Viral O‐GalNAc peptide epitopes: a novel potential target in viral envelope glycoproteins

Olofsson, Sigvard; Blixt, Ola; Bergström, Tomas; Frank, Martin; Wandall, Hans H.

doi:10.1002/rmv.1859

Cited by 17 publications

(24 citation statements)

References 80 publications

(120 reference statements)

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“…In a similar way, O-glycans have also been suggested to shield immunodominant epitopes in herpesviruses (91,92). Nevertheless, there is limited information regarding the impact of individual O-glycans on shielding underlying peptide epitopes or the capacity of host immunity to present and recognize glycosylated antigens (93,94 (18), mouse immune sera were able to detect and neutralize virus produced in mammalian cells, suggesting either that the epitope recognition is not affected by adjacent glycosylation or that putative O-linked glycans only partly occupy and protect the epitopes. We also found O-glycans located within a neutralizing peptide epitope at the N terminus of HCMV gH (S31) (119) and a discontinuous neutralizing antibody epitope on VZV gH (Ser 177 and Thr 184 ) (120).…”

Section: Discussionmentioning

confidence: 99%

“…We have recently developed a quantitative differential O-glycoproteomic strategy to address non-redundant contributions of individual GalNAc transferase isoforms to the O-glycosylation capacity of a cell (90), and this could be applied to address changes in viral O-glycosylation between clinical isolates or samples propagated in different cell types. Because O-glycans may affect immunity by shielding protein epitopes or introducing glycopeptide epitopes (91)(92)(93), it is important to consider O-glycans in the context of vaccine design. It is also important to consider O-glycans for innate immune targeting ligands to augment immunity (33,94).…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Global Mapping of O-Glycosylation of Varicella Zoster Virus, Human Cytomegalovirus, and Epstein-Barr Virus

Bagdonaite

Nordén

Joshi

et al. 2016

Journal of Biological Chemistry

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Herpesviruses are among the most complex and widespread viruses, infection and propagation of which depend on envelope proteins. These proteins serve as mediators of cell entry as well as modulators of the immune response and are attractive vaccine targets. Although envelope proteins are known to carry glycans, little is known about the distribution, nature, and functions of these modifications. This is particularly true for O-glycans; thus we have recently developed a "bottom up" mass spectrometry-based technique for mapping O-glycosylation sites on herpes simplex virus type 1. We found wide distribution of O-glycans on herpes simplex virus type 1 glycoproteins and demonstrated that elongated O-glycans were essential for the propagation of the virus. Here, we applied our proteome-wide discovery platform for mapping O-glycosites on representative and clinically significant members of the herpesvirus family: varicella zoster virus, human cytomegalovirus, and Epstein-Barr virus. We identified a large number of O-glycosites distributed on most envelope proteins in all viruses and further demonstrated conserved patterns of O-glycans on distinct homologous proteins. Because glycosylation is highly dependent on the host cell, we tested varicella zoster virus-infected cell lysates and clinically isolated virus and found evidence of consistent O-glycosites. These results present a comprehensive view of herpesvirus O-glycosylation and point to the widespread occurrence of O-glycans in regions of envelope proteins important for virus entry, formation, and recognition by the host immune system. This knowledge enables dissection of specific functional roles of individual glycosites and, moreover, provides a framework for design of glycoprotein vaccines with representative glycosylation.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Global Mapping of O-Glycosylation of Varicella Zoster Virus, Human Cytomegalovirus, and Epstein-Barr Virus

Bagdonaite

Nordén

Joshi

et al. 2016

Journal of Biological Chemistry

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“…This strategy was based on the following considerations: (i) The two most efficient subunit vaccines to enveloped viruses, i.e., VZV gE against HZ and HBsAg against HBV infection, are both based on viral glycoproteins containing MLDs decorated by numerous short O-linked glycans [15,16,24,25]. (ii) Short O-linked glycans, together with their surrounding oligopeptide environment, constitute immunoreactive epitopes in viral glycoproteins with MLD [21,34]. (iii) In contrast to N-linked glycans being added by one single enzyme species, strongly homologous over the entire animal kingdom, the initiation of O-linked glycans is carried out by twenty different isotypes of GalNAc transferases, differing in target specificity [27,35].…”

Section: Discussionmentioning

confidence: 99%

“…Owing to viral interference with O-glycan elongation, much of these carbohydrate components of viral MLDs are composed of shorter glycans, including O-linked GalNAc monosaccharides (Tn antigen) and GalGalNAc disaccharides (T antigen) [16,19,20]. Recently, we established that such truncated O-linked glycans, in combination with the proximate peptide sequence at the glycosylation site, forms a novel variant of viral B cell epitope to which prominent antibody responses are raised in the infected patient [21,22,23]. Importantly, it was shown that a single GalNAc unit and each specific amino acid of the glycopeptide stretch contributed equally well to the B cell epitope specificity and avidity towards the cognate antibody [22,23].…”

Section: Introductionmentioning

confidence: 99%

Recombinant Glycoprotein E of Varicella Zoster Virus Contains Glycan-Peptide Motifs That Modulate B Cell Epitopes into Discrete Immunological Signatures

