Polyprenyl phosphates induce a high humoral and cellular response to immunization with recombinant proteins of the replicative complex of the hepatitis C virus

2018 ◽  
Vol 482 (4) ◽  
pp. 459-462 ◽  
Author(s):  
O. Masalova ◽  
◽  
E. Lesnova ◽  
A. Onishchuk ◽  
A. Ivanova ◽  
...  
Keyword(s):  
Hepatitis C Virus ◽  
Hepatitis C ◽  
Recombinant Proteins ◽  
Cellular Response ◽  
Polyprenyl Phosphates ◽  
Humoral And Cellular Response

scholarly journals NOD1 Participates in the Innate Immune Response Triggered by Hepatitis C Virus Polymerase

2016 ◽  
Vol 90 (13) ◽  
pp. 6022-6035 ◽  
Author(s):  
Serena Vegna ◽  
Damien Gregoire ◽  
Marie Moreau ◽  
Patrice Lassus ◽  
David Durantel ◽  
...  
Keyword(s):  
Pattern Recognition ◽  
Innate Immunity ◽  
Hepatitis C Virus ◽  
Hepatitis C ◽  
Inflammatory Response ◽  
Cellular Response ◽  
Pattern Recognition Receptors ◽  
Type I ◽  
Mitochondrial Antiviral Signaling

ABSTRACTHepatitis C virus (HCV) triggers innate immunity signaling in the infected cell. Replication of the viral genome is dispensable for this phenotype, and we along with others have recently shown that NS5B, the viral RNA-dependent RNA polymerase, synthesizes double-stranded RNA (dsRNA) from cellular templates, thus eliciting an inflammatory response, notably via activation of type I interferon and lymphotoxin β. Here, we investigated intracellular signal transduction pathways involved in this process. Using HepaRG cells, a model that largely recapitulates thein vivocomplexities of the innate immunity receptor signaling, we have confirmed that NS5B triggered increased expression of the canonical pattern recognition receptors (PRRs) specific for dsRNA, namely, RIG-I, MDA5, and Toll-like receptor 3 (TLR3). Unexpectedly, intracellular dsRNA also led to accumulation of NOD1, a receptor classically involved in recognition of bacterial peptidoglycans. NOD1 activation, confirmed by analysis of its downstream targets, was likely due to its interaction with dsRNA and was independent of RIG-I and mitochondrial antiviral signaling protein (MAVS/IPS-1/Cardif/VISA) signaling. It is likely to have a functional significance in the cellular response in the context of HCV infection since interference with the NOD1 pathway severely reduced the inflammatory response elicited by NS5B.IMPORTANCEIn this study, we show that NOD1, a PRR that normally senses bacterial peptidoglycans, is activated by HCV viral polymerase, probably through an interaction with dsRNA, suggesting that NOD1 acts as an RNA ligand recognition receptor. In consequence, interference with NOD1-mediated signaling significantly weakens the inflammatory response to dsRNA. These results add a new level of complexity to the understanding of the cross talk between different classes of pattern recognition receptors and may be related to certain complications of chronic hepatitis C virus infection.

scholarly journals The Investigation of the Prospects for Using NS4A Antigen of Hepatitis C Virus for the Development of Recombinant Mosaic Vaccines with Self-Adjuvant Properties

2017 ◽  
Vol 16 (1) ◽  
pp. 69-74
Author(s):  
V. V. Koupriyanov ◽  
L. I. Nikolaeva ◽  
A. A. Zykova ◽  
P. I. Makhnovskiy
Keyword(s):  
Hepatitis C Virus ◽  
Hepatitis C ◽  
Genetic Engineering ◽  
Recombinant Protein ◽  
Recombinant Proteins ◽  
Interleukin 2 ◽  
High Expression ◽  
Subsequent Work ◽  
Atomic Force ◽  
High Expression Level

