must be precise, and the simultaneous use of
sequential immunogens for multiple bnAb
typesalsorunstheriskofleadingsomebnAb
B cell lineages off track. Thus, multiple rounds
of iterative immunogen design will be needed
to develop complex sequential immunogens
that can induce a polyclonal bnAb response.
Finally, this study demonstrates proof-of-
concept for targeted selection of improb-
able mutations to improve antibody affinitymaturation. Although HIV-1 was the focus
of this study, this strategy of selection of
specific antibody nucleotides by immunogen
design can be applied to B cell lineages tar-
geting other pathogenswhere guided affinitySaunderset al.,Science 366 , eaay7199 (2019) 6 December 2019 11 of 17
Fig. 6. CH505 M5.G458Y SOSIP nanoparticle immunization of rhesus
macaques elicits serum autologous neutralizing antibodies against the
CD4 binding site.(A) Two-dimensional class averages of negative-stain electron
microscopy images of GnTI–/–cell–expressed M5.G458Y-stabilized SOSIP
gp140 ferritin nanoparticles. (B) Rhesus macaque immunization with M5.G458Y-
stabilized SOSIP gp140 ferritin nanoparticles enriched for Man 5 GlcNAc 2 glycans.
(C) Plasma IgG binding over time to GnTI–/–cell–expressed M5.G458Y-stabilized
SOSIP gp140. (D) Serum neutralization kinetics against GnTI–/–cell–produced
HIV-1 CH505 M5.G458Y virus. Each curve represents an individual macaque.
(E) Macaque serum after three immunizations neutralizes Man 5 GlcNAc 2 -
enriched and heterogeneously glycosylated CH505 M5.G458Y virus. Horizontal
bars are the geometric mean for the group (n= 4). SVA is a negative
control murine leukemia virus. (F) Macaque serum after three immunizations
neutralizes Man 5 GlcNAc 2 -enriched CH505 M5.G458Y, M5, and TF viruses.
Horizontal bars are the geometric mean for the group (n= 4). (G) Geometric
mean of the neutralization titers of all four animals against CH505 M5.G458Y
wild-type and CD4bs CH235 bnAb knockout (N280D) viruses over time.
(H) Individual macaque neutralization titers after three immunizations against
M5.G458Y wild-type and CD4bs CH235 bnAb knockout (N280D) viruses.
Horizontal bars are the geometric mean for the group (n=4).(I) Fold decrease
in week 10 neutralization ID 50 titers shown in (H) for each macaque upon
mutation of the CD4 binding site with a N280D mutation.RESEARCH | RESEARCH ARTICLE
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