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INTRODUCTION: Establishing the safety and immunogenicity of a hepatitis E virus vaccine in multiple populations could facilitate broader access and prevent maternal and infant mortality. METHODS: We conducted a phase 1, randomized, double-blinded, placebo-controlled (4:1 vaccine: placebo) trial of 30 µg HEV-239 (Hecolin®, Xiamen Innovax Biotech Company Limited, China) administered intramuscularly in healthy US adults aged 18-45 years. Participants were vaccinated on days 1, 29, and 180. Participants reported solicited local and systemic reactions for 7 days following vaccination and were followed through 12 months after enrollment for safety and immunogenicity (IgG, IgM). RESULTS: Solicited local and systemic reactions between treatment and placebo group were similar and overall mild. No participants experienced serious adverse events related to HEV-239. All participants receiving HEV-239 seroconverted at one month following the first dose and remained seropositive throughout the study. HEV-239 elicited a robust hepatitis E IgG response that peaked one month following the second dose (Geometric Mean Concentration (GMC) 6.16; 95% CI 4.40-8.63), was boosted with the third dose (GMC 11.50; 95% CI 7.90-16.75) and persisted through 6 months. CONCLUSIONS: HEV-239 is safe and elicits a durable immune response through at least 6 months after the third dose in healthy US adults. CLINICAL TRIALS REGISTRATION: NCT03827395. Safety Study of Hepatitis E Vaccine (HEV239) - Full Text View - ClinicalTrials.gov.
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BACKGROUND: Hepatitis E virus (HEV) is a leading cause of acute hepatitis. The long-term efficacy of a hepatitis E vaccine needs to be determined. METHODS: In an initial efficacy study, we randomly assigned healthy adults 16 to 65 years of age to receive three doses of either a hepatitis E vaccine (vaccine group; 56,302 participants) or a hepatitis B vaccine (control group; 56,302 participants). The vaccines were administered at 0, 1, and 6 months, and the participants were followed for 19 months. In this extended follow-up study, the treatment assignments of all participants remained double-blinded, and follow-up assessments of efficacy, immunogenicity, and safety were continued for up to 4.5 years. RESULTS: During the 4.5-year study period, 60 cases of hepatitis E were identified; 7 cases were confirmed in the vaccine group (0.3 cases per 10,000 person-years), and 53 cases in the control group (2.1 cases per 10,000 person-years), representing a vaccine efficacy of 86.8% (95% confidence interval, 71 to 94) in the modified intention-to-treat analysis, rather than (95% confidence interval, 71 to 84) [corrected]. Of the participants who were assessed for immunogenicity and were seronegative at baseline, 87% of those who received three doses of the hepatitis E vaccine maintained antibodies against HEV for at least 4.5 years; HEV antibody titers developed in 9% in the control group. The rate of adverse events was similar in the two groups. CONCLUSIONS: Immunization with this hepatitis E vaccine induced antibodies against HEV and provided protection against hepatitis E for up to 4.5 years. (Funded by the Chinese Ministry of Science and Technology and others; ClinicalTrials.gov number, NCT01014845.).
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Virus de la Hepatitis E/inmunología , Hepatitis E/prevención & control , Vacunas contra Hepatitis Viral/inmunología , Adolescente , Adulto , Anciano , Método Doble Ciego , Femenino , Genotipo , Anticuerpos Antihepatitis/sangre , Hepatitis E/inmunología , Virus de la Hepatitis E/genética , Humanos , Inmunoglobulina G/sangre , Masculino , Persona de Mediana Edad , Factores de Tiempo , Vacunas contra Hepatitis Viral/efectos adversos , Adulto JovenRESUMEN
Laboratory diagnosis of acute infection of hepatitis E virus (HEV) is commonly based on the detection of HEV RNA, IgM and/or rising IgG levels. However, the profile of these markers when the patients present have not been well determined. To clarify the extent of misdiagnosed sporadic hepatitis E in the initial laboratory detection, serial sera of 271 sporadic acute hepatitis cases were collected, detected and the dynamics of each acute marker during the illness course were analyzed. 91 confirmed cases of hepatitis E were identified based on the presentation of HEV RNA, IgM or at least 4 fold rising of IgG levels. 21 (23.1%) hepatitis E cases were false negative for the viral RNA and 40 (44.0%) for rising IgG, because occurrence of these markers were confined to acute phase of infection and viremia had already subsided and antibody level peaked when these patients presented. IgM was detected in 82 (90.1%) cases. It is the most prevalent of the three markers, because the antibody persisted until early convalescence. Nine cases negative for IgM were positive for rising IgG and one was also positive for the viral RNA; all of these nine cases showed high avid IgG in their acute phase sera, which indicated re-infection. In summary, it is not practicable to determine the true occurrence of sporadic hepatitis E. Nevertheless, it could be closely approximated by approach using a combination of all three acute markers.
