RESUMEN
Immunity acquired from infection or vaccination protects humans from symptomatic hepatitis E. However, whether the risk of hepatitis E virus (HEV) infection is reduced by the immunity remains unknown. To understand this issue, a cohort with 12 409 participants randomized to receive the hepatitis E vaccine Hecolin(®) or placebo were serologically followed up for 2 years after vaccination. About half (47%) of participants were initially seropositive. A total of 139 infection episodes, evidenced by four-fold or greater rise of anti-HEV level or positive seroconversion, occurred in participants who received three doses of treatment. Risk of infection was highest among the baseline seronegative placebo group participants (2.04%). Pre-existing immunity and vaccine-induced immunity lower the risk significantly, to 0.52% and 0.30%, respectively. In conclusion, both vaccine-induced and naturally acquired immunity can effectively protect against HEV infection.
Asunto(s)
Virus de la Hepatitis E/inmunología , Hepatitis E/inmunología , Hepatitis E/prevención & control , Vacunas Sintéticas/inmunología , Vacunas contra Hepatitis Viral/inmunología , Adolescente , Adulto , Estudios de Cohortes , Femenino , Estudios de Seguimiento , Hepatitis E/epidemiología , Humanos , Masculino , Persona de Mediana Edad , Placebos/administración & dosificación , Medición de Riesgo , Vacunas Sintéticas/administración & dosificación , Vacunas contra Hepatitis Viral/administración & dosificación , Adulto JovenRESUMEN
DDX3 is a human RNA helicase with plethoric functions. Our previous studies have indicated that DDX3 is a transcriptional regulator and functions as a tumor suppressor. In this study, we use a bicistronic reporter to demonstrate that DDX3 specifically represses cap-dependent translation but enhances hepatitis C virus internal ribosome entry site-mediated translation in vivo in a helicase activity-independent manner. To elucidate how DDX3 modulates translation, we identified translation initiation factor eukaryotic initiation factor 4E (eIF4E) as a DDX3-binding partner. Interestingly, DDX3 utilizes a consensus eIF4E-binding sequence YIPPHLR to interact with the functionally important dorsal surface of eIF4E in a similar manner to other eIF4E-binding proteins. Furthermore, cap affinity chromatography analysis suggests that DDX3 traps eIF4E in a translationally inactive complex by blocking interaction with eIF4G. Point mutations within the consensus eIF4E-binding motif in DDX3 impair its ability to bind eIF4E and result in a loss of DDX3's regulatory effects on translation. All these features together indicate that DDX3 is a new member of the eIF4E inhibitory proteins involved in translation initiation regulation. Most importantly, this DDX3-mediated translation regulation also confers the tumor suppressor function on DDX3. Altogether, this study demonstrates regulatory roles and action mechanisms for DDX3 in translation, cell growth and likely viral replication.
Asunto(s)
ARN Helicasas DEAD-box/fisiología , Factor 4E Eucariótico de Iniciación/metabolismo , Hepacivirus/fisiología , Proteínas de Unión a Caperuzas de ARN/metabolismo , ARN Helicasas DEAD-box/genética , Humanos , Mutación Puntual , Biosíntesis de Proteínas , Ribosomas/fisiología , Replicación Viral/fisiologíaRESUMEN
The protein Cdc13p binds telomeres in vivo and is essential for the maintenance of the telomeres of Saccharomyces cerevisiae. In addition, Cdc13p is known to bind single-stranded TG(1-3) DNA in vitro. Here we have shown that Cdc13p also binds DNA quadruplex, G-quartet, formed by TG(1-3) DNA. Moreover, the binding of Cdc13p causes a partial denaturing of the G-quartet DNA. Formation of DNA quadruplexes may involve the intermolecular association of TG(1-3) DNA and inhibit the extension of telomeres by telomerase. Thus, our finding suggests that Cdc13p may disrupt telomere association and facilitate telomere replication.
