scholarly journals Modification of Asparagine-Linked Glycan Density for the Design of Hepatitis B Virus Virus-Like Particles with Enhanced Immunogenicity

2015 ◽  
Vol 89 (22) ◽  
pp. 11312-11322 ◽  
Author(s):  
Michiko Hyakumura ◽  
Renae Walsh ◽  
Morten Thaysen-Andersen ◽  
Natalie J. Kingston ◽  
Mylinh La ◽  
...  
Keyword(s):  
Hepatitis B Virus ◽  
Hepatitis B ◽  
Immune Responses ◽  
Envelope Proteins ◽  
Wild Type ◽  
Virus Envelope ◽  
Virus Like Particles ◽  
Loop Region ◽  
Glycosylation Sites ◽  
B Virus

ABSTRACTThe small envelope proteins (HBsAgS) derived from hepatitis B virus (HBV) represent the antigenic components of the HBV vaccine and are platforms for the delivery of foreign antigenic sequences. To investigate structure-immunogenicity relationships for the design of improved immunization vectors, we have generated biochemically modified virus-like particles (VLPs) exhibiting glycoengineered HBsAgS. For the generation of hypoglycosylated VLPs, the wild-type (WT) HBsAgS N146 glycosylation site was converted to N146Q; for constructing hyperglycosylated VLPs, potential glycosylation sites were introduced in the HBsAgS external loop region at positions T116 and G130 in addition to the WT site. The introduced T116N and G130N sites were utilized as glycosylation anchors resulting in the formation of hyperglycosylated VLPs. Mass spectroscopic analyses showed that the hyperglycosylated VLPs carry the same types of glycans as WT VLPs, with minor variations regarding the degree of fucosylation, bisectingN-acetylglucosamines, and sialylation. Antigenic fingerprints for the WT and hypo- and hyperglycosylated VLPs using a panel of 19 anti-HBsAgS monoclonal antibodies revealed that 15 antibodies retained their ability to bind to the different VLP glyco-analogues, suggesting that the additionalN-glycans did not shield extensively for the HBsAgS-specific antigenicity. Immunization studies with the different VLPs showed a strong correlation betweenN-glycan abundance and antibody titers. The T116N VLPs induced earlier and longer-lasting antibody responses than did the hypoglycosylated and WT VLPs. The ability of nonnative VLPs to promote immune responses possibly due to differences in their glycosylation-related interaction with cells of the innate immune system illustrates pathways for the design of immunogens for superior preventive applications.IMPORTANCEThe use of biochemically modified, nonnative immunogens represents an attractive strategy for the generation of modulated or enhanced immune responses possibly due to differences in their interaction with immune cells. We have generated virus-like particles (VLPs) composed of hepatitis B virus envelope proteins (HBsAgS) with additionalN-glycosylation sites. Hyperglycosylated VLPs were synthesized and characterized, and the results demonstrated that they carry the same types of glycans as wild-type VLPs. Comparative immunization studies demonstrated that the VLPs with the highestN-glycan density induce earlier and longer-lasting antibody immune responses than do wild-type or hypoglycosylated VLPs, possibly allowing reduced numbers of vaccine injections. The ability to modulate the immunogenicity of an immunogen will provide opportunities to develop optimized vaccines and VLP delivery platforms for foreign antigenic sequences, possibly in synergy with the use of suitable adjuvanting compounds.

scholarly journals Analysis of the Cytosolic Domains of the Hepatitis B Virus Envelope Proteins for Their Function in Viral Particle Assembly and Infectivity

2006 ◽  
Vol 80 (24) ◽  
pp. 11935-11945 ◽  
Author(s):  
Matthieu Blanchet ◽  
Camille Sureau
Keyword(s):  
Hepatitis B Virus ◽  
Hepatitis B ◽  
Self Assembly ◽  
Inner Core ◽  
Envelope Proteins ◽  
Endoplasmic Reticulum Membrane ◽  
Virus Envelope ◽  
Particle Assembly ◽  
B Virus

