A. MPER (2F5 epitope)-specific serum IgG subclass levels in 2F5
complete KI mice from placebo (saline-injected) or experimental (JRFL primed,
TLR agonist-MPER peptide-liposome conjugate boosted) immunization groups,
calculated as described in Materials and Methods. B. Total or
MPER-specific IgM and IgG subclass splenic ASC responses in immunization 2F5
complete KI groups. Shown is ELISPOT data calculated from dilution curve
platings (as shown in Fig.
S2A), from two independent experiments, each with ≥3
mice/group, graphically represented either as total ASCs for each isotype
(left) or as MPER+ B-cells, normalized as
frequency of all ASCs (right). C. Kinetics and
potency of serum neutralizing antibody induction in vaccinated 2F5 complete KI
mice. Top panels: GMTs of reciprocal dilutions of sera to
inhibit HIV-1 B.MN.3 at 50 or 80% levels in the TZM-b/l neutralization assay
(12,31), Bottom panel: corresponding MPER
(2F5)-specific Ig levels from individual mice required. To demonstrate
neutralization potency, GMTs of sera from WT mice spiked with 50,100, or 1000
μg/ml of m2F5 are also shown, with shaded boxes comparing control vs.
vaccinated sera GMTs at similar MPER-specific Ig concentrations. Data are from
two immunization studies, each with sera from ≥3 individual mice/time
point assayed. D. Relative HIV-1 neutralization activity of serum
Igs elicited by placebo or experimentally vaccinated WT, 2F5
VH+/+ KI, and 2F5 complete KI strains. Neutralization
curves of serum were taken from vaccinated WT, 2F5 VH+/+
KI, and 2F5 complete KI groups (≥three mice/group) at peak
MPER+ serum Ig induction (immunized 6 times), using the HIV-1
B.MN.3 isolate in the TZM-b/l neutralization assay.