Asunto(s)
Ciclina B/metabolismo , ADN/química , Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/metabolismo , Alquilantes/farmacología , Animales , Baculoviridae , Línea Celular , ADN/metabolismo , Escherichia coli/metabolismo , Insectos , Mutágenos/farmacología , Conformación de Ácido Nucleico , Plásmidos/metabolismo , Unión Proteica , Desnaturalización Proteica , Ésteres del Ácido Sulfúrico/metabolismo , Telómero/metabolismoRESUMEN
There is a strong association between 2 SEN virus (SENV) variants (SENV-D and SENV-H) and transfusion-associated non-A-E hepatitis. In total, 200 subjects from a Japanese region where hepatitis C virus (HCV) is highly endemic and 194 persons from a contiguous area where HCV is not endemic were tested for SENV-D and SENV-H DNA by polymerase chain reaction. SENV DNA was detected equally in subjects from each area (56% prevalence in the area of high endemicity vs. 61% in the nonendemic area). Age-specific prevalence of SENV was similar to that of TT virus, with equal distribution at all ages in both areas; HCV was predominant in the elderly population. Alanine aminotransferase levels were significantly associated with HCV viremia but not with SENV viremia. SENV is a common infection that appears to have transmission routes and age-related prevalence that are distinct from those of HCV. No evidence was found that SENV caused hepatitis or worsened the course of hepatitis C.
Asunto(s)
Infecciones por Virus ADN/epidemiología , Hepatitis C/epidemiología , Adolescente , Adulto , Factores de Edad , Anciano , Alanina Transaminasa/sangre , Infecciones por Virus ADN/complicaciones , Infecciones por Virus ADN/transmisión , ADN Viral/sangre , Femenino , Hepatitis C/complicaciones , Hepatitis C/virología , Humanos , Japón/epidemiología , Masculino , Persona de Mediana Edad , Prevalencia , Factores de Riesgo , ViremiaRESUMEN
The existence of the newly discovered SEN virus (SENV) was investigated in 379 Japanese patients with liver diseases and in 277 blood donors, to determine whether SENV is associated with non-A-E hepatitis. SENV DNA was detected by seminested polymerase chain reaction, with primers directed to 2 SENV strains: SENV-H and SENV-D. SENV was detected in 7 (32%) of 22 patients with fulminant hepatitis, in 15 (17%) of 86 patients with acute hepatitis, in 38 (27%) of 139 patients with chronic hepatitis, in 29 (31%) of 93 patients with liver cirrhosis, in 5 (33%) of 15 patients with autoimmune hepatitis, in 11 (46%) of 24 patients with primary biliary cirrhosis, and in 27 blood donors (10%). Infection occurred more frequently in patients with liver diseases than in blood donors; however, there were no significant differences in SENV-positive rates between patients with non-A-C hepatitis and those with acute or chronic hepatitis due to known hepatitis virus or nonviral liver disease. This study did not suggest SENV as a possible causative agent of non-A-C hepatitis.
Asunto(s)
Donantes de Sangre , Infecciones por Virus ADN/epidemiología , Virus ADN/aislamiento & purificación , Hepatopatías/epidemiología , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Estudios Transversales , Infecciones por Virus ADN/virología , Virus ADN/genética , ADN Viral/análisis , Humanos , Japón/epidemiología , Hepatopatías/virología , Masculino , Persona de Mediana Edad , Reacción en Cadena de la Polimerasa/métodos , PrevalenciaRESUMEN
This study examines the relationship between HCV-RNA levels and disease severity in 60 individuals with chronic hepatitis C virus infection. HCV-RNA levels were quantified by the branched DNA (bDNA) assay in 445 samples (median: eight samples per patient) obtained over a median of 40.4 months (95% confidence interval (CI): 37.0-42.5). The median log HCV-RNA level was 6.77 (95% CI: 6.62-6.92) molecular equivalents/mL (MEQ/mL). The median log range of HCV-RNA levels in individual patients over the course of the study was 0.89 (95% CI: 0.69-1.16). HCV-RNA level varied over time by less than one log in 62% of patients, by 1-1.5 logs in 22% and by greater than 1.5 logs in only 17%. Univariate analysis, revealed an inverse association between HCV-RNA levels and ALT levels (P=0.037). Univariate and logistic regression analysis showed no significant association between HCV-RNA levels and either the degree of inflammation or fibrosis. In contrast, there was a significant positive association between alanine aminotransferase (ALT) levels and histological activity especially in individuals with ALTs> 100 IU/L. Hence, HCV-RNA levels: (i) almost always fell within the dynamic range of the bDNA assay; (ii) were stable in asymptomatic chronically infected patients, with only a small proportion of patients exceeding a range of 1.5 logs; (iii) did not correlate with either the extent of inflammation or degree of fibrosis. In contrast, there was a strong association between ALT level and the histological severity of liver disease.