Nordén

Nilsson

Samuelsson

et al. 2019

IJMS

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A recombinant subunit vaccine (Shingrix®) was recently licensed for use against herpes zoster. This vaccine is based on glycoprotein E (gE) of varicella zoster virus (VZV), the most abundantly expressed protein of VZV, harboring sites for N- and O-linked glycosylation. The subunit vaccine elicits stronger virus-specific CD4+ T cell response as well as antibody B cell response to gE, compared to the currently used live attenuated vaccine (Zostavax®). This situation is at variance with the current notion since a live vaccine, causing an active virus infection, should be far more efficient than a subunit vaccine based on only one single viral glycoprotein. We previously found gE to be heavily glycosylated, not least by numerous clustered O-linked glycans, when it was produced in human fibroblasts. However, in contrast to Zostavax®, which is produced in fibroblasts, the recombinant gE of Shingrix® is expressed in Chinese hamster ovary (CHO) cells. Hence, the glycan occupancy and glycan structures of gE may differ considerably between the two vaccine types. Here, we aimed at (i) defining the glycan structures and positions of recombinant gE and (ii) identifying possible features of the recombinant gE O-glycosylation pattern contributing to the vaccine efficacy of Shingrix®. Firstly, recombinant gE produced in CHO cells (“Shingrix situation”) is more scarcely decorated by O-linked glycans than gE from human fibroblasts (“Zostavax situation”), with respect to glycan site occupancy. Secondly, screening of immunodominant B cell epitopes of gE, using a synthetic peptide library against serum samples from VZV-seropositive individuals, revealed that the O-linked glycan signature promoted binding of IgG antibodies via a decreased number of interfering O-linked glycans, but also via specific O-linked glycans enhancing antibody binding. These findings may, in part, explain the higher protective efficacy of Shingrix®, and can also be of relevance for development of subunit vaccines to other enveloped viruses.

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“…Furthermore, viral glycoproteins can be used as targets for vaccines and antiviral therapies (41)(42)(43)(44).…”

Section: Viral Protein Glycosylationmentioning

confidence: 99%

Structural characterisation of glycosylation of paramyxovirus attachment and fusion proteins by mass spectrometry

Pegg¹

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The family Paramyxoviridae (paramyxovirus) contains several significant human and animal pathogens.Represented within this family are human respiratory syncytial virus (hRSV) and Newcastle disease virus (NDV). The former contributes significantly to severe respiratory tract disease in infants, children and immunocompromised individuals. At present, highly efficacious therapeutics or safe and effective vaccines are not available for hRSV. NDV is the causative agent of Newcastle disease, afflicting a wide range of avian species. The desire to study NDV is due not only to the significant economic impact it has on the poultry industry worldwide, but also its potential use as an oncolytic agent and vaccine vector in humans and animals. Additionally, findings on NDV may be translated to closely related viruses that cause disease in humans, such as parainfluenza viruses.As outlined in Chapter 1 of this thesis, the infectious processes of all members of this family are driven by two major membrane glycoproteins whose ectodomains project from the viral envelope: the fusion (F) and attachment glycoproteins. The F glycoprotein is responsible for viral entry by means of fusion with host cell membranes while the variable attachment glycoproteins, haemagglutinin, haemagglutininneuraminidase (HN) and major surface glycoprotein (G), are involved in viral attachment to host cells.Previous studies have shown that altering the glycosylation profile of these proteins can modulate the ability of the virus to infect host cells and stimulate the host immune system. As yet, site-specific glycan heterogeneity of hRSV and NDV surface glycoproteins has not been defined at a chemical level. As described in Chapter 2, reverse-phase liquid chromatography tandem mass spectrometry (MS) strategies were implemented to characterise intact glycopeptides from the attachment and F glycoproteins of hRSV and NDV. Site-occupancy and monosaccharide compositions at a given site were determined using collision-induced dissociation, higher-energy collision dissociation, electron-transfer dissociation and electron-transfer dissociation combined with collision dissociation. As described in Chapter 3, a spectral processing program was developed called OxoExtract to aid identification of intact glycopeptides that were fragmented with higher-energy collision dissociation.The F and HN proteins of NDV were derived from virions propagated in embryonic eggs. Analyses of HN in Chapter 4 revealed high mannose N-linked glycans and complex or hybrid N-linked glycans that were variably fucosylated, sialylated and sulfated or phosphorylated. In total 63, 58, and 37 glycans were identified at sites N341, N433 and N481, respectively. In addition, a previously undocumented Olinked glycosylation site was identified in the stalk domain of the protein. Observed glycans from NDV F described in Chapter 5 were mainly high mannose, containing variations of five to nine mannose ii residues across four sites, N85, N191, N366 and N471. There was also evidence of fucosylated c...

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Viral O‐GalNAc peptide epitopes: a novel potential target in viral envelope glycoproteins

Cited by 17 publications

References 80 publications

Global Mapping of O-Glycosylation of Varicella Zoster Virus, Human Cytomegalovirus, and Epstein-Barr Virus

Global Mapping of O-Glycosylation of Varicella Zoster Virus, Human Cytomegalovirus, and Epstein-Barr Virus

Recombinant Glycoprotein E of Varicella Zoster Virus Contains Glycan-Peptide Motifs That Modulate B Cell Epitopes into Discrete Immunological Signatures

Structural characterisation of glycosylation of paramyxovirus attachment and fusion proteins by mass spectrometry

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