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Biomarcadores/sangre , Hepatitis E/sangre , Enfermedad Aguda , Secuencia de Bases , Cartilla de ADN , Progresión de la Enfermedad , Ensayo de Inmunoadsorción Enzimática , Virus de la Hepatitis E/genética , Humanos , ARN Viral/sangreRESUMEN
Hemagglutinin (HA), the major antigen on the surface of influenza viruses, is the primary target for neutralizing antibodies and vaccine design. However, frequent mutations in this gene allow the virus to evade host immune responses and conventional prophylaxis and treatment. In this report, we humanized 4D1 and 10F7 mouse monoclonal antibodies (mAbs) that, we had previously shown to display broad-spectrum neutralization to avian H5N1 virus. The genes of variable (V) regions of 4D1 and 10F7 mAbs were combined with constant region of human antibody to construct the chimeric antibodies (cAbs). The results of hemagglutinin inhibition (HI) and neutralization assays showed that 4D1 and 10F7 cAbs were functional and retained broad-spectrum reactivity. Antibody competitive ELISA and affinity tests indicated that the cAbs recognized the same epitope as the parent mAbs with similar affinity. In animal experiments, the 10F7 cAb showed full protection against lethal challenge of highly virulent avian H5N1 virus, A/BH Goose/QH/15C/2005, in all infected mice. These humanized broad-spectrum antibodies may be potentially important for the control of both current and future antigenic variants of H5N1 virus.
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Anticuerpos Monoclonales/inmunología , Anticuerpos Neutralizantes/inmunología , Anticuerpos Antivirales/inmunología , Antivirales/inmunología , Subtipo H5N1 del Virus de la Influenza A/inmunología , Animales , Anticuerpos Monoclonales/genética , Anticuerpos Monoclonales/uso terapéutico , Anticuerpos Neutralizantes/genética , Anticuerpos Neutralizantes/uso terapéutico , Anticuerpos Antivirales/genética , Anticuerpos Antivirales/uso terapéutico , Antivirales/uso terapéutico , Peso Corporal , Mapeo Epitopo , Pruebas de Inhibición de Hemaglutinación , Humanos , Regiones Constantes de Inmunoglobulina/genética , Región Variable de Inmunoglobulina/genética , Gripe Humana , Ratones , Pruebas de Neutralización , Infecciones por Orthomyxoviridae/tratamiento farmacológico , Infecciones por Orthomyxoviridae/patología , Análisis de SupervivenciaRESUMEN
A novel immunoassay that detects simultaneously hepatitis B virus (HBV) PreS1 and/or core-related antigens was developed and evaluated for its potential for detecting hepatitis B surface antigen (HBsAg) variants. The detection limits of the assay was 10(2.9+/-0.5)copies/mL (mean+/-SD) for HBsAg-positive sera with different genotypes, and 10(3.5+/-1.2)copies/mL for HBsAg variants sera. The specificity of the assay was 99.9% (95% CI: 99.7-99.9%, 4551 healthy individuals). The sensitivities were 93.9% (95% CI: 92.8-94.9%), 59.3% (95% CI: 38.7-77.6%) and 80% (95% CI: 44.4-97.5%) in three independent groups which include: 2065 hepatitis patients, 27 patients with occult hepatitis B and 10 HBsAg variants, respectively. In addition, a novel premature stop code mutation at position 112 of HBsAg was observed in two patients with chronic hepatitis B with different genotypes.