The aim of this study was to design promising variants of recombinant proteins based on NS4A antigen of hepatitis C virus (HCV) for subsequent work on the creation of a mosaic recombinant vaccine against hepatitis C. Methods. The recombinant proteins, containing different fragments of NS4A (belong to HCV subtype 1b) and murine interleukin-2, were prepared by genetic engineering approaches, using vectors pQE30 and pQE60 for E. coli. The size of the recombinant protein particles were evaluated by atomic force microscopy. Immunogenicity of these recombinant proteins was tested for Balb/c mice. The murine sera were analyzed by enzyme immunoassay. The recombinant proteins were also tested by immunoblotting with human sera specific to HCV antigens. Results. Six variants of recombinant genetic engineering constructions based on NS4A antigen of hepatitis C virus were designed. In the first variant amino acid sequence of NS4A was inserted using vector pQE60 into the immunodominant loop of HBc protein (core protein of hepatitis B virus). However, further analysis of the product showed the absence of virus-like particles in it. The following three constructs (with glycine linker 19s), without it and N-truncated NS4A) were done using vector pQE30. Only N-truncated NS4а product had a high expression level. Then new protein, consisted of NS4A and N-truncated murine interleukin-2 (IL-2), was obtained to enhance immunogenicity. It is known that IL-2 has adjuvant property. The new product (NS4a-IL-2) is well expressed, but it is accumulated in inclusion bodies. It was extracted with 7M guanidine chloride, purified on a Ni-sorbent and dialyzed in PBS. A shortened version of NS4A (ANS4a-IL-2) was also obtained with a high expression level. Taking in account that increasing the repetition of antigenic regions in recombinant constructs can enhance their immunogenicity, we obtained a recombinant protein comprising three repeat of NS4A. But its efficiency of expression was low. The construction NS4a had very poor immunogenicity, but NS4a-IL-2 (which contains the full length NS4A) displayed the best one for Balb/c mice. As it was shown earlier the immunogenicity of the protein preparation is dependent on the presence of aggregates, so we investigated our recombinant proteins for the presence of protein aggregates by atomic force microscopy.The presence of the particles with size of 6 - 8 nm was revealed in solution of NS4a-IL-2. Conclution. Only ANS4a-IL-2 and ANS4a-IL-2 of the six constructs had high expression and antigenic properties. And only NS4a-IL-2 possessed the high immunogenic property. So, this construction can be used for subsequent work on the creation of a mosaic recombinant vaccine against hepatitis C.

IV. Pathogenetic mechanisms involved in hepatitis C virus-induced liver diseases

1998 ◽  
Vol 275 (6) ◽  
pp. G1217-G1220 ◽  
Author(s):  
Johnson Yiu-Nam Lau
Keyword(s):  
Hepatitis C Virus ◽  
Hepatitis C ◽  
Liver Damage ◽  
Cellular Response ◽  
High Dose ◽  
Acute Hepatitis C ◽  
Pathogenetic Mechanisms ◽  
Transplant Patients ◽  
Hepatocellular Damage ◽  
High Level

The pathogenetic mechanisms for liver damage in acute hepatitis C are not clear, but a host immune cellular response may be involved. In chronic hepatitis C, there is strong evidence that host cellular immune response is involved in the control of viral replication and contributes to hepatocellular damage. As hepatitis C virus infection persists, continuous liver damage and regeneration, together with enhanced fibrogenesis, may eventually lead to cirrhosis in a proportion of patients. Transplant patients on high-dose immunosuppression may have high-level intrahepatic hepatitis C viral expression, and, in this setting, the virus may induce direct cytopathic liver damage.

scholarly journals Enhancing B- and T-Cell Immune Response to a Hepatitis C Virus E2 DNA Vaccine by Intramuscular Electrical Gene Transfer

2000 ◽  
Vol 74 (24) ◽  
pp. 11598-11607 ◽  
Author(s):  
Silvia Zucchelli ◽  
Stefania Capone ◽  
Elena Fattori ◽  
Antonella Folgori ◽  
Annalise Di Marco ◽  
...  
Keyword(s):  
T Cells ◽  
Electrical Stimulation ◽  
Hepatitis C Virus ◽  
Hepatitis C ◽  
Gene Transfer ◽  
Cellular Response ◽  
Low Voltage ◽  
Mouse Tissue ◽  
Cell Immune Response ◽  
Electric Pulses

