Expression-System-Dependent Modulation of HIV-1 Envelope Glycoprotein Antigenicity and Immunogenicity

被引：65

作者：

Kong, Leopold ^{[1
,2
]}

Sheppard, Neil C. ^{[1
]}

Stewart-Jones, Guillaume B. E. ^{[3
]}

Robson, Cynthia L. ^{[1
]}

Chen, Hongying ^{[4
]}

Xu, Xiaodong ^{[4
]}

Krashias, George ^{[1
]}

Bonomelli, Camille ^{[5
]}

Scanlan, Christopher N. ^{[5
]}

Kwong, Peter D. ^{[2
]}

Jeffs, Simon A. ^{[6
]}

Jones, Ian M. ^{[4
]}

Sattentau, Quentin J. ^{[1
]}

机构：

[1] Univ Oxford, Sir William Dunn Sch Pathol, Oxford OX1 3RE, England

[2] NIAID, Vaccine Res Ctr, NIH, Bethesda, MD 20892 USA

[3] Univ Oxford, Wellcome Trust Ctr Human Genet, Oxford OX3 7BN, England

[4] Univ Reading, Sch Biol Sci, Reading RG6 6UR, Berks, England

[5] Univ Oxford, Dept Biochem, Glycobiol Inst, Oxford OX1 3QU, England

[6] Univ London Imperial Coll Sci Technol & Med, Div Med, Wright Fleming Inst, London W2 1PG, England

来源：

JOURNAL OF MOLECULAR BIOLOGY | 2010年 / 403卷 / 01期

基金：

英国医学研究理事会;

关键词：

computational modeling; HIV-1; gp120; mass spectrometry; B-cell immunogenicity; glycan type; HUMAN-IMMUNODEFICIENCY-VIRUS; SITE-SPECIFIC ANALYSIS; INFECTED INSECT CELLS; HAMSTER OVARY CELLS; COMPLEX N-GLYCANS; MONOCLONAL-ANTIBODIES; HTLV-III; GLYCOSYLATION SITES; LINKED GLYCANS; NMR SYSTEM;

D O I：

10.1016/j.jmb.2010.08.033

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Recombinant expression systems differ in the type of glycosylation they impart on expressed antigens such as the human immunodeficiency virus type 1 (HIV-1) envelope glycoproteins, potentially affecting their biological properties. We performed head-to-head antigenic, immunogenic and molecular profiling of two distantly related Env surface (gp120) antigens produced in different systems: (a) mammalian (293 FreeStyle (TM) cells; 293F) cells in the presence of kifunensine, which impart only high-mannose glycans; (b) insect cells (Spodoptera frugiperda, Sf9), which confer mainly paucimannosidic glycans; (c) Sf9 cells recombinant for mammalian glycosylation enzymes (Sf9 Mimic (TM)), which impart high-mannose, hybrid and complex glycans without sialic acid; and (d) 293F cells, which impart high-mannose, hybrid and complex glycans with sialic acid. Molecular models revealed a significant difference in gp120 glycan coverage between the Sf9-derived and wild-type mammalian-cell-derived material that is predicted to affect ligand binding sites proximal to glycans. Modeling of solvent-exposed surface electrostatic potentials showed that sialic acid imparts a significant negative surface charge that may influence gp120 antigenicity and immunogenicity. Gp120 expressed in systems that do not incorporate sialic acid displayed increased ligand binding to the CD4 binding and CD4-induced sites compared to those expressed in the system that do, and imparted other more subtle differences in antigenicity in a gp120 subtype-specific manner. Non-sialic-acid-containing gp120 was significantly more immunogenic than the sialylated version when administered in two different adjuvants, and induced higher titers of antibodies competing for CD4 binding site ligand-gp120 interaction These findings suggest that non-sialic-acid-imparting systems yield gp120 immunogens with modified antigenic and immunogenic properties, considerations that should be considered when selecting expression systems for glycosylated antigens to be used for structure-function studies and for vaccine use. (C) 2010 Elsevier Ltd All rights reserved

引用

页码：131 / 147

页数：17