The W100 pocket on HIV-1 gp120 penetrated by b12 is not a target for other CD4bs monoclonal antibodies
© Dueñas-Decamp et al; licensee BioMed Central Ltd. 2012
Received: 19 July 2011
Accepted: 27 January 2012
Published: 27 January 2012
The conserved CD4 binding site (CD4bs) on HIV-1 gp120 is a major target for vaccines. It is a priority to determine sites and structures within the CD4bs that are important for inclusion in vaccines. We studied a gp120 pocket penetrated by W100 of the potent CD4bs monoclonal antibody (mab), b12. We compared HIV-1 envelopes and corresponding mutants that carried blocked W100 pockets to evaluate whether other CD4bs mabs target this site.
All CD4bs mabs tested blocked soluble CD4 binding to gp120 consistent with their designation as CD4bs directed antibodies. All CD4bs mabs tested neutralized pseudovirions carrying NL4.3 wild type (wt) envelope. However, only b12 failed to neutralize pseudoviruses carrying mutant envelopes with a blocked W100 pocket. In addition, for CD4bs mabs that neutralized pseudovirions carrying primary envelopes, mutation of the W100 pocket had little or no effect on neutralization sensitivity.
Our data indicate that the b12 W100 pocket on gp120 is infrequently targeted by CD4bs mabs. This site is therefore not a priority for preservation in vaccines aiming to elicit antibodies targeting the CD4bs.
CD4 binding site monoclonal antibodies investigated
Polymun Sci. GmbH
NIH AIDS Rea. Pr.2
NIH AIDS Rea. Pr.
NIH AIDS Rea. Pr.
Recently, further CD4bs mabs, e.g. 3BNC60  and VRC-PG04  have been reported [13–15]. Like VRC01, VRC03, and HJ16 (studied here), they were derived from so called 'elite neutralizers', HIV-1+ subjects who carried potent neutralizing antbodies active against diverse viral strains. These new mabs are highly potent and have binding specificities that focus on similar gp120 residues to that of VRC01 , which (as confirmed here) does not penetrate the W100 pocket on gp120. The crystal structure of mab 3BNC60 verifies a structure where predicted gp120 contact residues are conserved with those of VRC01 , while a structure of mab VRC-PG04 complexed with a gp120 core shows the same for that mab and confirms no interaction with the W100 pocket on gp120. Diskin et al. also reported that NIH45-46 conferred even more potent neutralization than VRC01. However, the increased potency of this mab was due to increased contact with the gp120 inner domain and bridging sheet determinants without targeting the W100 pocket .
In summary, we investigated whether the W100 pocket on gp120 (a critical target for mab b12) is required for HIV-1 neutralization by other CD4bs mabs. We found that neutralization of pseudovirions carrying primary or T-cell line adapted HIV-1 envelopes was either unaffected or only marginally shifted by the presence of a mutated W100 pocket. These observations contrasted with neutralization by b12, which was severely curtailed by the presence of a mutated W100 pocket for all six HIV-1 envelopes tested. Our data indicate that the W100 pocket on gp120 is not a frequent target among CD4bs mabs and is not required for potent neutralization of diverse primary strains of HIV-1 via the CD4bs.
Availability of supporting data
The data sets supporting the results of this article are available in the Dryad repository http://dx.doi.org/10.5061/dryad.66h5g23t.
List of abbreviations
We thank Paul Peters for his insightful comments and input for this study. We also thank John Mascola (Vaccine Research Center, NIAID) and Robin Weiss (UCL, London) for supportive comments and for help in obtaining mabs VRC01, VRC03 and HJ16 respectively. We thank James Robinson (Tulane University School of Medicine) for providing mab 17b. Our work was supported by NIH grants R01s MH64408, AI089334 and P01 AI082274. We also wish to thank the University of Massachusetts Medical School Center for AIDS Research (CFAR), the NIH AIDS Research and Reference Reagent Program, the Vaccine Research Center, NIH and the Centre for AIDS Reagents, NIBSC, UK, for services and reagents.
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