Asunto(s)
Hepacivirus/genética , Hepatitis C Crónica/virología , ARN Viral/sangre , Adulto , Alanina Transaminasa/metabolismo , Femenino , Genotipo , Hepacivirus/aislamiento & purificación , Hepatitis C Crónica/sangre , Hepatitis C Crónica/fisiopatología , Humanos , Hígado/enzimología , Hígado/patología , Cirrosis Hepática/sangre , Cirrosis Hepática/virología , Masculino , Persona de Mediana Edad , Análisis de RegresiónRESUMEN
Production of recombinant proteins can be valuable in studying their biological functions. However, recombinant proteins expressed in Escherichia coli sometimes form undesirable insoluble aggregates. Solubilization and renaturation of these aggregates becomes a problem that one needs to solve. Here we used recombinant Cdc13(451-693)p as example to show the presence of l-arginine during renaturation greatly enhanced the renaturation efficiency. Cdc13p is the single-stranded telomere-binding protein of yeast Saccharomyces cerevisiae. The telomere-binding domain has been mapped within amino acids 451-693 of Cdc13p, Cdc13(451-693)p. Recombinant Cdc13(451-693)p was expressed in E. coli as insoluble protein aggregates. Purification of insoluble Cdc13(451-693)p was achieved by denaturing the protein with 6 M guanidine-HCl and followed by Ni-nitrilotriacetic acid agarose column chromatography. Renaturation of Cdc13(451-693)p to the active form was achieved by dialyzing denatured protein in the presence of l-arginine. Moreover, the presence of l-arginine was also helped in maintaining the telomere-binding activity of Cdc13(451-693)p. Taking together, l-arginine might have a general application in renaturation of insoluble aggregates.
Asunto(s)
Arginina/farmacología , Proteínas de Unión al ADN/metabolismo , Saccharomyces cerevisiae/metabolismo , Secuencia de Bases , Cartilla de ADN , Proteínas de Unión al ADN/química , Renaturación de Proteína , Proteínas Recombinantes/química , Proteínas Recombinantes/metabolismoRESUMEN
SEN virus (SEN-V) is a recently identified single-stranded, circular DNA virus. Two SEN-V variants (SENV-D and SENV-H) were assayed by polymerase chain reaction (PCR) to investigate their role in the causation of transfusion-associated non-A to E hepatitis. The incidence of SEN-V infection after transfusion was 30% (86 of 286) compared with 3% (3 of 97) among nontransfused controls (P < .001). Transfusion risk increased with the number of units transfused (P < .0001) and donor-recipient linkage for SEN-V was shown by sequence homology. The prevalence of SEN-V in 436 volunteer donors was 1.8%. Among patients with transfusion-associated non-A to E hepatitis, 11 of 12 (92%) were infected with SEN-V at the time of transfusion compared with 55 of 225 (24%) identically followed recipients who did not develop hepatitis (P < .001). No effect of SEN-V on the severity or persistence of coexistent hepatitis C virus (HCV) infection was observed. In 31 infected recipients, SEN-V persisted for greater than 1 year in 45% and for up to 12 years in 13%. SEN-V-specific RNA (a possible replicative intermediate) was recovered from liver tissue. In summary, SENV-D and -H were present in nearly 2% of US donors, and were unequivocally transmitted by transfusion and frequently persisted. The strong association of SEN-V with transfusion-associated non-A to E hepatitis compared with controls raises the possibility, but does not establish that SEN-V might be a causative agent of posttransfusion hepatitis. The vast majority of SEN-V-infected recipients did not develop hepatitis.
Asunto(s)
Infecciones por Virus ADN/complicaciones , Virus ADN , Hepatitis Viral Humana/etiología , Hepatitis Viral Humana/virología , Reacción a la Transfusión , Alanina Transaminasa/sangre , Donantes de Sangre , Enfermedad Crónica , Virus ADN/genética , Virus ADN/aislamiento & purificación , Variación Genética , Hepatitis Viral Humana/fisiopatología , Humanos , Incidencia , Hígado/virología , Datos de Secuencia Molecular , Pacientes , Índice de Severidad de la Enfermedad , Donantes de Tejidos , Viremia/sangreRESUMEN
Cdc13p is a single strand telomere-binding protein of Saccharomyces cerevisiae; its telomere-binding region is within amino acids 451-693, Cdc13(451-693)p. In this study, we used purified Cdc13p and Cdc13(451-693)p to characterize their telomere binding activity. We found that the binding specificity of single-stranded TG(1-3) DNA by these two proteins is similar. However, the affinity of Cdc13(451-693)p to DNA was slightly lower than that of Cdc13p. The binding of telomeric DNA by these two proteins was disrupted at NaCl concentrations higher than 0.3 m, indicating that electrostatic interaction contributed significantly to the binding process. Because both proteins bound to strand TG(1-3) DNA positioned at the 3' end, the 5' end, or in the middle of the oligonucleotide substrates, our results indicated that the location of TG(1-3) in single-stranded DNA does not appear to be important for Cdc13p binding. Moreover, using DNase I footprint analysis, the structure of the telomeric DNA complexes of Cdc13p and Cdc13(451-693)p was analyzed. The DNase I footprints of these two proteins to three different telomeric DNA substrates were virtually identical, indicating that the telomere contact region of Cdc13p is within Cdc13(451-693)p. Together, the binding properties of Cdc13p and its binding domain support the theory that the specific binding of Cdc13p to telomeres is an important feature of telomeres that regulate telomerase access and/or differentiate natural telomeres from broken ends.
