Glycan-Dependent Immunogenicity of Recombinant Soluble Trimeric Hemagglutinin

Vries, Robert P. de; Smit, Cornelis H.; Bruin, Erwin de; Rigter, Alan; Vries, Erik de; Cornelissen, Lisette A. H. M.; Eggink, Dirk; Chung, Nancy P. Y.; Moore, John P.; Sanders, Rogier W.; Hokke, Cornelis H.; Rottier, Peter J. M.; Haan, Cornelis A. M. de

doi:10.1128/jvi.01084-12

Cited by 63 publications

(85 citation statements)

References 37 publications

(68 reference statements)

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“…10). However, these results conflict with one report of mannose-terminated rH5HA protein N-glycans (but not complex-type N-glycans) suppressing CpG-induced IFN-␣ release in pDCs via binding with BDCA-2, a C-type lectin receptor (36). It has also been reported that the mannose moieties of HIV-1 gp120 glycoproteins induced immunosuppressive hIL-10 expression in human mDCs via the DC-SIGN of C-type lectin receptors (37).…”

Section: Discussioncontrasting

confidence: 53%

Influenza Virus Hemagglutinin Glycoproteins with Different N-Glycan Patterns Activate Dendritic Cells In Vitro

Liu

Lin

Spearman

et al. 2016

J Virol

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Influenza virus hemagglutinin (HA) N-glycans play important regulatory roles in the control of virus virulence, antigenicity, receptor-binding specificity, and viral escape from the immune response. Considered essential for controlling innate and adaptive immune responses against influenza virus infections, dendritic cells (DCs) trigger proinflammatory and adaptive immune responses in hosts. In this study, we engineered Chinese hamster ovary (CHO) cell lines expressing recombinant HA from pandemic H1, H5, and H7 influenza viruses. rH1HA, rH5HA, and rH7HA were obtained as wild-type proteins or in the presence of kifunensine (KIF) or further with endo-␤-N-acetylglucosaminidase-treated KIF (KIF؉E) to generate single-N-acetylglucosamine (GlcNAc) N-glycans consisting of (i) terminally sialylated complex-type N-glycans, (ii) high-mannose-type N-glycans, and (iii) single-GlcNAc-type N-glycans. Our results show that high-mannose-type and single-GlcNAc-type N-glycans, but not complex-type N-glycans, are capable of inducing more active hIL12 p40, hIL12 p70, and hIL-10 production in human DCs. Significantly higher HLA-DR, CD40, CD83, and CD86 expression levels, as well reduced endocytotic capacity in human DCs, were noted in the high-mannose-type rH1HA and single-GlcNAc-type rH1HA groups than in the complex-type N-glycan rH1HA group. Our data indicate that native avian rHA proteins (H5N1 and H7N9) are more immunostimulatory than human rHA protein (pH1N1). The high-mannose-type or single-GlcNAc-type N-glycans of both avian and human HA types are more stimulatory than the complex-type N-glycans. HA-stimulated DC activation was accomplished partially through a mannose receptor(s). These results provide more understanding of the contribution of glycosylation of viral proteins to the immune responses and may have implications for vaccine development. IMPORTANCEInfluenza viruses trigger seasonal epidemics or pandemics with mild-to-severe consequences for human and poultry populations. DCs are the most potent professional antigen-presenting cells, which play a crucial role in the link between innate and adaptive immunity. In this study, we obtained stable-expression CHO cells to produce rH1HA, rH5HA, and rH7HA proteins containing distinct N-glycan patterns. These rHA proteins, each with a distinct N-glycan pattern, were used to investigate interactions with mouse and human DCs. Our data indicate that native avian rHA proteins (H5N1 and H7N9) are more immunostimulatory than human rHA protein (pH1N1). High-mannose-type and single-GlcNAc-type N-glycans were more effective than complex-type N-glycans in triggering mouse and human DC activation and maturation. We believe these results provide some useful information for influenza vaccine development regarding how influenza virus HA proteins with different types of N-glycans activate DCs.