ABSTRACT The hepatitis B virus (HBV) envelope proteins have the ability to assemble three types of viral particles, (i) the empty subviral particles (SVPs), (ii) the mature HBV virions, and (iii) the hepatitis delta virus (HDV) particles, in cells that are coinfected with HBV and HDV. To gain insight into the function of the HBV envelope proteins in morphogenesis of HBV or HDV virions, we have investigated subdomains of the envelope proteins that have been shown or predicted to lie at the cytosolic face of the endoplasmic reticulum membrane during synthesis, a position prone to interaction with the inner core structure. These domains, referred to here as cytosolic loops I and II (CYL-I and -II, respectively), were subjected to mutagenesis. The mutations were introduced in the three HBV envelope proteins, designated small, middle, and large (S-HBsAg, M-HBsAg, and L-HBsAg, respectively). The mutants were expressed in HuH-7 cells to evaluate their capacity for self-assembly and formation of HBV or HDV virions when HBV nucleocapsid or HDV ribonucleoprotein, respectively, was provided. We found that SVP-competent CYL-I mutations between positions 23 and 78 of the S domain were permissive to HBV or HDV virion assembly. One mutation (P29A) was permissive for synthesis of the S- and M-HBsAg but adversely affected the synthesis or stability of L-HBsAg, thereby preventing the assembly of HBV virions. Furthermore, using an in vitro infection assay based on the HepaRG cells and the HDV model, we have shown that particles coated with envelope proteins bearing CYL-I mutations were fully infectious, hence indicating the absence of an infectivity determinant in this region. Finally, we demonstrated that the tryptophan residues at positions 196, 199, and 201 in CYL-II, which were shown to exert a matrix function for assembly of HDV particles (I. Komla-Soukha and C. Sureau, J. Virol. 80:4648-4655, 2006), were dispensable for both assembly and infectivity of HBV virions.

scholarly journals Inducing oral immune regulation of hepatitis B virus envelope proteins suppresses the growth of hepatocellular carcinoma in mice

Cancer ◽  
2002 ◽  
Vol 94 (2) ◽  
pp. 406-414 ◽  
Author(s):  
Israel Gotsman ◽  
Ruslana Alper ◽  
Athalia Klein ◽  
Elazar Rabbani ◽  
Dean Engelhardt ◽  
...  
Keyword(s):  
Hepatocellular Carcinoma ◽  
Hepatitis B Virus ◽  
Hepatitis B ◽  
Immune Regulation ◽  
Envelope Proteins ◽  
Virus Envelope ◽  
B Virus

scholarly journals T Cells Expressing a Chimeric Antigen Receptor That Binds Hepatitis B Virus Envelope Proteins Control Virus Replication in Mice

2013 ◽  
Vol 145 (2) ◽  
pp. 456-465 ◽  
Author(s):  
Karin Krebs ◽  
Nina Böttinger ◽  
Li–Rung Huang ◽  
Markus Chmielewski ◽  
Silke Arzberger ◽  
...  
Keyword(s):  
T Cells ◽  
Hepatitis B Virus ◽  
Hepatitis B ◽  
Virus Replication ◽  
Chimeric Antigen Receptor ◽  
Antigen Receptor ◽  
Envelope Proteins ◽  
Virus Envelope ◽  
B Virus ◽  
Control Virus

scholarly journals Role of N Glycosylation of Hepatitis B Virus Envelope Proteins in Morphogenesis and Infectivity of Hepatitis Delta Virus

2003 ◽  
Vol 77 (9) ◽  
pp. 5519-5523 ◽  
Author(s):  
Camille Sureau ◽  
Chantal Fournier-Wirth ◽  
Patrick Maurel
Keyword(s):  
Hepatitis B Virus ◽  
Hepatitis B ◽  
Hepatitis Delta Virus ◽  
Envelope Proteins ◽  
Virus Envelope ◽  
Hepatitis Delta ◽  
B Virus ◽  
L Proteins ◽  
Delta Virus

