Identification of an N-linked glycan in the V1-loop of HIV-1 gp120 influencing neutralization by anti-V3 antibodies and soluble CD4

Gram, Gregers J.; Hemming, Anna; Bolmstedt, Anders; Jansson, Bjarne; Olofsson, Sigvard; Åkerblom, Lennart; Nielsen, Jens; Hansen, Jan Erik

doi:10.1007/bf01310789

Cited by 46 publications

(45 citation statements)

References 21 publications

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“…Plasmid DNA was isolated (Qiagen) and transfected into the CD4 + cell line H9 using a modified DEAE-dextran method as described elsewhere (Gram et al, 1994). Supernatants were harvested at the peak of antigen production, filtered (0-45 ~tm), divided into aliquots and stored at -8 0 °C until use.…”

Section: Nf-a-920smentioning

confidence: 99%

Rapid selection for an N-linked oligosaccharide by monoclonal antibodies directed against the V3 loop of human immunodeficiency virus type 1

Schønning

Jansson

Olofsson

et al. 1996

Journal of General Virology

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The V3 loop of the human immunodeficiency virus (HIV) surface protein, gpl20, constitutes a principal neutralizing determinant. HIV strains lacking a naturally conserved N-linked oligosaccharide (at position 306) within the V3 loop are highly sensitive to neutralization. We subjected molecular clones of HIVLA ~ lacking this 3°6N-glycan to in vitro immune selection with MAbs directed against the V3 loop. In all, ten clones were characterized, and all proved resistant to V3-directed neutralization. Sequencing of the V3 loop revealed that six of the clones had become resistant at least partly by reacquisition of the 3°6N-glycan. Only two of the clones possessed mutations within the binding site of the antibody itself, while the two remaining clones did not display changes within the V3 loop itself. Thus, HIV strains lacking the ~°6N-glycan primarily develop resistance to V3-directed neutralization through acquisition of the specific oligosaccharide. This demonstrates that protein glycosylation can be a primary modifier of virus antigenicity of possible importance for the interaction of HIV with the host immune response.

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Section: Nf-a-920smentioning

confidence: 99%

Rapid selection for an N-linked oligosaccharide by monoclonal antibodies directed against the V3 loop of human immunodeficiency virus type 1

Schønning

Jansson

Olofsson

et al. 1996

Journal of General Virology

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“…Just as the acquisition of particular N-linked sites decreases neutralization sensitivity, elimination of N-linked sites at the same or nearby locations has been shown to increase neutralization sensitivity within both HIV-1 and SIV strains, particularly in the V1/V2 and V3 loops (4,16,27,34,45,48,51). Less is known with regard to the effects of mutagenesis of glycosylation sites outside of the V1/V2 and V3 loops on neutralization sensitivity of SIV or HIV.…”

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confidence: 99%

“…Loss of particular glycans can also affect viral infectivity, possibly through structural alterations that influence the ability of the glycoprotein to bind its receptors, monomer interactions within the trimer, or interactions of the surface and transmembrane fusion proteins (31,42,55). However, many single and multiple glycosylation sites have been shown to be dispensable to viral replication within HIV-1 gp41 (11,23) and gp120 of both HIV-1 (4,16,25,26,34,51) and simian immunodeficiency virus (SIV) (39,47,48). The dispensability of some of these N-linked glycans to viral replication and the greater sensitivity of some mutants missing glycan attachment sites to antibodymediated neutralization (4,16,27,34,36,45,48,51) suggest that these glycans may also serve to shield the spike from recognition by antibodies.…”

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confidence: 99%

Identification of Two N-Linked Glycosylation Sites within the Core of the Simian Immunodeficiency Virus Glycoprotein Whose Removal Enhances Sensitivity to Soluble CD4

Pikora

Wittish

Desrosiers

2005

J Virol

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Using PCR mutagenesis to disrupt the NXT/S N-linked glycosylation motif of the Env protein, we created 27 mutants lacking 1 to 5 of 14 N-linked glycosylation sites within regions of gp120 lying outside of variable loops 1 to 4 within simian immunodeficiency virus strain 239 (SIV239). Of 18 mutants missing N-linked glycosylation sites predicted to lie within 10 Å of CD4 contact sites, the infectivity of 12 was sufficient to measure sensitivity to neutralization by soluble CD4 (sCD4), pooled immune sera from SIV239-infected rhesus macaques, and monoclonal antibodies known to neutralize certain derivatives of SIV239. Three of these 12 mutants (g3, lacking the 3rd glycan at position 79; g11, lacking the 11th glycan at position 212; and g3,11, lacking both the 3rd and 11th glycans) were approximately five times more sensitive to neutralization by sCD4 than wild-type (WT) SIV239. However, these same mutants were no more sensitive to neutralization than WT by pooled immune sera. The other 9 of 12 replication-competent mutants in this group were no more sensitive to neutralization than the WT by any of the neutralizing reagents. Six of the nine mutants that did not replicate appreciably had three or more glycosylation sites eliminated; the other three replication-deficient strains involved mutation of site 15. Our results suggest that elimination of glycan attachment sites 3 and 11 enhanced the exposure of contact residues for CD4. Thus, glycans at positions 3 and 11 of SIV239 gp120 may be particularly important for shielding the CD4-binding site from antibody recognition.