Asunto(s)
Ciclina B/metabolismo , ADN/metabolismo , Saccharomyces cerevisiae/metabolismo , Telómero/metabolismo , Animales , Baculoviridae , Secuencia de Bases , Línea Celular , Huella de ADN , Escherichia coli , Datos de Secuencia Molecular , Unión Proteica , Proteínas Recombinantes/aislamiento & purificación , Proteínas Recombinantes/metabolismo , SpodopteraRESUMEN
A new group of transmissible single-stranded (ss) DNA viruses (SENV) distantly related to the large TT virus (TTV) family was recently identified. Eight different SENV isolates have been found, some with an association with posttransfusion hepatitis. A phylogenetic analysis of near-complete open-reading frame 1, including conserved motifs and excluding recombinant regions, was performed. The analysis used TTV-like minivirus as an outgroup, to determine a root of the phylogenetic tree, and compared 8 SENV isolates, 6 prototype TTV isolates, and 7 TTV variants (including SANBAN, TUS01, PMV, and YONBAN). Four distinct clusters separated by a bootstrap value of 100% were observed. YONBAN isolates formed a distinct outer group, representing the earliest recognized phylogenetic divergence (group 1). Prototype TTV formed group 2, PMV formed group 3, and SENV, SANBAN, and TUS01 isolates formed group 4, the most recently evolved group. This taxonomic classification suggests that these circular ssDNA viruses probably evolved from a common ancestor virus.
Asunto(s)
Infecciones por Virus ADN/virología , Virus ADN/genética , Evolución Molecular , Genoma Viral , Torque teno virus/genética , Secuencia de Aminoácidos , Virus ADN/clasificación , ADN de Cadena Simple/genética , ADN Viral/sangre , Humanos , Datos de Secuencia Molecular , Sistemas de Lectura Abierta/genética , Filogenia , Alineación de Secuencia , Análisis de Secuencia de ADN , Torque teno virus/clasificaciónRESUMEN
BACKGROUND: Infection with hepatitis G virus (HGV), also known as GB virus C, is prevalent but is not known to be associated with any chronic disease. Infection with HGV may affect the risk for AIDS in HIV-infected persons. OBJECTIVE: To compare AIDS-free survival in patients with and those without HGV infection during 16 years of follow-up after HIV seroconversion. DESIGN: Subanalysis of a prospective cohort study. SETTING: Comprehensive hemophilia treatment centers in the United States and Europe. PATIENTS: 131 patients with hemophilia who became HIV-positive between 1978 and 1985. MEASUREMENTS: Age, CCR5 genotype, HIV and HCV viral loads, CD4+ and CD8+ lymphocyte counts, and 12-year AIDS-free survival by HGV positivity (viremia [RNA] or anti-E2 antibodies). RESULTS: Compared with HGV-negative patients, the 60 HGV-positive patients (46%), including 22 who were positive for HGV RNA, had higher CD4+ lymphocyte counts (difference, 211 cells/mm3 [95% Cl, 88 to 333 cells/mm3]) and 12-year AIDS-free survival rates (68% compared with 40%; rate difference, 1.9 per 100 person-years [Cl, -0.3 to 4.2 per 100 person-years]), despite similar ages and HIV viral loads. In multivariate proportional hazards models, risk for AIDS was 40% lower for HGV-positive patients independent of age, HIV and HCV viral loads, CD4+ and CD8+ lymphocyte counts, and CCR5 genotype. CONCLUSIONS: Patients with past or current HGV infection have higher CD4+ lymphocyte counts and better AIDS-free survival rates. The mechanism of this association is unknown.
