A novel, glycan-dependent epitope in the V2 domain of human immunodeficiency virus type 1 gp120 is recognized by a highly potent, neutralizing chimpanzee monoclonal antibody

Warrier, Sujata V.; Pinter, Abraham; Honnen, William J.; Girard, Marc; Muchmore, Elizabeth; Tilley, Shermaine A.

doi:10.1128/jvi.68.7.4636-4642.1994

Cited by 55 publications

(32 citation statements)

References 47 publications

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“…Interestingly, MAbs 12b, 60b, and 74 demonstrated poor binding to cell surface-expressed gp120 (at 50 g/ml), giving maximal mean fluorescence intensities of 15.7, 9.3, and 7.4, respectively, compared with 197.9 for MAb 10/76b, which recognizes an epitope (aa 162 to 170) in the N-terminal region of V2. Recently, Warrier et al reported that a chimpanzee MAb, C108G, which recognizes a glycan-dependent epitope within aa 162 to 169 and is capable of cross-competing with all of our group F MAbs for gp120 binding (data not shown), was able to neutralize HXB2 at 0.1 g/ml (38). These data further support the conclusion that the N-terminal region of V2 on oligomeric gp120 is well exposed and capable of binding neutralizing antibodies.…”

Section: Discussionsupporting

confidence: 86%

“…A number of human and rat monoclonal antibodies (MAbs) have been reported to bind to regions overlapping the CD4-binding site (4,10,23,30,37) and are thought to block infectivity by inhibiting gp120-CD4 interaction. Recently, a number of neutralizing MAbs mapping to the V2 region of HIV-1 gp120 and to the corresponding region of simian immunodeficiency virus gp130 have been reported (2,6,9,13,14,22,38), implying that this variable loop may also have a functional role in the entry of viruses into target cells. The V2 region has been reported to interact with the V3 region in determining macrophage tropism and sensitivity to neutralization by soluble CD4 (sCD4) (1,5,15,39).…”

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confidence: 99%

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Identification and characterization of monoclonal antibodies specific for polymorphic antigenic determinants within the V2 region of the human immunodeficiency virus type 1 envelope glycoprotein

Shotton

Arnold

Sattentau

et al. 1995

J Virol

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We have identified six monoclonal antibodies (MAbs) mapping to both linear and conformation-dependent epitopes within the V2 region of the human immunodeficiency virus type 1 clone HXB10. Three of the MAbs (12b, 66c, and 66a) were able to neutralize the molecular clones HXB10 and HXB2, with titers in the range of 9.5 to 20.0 g/ml. MAbs mapping to the crown of the V2 loop (12b, 60b, and 74) bound poorly to cell surface-expressed oligomeric gp120, suggesting an explanation for the poor or negligible neutralizing activity of MAbs to this region. In contrast, MAbs 12b and 60b demonstrated good reactivity with recombinant gp120 in an enzyme-linked immunosorbent assay format, suggesting differential epitope exposure between the recombinant and native forms of gp120. Cross-competition analysis of these MAbs and additional V1V2 MAbs for gp120 binding enabled us to assign the MAbs to six groups (A to F). Selection of neutralization escape mutants with MAbs 10/76b and 11/68b, belonging to nonoverlapping competition groups, identified amino acid changes at residues 165 (I to T) and 185 (D to N), respectively. Interestingly, these escape variants remained sensitive to neutralization by the nonselecting V2 MAbs. All MAbs demonstrated good recognition of IIIB viral gp120 yet failed to neutralize nonclonal stocks of IIIB. In addition, MAbs 12b and 62c bound MN and RF viral gp120, respectively, yet failed to neutralize the respective isolates. Cloning and expression of a library of gp120 and V1V2 fragments from IIIB-, MN-, and RF-infected H9 cultures identified a number of polymorphic sites, resulting in antigenic variation and subsequent loss of V2 MAb recognition. In contrast, the V3 region from the clones of the same isolates showed no amino acid changes, suggesting that the V2 region is polymorphic in long-term-passaged laboratory isolates and may account for the reduced antibody recognition observed.* Corresponding author. 222 MATERIALS AND METHODS Sources of reagents.The following gp120 MAbs were used for neutralization and/or gp120-binding studies: 11/65a and 33/79, specific for amino acids (aa) 102 to 121 within C1; 11/4c, 11/41e, and 10/76b, specific for aa 162 to 171 (22); 11/68b, CRA-3, CRA-4, and CRA-5, specific for conserved conformation-dependent epitopes within V2 (22); 39.13g, specific for a conserved conformation-dependent epitope involved in CD4 binding (4, 23); 38.1a, specific for aa 430 to 447 (4, 21); 41.1, specific for a conformation-dependent epitope within V3 (19); and MAb 11/85b, mapping to aa 311 to 321 in V3 (19).Recombinant CHO-expressed gp120 and peptides 740.13 (GEIKNCSFNIST SIRGKVQK), 740.14 (STSIRGKVQKEYAFFYKLDI), and 740.15 (EYAFFY KLDIIPIDNDTTSY) are members of the full set of gp120 overlapping peptides, which were obtained from the Medical Research Council AIDS Directed Programme. The control Raf-c peptide (IVQQFGYQRRASDDGKLTD) was a gift from C. Marshall, Institute of Cancer Research, London, United Kingdom. IIIB, MN, and RF viral stocks were obtained from the Medical Research Council AIDS Dir...

