Single-cell sorting identifies 11 cross-clade–neutralizing lineages from 10 SHIV-infected rhesus macaques with HIV-1 V2 apex–directed heterologous neutralization breadth. (a) Schematic for the present study. (b) Macaque host ID, infecting SHIV strain, and immunogenetics of a representative mAb from each of the 11 rhesus lineages reported in this study. (c) Neutralization breadth and potency of representative monoclonal lineage antibodies. Left: Neutralization activity against a 19-member panel of cross-clade tier-2 HIV-1 strains and a simian immunodeficiency virus infecting chimpanzees (SIVcpz), MT145K, which has been shown to bear the conserved HIV-1 V2 apex epitope. Data are reported as IC50 titer (µg/ml) and colored according to the legend. Bold boxes demarcate activity against autologous virus; for example, 6070-a.01 was isolated from an animal infected with an SHIV bearing the HIV-1 CH505.TF Env (SHIV.CH505). All small panel neutralization experiments were performed in duplicate and repeated twice. Right: Neutralization breadth and geomean IC50 against one or two large cross-clade panels of HIV-1 strains. # denotes activity against 119 viruses (Seaman panel); & denotes activity against 208 viruses (VRC panel). Bottom: Previously published rhesus and human V2 apex broadly neutralizing antibodies are included below the gray row for comparison; human antibodies are denoted with *. IC50 data for these antibodies are shown in italics when obtained from their respective publications. 119 virus panel data (#) for CH01, VRC26.08, and PCT64-35M were derived from CATNAP (https://www.hiv.lanl.gov/components/sequence/HIV/neutralization/). All large panel neutralization assays were performed in duplicate. (d) SHM versus antibody breadth on a 119-isolate panel is shown for representative antibodies, with V2 apex rhesus antibodies in red. (e) SHM versus antibody category is shown for antibodies with over 30% breadth on a 208-strain HIV-1 panel. V2 apex antibodies have SHM levels lower than other categories, though similar to those of the MPER category. Notably, V033-a.01 with 37% breadth showed substantially lower SHM.
Figure 1.

Single-cell sorting identifies 11 cross-clade–neutralizing lineages from 10 SHIV-infected rhesus macaques with HIV-1 V2 apex–directed heterologous neutralization breadth. (a) Schematic for the present study. (b) Macaque host ID, infecting SHIV strain, and immunogenetics of a representative mAb from each of the 11 rhesus lineages reported in this study. (c) Neutralization breadth and potency of representative monoclonal lineage antibodies. Left: Neutralization activity against a 19-member panel of cross-clade tier-2 HIV-1 strains and a simian immunodeficiency virus infecting chimpanzees (SIVcpz), MT145K, which has been shown to bear the conserved HIV-1 V2 apex epitope. Data are reported as IC50 titer (µg/ml) and colored according to the legend. Bold boxes demarcate activity against autologous virus; for example, 6070-a.01 was isolated from an animal infected with an SHIV bearing the HIV-1 CH505.TF Env (SHIV.CH505). All small panel neutralization experiments were performed in duplicate and repeated twice. Right: Neutralization breadth and geomean IC50 against one or two large cross-clade panels of HIV-1 strains. # denotes activity against 119 viruses (Seaman panel); & denotes activity against 208 viruses (VRC panel). Bottom: Previously published rhesus and human V2 apex broadly neutralizing antibodies are included below the gray row for comparison; human antibodies are denoted with *. IC50 data for these antibodies are shown in italics when obtained from their respective publications. 119 virus panel data (#) for CH01, VRC26.08, and PCT64-35M were derived from CATNAP (https://www.hiv.lanl.gov/components/sequence/HIV/neutralization/). All large panel neutralization assays were performed in duplicate. (d) SHM versus antibody breadth on a 119-isolate panel is shown for representative antibodies, with V2 apex rhesus antibodies in red. (e) SHM versus antibody category is shown for antibodies with over 30% breadth on a 208-strain HIV-1 panel. V2 apex antibodies have SHM levels lower than other categories, though similar to those of the MPER category. Notably, V033-a.01 with 37% breadth showed substantially lower SHM.

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