Asunto(s)
Infecciones por VIH/complicaciones , Hemofilia A/complicaciones , Hepatitis Viral Humana/complicaciones , Síndrome de Inmunodeficiencia Adquirida/etiología , Relación CD4-CD8 , Progresión de la Enfermedad , Europa (Continente) , Flaviviridae , Estudios de Seguimiento , Eliminación de Gen , Genotipo , Humanos , Masculino , Modelos de Riesgos Proporcionales , Estudios Prospectivos , Receptores CCR5/genética , Factores de Riesgo , Tasa de Supervivencia , Estados Unidos , Carga ViralRESUMEN
BACKGROUND: Hepatitis G virus (HGV), also known as GB virus C, is a newly discovered Flavivirus that is transmissible by blood transfusion and other possible routes. OBJECTIVE: To study the risk of sexual transmission of HGV in female sexual partners of men with hemophilia (n = 161 couples). METHODS: Blood samples obtained from 11 medical centers were analyzed for (1) HGV RNA by polymerase chain reaction; (2) antibodies to HGV by enzyme immunoassay; and (3) other viruses and T-cell counts by routine laboratory tests. Subjects completed a questionnaire that assessed sexual intercourse frequency, number of sexual partners, condom usage, sexually transmitted diseases, illicit drug usage, and needlestick or broken-glass injuries. RESULTS: The HGV infection (RNA +/- antibody positive) prevalence was 48% among men and 21% among women. Prevalence of hepatitis C virus, hepatitis B virus, and HIV among men was 99%, 94%, and 86%, compared with 3%, 11%, and 12% among women, respectively. The odds ratio for HGV infection for women with an HGV-positive male sexual partner was 2.14 (P = 0.06) without adjustment, and 2.77 (P = 0.03) with adjustment for other variables, none of which were independently significant. CONCLUSION: These results suggest a low level of HGV sexual transmission.
Asunto(s)
Flaviviridae , Hemofilia A/complicaciones , Hepatitis Viral Humana/etiología , Hepatitis Viral Humana/transmisión , Parejas Sexuales , Enfermedades de Transmisión Sexual/etiología , Enfermedades de Transmisión Sexual/transmisión , Anticuerpos Antivirales/análisis , Austria , Femenino , Flaviviridae/genética , Flaviviridae/inmunología , Grecia , Hepatitis Viral Humana/sangre , Hepatitis Viral Humana/inmunología , Humanos , Técnicas para Inmunoenzimas , Modelos Logísticos , Masculino , Reacción en Cadena de la Polimerasa , ARN Viral/análisis , Factores de Riesgo , Estudios Seroepidemiológicos , Conducta Sexual , Enfermedades de Transmisión Sexual/sangre , Enfermedades de Transmisión Sexual/inmunología , Abuso de Sustancias por Vía Intravenosa/complicaciones , Encuestas y Cuestionarios , Estados UnidosRESUMEN
Mycoplasma firmentans is suspected in the development of 'Gulf War illness' in veterans of Operation Desert Storm. We conducted a matched case-control study for the prevalence of M. firmentans-specific antibodies before and after the operation, as well as seroconversion rates in veterans with and without complaints of 'Gulf War illness'. Cases consisted of Gulf War veterans, who complained of various illnesses and were enrolled in the second phase of the health evaluation by the Army Comprehensive Clinical Examination Program (CCEP). Controls were selected from Gulf War veterans who did not participate in the registry and did not request a health evaluation by the CCEP. Before operation deployment, 34 out of 718 of the cases (48%) and 116 out of 2233 of the controls (5.2%) tested positive for M. fermentans-specific antibodies. There was no difference in rates of seroconversion between cases and controls (1.1 vs. 1.2%) to M. fermentans during Operation Desert Storm. Thus, there is no serological evidence that suggests infectionby M. fermentans is associated with development of 'Gulf War illness'.