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Antígenos de Superficie de la Hepatitis B/sangre , Hepatitis B Crónica/diagnóstico , Virología/métodos , Adolescente , Adulto , Niño , Codón sin Sentido , Femenino , Antígenos del Núcleo de la Hepatitis B/sangre , Antígenos de Superficie de la Hepatitis B/genética , Hepatitis B Crónica/genética , Hepatitis B Crónica/inmunología , Humanos , Inmunoensayo/métodos , Masculino , Persona de Mediana Edad , Mutación Missense , Sensibilidad y Especificidad , Adulto JovenRESUMEN
Several peptide mimics of a conserved H5N1 avian influenza virus neutralization site recognized by 8H5 mAb have been reported previously. In this study, the secondary and possibly higher structural orders of the peptide mimics 122 and 125 were investigated and found to be closely related to the specific binding with 8H5 mAb. These two peptide mimics were fused to three different carrier proteins, and the antibody binding activities were recovered in 4 of the 11 fusion proteins. HEV structural protein p239 and HBc were more suitable than the outer membrane protein T47 of the Treponema pallidum particle for the recovery of reactivity. The increase in the copy number of peptide mimics was important for the recovery of antibody-binding activity and the interaction between peptide and carrier protein may affect the spatial structure of both the peptide and the carrier protein. These results are likely to be of relevance for conformational peptide mimics in diagnostic tests, vaccine and inhibitors.
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Anticuerpos Monoclonales/inmunología , Subtipo H5N1 del Virus de la Influenza A/inmunología , Péptidos/química , Proteínas Recombinantes de Fusión/inmunología , Sitios de Unión de Anticuerpos , Subtipo H5N1 del Virus de la Influenza A/química , Subtipo H5N1 del Virus de la Influenza A/genética , Pruebas de Neutralización , Péptidos/genética , Péptidos/inmunología , Proteínas Recombinantes de Fusión/química , Proteínas Recombinantes de Fusión/genéticaRESUMEN
A point prevalence study of hepatitis E virus (HEV) in Chinese blood donors was conducted, and the prevalences of antibodies against HEV immunoglobulin G (IgG) and IgM among Chinese blood donors were 32.60% and 0.94%, respectively. HEV viremia was 0.07%.
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Virus de la Hepatitis E/genética , Virus de la Hepatitis E/aislamiento & purificación , Hepatitis E/epidemiología , Hepatitis E/virología , Adolescente , Adulto , Donantes de Sangre , China/epidemiología , Análisis por Conglomerados , Femenino , Anticuerpos Antihepatitis/sangre , Hepatitis E/inmunología , Virus de la Hepatitis E/inmunología , Humanos , Inmunoglobulina G/sangre , Masculino , Persona de Mediana Edad , Datos de Secuencia Molecular , Filogenia , ARN Viral/sangre , ARN Viral/genética , Análisis de Secuencia de ADN , Homología de Secuencia , Estudios Seroepidemiológicos , Adulto JovenRESUMEN
Hepatitis E virus (HEV), a non-enveloped, positive-stranded RNA virus, is transmitted in a faecal-oral manner, and causes acute liver diseases in humans. The HEV capsid is made up of capsomeres consisting of homodimers of a single structural capsid protein forming the virus shell. These dimers are believed to protrude from the viral surface and to interact with host cells to initiate infection. To date, no structural information is available for any of the HEV proteins. Here, we report for the first time the crystal structure of the HEV capsid protein domain E2s, a protruding domain, together with functional studies to illustrate that this domain forms a tight homodimer and that this dimerization is essential for HEV-host interactions. In addition, we also show that the neutralizing antibody recognition site of HEV is located on the E2s domain. Our study will aid in the development of vaccines and, subsequently, specific inhibitors for HEV.
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Proteínas de la Cápside/química , Virus de la Hepatitis E/química , Interacciones Huésped-Parásitos/fisiología , Multimerización de Proteína , Antígenos Virales/química , Antígenos Virales/genética , Antígenos Virales/inmunología , Secuencia de Bases , Western Blotting , Proteínas de la Cápside/genética , Proteínas de la Cápside/inmunología , Electroforesis en Gel de Poliacrilamida , Virus de la Hepatitis E/genética , Virus de la Hepatitis E/inmunología , Datos de Secuencia Molecular , Mutación , Estructura Cuaternaria de ProteínaRESUMEN
BACKGROUND: Passive immunization with human H5 antisera or H5-specific monoclonal antibodies (MAbs) has potential as an effective treatment for acute H5N1 influenza virus infection, but its efficacy against antigenically diverse H5N1 viruses is unconfirmed. METHODS: Cross-protection against antigenically diverse H5N1 strains with H5-specific MAbs, generated by successive immunization of antigenically distinct strains, was evaluated in mice. RESULTS: A panel of 52 broadly cross-reactive H5 specific MAbs were generated and characterized. One of these MAbs, 13D4, has been demonstrated to protect mice against lethal challenge by 4 H5N1 strains representing the current major genetic populations, clades 1, 2.1, 2.2, and 2.3, even at a stage of infection when H5N1 virus has disseminated beyond the pulmonary system. Complete neutralization of virus in lung tissue of infected animals was observed 24 h after treatment with 13D4. Mapping of this MAb with escape mutants showed it to bind to 2 conserved, possibly critical, sites of H5N1 hemagglutinin, 152 and 182. CONCLUSION: Generation of broadly cross-protective MAbs against H5N1 influenza virus may be optimized by selecting MAbs that target conserved sites in hemagglutinin. H5 MAbs such as 13D4 may prove to have therapeutic value in controlling infection due to current and future H5N1 variants.