ABSTRACT Hepatitis delta virus (HDV) particles are coated with the large (L), middle (M), and small (S) hepatitis B virus envelope proteins. In the present study, we constructed glycosylation-defective envelope protein mutants and evaluated their capacity to assist in the maturation of infectious HDV in vitro. We observed that the removal of N-linked carbohydrates on the S, M, and L proteins was tolerated for the assembly of subviral hepatitis B virus (HBV) particles but was partially inhibitory for the formation of HDV virions. However, when assayed on primary cultures of human hepatocytes, virions coated with S, M, and L proteins lacking N-linked glycans were infectious. Furthermore, in the absence of M, HDV particles coated with nonglycosylated S and L proteins retained infectivity. These results indicate that carbohydrates on the HBV envelope proteins are not essential for the in vitro infectivity of HDV.

scholarly journals Distinctive properties of the hepatitis B virus envelope proteins.

1988 ◽  
Vol 62 (2) ◽  
pp. 407-416 ◽  
Author(s):  
K L Molnar-Kimber ◽  
V Jarocki-Witek ◽  
S K Dheer ◽  
S K Vernon ◽  
A J Conley ◽  
...  
Keyword(s):  
Hepatitis B Virus ◽  
Hepatitis B ◽  
Envelope Proteins ◽  
Virus Envelope ◽  
B Virus

scholarly journals Exogenous hepatitis B virus envelope proteins induce endoplasmic reticulum stress: involvement of cannabinoid axis in liver cancer cells

Oncotarget ◽  
2016 ◽  
Vol 7 (15) ◽  
pp. 20312-20323 ◽  
Author(s):  
Roberta Montalbano ◽  
Birgit Honrath ◽  
Thaddeus Till Wissniowski ◽  
Moritz Elxnat ◽  
Silvia Roth ◽  
...  
Keyword(s):  
Endoplasmic Reticulum ◽  
Hepatitis B Virus ◽  
Liver Cancer ◽  
Hepatitis B ◽  
Endoplasmic Reticulum Stress ◽  
Cancer Cells ◽  
Envelope Proteins ◽  
Virus Envelope ◽  
Liver Cancer Cells ◽  
B Virus

scholarly journals Immunological Analysis of the Hepatitis B Virus “a” Determinant Displayed on Chimeric Virus-Like Particles of Macrobrachium rosenbergii Nodavirus Capsid Protein Produced in Sf9 Cells

Vaccines ◽  
2020 ◽  
Vol 8 (2) ◽  
pp. 275
Author(s):  
Nathaniel Nyakaat Ninyio ◽  
Kok Lian Ho ◽  
Hui Kian Ong ◽  
Chean Yeah Yong ◽  
Hui Yee Chee ◽  
...  
Keyword(s):  
Hepatitis B Virus ◽  
Hepatitis B ◽  
Immune Responses ◽  
Natural Killer ◽  
Capsid Protein ◽  
Macrobrachium Rosenbergii ◽  
Hepatitis B Vaccine ◽  
Chimeric Virus ◽  
Virus Like Particles ◽  
B Virus

Chimeric virus-like particles (VLPs) have been widely exploited for various purposes including their use as vaccine candidates, particularly due to their ability to induce stronger immune responses than VLPs consisting of single viral proteins. In the present study, VLPs of the Macrobrachium rosenbergii nodavirus (MrNV) capsid protein (Nc) displaying the hepatitis B virus “a” determinant (aD) were produced in Spodoptera frugiperda (Sf9) insect cells. BALB/c mice immunised with the purified chimeric Nc-aD VLPs elicited a sustained titre of anti-aD antibody, which was significantly higher than that elicited by a commercially available hepatitis B vaccine and Escherichia coli-produced Nc-aD VLPs. Immunophenotyping showed that the Sf9-produced Nc-aD VLPs induced proliferation of cytotoxic T-lymphocytes and NK1.1 natural killer cells. Furthermore, enzyme-linked immunospot (ELISPOT)analysis showed the presence of antibody-secreting memory B cells in the mice splenocytes stimulated with the synthetic aD peptide. The significant humoral, natural killer cell and memory B cell immune responses induced by the Sf9-produced Nc-aD VLPs suggest that they present good prospects for use as a hepatitis B vaccine candidate.

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