Asunto(s)
Anticuerpos Antibacterianos/análisis , Infecciones por Mycoplasma/complicaciones , Mycoplasma fermentans/inmunología , Síndrome del Golfo Pérsico/microbiología , Adolescente , Adulto , Estudios de Casos y Controles , Femenino , Humanos , Masculino , Persona de Mediana Edad , Personal Militar , Infecciones por Mycoplasma/diagnóstico , Infecciones por Mycoplasma/inmunología , Mycoplasma fermentans/patogenicidad , Síndrome del Golfo Pérsico/etiología , Pruebas SerológicasRESUMEN
Excluding acute hepatic failure caused by drugs, the etiology of fulminant hepatitis (FH) remains unknown in many patients. There are conflicting data about a possible pathogenic role for the hepatitis G virus (HGV) in patients with cryptogenic fulminant hepatitis (non-A-E FH). We investigated the presence of circulating HGV in 36 patients with well-documented non-A-E fulminant and 5 patients with subfulminant hepatitis from 3 geographic locations in the United States. Serum HGV RNA was determined by reverse transcriptase-polymerase chain reaction using primers from the NS5 region of the HGV genome. HGV RNA was also measured before and after liver transplantation in 5 patients and at different time points in 7 patients. Serum samples were recoded and reanalyzed for HGV RNA using different primer sets to assess the validity of the HGV RNA assay. HGV was present in serum of 14 of the 36 patients (38.8%) with non-A-E fulminant hepatitis. Twenty percent of patients from the Northeast, 11% of the patients from the Southeast, and 50% from the Mid-Atlantic regions of the United States had circulating HGV RNA. The use of therapeutic blood products was significantly associated with the presence of serum HGV RNA (P <.02). Retesting for HGV RNA with different primers was positive in all but 1 case. HGV RNA is not causally related to non-A-E fulminant hepatitis. The finding of HGV RNA in serum from these patients is likely related to the administration of blood product transfusion after the onset of fulminant hepatitis.
Asunto(s)
Flaviviridae/aislamiento & purificación , Encefalopatía Hepática/virología , Hepatitis Viral Humana/virología , Adolescente , Adulto , Femenino , Encefalopatía Hepática/sangre , Hepatitis Viral Humana/sangre , Hepatitis Viral Humana/complicaciones , Humanos , Masculino , Persona de Mediana Edad , ARN Viral/sangre , Reacción a la TransfusiónRESUMEN
Hepatitis C Virus (HCV) causes most cases of posttransfusion hepatitis. Chronic HCV infection is highly related to chronic hepatitis, cirrhosis and hepatocellular carcinoma. Current therapies are only minimally effective and no vaccine has been developed. DNA-based immunization could be of prophylactic and therapeutic value for HCV infection. By intramuscular inoculation in BALB/c mice with an HCV recombinant plasmid pCI-HCV-C, we found significant levels of IgM antibody, but no significant IgG rise. After boost the immunized mice with recombinant HCV-core protein (cp1-10; 1-164aa), the anticore IgG, verified by Western-blotting, rose rapidly, which was two weeks earlier than that with control plasmid. Spleen cells from pCI-HCV-C immunized mice gave higher proliferation index (PI) than control (P < 0.05). The PI of cp1-10 boosted mice was even higher. Proliferation blocking assay with mAb proved the responding cell to be of CD4+ CD8- phenotype, supporting specific priming of T helper cells. A 51Cr-releasing CTL assay specific for HCV-core was developed, and a specific CTL response against HCV-core was demonstrated in both pCI-HCV-C immunized mice and mice boosted with cp1-10. Strong cytotoxic activity against peptide-pulsed p815 cells (H-2d), but not EL-4 cells (H-2b), suggested MHC class I restriction of the CTL activity. Blocking of CTL with mAb proved the effector cells to be of CD4- CD8+. Three CTL epitopes in HCV-core protein were demonstrated. We failed to detect CTL when immunized only with core protein. The results suggested that vaccination with HCV-core derived DNA sequences could be an effective method to induce humoral and cellular immune responses to HCV.
