Figure 3.

Characteristics of HIV-1 Env glycan-reactive neutralizing Abs in the B cell repertoire of naive humans

(A) Repertoire analysis of blood-derived Env glycan-reactive B cells in nine HIV-1 naive individuals. Enriched B cells were studied via different sort strategies listed that used fluorophore-labeled Man₉-V3 or soluble stabilized recombinant HIV-1 Env trimers.

(B) Immunogenetics of candidate Env glycan-reactive B cells (N = 37) that were isolated from individuals listed in (A).

(C) Repertoire of Env glycan-reactive B cells with IgM⁺IgD⁺CD27⁺ phenotype among six individuals. Flow cytometry data were analyzed from B cells sorted via strategies listed in (A); strategy “c” was used for LKP04.

(D–F) NSEM 2D class averages of DH1005 Ab with I-shaped Ab conformations denoted by “I” in image. 2D class averages of DH1005-SOSIP complex shows Fab-dimer binding to soluble stabilized recombinant HIV-1 Env trimer. Fc domain is not visible here because it lies outside the circular mask used during class averaging. Shown also are 2D class averages of DH1009 (E) and DH1010 (F) that displayed I-shaped Abs.

(G) Seven Env glycan-reactive mAbs were tested in ELISA for binding Man₉-V3 and non-glycosylated aglycone V3 peptide and heat-killed yeast antigens. Data shown are from a representative ELISA. Reference mAbs included peptide-reactive DH1013, Env glycan-reactive DH717.2 and 2G12, trimer-reactive PGT151, and negative control CH65 Abs.

(H) Only DH1005 neutralized multi-clade env-pseudotyped HIV-1 bearing Envs with Man₉-enriched glycans [Kif]. Neutralization titers are representative of two assays in TZM-bl cells, and neutralization titers were reported as IC₅₀ in μg/mL. Control mAbs were 2G12 and CH65.

See also Data S3 and STAR Methods.