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4.
J Virol ; 98(6): e0029524, 2024 Jun 13.
Artículo en Inglés | MEDLINE | ID: mdl-38712945

RESUMEN

Hepatitis E virus is a single-strand, positive-sense RNA virus that can lead to chronic infection in immunocompromised patients. Virus-host recombinant variants (VHRVs) have been described in such patients. These variants integrate part of human genes into the polyproline-rich region that could introduce new post-translational modifications (PTMs), such as ubiquitination. The aim of this study was to characterize the replication capacity of different VHRVs, namely, RNF19A, ZNF787, KIF1B, EEF1A1, RNA18, RPS17, and RPL6. We used a plasmid encoding the Kernow strain, in which the fragment encoding the S17 insertion was deleted (Kernow p6 delS17) or replaced by fragments encoding the different insertions. The HEV RNA concentrations in the supernatants and the HepG2/C3A cell lysates were determined via RT-qPCR. The capsid protein ORF2 was immunostained. The effect of ribavirin was also assessed. The HEV RNA concentrations in the supernatants and the cell lysates were higher for the variants harboring the RNF19A, ZNF787, KIF1B, RPS17, and EEF1A1 insertions than for the Kernow p6 del S17, while it was not with RNA18 or RPL6 fragments. The number of ORF2 foci was higher for RNF19A, ZNF787, KIF1B, and RPS17 than for Kernow p6 del S17. VHRVs with replicative advantages were less sensitive to the antiviral effect of ribavirin. No difference in PTMs was found between VHRVs with a replicative advantage and those without. In conclusion, our study showed that insertions did not systematically confer a replicative advantage in vitro. Further studies are needed to determine the mechanisms underlying the differences in replicative capacity. IMPORTANCE: Hepatitis E virus (HEV) is a major cause of viral hepatitis. HEV can lead to chronic infection in immunocompromised patients. Ribavirin treatment is currently used to treat such chronic infections. Recently, seven virus-host recombinant viruses were characterized in immunocompromised patients. These viruses have incorporated a portion of a human gene fragment into their genome. We studied the consequences of these insertions on the replication capacity. We found that these inserted fragments could enhance virus replication for five of the seven recombinant variants. We also showed that the recombinant variants with replicative advantages were less sensitive to ribavirin in vitro. Finally, we found that the mechanisms leading to such a replicative advantage do not seem to rely on the post-translational modifications introduced by the human gene fragment that could have modified the function of the viral protein. The mechanisms involved in improving the replication of such recombinant viruses remain to be explored.


Asunto(s)
Virus de la Hepatitis E , Interacciones Microbiota-Huesped , Recombinación Genética , Humanos , Antivirales/farmacología , Células Hep G2 , Hepatitis E/genética , Hepatitis E/virología , Virus de la Hepatitis E/clasificación , Virus de la Hepatitis E/efectos de los fármacos , Virus de la Hepatitis E/genética , Virus de la Hepatitis E/crecimiento & desarrollo , Procesamiento Proteico-Postraduccional , Ribavirina/farmacología , ARN Viral/genética , ARN Viral/metabolismo , Replicación Viral/efectos de los fármacos , Replicación Viral/genética , Interacciones Microbiota-Huesped/genética , Ubiquitinación/genética , Plásmidos/genética
5.
Am J Trop Med Hyg ; 110(6): 1210-1213, 2024 Jun 05.
Artículo en Inglés | MEDLINE | ID: mdl-38653231

RESUMEN

We explored the association between serological status for hepatitis E and neurocysticercosis (NCC) in neurologic patients attending a national neurological referral center in Lima, Perú, between the years 2008 and 2012. Anti-hepatitis E antibodies were evaluated in patients with and without NCC, and a control group of rural general population. Anti-hepatitis E IgG was found in 23.8% of patients with NCC, compared with 14.3% in subjects without NCC from a general rural population (P = 0.023) and 14.4% in subjects with neurological complaints without NCC (P = 0.027). Seropositive patients had a median age of 44 years compared with 30 years in seronegative patients (P <0.001). No significant differences in sex, region of residence, or liver enzyme values were found. Seropositivity to hepatitis E was frequent in this Peruvian population and higher in patients with NCC, suggesting shared common routes of infection.


Asunto(s)
Virus de la Hepatitis E , Hepatitis E , Neurocisticercosis , Humanos , Neurocisticercosis/epidemiología , Neurocisticercosis/inmunología , Neurocisticercosis/complicaciones , Masculino , Adulto , Femenino , Hepatitis E/epidemiología , Hepatitis E/inmunología , Virus de la Hepatitis E/inmunología , Persona de Mediana Edad , Perú/epidemiología , Adulto Joven , Prevalencia , Inmunoglobulina G/sangre , Anticuerpos Antihepatitis/sangre , Estudios Seroepidemiológicos , Adolescente , Anciano
6.
J Virol Methods ; 327: 114920, 2024 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-38574772

RESUMEN

INTRODUCTION: We evaluated the performance of the automated Altostar HEV RNA platform for detecting HEV RNA. METHODS AND RESULTS: Clinical performance was determined by testing 81 plasma samples and 10 fecal samples manually quantified previously with the Realstar RT-PCR assay using the Magnapure instrument for extraction. The assays were concordant for 79/81 plasma samples (97.5%) and 10/10 (100%) fecal samples. The two plasma samples that tested negative with the Altostar assay had a very low HEV RNA concentration (1.6 and 1.4 log10 IU/ml). Quantitative results obtained with the automated platform and the manual workflow were highly correlated (ρ= 0.98, p<0.01). The intra-run and inter-run standard deviation were 0.09 IU/ml and 0.13 IU/ml respectively. The assay was linear from 2 to 6 log IU/ml. The limit of detection determined by Probit analysis with the WHO HEV RNA standard was 7.6 [95% CI: 4.4-52.5] IU/ml. CONCLUSIONS: The Altostar platform enables highly accurate testing for the detection of HEV RNA in stool and the quantification of HEV RNA in plasma. This allowed us to shorten turnaround times and to save time for the technical staff.


Asunto(s)
Automatización de Laboratorios , Heces , Virus de la Hepatitis E , Hepatitis E , ARN Viral , Heces/virología , Humanos , ARN Viral/aislamiento & purificación , ARN Viral/sangre , ARN Viral/análisis , ARN Viral/genética , Virus de la Hepatitis E/aislamiento & purificación , Virus de la Hepatitis E/genética , Hepatitis E/diagnóstico , Hepatitis E/virología , Hepatitis E/sangre , Sensibilidad y Especificidad , Plasma/virología , Técnicas de Diagnóstico Molecular/métodos
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