Asunto(s)
Anticuerpos contra la Hepatitis C/biosíntesis , Antígenos de la Hepatitis C/inmunología , Vacunas de ADN/inmunología , Proteínas del Núcleo Viral/inmunología , Vacunas contra Hepatitis Viral/inmunología , Animales , Antígenos CD8/inmunología , Células CHO , Cricetinae , Femenino , Anticuerpos contra la Hepatitis C/sangre , Antígenos de la Hepatitis C/biosíntesis , Antígenos de la Hepatitis C/genética , Sueros Inmunes/farmacología , Inmunoglobulina G/sangre , Inmunoglobulina M/sangre , Inyecciones Intramusculares , Activación de Linfocitos/inmunología , Ratones , Ratones Endogámicos BALB C , Fragmentos de Péptidos/biosíntesis , Fragmentos de Péptidos/genética , Fragmentos de Péptidos/inmunología , Plásmidos/genética , Plásmidos/inmunología , Proteínas Recombinantes/genética , Proteínas Recombinantes/inmunología , Linfocitos T Citotóxicos/inmunología , Transfección , Vacunas de ADN/genética , Proteínas del Núcleo Viral/genética , Vacunas contra Hepatitis Viral/genéticaRESUMEN
TT virus (TTV) has been proposed as the causative agent of non-A to E hepatitis. We studied the association between TTV viremia and biochemical evidence of hepatitis in blood donors and prospectively-followed patients. TTV was found in 7.5% of 402 donors and in 11.0% of 347 patients before transfusion. The rate of new TTV infections was 4.7% in 127 nontransfused, and 26.4% in 182 transfused patients (P <.0001). The risk of infection increased with the number of units transfused (P <.0001). The rate of new TTV infections in 13 patients with non-A to E hepatitis (23.2%) was almost identical to the rate in 124 patients who were transfused, but did not develop hepatitis (21.8%). Of 45 patients with acute hepatitis C, 40.0% were simultaneously infected with TTV. TTV did not worsen the biochemical severity (mean ALT: 537 in TTV+; 550 in TTV-) or persistence of hepatitis C. In non-A to E cases, the mean ALT was 182 in those TTV-positive and 302 in TTV-negatives. No consistent relationship between alanine transaminase level and TTV DNA level was observed in 4 patients with long-term, sequential samples. Of 21 viremic subjects, 67% cleared TTV within 5 years (38% in 1 year); 33% were viremic throughout follow-up extending to 22 years. We conclude that TTV is a very common, often persistent infection that is transmitted by transfusion and by undefined nosocomial routes. We found no association between TTV and non-A to E hepatitis and no effect of TTV on the severity or duration of coexistent hepatitis C. TTV may not be a primary hepatitis virus.
Asunto(s)
Donantes de Sangre , Virus ADN/aislamiento & purificación , ADN Viral/sangre , Hepatitis Viral Humana/epidemiología , Hepatitis Viral Humana/transmisión , Procedimientos Quirúrgicos Torácicos/efectos adversos , Reacción a la Transfusión , District of Columbia/epidemiología , Anticuerpos Antihepatitis/sangre , Humanos , National Institutes of Health (U.S.) , Complicaciones Posoperatorias/virología , ARN Viral/sangre , Cruz Roja , Estados UnidosRESUMEN
Serologic, biochemical, and molecular analyses were used to study hepatitis G virus (HGV), antibody to the HGV envelope protein (anti-E2), risk factors, clinical significance, and the impact of HGV on coexistent hepatitis C virus (HCV). Among 329 donors with confirmed HCV infection, 12% were HGV RNA-positive and 44% were anti-E2-positive (total exposure, 56%). HGV RNA and anti-E2 were mutually exclusive except in 9 donors (1.5%); 8 of 9 subsequently lost HGV RNA but anti-E2 persisted. HGV had little impact on alanine aminotransferase, aspartate aminotransferase, or gamma-glutamyl transpeptidase in donors with HGV infection alone or those coinfected with HCV. A multivariate analysis showed that intravenous drug abuse was the leading risk factor for HGV transmission, followed by blood transfusion, snorting cocaine, imprisonment, and a history of sexually transmitted diseases. In summary, HGV and HCV infections were frequently associated and shared common parenteral risk factors; HGV did not appear to cause hepatitis or to worsen the course of coexistent hepatitis C.
Asunto(s)
Flaviviridae/aislamiento & purificación , Anticuerpos Antihepatitis/sangre , Hepatitis C/complicaciones , Hepatitis Viral Humana/complicaciones , Hepatitis Viral Humana/transmisión , ARN Viral/sangre , Adulto , Donantes de Sangre , Digoxigenina , Femenino , Flaviviridae/genética , Flaviviridae/inmunología , Hepacivirus/inmunología , Hepatitis C/transmisión , Hepatitis C/virología , Hepatitis Viral Humana/inmunología , Hepatitis Viral Humana/virología , Humanos , Pruebas de Función Hepática , Masculino , Persona de Mediana Edad , Reacción en Cadena de la Polimerasa , Factores de Riesgo , Proteínas del Envoltorio Viral/inmunologíaRESUMEN
Mycoplasma fermentans is currently being examined as an agent potentially associated with human disease. Several strains of M. fermentans were isolated from patients with respiratory tract disease and AIDS. Two of these clinical strains, M64 and SK6, were triple-filter-cloned and designated as the parental clones in this study. Genomic DNA of randomly picked subclones in four and five subsequent generations passed from the parental M64 and SK6 clones were analyzed by using a radiolabeled M. fermentans-specific insertion sequence (IS)-like element as the probe. The hybridization patterns of DNA restriction fragments revealed high frequencies of chromosomal changes accompanied with excision or new insertion of the IS-like element in M. fermentans chromosome. The findings indicate M. fermentans has an effective mechanism(s) to produce a rapid gene rearrangement that may be mediated by one or more copies of the IS-like element.