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Anticuerpos Monoclonales/inmunología , Epítopos/inmunología , Subtipo H5N1 del Virus de la Influenza A/inmunología , Vacunas contra la Influenza/uso terapéutico , Gripe Aviar/inmunología , Gripe Humana/inmunología , Gripe Humana/patología , Animales , Aves/virología , Peso Corporal , Embrión de Pollo/virología , Secuencia Conservada , Reacciones Cruzadas , Pruebas de Inhibición de Hemaglutinación , Humanos , Subtipo H5N1 del Virus de la Influenza A/clasificación , Subtipo H5N1 del Virus de la Influenza A/genética , Subtipo H5N1 del Virus de la Influenza A/aislamiento & purificación , Gripe Humana/mortalidad , Ratones , Ratones Endogámicos BALB CRESUMEN
AIMS: To evaluate the specific T cell response together with IgM anti-hepatitis-E-virus (HEV) antibodies in acute hepatitis E (HE) patients. METHODS: Blood samples were collected from 11 HE patients every week and assayed for routine blood investigation after onset of disease until their convalescence. Peripheral blood mononuclear cells were separated from some of the blood samples (1-3 samples per patient) and tested for specific T cell response by enzyme-linked immunosorbent spot assay and IgM anti-hepatitis E virus by enzyme-linked immunosorbent assay. RESULTS: A particulate HEV capsid protein, HEV 239, effectively stimulated the response of T cells from HE patients infected by type 1 or type 4 HEV. In acute HE, a burst of HEV-specific cellular immune response occurred, which decreased along with the decreasing IgM anti-HEV antibody titre and normalization of liver function. CONCLUSIONS: HEV open reading frame 2 amino acids 368-606 can effectively stimulate the HEV-specific T cell response in vitro; the specific T cell response decreases along with convalescence and may play a role in the pathogenesis of acute HE and recovery.
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Anticuerpos Antivirales/sangre , Hepatitis E/inmunología , Inmunoglobulina M/sangre , Linfocitos T/inmunología , Adulto , Anciano , Animales , Células Cultivadas , Ensayo de Inmunoadsorción Enzimática , Femenino , Humanos , Hígado/fisiopatología , Pruebas de Función Hepática , Estudios Longitudinales , Masculino , Persona de Mediana EdadRESUMEN
Recent studies demonstrate that Th1-type immune responses against a broad spectrum of hepatitis C virus (HCV) gene products are crucial to the resolution of acute HCV infection. We investigated new vaccine approaches to augment the strength of HCV-specific Th1-type immune responses. ELISPOT assay revealed that single or multiple protein immunization using both CpG ODN and Montanide ISA 720 as adjuvants induced much stronger IFN-gamma-producing Th1 responses against core, NS3 and NS5b targets than did the formulation without these adjuvants. Protein vaccination using CpG ODN and Montanide ISA 720 as adjuvants also greatly enhanced humoral responses to HCV core, E1/E2 and NS3. When specific IgG isotypes were assayed, protein immunization using CpG ODN and Montanide ISA 720 as adjuvants produced higher titers of IgG2a dominant antibodies than did protein immunization alone, indicating a more Th1-biased pathway. This increase in IgG2a is consistent with the induction of Th1 cells secreting IFN-gamma demonstrated by ELISPOT assay. In conclusion, protein immunization using CpG ODN and Montanide ISA 720 as adjuvants greatly enhanced cellular (Th1 type) as well as humoral immune responses against HCV in Balb/c mice. The use of adjuvants appears critical to the induction of Th1 immune responses during HCV vaccination with recombinant proteins.