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Section: Discussionsupporting

confidence: 86%

mentioning

confidence: 99%

Identification and characterization of monoclonal antibodies specific for polymorphic antigenic determinants within the V2 region of the human immunodeficiency virus type 1 envelope glycoprotein

Shotton

Arnold

Sattentau

et al. 1995

J Virol

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“…Figure 6 shows that binding of 42F was retained after reduction of disulfide bonds and after reduction and deglycosylation of rgp160 LAI with N-glycosidase F. The successful reduction of gp160 disulfide bonds in this experiment was documented by loss of 1125H reactivity, since the epitope of 1125H is dependent on disulfide bonds (38). The deglycosylation of gp160 was demonstrated by a marked decrease in its molecular weight as well as loss of C108G reactivity; the epitope of C108G is glycan dependent (41). Identical immunoblot results were obtained with 43F (data not shown).…”

Section: Resultsmentioning

confidence: 77%

“…Competition assays. These assays were performed by ELISA on rgp160 LAI , using biotinylated 42F and various competing unlabeled MAbs as described previously (28,38,41).…”

Section: Hiv-1 Strains Hiv-1-infected Cells and Recombinant Hiv-1 Ementioning

confidence: 99%

“…rgp160 LAI was treated with dithiothreitol or with dithiothreitol and N-glycosidase F (Boehringer Mannheim, Indianapolis, Ind. ), and immunoblot strips of untreated and treated rgp160 LAI were prepared essentially as described previously (41). The strips were reacted with anti-Env MAbs followed by development with recombinant protein G conjugated to alkaline phosphatase (1/250 dilution; Zymed, South San Francisco, Calif.).…”

Section: Hiv-1 Strains Hiv-1-infected Cells and Recombinant Hiv-1 Ementioning

confidence: 99%

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A novel antibody-dependent cellular cytotoxicity epitope in gp120 is identified by two monoclonal antibodies isolated from a long-term survivor of human immunodeficiency virus type 1 infection