Asunto(s)
Cromosomas Bacterianos/genética , Reordenamiento Génico , Mycoplasma fermentans/genética , Southern Blotting , Elementos Transponibles de ADN , ADN Bacteriano , Reordenamiento Génico/genética , HumanosRESUMEN
Chronic persistent infections by mycoplasmas induced malignant transformation of C3H mouse embryo cells that normally had never been reported to undergo spontaneous transformation. This mycoplasma-mediated oncogenic process had a long latency (more than 7 weeks of continuous mycoplasmal infection) and showed a multistage progression characterized by reversibility (at least up to 11 weeks of mycoplasmal infection) and irreversibility of malignant properties upon removal of the mycoplasma from culture. Further prolonged infections (18 weeks) by Mycoplasma fermentans or M. penetrans resulted in permanent transformation of these C3H cells that no longer required the continued presence of the transformation-inducing mycoplasmas in cultures to retain their malignant properties. Previous studies of viral oncogenesis revealed that virus-transformed cells always had viral gene(s) present. Integration of viral gene(s) apparently played an important role in the process of oncogenesis. In this study, we examined if the continued presence of any mycoplasmal gene(s) in mammalian cells, in whatever form, was also crucial in causing malignant cell transformation. Representational difference analysis (RDA) was a recently developed powerful technique to compare differences between two complex genomes. In the RDA system, subtractive and kinetic enrichment was used to purify and isolate restriction endonuclease gene fragment(s) of mycoplasmal origin, presumably present only in mycoplasma-transformed C3H cells, but not in nonmycoplasma-exposed control C3H cells. After three rounds of subtractive hybridization following PCR enrichment for each of three different restriction enzymes DNA digests, no gene fragment of mycoplasmal origin was amplified or identified in the permanently transformed C3H cells. Differing from tumorigenesis in animal cells induced by most oncogenic viruses or in plant cells induced by Agrobacteria, mycoplasmas evidently did not cause malignant transformation by integrating their gene(s) into the mammalian cell genome.
Asunto(s)
Transformación Celular Neoplásica/genética , ADN Bacteriano/análisis , Regulación Bacteriana de la Expresión Génica/genética , Genes Bacterianos/genética , Mycoplasma fermentans/genética , Mycoplasma penetrans/genética , Animales , Línea Celular Transformada , Cartilla de ADN/química , Electroforesis en Gel de Agar , Amplificación de Genes , Ratones , Ratones Endogámicos C3H , Mycoplasma fermentans/fisiología , Mycoplasma penetrans/fisiología , Hibridación de Ácido Nucleico/métodos , Reacción en Cadena de la PolimerasaRESUMEN
OBJECTIVES: The hepatitis G virus (HGV) is a newly described flavivirus that affects a high proportion of patients with chronic viral hepatitis: our objective was to determine what role HGV might play in the course of disease. METHODS: We evaluated stored serum samples from 108 patients with chronic hepatitis B and 99 patients with chronic hepatitis C who participated in trials of alpha-interferon or ribavirin for the presence of hepatitis B virus (HBV) DNA and hepatitis C virus (HCV) RNA by branched DNA and for the presence of HGV RNA by polymerase chain reaction (PCR), using primers from the NS5 region of the genome. RESULTS: Initially, 20 (19%) patients with hepatitis B and 11 (11%) with hepatitis C had HGV RNA in their serum. Patients with and without HGV infection were similar with regard to clinical features, laboratory tests, and hepatic histology. HGV RNA levels fell during interferon therapy and became undetectable in those receiving the highest doses; however, HGV RNA levels returned to pretreatment values when therapy was stopped. With ribavirin therapy, HGV RNA levels did not change. Two- to 12-yr follow-up serum samples were available from 17 initially HGV RNA-positive patients, of whom only 10 (59%) were still positive. CONCLUSIONS: HGV infection is common among patients with chronic hepatitis B and C but has little effect on the short-term course of disease or response to therapy. HGV RNA levels are suppressed but not eradicated by alpha-interferon and are unaffected by ribavirin treatment. Spontaneous loss of HGV RNA occurs over time in a proportion of patients.