title: HIV-1 Glycan Density Drives the Persistence of the Mannose Patch within an Infected Individual.
abstract: The HIV envelope glycoprotein (Env) is extensively modified with host-derived N-linked glycans. The high density of glycosylation on the viral spike limits enzymatic processing, resulting in numerous underprocessed oligomannose-type glycans. This extensive glycosylation not only shields conserved regions of the protein from the immune system but also acts as a target for anti-HIV broadly neutralizing antibodies (bnAbs). In response to the host immune system, the HIV glycan shield is constantly evolving through mutations affecting both the positions and numbers of potential N-linked glycosylation sites (PNGSs). Here, using longitudinal Env sequences from a clade C-infected individual (CAP256), we measured the impact of the shifting glycan shield during HIV infection on the abundance of oligomannose-type glycans. By analyzing the intrinsic mannose patch from a panel of recombinant CAP256 gp120s displaying high protein sequence variability and changes in PNGS number and positioning, we show that the intrinsic mannose patch persists throughout the course of HIV infection and correlates with the number of PNGSs. This effect of the glycan density on the processing state was also supported by the analysis of a cross-clade panel of recombinant gp120 glycoproteins. Together, these observations underscore the importance of glycan clustering for the generation of carbohydrate epitopes for anti-HIV bnAbs. The persistence of the intrinsic mannose patch over the course of HIV infection further highlights this epitope as an important target for HIV vaccine strategies.
  
  
  OBJECTIVE
  Development of an HIV vaccine is critical for control of the HIV pandemic, and elicitation of broadly neutralizing antibodies (bnAbs) is likely to be a key component of a successful vaccine response. The HIV envelope glycoprotein (Env) is covered in an array of host-derived N-linked glycans often referred to as the glycan shield. This glycan shield is a target for many of the recently isolated anti-HIV bnAbs and is therefore under constant pressure from the host immune system, leading to changes in both glycan site frequency and location. This study aimed to determine whether these genetic changes impacted the eventual processing of glycans on the HIV Env and the susceptibility of the virus to neutralization. We show that despite this variation in glycan site positioning and frequency over the course of HIV infection, the mannose patch is a conserved feature throughout, making it a stable target for HIV vaccine design.
authors.affiliation: Department of Infectious Diseases, Faculty of Life Sciences and Medicine, King's College London, London, United Kingdom.
authors.forename: Karen P
authors.initials: KP
authors.lastname: Coss
authors.affiliation: Oxford Glycobiology Institute, Department of Biochemistry, University of Oxford, Oxford, United Kingdom.
authors.forename: Snezana
authors.initials: S
authors.lastname: Vasiljevic
authors.affiliation: Oxford Glycobiology Institute, Department of Biochemistry, University of Oxford, Oxford, United Kingdom.
authors.forename: Laura K
authors.initials: LK
authors.lastname: Pritchard
authors.affiliation: Department of Infectious Diseases, Faculty of Life Sciences and Medicine, King's College London, London, United Kingdom.
authors.forename: Stefanie A
authors.initials: SA
authors.lastname: Krumm
authors.affiliation: Oxford Glycobiology Institute, Department of Biochemistry, University of Oxford, Oxford, United Kingdom.
authors.forename: Molly
authors.initials: M
authors.lastname: Glaze
authors.affiliation: Department of Virology, University of the Witwatersrand, Johannesburg, South Africa.|National Institute for Communicable Diseases (NICD) of the National Health Laboratory Service (NHLS), Johannesburg, South Africa.|Centre for the AIDS Programme of Research in South Africa (CAPRISA), University of KwaZulu Natal, Durban, South Africa.
authors.forename: Sharon
authors.initials: S
authors.lastname: Madzorera
authors.affiliation: Department of Virology, University of the Witwatersrand, Johannesburg, South Africa.|National Institute for Communicable Diseases (NICD) of the National Health Laboratory Service (NHLS), Johannesburg, South Africa.|Centre for the AIDS Programme of Research in South Africa (CAPRISA), University of KwaZulu Natal, Durban, South Africa.
authors.forename: Penny L
authors.initials: PL
authors.lastname: Moore
authors.affiliation: Oxford Glycobiology Institute, Department of Biochemistry, University of Oxford, Oxford, United Kingdom max.crispin@bioch.ox.ac.uk katie.doores@kcl.ac.uk.
authors.forename: Max
authors.initials: M
authors.lastname: Crispin
authors.affiliation: Department of Infectious Diseases, Faculty of Life Sciences and Medicine, King's College London, London, United Kingdom max.crispin@bioch.ox.ac.uk katie.doores@kcl.ac.uk.
authors.forename: Katie J
authors.initials: KJ
authors.lastname: Doores
mesh_terms: D057134:Antibodies, Neutralizing / Q000096:biosynthesis* / Q000737:chemistry; D002236:Carbohydrate Conformation; D003001:Cloning, Molecular; D018604:Epitope Mapping; D000939:Epitopes / Q000737:chemistry / Q000276:immunology; D015967:Gene Expression Regulation, Viral; D006031:Glycosylation; D057809:HEK293 Cells; D015483:HIV Antibodies / Q000096:biosynthesis* / Q000737:chemistry; D015699:HIV Envelope Protein gp120 / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D015658:HIV Infections / Q000821:virology; D015497:HIV-1 / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D054884:Host-Pathogen Interactions; D006801:Humans; D008358:Mannose / Q000737:chemistry / Q000276:immunology*; D000072417:Protein Domains; D011499:Protein Processing, Post-Translational; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology
chemical_list: D057134:Antibodies, Neutralizing; D000939:Epitopes; D015483:HIV Antibodies; D015699:HIV Envelope Protein gp120; D011994:Recombinant Proteins; D008358:Mannose