ABSTRACT We describe an improved genetic immunization strategy for eliciting a full spectrum of anti-hepatitis C virus (HCV) envelope 2 (E2) glycoprotein responses in mammals through electrical gene transfer (EGT) of plasmid DNA into muscle fibers. Intramuscular injection of a plasmid encoding a cross-reactive hypervariable region 1 (HVR1) peptide mimic fused at the N terminus of the E2 ectodomain, followed by electrical stimulation treatment in the form of high-frequency, low-voltage electric pulses, induced more than 10-fold-higher expression levels in the transfected mouse tissue. As a result of this substantial increment of in vivo antigen production, the humoral response induced in mice, rats, and rabbits ranged from 10- to 30-fold higher than that induced by conventional naked DNA immunization. Consequently, immune sera from EGT-treated mice displayed a broader cross-reactivity against HVR1 variants from natural isolates than sera from injected animals that were not subjected to electrical stimulation. Cellular response against E2 epitopes specific for helper and cytotoxic T cells was significantly improved by EGT. The EGT-mediated enhancement of humoral and cellular immunity is antigen independent, since comparable increases in antibody response against ciliary neurotrophic factor or in specific anti-human immunodeficiency virus type 1 gag CD8+ T cells were obtained in rats and mice. Thus, the method described potentially provides a safe, low-cost treatment that may be scaled up to humans and may hold the key for future development of prophylactic or therapeutic vaccines against HCV and other infectious diseases.

Virus-Specific Cellular Response in Hepatitis C Virus Infection

2015 ◽  
Vol 64 (2) ◽  
pp. 101-110 ◽  
Author(s):  
Justyna Kaźmierczak ◽  
Kamila Caraballo Cortes ◽  
Iwona Bukowska-Ośko ◽  
Marek Radkowski
Keyword(s):  
Hepatitis C Virus ◽  
Hepatitis C ◽  
Virus Infection ◽  
Cellular Response ◽  
Hepatitis C Virus Infection

scholarly journals Induction of Hepatitis C Virus E1 Envelope Protein-Specific Immune Response Can Be Enhanced by Mutation of N-Glycosylation Sites

2001 ◽  
Vol 75 (24) ◽  
pp. 12088-12097 ◽  
Author(s):  
A. Fournillier ◽  
C. Wychowski ◽  
D. Boucreux ◽  
T. F. Baumert ◽  
J.-C. Meunier ◽  
...  
Keyword(s):  
Hepatitis C Virus ◽  
Hepatitis C ◽  
Cellular Response ◽  
Humoral Response ◽  
Glycosylation Site ◽  
Immune Recognition ◽  
E1 Protein ◽  
Glycosylation Sites ◽  
Antibody Titers

ABSTRACT Deglycosylation of viral glycoproteins has been shown to influence the number of available epitopes and to modulate immune recognition of antigens. We investigated the role played by N-glycans in the immunogenicity of hepatitis C virus (HCV) E1 envelope glycoprotein, a naturally poor immunogen. Eight plasmids were engineered, encoding E1 protein mutants in which the four N-linked glycosylation sites of the protein were mutated separately or in combination. In vitro expression studies showed an influence of N-linked glycosylation on expression efficiency, instability, and/or secretion of the mutated proteins. Immunogenicity of the E1 mutants was studied in BALB/c mice following intramuscular and intraepidermal injection of the plasmids. Whereas some mutations had no or only minor effects on the antibody titers induced, mutation of the fourth glycosylation site (N4) significantly enhanced the anti-E1 humoral response in terms of both seroconversion rates and antibody titers. Moreover, antibody induced by the N4 mutant was able to recognize HCV-like particles with higher titers than those induced by the wild-type construct. Epitope mapping indicated that the E1 mutant antigens induced antibody directed at two major domains: one, located at amino acids (aa) 313 to 332, which is known to be reactive with sera from HCV patients, and a second one, located in the N-terminal domain of E1 (aa 192 to 226). Analysis of the induced immune cellular response confirmed the induction of gamma interferon-producing cells by all mutants, albeit to different levels. These results show that N-linked glycosylation can limit the antibody response to the HCV E1 protein and reveal a potential vaccine candidate with enhanced immunogenicity.

Sign in / Sign up

Export Citation Format

Share Document