Expression and immunological evaluation of the <i>Escherichia coli</i> ‐derived hepatitis C virus envelope E1 protein

Lorenzo, Lazaro; García, Odalys Águila; Acosta‐Rivero, Nelson; Dueñas‐Carrera, Santiago; Martínez, Gillian; Alvarez-Obregón, Julio César; Pichardo, Dagmara; Ramos, Alberto; Guerra, Ignacio Ruiz; Morales, Juan

doi:10.1042/ba20000040

Cited by 17 publications

(13 citation statements)

References 28 publications

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“…For immunoblotting the samples were applied to a nitrocellulose membrane and binding of antibodies was detected as previously described (12). In brief, the membrane was blocked for 1 h at room temperature with phosphate-buffered saline solution (PBS) (0.1 M NaCl, 2 mM KCl, 10 mM Na 2 HPO 4 , 1 mM KH 2 PO 4 pH 7.4) containing 5% skim milk, then incubated either with mAb SS-HepC.1 or human serum for 1 h at 37°C.…”

Section: Antibodiesmentioning

confidence: 99%

Characterization of the HCV Core Virus-like Particles Produced in the Methylotrophic Yeast Pichia pastoris

Acosta‐Rivero

Aguilar

Musacchio

et al. 2001

Biochemical and Biophysical Research Communications

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Section: Antibodiesmentioning

confidence: 99%

Characterization of the HCV Core Virus-like Particles Produced in the Methylotrophic Yeast Pichia pastoris

Acosta‐Rivero

Aguilar

Musacchio

et al. 2001

Biochemical and Biophysical Research Communications

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“…HCV protein variants Co.120 (Co), E1.340, and NS3 are HCV Core, E1, and NS3 protein variants encompassing amino acids 1 to 120, 192 to 340, and 1192 to 1457 of the HCV polyprotein from a genotype 1b Cuban isolate, respectively. These proteins were obtained from recombinant Escherichia coli cells and purified by a combination of washed pellet procedures and gel-filtration chromatography to 95% purity as described previously (25)(26)(27). E2.680 is an HCV E2 protein variant encompassing amino acids 384 to 680 of the viral polyprotein, obtained from recombinant Pichia pastoris cells.…”

Section: Methodsmentioning

confidence: 99%

Protective T Cell and Antibody Immune Responses against Hepatitis C Virus Achieved Using a Biopolyester-Bead-Based Vaccine Delivery System

Martínez‐Donato

Piniella

Aguilar

et al. 2016

Clin Vaccine Immunol

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Hepatitis C virus (HCV) infection is a major worldwide problem. Chronic hepatitis C is recognized as one of the major causes of cirrhosis, hepatocellular carcinoma, and liver failure. Although new, directly acting antiviral therapies are suggested to overcome the low efficacy and adverse effects observed for the current standard of treatment, an effective vaccine would be the only way to certainly eradicate HCV infection. Recently, polyhydroxybutyrate beads produced by engineered Escherichia coli showed efficacy as a vaccine delivery system. Here, an endotoxin-free E. coli strain ( Hepatitis C virus (HCV) is an etiologic agent of chronic hepatitis C (1). Chronic HCV infection affects more than 170 million people worldwide and is responsible for approximately 350,000 deaths each year (2). Viral exposure results in acute disease in a small proportion of cases, while the majority (80%) progress to chronic infection, causing liver inflammation that slowly progresses to cirrhosis, liver failure, hepatocellular carcinoma, and death (3).Despite a substantial decline in HCV transmission due to improved prevention strategies and the introduction of new and powerful targeted therapies, hepatitis C remains a huge health problem, justifying further endeavors to develop new vaccines. Indeed, the pool of asymptomatic chronic HCV carriers who represent an infectious reservoir will remain substantial for many years. Less than 30% of patients with chronic hepatitis C are aware of the infection, and only about 10% of patients are currently treated (4, 5). Therefore, even if new antivirals could cure 90% of patients, there would still be a considerable percentage of patients who would be excluded (6). Hence, development of a vaccine to prevent infection or to at least prevent progression to chronicity represents a significant unmet medical need and is of high priority.Since 1% of infected patients show an immune response clearing the infection and the rate of spontaneous resolution is higher in the case of reinfected patients, this demonstrates that the induction of protective immunity which prevents development of chronic disease is a feasible goal for the development of a preventive vaccine against HCV (7,8).The role of HCV-specific T cell responses in the outcome of primary HCV infection has been widely studied, although a single correlate of protection has not been determined. However, it is known that this type of immune response is a determinant in the clearance of the virus. Comparative studies in humans and chimpanzees have shown that widespread and long-lasting CD8 ϩ and CD4 ϩ T cell responses against multiple HCV regions are linked to spontaneous viral clearance (9, 10).However, there is also strong evidence that rapid induction of high-titer cross-neutralizing antibodies targeting HCV envelope proteins correlates with viral clearance and protects against reinfection (11, 12). Therefore, an optimal HCV vaccine probably needs to elicit broad cross-reactive cellular immune responses together with cross-neutralizing antibo...

