. Most HAV strains have no detectable cytopathic effect in cell culture and no apparent effect on cell growth or metabolism (16), and HAV-infected cells are lysed by cytotoxic T cells isolated from the liver of acutely infected patients (30,31). Clinical studies have suggested that host factors such as age and underlying liver disease were involved in the severity of liver diseases (32, 33) and that the host immune response also played a role in the fulminant forms of hepatitis A, as evidenced by markedly low viral loads (26).Nevertheless, the existence of viral determinants of hepatitis A severity is suggested by both experimental and clinical studies. Indeed, mutations within the VP1-2A and 2C genes have been shown to enhance virulence in tamarins (9). It has also been suggested that 5Ј UTR mutations associated with viral adaptation to cell culture were also responsible for viral attenuation in vivo (15). The 5Ј UTR of HAV is about 735 nucleotides long and is considered the most conserved region of the genome. The 5Ј UTR is involved in genome replication and translation initiation. Folding predictions and biochem...

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Section: Resultssupporting

confidence: 54%

mentioning

confidence: 99%

Nucleotide Variability and Translation Efficiency of the 5′ Untranslated Region of Hepatitis A Virus: Update from Clinical Isolates Associated with Mild and Severe Hepatitis

Mackiewicz

Cammas

Desbois

et al. 2010

J Virol

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“…Total RNA was extracted with TRIzol (Invitrogen), and HAV RNA was reverse transcribed at 50°C for 1 h using 2.5 mM oligo(dT) [15][16][17][18] and 10 U/l SuperScript III (Invitrogen). Real-time PCR with 0.5 M primers specific for HAV 3C (sense, 5Ј-ACTTTGGAAATAGCAGGACT GG-3Ј; antisense, 5Ј-AGATTGATTCGATGAAACC-3Ј) was performed by using SYBR Green supermix (Bio-Rad) in a Bio-Rad iCycler.…”

Section: Cells and Virusesmentioning

confidence: 99%

“…HAV causes a self-limiting infection of the liver with usually mild or no symptoms in young patients, whereas adult patients might suffer from severe symptoms. In immune-compromised or chronically infected patients, fulminant hepatic failure is often associated with a high mortality rate (15,52). Albeit vaccines against hepatitis A virus can prevent the disease (3,14,31), they are futile for fulminant hepatitis.…”

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confidence: 99%

Silencing of Hepatitis A Virus Infection by Small Interfering RNAs

Kusov

Palmenberg

Sgro

et al. 2006

J Virol

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Infection by hepatitis A virus (HAV) can cause acute hepatitis and, rarely, fulminant liver failure, in particular in patients chronically infected with hepatitis C virus. Based on our previous observation that small interfering RNAs (siRNAs) can silence translation and replication of the firefly luciferase-encoding HAV replicon, we now exploited this technology to demonstrate the effect of siRNAs on viral infection in Huh-7 cells. Freshly and persistently infected cells were transfected with siRNAs targeting various sites in the HAV nonstructural genes. Compared to a single application, consecutive siRNA transfections targeting multiple sequences in the viral genome resulted in a more efficient and sustained silencing effect than a single transfection. In most instances, multiple applications of a single siRNA led to the emergence of viral escape mutants with mutated target sites that rendered these genomes resistant to RNA interference (RNAi). Efficient and sustained suppression of the viral infectivity was achieved after consecutive applications of an siRNA targeting a computer-predicted hairpin structure. This siRNA holds promise as a therapeutic tool for severe courses of HAV infection. In addition, the results provide new insight into the structural bases for sequencespecific RNAi.Hepatitis A virus (HAV) has a single-stranded RNA genome of 7.5 kb with positive polarity. As a member of the picornavirus family, it is unique in its biological properties, in particular, in its highly protracted replicative cycle in cell cultures and inability to shut off the host cell metabolism. HAV causes a self-limiting infection of the liver with usually mild or no symptoms in young patients, whereas adult patients might suffer from severe symptoms. In immune-compromised or chronically infected patients, fulminant hepatic failure is often associated with a high mortality rate (15, 52). Albeit vaccines against hepatitis A virus can prevent the disease (3, 14, 31), they are futile for fulminant hepatitis. Therefore, therapeutic intervention strategies to prevent progression to a life-threatening liver disease are still in need. Given that the rapid degradation of the HAV genome can ameliorate viral infection and thus prevent fulminant failure, RNA interference (RNAi) based on sequence-specific genome degradation may present a novel and specific therapeutic approach in preventing severe disease progression.Initially demonstrated in Caenorabditis elegans and Drosophila melanogaster, RNAi is now recognized as a valuable tool to silence viral infections (reviewed in references 22, 48, and 50). Among the numerous viruses that have been successfully silenced by RNAi, there are five representatives of the picornavirus family: poliovirus (19,20), human rhinovirus (45), enterovirus 71 (37), foot-and-mouth disease virus (7, 9, 36), and coxsackievirus B3 (1, 40). Recently, we have shown that genome replication of an HAV replicon can be efficiently inhibited by small interfering RNAs (siRNAs) targeting either coding or noncoding regions...

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“…Viral hepatitis A is one of the most important causes of ALF in children, besides hepatitis B, D and E, Epstein-Barr virus, cytomegalovirus, parvovirus and acetaminophen overdoses. Even though there is evidence of specific genetic features in viruses found in ALF patients [2], the reason why some hepatitis A patients present ALF, whereas most of them present a self-limited picture, remains known.…”

mentioning

confidence: 99%

Acute liver failure complicating viral hepatitis A

Diniz-Santos¹,

Melo²,

Melo³

et al. 2004

Braz J Infect Dis

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Hepatitis A is one of the most frequent infectious liver diseases affecting children worldwide. The disease is usually mild and self-limited, and complications are very rare. Nevertheless, hepatitis A can sometimes cause acute liver failure (ALF), a severe, life-threatening condition. Herein is reported a case of a child who presented ALF during a course of hepatitis A. The need for early identification of possible ALF cases among hepatitis A patients, and for effective ways of evaluating such a possibility, are discussed. We also emphasize the importance of prevention measures, especially vaccination.

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Analysis of full-length hepatitis A virus genome in sera from patients with fulminant and self-limited acute type A hepatitis

Cited by 89 publications

References 23 publications

Nucleotide Variability and Translation Efficiency of the 5′ Untranslated Region of Hepatitis A Virus: Update from Clinical Isolates Associated with Mild and Severe Hepatitis

Nucleotide Variability and Translation Efficiency of the 5′ Untranslated Region of Hepatitis A Virus: Update from Clinical Isolates Associated with Mild and Severe Hepatitis

Silencing of Hepatitis A Virus Infection by Small Interfering RNAs

Acute liver failure complicating viral hepatitis A

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