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Adyuvantes Inmunológicos/farmacología , Hepacivirus/inmunología , Antígenos de la Hepatitis C/inmunología , Manitol/análogos & derivados , Ácidos Oléicos/farmacología , Oligodesoxirribonucleótidos/farmacología , Células TH1/inmunología , Vacunas contra Hepatitis Viral/farmacología , Proteínas no Estructurales Virales/inmunología , Animales , Formación de Anticuerpos/inmunología , Ensayo de Inmunoadsorción Enzimática , Escherichia coli/metabolismo , Femenino , Anticuerpos Antihepatitis/análisis , Anticuerpos Antihepatitis/biosíntesis , Inmunidad Celular/inmunología , Factores Inmunológicos/análisis , Factores Inmunológicos/biosíntesis , Interferón gamma/biosíntesis , Interleucina-4/biosíntesis , Manitol/farmacología , Ratones , Ratones Endogámicos BALB C , Vacunas Sintéticas/biosíntesis , Vacunas Sintéticas/inmunología , Vacunas contra Hepatitis Viral/química , Vacunas contra Hepatitis Viral/inmunología , Proteínas no Estructurales Virales/biosíntesisRESUMEN
Double-stranded RNA is produced during virus replication and, together with the viral antigen, is responsible for inducing host antivirus immunity. The hepatitis C virus (HCV) non-structural protein-3 (NS3) has been implicated in the immune evasion of HCV, and is one of the prime targets for inducing immunity against HCV infection. Mice were immunized with recombinant NS3 protein (rNS3) and poly (I:C) emulsified in Montanide ISA 720 (M720). Cytokine production was assayed by enzyme-linked immunospot assay, and CD4(+) IFN-gamma(+) T helper (Th) cells or CD8(+) IFN-gamma(+) cytotoxic T lymphocytes were detected by flow cytometry. Anti-NS3 titre and immunoglobulin G2a (IgG2a) and IgG1 levels were monitored by enzyme-linked immunosorbent assay. Administration of rNS3 formulated in poly (I:C) and M720 induced anti-NS3 titres with a predominantly IgG2a isotype comparable to those induced by rNS3 in CpG-ODN and M720. The cytokine profiles showed that this formulation induced a Th1-biased immune response with several-fold more interferon-gamma (IFN-gamma)-producing cells than interleukin-4-producing cells. In contrast, rNS3 in M720 induced a Th2-biased immune response. The frequency of IFN-gamma-producing CD4(+) and CD8(+) cells induced by rNS3 in poly (I:C) and M720 was significantly higher than that induced by rNS3, rNS3 in M720, or rNS3 in poly (I:C), and was comparable to that induced by rNS3 in CpG-ODN with M720. The antigen-specific CD8(+) T-cell immune response persisted for up to 7 months after immunization. In conclusion, poly (I:C) with rNS3 in M720 can elicit a strong and persistent Th1-biased immune response and a cytotoxic T-lymphocyte response through cross-priming in mice. This study highlighted a promising formulation for inducing an efficient cellular immune response against HCV that has potential for HCV vaccine development.
