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. 2021 Jan 11;118(3):e2018027118. doi: 10.1073/pnas.2018027118

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Copyright © 2021 the Author(s). Published by PNAS.

This open access article is distributed under Creative Commons Attribution-NonCommercial-NoDerivatives License 4.0 (CC BY-NC-ND).

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Fig. 4. — Antisera from guinea pigs immunized with an NHR-based vaccine candidate, (ccIZN36)₃, neutralize multiple tier-2 HIV-1 strains in TZM-bl/FcγRI cells. (A) (ccIZN36)₃ contains 36 residues from the NHR region of gp41 (HXB2) and is stabilized by disulfide bonds and a coiled-coil domain (7). (B) ID₅₀ titers (serum dilution) were determined for antisera against viruses pseudotyped with Env from various HIV-1 strains measured in TZM-bl cells not expressing (Top; closed) or expressing (Bottom; open) FcγRI using a validated neutralization assay (63–66). V570A is a mutant of the HXB2 strain that is more sensitive to antibodies that target the PHI (7), and P.I. denotes preimmune antisera tested against this strain. Preimmune antiserum from each animal was tested against all viruses and did not have detectable neutralization in any of the strains (ID₅₀ titers were below the limit of quantitation, which was 100 for TZM-bl/FcγRI assays with V570A antisera and 10 for all others). Each data point represents the ID₅₀ value of antiserum from a single guinea pig after a prime and two boosts with (ccIZN36)₃.