Alsmadi¹,

Herz²,

Murphy³

et al. 1997

J Virol

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Two monoclonal antibodies (MAbs), 42F and 43F, were isolated some 14 months apart from a single long-term survivor of human immunodeficiency virus type 1 (HIV-1) infection. These MAbs were found to be indistinguishable in terms of their isotypes, specificities, affinities, and biological activities. Both 42F and 43F directed substantial antibody-dependent cellular cytotoxicity (ADCC) against cells infected with four divergent lab-adapted strains of HIV-1, but no neutralizing activity against these strains was detectable. The ability of MAbs 42F and 43F, as well as that of MAbs against two other gp120 epitopes, to direct ADCC against uninfected CD4 ؉ cells to which recombinant gp120 SF2 had been adsorbed (i.e., "innocent bystanders") was demonstrated to be less efficient by at least an order of magnitude than their ability to direct ADCC against HIV-1-infected cells. Flow cytometry analyses showed that 42F and 43F also bind to native primary isolate Envs from clades B and E expressed on cell surfaces. By direct binding and competition assays, it was demonstrated that the 42F/43F epitope lies in a domain of gp120 outside the previously described CD4-binding site and V3 loop ADCC epitope clusters. Immunoblot analysis revealed that the 42F/43F epitope is not dependent on disulfide bonds or N-linked glycans in gp120. Epitope mapping of 42F and 43F by binding to linear peptides demonstrated specificity of these MAbs for a sequence of 10 amino acids in the C5 domain comprising residues 491 to 500 (Los Alamos National Laboratory numbering for the HXB2 strain). Thus, 42F and 43F define a new ADCC epitope in gp120. Because of the relative conservation of this epitope and the fact that it appears to have been significantly immunogenic in the individual from which these MAbs were derived, it may prove to be a useful component of HIV vaccines. Furthermore, these MAbs may be used as tools to probe the potential importance of ADCC as an antiviral activity in HIV-1 infection.

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Temporal development and prognostic value of antibody response to the major neutralizing epitopes of gp120 during HIV-1 infection

et al. 1997

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Our objective was to analyse the humoral response to the major neutralizing epitopes of gp120. The kinetics of the appearance of antibodies directed to the V3 region (V3 Abs) and antibodies directed to the CD4 binding site (CD4BS Abs) were compared in sequential sera from 20 seroconverters. V3 Abs were titrated using 2 different indirect EIAs with synthetic oligopeptides coated on the solid phase. The sequences of the oligopeptides used were those of the MN isolate or a mixture of the consensus sequences of the 5 major HIV-1 subtypes (A-E). CD4BS Abs titers were determined using an EIA in which serum antibodies compete with a labeled human monoclonal antibody, F105, whose corresponding epitope overlaps the conformation-dependent CD4BS, for binding to purified recombinant gp120 coated on a solid phase. The prognostic value of both antibodies was analyzed in a longitudinal study of 60 HIV-1 infected patients (17 nonprogressors and 43 progressors). Eighty-five percent and 70% of HIV sero-converters were positive for V3 Abs and CD4BS Abs, respectively, during the observation period. V3 Abs were detected first in the majority of the patients (mean delay of appearance, 1.22 +/- 0.96 months vs. 4.81 +/- 2.05 months for CD4BS Abs). Both categories of antibodies appeared simultaneously in 4 patients (20%). No prognostic value could be attributed to these antibodies. Our data confirm that V3 Abs and CD4BS Abs appear with some delay after primary infection, suggesting that they do not play a large or early role in the rapid clearance of viremia in primary HIV-1 infection. These antibodies were not associated with progression to symptomatic infection and are thus of no value for surveillance in HIV-1 infected patients.

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A novel, glycan-dependent epitope in the V2 domain of human immunodeficiency virus type 1 gp120 is recognized by a highly potent, neutralizing chimpanzee monoclonal antibody

Cited by 55 publications

References 47 publications

Identification and characterization of monoclonal antibodies specific for polymorphic antigenic determinants within the V2 region of the human immunodeficiency virus type 1 envelope glycoprotein

Identification and characterization of monoclonal antibodies specific for polymorphic antigenic determinants within the V2 region of the human immunodeficiency virus type 1 envelope glycoprotein

A novel antibody-dependent cellular cytotoxicity epitope in gp120 is identified by two monoclonal antibodies isolated from a long-term survivor of human immunodeficiency virus type 1 infection

Temporal development and prognostic value of antibody response to the major neutralizing epitopes of gp120 during HIV-1 infection

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