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ARN Bicatenario/inmunología , Vacunas contra Hepatitis Viral/inmunología , Proteínas no Estructurales Virales/inmunología , Adyuvantes Inmunológicos , Animales , Anticuerpos Antivirales/biosíntesis , Relación Dosis-Respuesta Inmunológica , Ensayo de Inmunoadsorción Enzimática/métodos , Femenino , Inmunidad Celular , Activación de Linfocitos , Ratones , Ratones Endogámicos BALB C , Fragmentos de Péptidos/inmunología , Polinucleótidos/inmunología , Proteínas Recombinantes/inmunología , Linfocitos T Citotóxicos/inmunología , Linfocitos T Colaboradores-Inductores/inmunología , Vacunas Sintéticas/inmunologíaRESUMEN
BACKGROUND: A prophylactic vaccine for hepatitis C virus (HCV) requires generation of strong humoral as well as CD4(+) and CD8(+) T cell responses. METHODS: The immunomodulatory effects of the combination of 2 adjuvants, synthetic oligodeoxynucleotides containing immunostimulatory cytosine-guanine dinucleotide motifs emulsified with Montanide ISA720 (M-ISA720/CpG), were investigated using the murine model. RESULTS: Administration of recombinant HCV (rHCV) nonstructural (NS) 3 and NS5B proteins plus M-ISA720/CpG (hereafter, "M-ISA720/CpG/rHCV protein") induced high anti-NS3 and anti-NS5B immunoglobulin (Ig) G titers, with the IgG2a isotype being predominant. NS3- and NS5B-specific interferon (IFN)- gamma - and interleukin-2-producing CD4(+) T cell responses, as assessed by enzyme-linked immunospot assay, were significantly more vigorous in mice immunized with M-ISA720/CpG/rHCV protein than in control mice immunized without adjuvant. NS3- and NS5B-specific IFN- gamma -producing CD8(+) T cell percentages, as measured by direct ex vivo intracellular cytokine staining assay, were, respectively, a mean+/-SD of 0.14% +/- 0.04% and 0.15% +/- 0.05% in mice immunized with M-ISA720/CpG/rHCV protein. Furthermore, boosting with recombinant NS3 expression plasmid DNA after priming with M-ISA720/CpG-adjuvanted rNS3 strikingly enhanced both CD4(+) and CD8(+) T cell responses. CONCLUSION: Immunization with M-ISA720/CpG/rHCV protein is capable of inducing potent humoral as well as HCV-specific T helper type 1-biased CD4(+) and CD8(+) T cell responses. A DNA boost after a protein prime--a reversal of the conventional approach--may provide an alternative path to the development of an effective HCV vaccine.
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Linfocitos T CD4-Positivos/efectos de los fármacos , Linfocitos T CD8-positivos/efectos de los fármacos , Manitol/análogos & derivados , Ácidos Oléicos/administración & dosificación , Oligodesoxirribonucleótidos/administración & dosificación , Vacunas contra Hepatitis Viral/administración & dosificación , Adyuvantes Inmunológicos , Animales , Emparejamiento Base , Linfocitos T CD4-Positivos/inmunología , Linfocitos T CD8-positivos/inmunología , Fosfatos de Dinucleósidos/administración & dosificación , Fosfatos de Dinucleósidos/química , Hepacivirus/inmunología , Manitol/administración & dosificación , Ratones , Oligodesoxirribonucleótidos/química , Vacunación , Vacunas Sintéticas/administración & dosificación , Vacunas Sintéticas/inmunología , Vacunas contra Hepatitis Viral/inmunologíaRESUMEN
To investigate the quantitative characteristics of humoral immunity in patients with hepatitis C, we established an enzyme-linked immunosorbent spot (ELISpot) assay for detection of anti-hepatitis C virus (HCV)-secreting B cells. Receiver operating characteristic curve analysis demonstrated 100% specificity and 58% to 92% sensitivity for detecting B-cell responses to NS5b, NS3, E2, and core antigens. The median sum of anti-HCV-secreting B cells to all HCV antigens tested was significantly higher in 39 patients with chronic hepatitis C (47.3 spot forming cells [SFCs]/10(6) peripheral blood mononuclear cells [PBMCs]) than in 9 recovered subjects (15.3 SFCs/10(6) PBMCs; P = .05) or 11 uninfected controls (5.3 SFCs/10(6) PBMCs; P < .001); the significant difference (P = .018) in chronic versus recovered patients was in reactivity to nonstructural antigens NS3 and NS5b. Anti-HCV immunoglubulin M (IgM)-secreting B cells were also readily detected and persisted decades into HCV infection; there was no difference in IgM-positive cells between chronic and recovered patients. ELISpot reactivity to genotype 1-derived antigens was equivalent in patients of genotypes 1, 2, and 3. There was significant correlation between the numbers of anti-HCV IgG-secreting B cells and serum aminotransferase and to the level of circulating antibody. In conclusion, ELISpot assays can be adapted to study B-cell as well as T-cell responses to HCV. Measurement at the single-cell level suggests that humoral immunity plays a minor role in recovery from HCV infection and that B-cell immunity is strongest in those with persistent infection.
