Fig. 2. |. Wild-type mouse immunization with RC1 elicits V3-glycan patch antibodies.

a, Immunization protocol. b,d,f,h, Representative ELISAs showing serum binding to indicated immunogens. Controls include naïve serum (red), purified PGT121 (green) and iGL-PGT121/10–1074 (black). b, iGL PGT121 KI mice⁹. d, f, h. Wild-type mice. c,e, Area under the curve (AUC) for ELISAs in b and d, respectively, but combined results from 2 experiments using 3 or 4 mice each. Each dot represents serum from one mouse. f, Binding to RC1 and RC1-glycanKO. g, AUC for RC1 vs RC1-glycan KO ELISAs from 7 experiments with 2 or 3 mice immunized with RC1. i, Ratio of the AUC for RC1 vs RC1-glycan KO ELISAs for wild-type mice immunized with RC1(7 experiments) or RC1–4fill (5 experiments). j, Pie charts show clonal expansion of RC1 binding germinal center B-cells. Colored slices are proportional to the number of clonal relatives. White indicates single IgV_H sequences. The number of heavy chains analyzed is indicated in the center. k, IgH nucleotide mutations from naïve and RC1 immunized mice in j. l, ELISA binding of representative mAbs from RC1-immunized mice to RC1 and RC1-glycanKO. m, ELISA binding of Ab275_MUR and Ab276_MUR to indicated Env proteins. Unpaired t-test in c, e, i. Data in c, g, i, k are mean.