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Section: Discussioncontrasting

confidence: 53%

Influenza Virus Hemagglutinin Glycoproteins with Different N-Glycan Patterns Activate Dendritic Cells In Vitro

Liu

Lin

Spearman

et al. 2016

J Virol

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“…4 Recent studies demonstrated that rHA engineered to form a stable trimeric configuration elicited a protective immune response in vaccinated animals. [5][6][7][8][9][10] Neutralizing antibody levels and protection from disease were enhanced in mice vaccinated with soluble rHA trimers as compared to animals vaccinated with rHA monomers, 5,8 indicating the importance of immunogens that mimic those expressed by the infectious agent. Along these lines, the composition and/or number of N-linked glycans on rHA trimers have been shown to modify the level of the protective antibody response.…”

Section: Ross Et Almentioning

confidence: 98%

“…Along these lines, the composition and/or number of N-linked glycans on rHA trimers have been shown to modify the level of the protective antibody response. 10,11 All of these studies indicate that rHA trimers could be an important component of a subunit vaccine strategy against influenza.…”

Section: Ross Et Almentioning

confidence: 99%

Hemagglutinin-based polyanhydride nanovaccines against H5N1 influenza elicit protective virus neutralizing titers and cell-mediated immunity

Ross¹,

Loyd²,

Wu³

et al. 2014

IJN

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H5N1 avian influenza is a significant global concern with the potential to become the next pandemic threat. Recombinant subunit vaccines are an attractive alternative for pandemic vaccines compared to traditional vaccine technologies. In particular, polyanhydride nanoparticles encapsulating subunit proteins have been shown to enhance humoral and cell-mediated immunity and provide protection upon lethal challenge. In this work, a recombinant H5 hemagglutinin trimer (H5 3 ) was produced and encapsulated into polyanhydride nanoparticles. The studies performed indicated that the recombinant H5 3 antigen was a robust immunogen. Immunizing mice with H5 3 encapsulated into polyanhydride nanoparticles induced high neutralizing antibody titers and enhanced CD4 + T cell recall responses in mice. Finally, the H5 3 -based polyanhydride nanovaccine induced protective immunity against a low-pathogenic H5N1 viral challenge. Informatics analyses indicated that mice receiving the nanovaccine formulations and subsequently challenged with virus were similar to naïve mice that were not challenged. The current studies provide a basis to further exploit the advantages of polyanhydride nanovaccines in pandemic scenarios.

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“…1C). However, because of limitations of the resolution of standard SDS-PAGE gels and expected variation in glycosylation site usage and glycan chain heterogeneity (25)(26)(27), it is not possible to determine whether each of the newly created glycosylation sites is efficiently glycosylated.…”

mentioning

confidence: 99%

“…A mammalian expression system was chosen because mammalian cells utilize large complex glycans, in contrast to other expression systems like insect cells (25). Naive 6-to 8-week-old BALB/c mice were vaccinated intramuscularly with 10 g of protein adjuvanted with 5 g of poly(I·C) (Invivogen) three times in 3-to 4-week intervals.…”

mentioning

confidence: 99%

Guiding the Immune Response against Influenza Virus Hemagglutinin toward the Conserved Stalk Domain by Hyperglycosylation of the Globular Head Domain

Eggink

Goff

Palese

2014

J Virol

Self Cite

162

137

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Influenza virus hemagglutinin consists of a highly variable and immunodominant head domain and a more conserved but immunosubdominant stalk domain. We introduced seven N-linked glycosylation sites in the hemagglutinin head domain to shield the immunodominant antigenic sites. The hyperglycosylated hemagglutinin enhanced stalk-directed seroreactivity while dampening the head response in immunized mice. Upon influenza virus challenge, mice vaccinated with the hyperglycosylated hemagglutinin were better protected against morbidity and mortality than mice receiving the wild-type hemagglutinin.

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Glycan-Dependent Immunogenicity of Recombinant Soluble Trimeric Hemagglutinin

Cited by 63 publications

References 37 publications

Influenza Virus Hemagglutinin Glycoproteins with Different N-Glycan Patterns Activate Dendritic Cells In Vitro

Influenza Virus Hemagglutinin Glycoproteins with Different N-Glycan Patterns Activate Dendritic Cells In Vitro

Hemagglutinin-based polyanhydride nanovaccines against H5N1 influenza elicit protective virus neutralizing titers and cell-mediated immunity

Guiding the Immune Response against Influenza Virus Hemagglutinin toward the Conserved Stalk Domain by Hyperglycosylation of the Globular Head Domain

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