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Linfocitos B/inmunología , Anticuerpos contra la Hepatitis C/biosíntesis , Hepatitis C Crónica/inmunología , Adulto , Anciano , Anciano de 80 o más Años , Femenino , Genotipo , Hepacivirus/clasificación , Hepacivirus/genética , Humanos , Inmunoglobulina G/biosíntesis , Inmunoglobulina M/biosíntesis , Masculino , Persona de Mediana Edad , Curva ROCRESUMEN
The regression of cirrhosis is associated with increased intrahepatic collagenolytic enzyme activity. We investigated whether collagenase supplementation via portal vein infusion can retard cirrhosis development and/or reverse cirrhosis. In all, 35 rabbits were initially assigned to study. However, because of high surgical mortality and infection, only 15 animals completed study. Four normal controls (group I) received olive oil subcutaneously (SC) for 12 weeks followed by normal saline portal perfusion for 12 weeks. Four (group II) received CCl(4) SC for 6 weeks followed by portal vein collagenase, 6 mg twice weekly, plus SC CCl(4) for 6 additional weeks and then killed. Four rabbits (group III) received CCl(4) SC for 12 weeks and then 6 mg of collagenase portally for 12 weeks, while three control rabbits (group IV) received CCl(4) for 12 weeks followed by saline for 12 weeks. After 12 weeks of CCl(4), liver hydroxyproline content of collagenase-treated group II (361.1+/-106.6 microg/g) was significantly reduced compared with group III+IV that had not yet received collagenase (589.0+/-162.9 microg/g; P<0.05). In the main comparison, hydroxyproline content of collagenase-treated group III (177.5+/-35.6 microg/g) was significantly decreased compared with saline-treated controls (446.3+/-150.1 microg/g; P<0.01). Further, liver histology showed complete regression of cirrhosis in the collagenase-treated animals. No toxicity of liver, kidney, lung, brain or heart was observed histologically. Anaphylaxis occurred in 2/35 original animals (one fatal). In conclusion, this study provides 'proof of principle' that collagenase portal administration can retard cirrhosis development and speed regression of established cirrhosis in the rabbit CCl(4) model. Potential application to humans is premature, but feasible, if these findings are confirmed in additional animal studies.
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Colagenasas/uso terapéutico , Cirrosis Hepática/tratamiento farmacológico , Animales , Tetracloruro de Carbono/toxicidad , Colágeno/metabolismo , Colagenasas/administración & dosificación , Infusiones Intravenosas , Hígado/metabolismo , Cirrosis Hepática/inducido químicamente , Cirrosis Hepática/metabolismo , Masculino , Vena Porta , ConejosRESUMEN
Mice were immunized intramuscularly with free recombinant hepatitis C virus (HCV) NS3 (non-structural protein 3) protein, liposomes encapsulating rNS3 or rNS3 and CpG mixture, liposomes co-encapsulating rNS3 and CpG or liposomes co-encapsulating rNS3 and GpC. Liposomes co-encapsulating rNS3 and CpG induced a much higher titre of anti-HCV NS3 IgG and the dominant IgG subtype was IgG2a. Liposomes co-encapsulating rNS3 and GpC also induced high levels of anti-HCV NS3 IgG antibody, but the dominant IgG subtype was still IgG1, the same as in free HCV/NS3 immunized mice. Liposomes encapsulating rHCV NS3 and the mixture of rHCV NS3 and CpG did not increase the antibody response but switched the IgG subtype. A cytokine profile analysis revealed that the levels of Th1 cytokines in the mice immunized with liposomes co-encapsulating rHCV NS3 and CpG were significantly higher than in other mice while the levels of Th2 cytokine were significantly lower than in the mice immunized with naked rNS3. IL-12 in the mice immunized with liposome-NS3-CpG was significantly higher than in other mice. In conclusion, liposomes co-encapsulating HCV NS3 and CpG are a good candidate vaccine to induce strong Th1 immune responses against hepatitis C viruses.
