3nv.2 triple immunogen elicits combined responses of IGT2 and RC1 immunogens in RMs. (A) Schematic and timeline of immunization regimens for RMs (n = 5 per group). (B–E) Serum neutralization ID50s against (B) IGT2, (C) IGT1, (D) RC1, and (E) 11MUTB pseudoviruses. The dotted line at y = 10² indicates the lowest dilution evaluated. Significance was demonstrated using an unpaired t test (P ≤ 0.05). (F) Serum ELISA binding to IGT1 and IGT1 CD4bs KO SOSIPs for RMs immunized with IGT2-mi3 or 3nV.2-mi3. All samples are from week 8 after prime immunizations. Significance was demonstrated using a paired t test (P ≤ 0.05). (G) Mouse serum neutralization ID50s against IGT2, IGT1, RC1, and 11MUTB pseudoviruses (n = 8). The dotted line at y = 10² indicates the lowest dilution evaluated. * denotes 0.01 < P ≤ 0.05, ** denotes 0.001 < P ≤ 0.01, and *** denotes 0.0001 < P ≤ 0.001.