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“…Recombinant Co.120 [18], E1.340 [19] and E2C [20] proteins were obtained from Escherichia coli with more than 85% purity by a combination of washed‐pellet procedures and gel‐filtration chromatography. Co.120 comprises the first 120 amino acids of the HCV core protein, whereas E1.340 and E2C encompass amino acids 192–340 and 384–605 in the viral polyprotein respectively.…”

Section: Methodsmentioning

confidence: 99%

Immunization with a recombinant fowlpox virus expressing a hepatitis C virus core–E1 polyprotein variant, protects mice and African green monkeys (Chlorocebus aethiops sabaeus) against challenge with a surrogate vaccinia virus

Alvarez‐Lajonchere

Amador-Cañizares

Frias

et al. 2008

Biotech and App Biochem

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HCV (hepatitis C virus) is a worldwide health problem nowadays. No preventive vaccine is available against this pathogen, and therapeutic treatments currently in use have important drawbacks, including limited efficacy. In the present work a recombinant fowlpox virus, FPCoE1, expressing a truncated HCV core-E1 polyprotein, was generated. FPCoE1 virus generally failed to elicit a humoral immune response against HCV antigens in BALB/c mice. By contrast, mice inoculated with FPCoE1 elicited a positive interferon-gamma secretion response against HCV core in ex-vivo ELISPOT (enzyme-linked immunospot) assays. Remarkably, mice inoculated with FPCoE1 significantly controlled viraemia in a surrogate challenge model with vvRE, a recombinant vaccinia virus expressing HCV structural antigens. In fact, 40% of the mice had no detectable levels of vvRE in their ovaries. Administration of FPCoE1 in vervet monkeys [Chlorocebus (formerly Cercophitecus) aethiops sabaeus] induced lymphoproliferative response against HCV core and E1 proteins in 50% of immunized animals. Monkeys immunized with FPCoE1 had no detectable levels of vvRE in their blood, whereas monkeys inoculated with FP9, the negative control virus, had detectable levels of vvRE in blood up to 7 days after challenge. In conclusion, recombinant fowlpox virus FPCoE1 is able to induce an anti-HCV immune response in mice and monkeys. This ability could be rationally employed to develop effective strategies against HCV infection by using FPCoE1 in combination with other vaccine candidates or antiviral treatments.

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Expression and immunological evaluation of the Escherichia coli ‐derived hepatitis C virus envelope E1 protein

Cited by 17 publications

References 28 publications

Characterization of the HCV Core Virus-like Particles Produced in the Methylotrophic Yeast Pichia pastoris

Characterization of the HCV Core Virus-like Particles Produced in the Methylotrophic Yeast Pichia pastoris

Protective T Cell and Antibody Immune Responses against Hepatitis C Virus Achieved Using a Biopolyester-Bead-Based Vaccine Delivery System

Immunization with a recombinant fowlpox virus expressing a hepatitis C virus core–E1 polyprotein variant, protects mice and African green monkeys (Chlorocebus aethiops sabaeus) against challenge with a surrogate vaccinia virus

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