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“…Patterns of addition and relocation of N-linked glycosylation sites during the course of HIV and SIV infection suggest an evolving "glycan shield" in response to antibody selection (4,8,26,33,38). Just as the acquisition of particular N-linked sites decreases neutralization sensitivity, the elimination of N-linked sites at the same or nearby locations has been shown to increase neutralization sensitivity for both HIV-1 and SIV (5,9,10,12,13,16,21,31,33). Reitter et al previously demonstrated that a mutation of specific Nlinked glycosylation sites in the V1-V2 region of gp120 of SIVmac239 results in replication-competent viruses capable of eliciting increased levels of antibodies with neutralizing activity against the parental wild-type strain SIVmac239 (32,33).…”

mentioning

confidence: 99%

“…The dispensability of some N-linked glycans for viral replication and the greater sensitivity of some glycan-deficient mutants to antibody-mediated neutralization suggest that these glycans may serve in part as barriers to shield the virus from effective antibody recognition (5,10,12,13,15,16,21,23,31,32,36). Variations in the number and location of glycosylation sites, particularly within the V1/V2 and V3 loops but also on the "silent face" of gp120, often correlate with altered sensitivity to neutralizing antibodies (1,6,11,21,22,34).…”

mentioning

confidence: 99%

Glycosylation of gp41 of Simian Immunodeficiency Virus Shields Epitopes That Can Be Targets for Neutralizing Antibodies

Yuste

Bixby

Lifson

et al. 2008

J Virol

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Human immunodeficiency virus type 1 and simian immunodeficiency virus possess three closely spaced, highly conserved sites for N-linked carbohydrate attachment in the extracellular domain of the transmembrane protein gp41. We infected rhesus monkeys with a variant of cloned SIVmac239 lacking the second and third sites or with a variant strain lacking all three of SIVmac239's glycosylation sites in gp41. For each mutation, asparagine (N) in the canonical N-X-S/T recognition sequence for carbohydrate attachment was changed to the structurally similar glutamine such that two nucleotide changes would be required for a reversion of the mutated codon. By 16 weeks, experimentally infected monkeys made antibodies that neutralized the mutant viruses to high titers. Such antibodies were not observed in monkeys infected with the parental virus. Thus, new specificities were revealed as a result of the carbohydrate attachment mutations, and antibodies of these specificities had neutralizing activity. Unlike monkeys infected with the parental virus, monkeys infected with the mutant viruses made antibodies that reacted with peptides corresponding to the sequences in this region. Furthermore, there was strong selective pressure for the emergence of variant sequences in this region during the course of infection. By analyzing the neutralization profiles of sequence variants, we were able to define three mutations (Q625R, K631N, and Q634H) in the region of the glycosylation site mutations that conferred resistance to neutralization by plasma from the monkeys infected with mutant virus. Based on the reactivity of antibodies to peptides in this region and the colocalization of neutralization escape mutations, we conclude that N-linked carbohydrates in the ectodomain of the transmembrane protein shield underlying epitopes that would otherwise be the direct targets of neutralizing antibodies.

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Identification of an N-linked glycan in the V1-loop of HIV-1 gp120 influencing neutralization by anti-V3 antibodies and soluble CD4

Cited by 46 publications

References 21 publications

Rapid selection for an N-linked oligosaccharide by monoclonal antibodies directed against the V3 loop of human immunodeficiency virus type 1

Rapid selection for an N-linked oligosaccharide by monoclonal antibodies directed against the V3 loop of human immunodeficiency virus type 1

Identification of Two N-Linked Glycosylation Sites within the Core of the Simian Immunodeficiency Virus Glycoprotein Whose Removal Enhances Sensitivity to Soluble CD4

Glycosylation of gp41 of Simian Immunodeficiency Virus Shields Epitopes That Can Be Targets for Neutralizing Antibodies

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