Fig. 2

Design and evaluation of the CD4-KO mutation. a The difference in computed Rosetta energy between trimers with glycine at position 473 and all 20 amino acids except cysteine at position 473 in three states: unbound, VRC01-bound, and CD4-bound. Energies are colored from white to red, with energies similar to glycine in white and unfavorable energies in red. b SPR binding of trimers to VRC01-IgG and CD4-IgG. IgGs were ligands and trimers were analytes. c Binding of different trimers to CD4⁺ T cells. Fluorescently labeled Env trimer probes were mixed with human PBMC. All panels gated on CD4⁺ T cells. Blue oval gate indicated probe positive population. A ‘no probe’ staining was used as a negative control to set the lower bound of the positive gate. d Frequency of Env trimer probe positive CD4⁺ T cells. N = 3 individual human donor samples for each probe. The data are representative of two experiments with a total of four independent samples