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Hepacivirus/inmunología , Oligodesoxirribonucleótidos/administración & dosificación , Células TH1/inmunología , Vacunas Sintéticas/inmunología , Vacunas contra Hepatitis Viral/inmunología , Proteínas no Estructurales Virales/inmunología , Animales , Citocinas/biosíntesis , Femenino , Anticuerpos contra la Hepatitis C/sangre , Inmunización , Inmunoglobulina G/sangre , Inmunoglobulina G/clasificación , Liposomas , Ratones , Ratones Endogámicos BALB C , Proteínas no Estructurales Virales/administración & dosificaciónRESUMEN
SEN viruses (SENV) are newly discovered blood-borne single-stranded circular DNA viruses that may play a role in liver disease. To date, no serologic assays are available for the detection of SENV antigens or antibodies. We report on a rapid and sensitive molecular assay for the detection of four SENV strains (SENV-A, -C, -D, -H). This method uses PCR with universal primers and microwell capture hybridization with type-specific probes. Cut-off points to define "infected" based on chemiluminescence readings were determined from a statistical mixture model applied to samples from 300 injection drug users (IDUs) in San Francisco. Based on the estimated cut-off points, we examined the prevalence of SENV infection among 232 healthy US blood donors and assessed sensitivity and specificity of the assay in a small validation sample of infected individuals with partial sequence information.
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Infecciones por Circoviridae/epidemiología , Infecciones por Circoviridae/virología , Circoviridae/genética , Circoviridae/aislamiento & purificación , Secuencia de Bases , Infecciones por Circoviridae/complicaciones , ADN Viral/genética , ADN Viral/aislamiento & purificación , Humanos , Modelos Biológicos , Modelos Estadísticos , Datos de Secuencia Molecular , Reacción en Cadena de la Polimerasa/métodos , Reacción en Cadena de la Polimerasa/estadística & datos numéricos , San Francisco/epidemiología , Sensibilidad y Especificidad , Homología de Secuencia de Ácido Nucleico , Abuso de Sustancias por Vía Intravenosa/complicaciones , Estados Unidos/epidemiologíaRESUMEN
SEN virus (SENV) has been tentatively linked to transfusion-associated non-A-E hepatitis. We investigated SENV's role in unexplained hepatitis in other settings. Polymerase chain reaction amplification was used to detect 2 SENV variants (SENV-D and SENV-H) in 1706 patients and control subjects. SENV was detected in 54 (22%) of 248 patients with acute or chronic non-A-E hepatitis, 9 (35%) of 26 patients with hepatitis-associated aplastic anemia, and 0 of 17 patients with cryptogenic acute liver failure, compared with 150 (24%) of 621 control subjects with liver disease and 76 (10%) of 794 healthy control subjects. When controlling for geographic region, the prevalence of SENV among case and control subjects was not significantly different. The severity of acute or chronic hepatitis A, B, or C was not influenced by coexisting SENV infection. No etiological role for SENV in the cause of cryptogenic hepatitis could be demonstrated.
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Anemia Aplásica/virología , Infecciones por Circoviridae/virología , Circoviridae/crecimiento & desarrollo , Hepatitis Crónica/virología , Fallo Hepático Agudo/virología , Adolescente , Adulto , Anciano , Anemia Aplásica/epidemiología , Niño , Circoviridae/genética , Infecciones por Circoviridae/epidemiología , ADN Viral/química , ADN Viral/genética , Femenino , Alemania/epidemiología , Grecia/epidemiología , Virus de Hepatitis/crecimiento & desarrollo , Hepatitis Crónica/epidemiología , Humanos , Japón/epidemiología , Fallo Hepático Agudo/epidemiología , Masculino , Persona de Mediana Edad , Reacción en Cadena de la Polimerasa , Prevalencia , Estados Unidos/epidemiologíaRESUMEN
SEN viruses (SENVs) are newly discovered bloodborne viruses that may play a role in liver disease. SENV strain prevalence was examined in a race/ethnicity-stratified sample of 531 injection drug users (IDUs) from the San Francisco Bay area. Weighted prevalences were as follows: SENV-A, 45.7%; SENV-C/H, 35.6%; and SENV-D, 10.3%. Infection was associated with a longer duration of injection drug use. SENV-A was more common in black subjects (adjusted odds ratio [OR(a)], 4.37; 95% confidence interval [CI], 2.65-7.21) and Hispanic subjects (OR(a), 2.30; 95% CI, 1.38-3.85) than in white and non-Hispanic subjects, and the pattern was similar for SENV-C/H. For SENV-D, prevalence was similar in black and white subjects, but lower in Hispanic subjects; infection was less common among women than men (OR(a), 0.32; 95% CI, 0.15-0.71) and more common among men with at least 1 recent male sex partner than among heterosexual men (OR(a), 7.05; 95% CI, 2.62-18.95). SENV strains are common among San Francisco Bay area IDUs, and prevalence varies demographically within this group.