Fig. 6 | Nature Communications

Fig. 6

From: Bacterially derived synthetic mimetics of mammalian oligomannose prime antibody responses that neutralize HIV infectivity

Fig. 6

Immunization with a bacterial mimic of oligomannose evokes HIV gp120-cross-reactive antibodies with tier 2 neutralizing activity in human-antibody transgenic OmniRats. Animals (n = 3) were primed and then boosted (at day 21) with lead conjugate NIT82B formulated in the MF59-like adjuvant Addavax. Sera were collected 6 days thereafter. a Binding of IgM and IgG antibody in pre-immune and immune sera to biotinylated NIT82B (5 µg ml−1) in ELISA. b Comparison of pre-immune and immune serum IgM binding to solubilized HIV gp120 from viruses 92RW020.2, JRFL, and JRCSF. Graphs in a and b depict mean values for the three serum samples, with error bars denoting the error from the mean. c Heat-inactivated sera (1 h, 56 °C) from unimmunized and immunized animals (n = 3 each) were assayed (1:10) against a panel of 7 (tier 2) subtype A, B, and C HIV strains and a VSV control virus. Results for the immune sera with the subtype A and B strains are from two independent experiments performed in duplicate or triplicate, while results for the immune sera with the subtype C strains are from a single experiment, performed in duplicate or triplicate. Results for the immune sera with VSV are from seven independent experiments, performed in duplicate or triplicate. Error bars represent the standard error from the mean. d Probing of the main serum specificity and factors hindering neutralizing activity. Each bar graph denotes the serum from each of the 3 immunized animals (orange: animal #13665; blue: animal #13667; grey: animal #13668). Left: Comparative assessment of serum neutralizing activity against JRCSF wild-type virus and mutant viruses N301A and S334A. Right: Serum neutralizing activity against JRFL wild-type virus to JRFL virus mutants with glycosylation sites missing in V1 (JRFL-T140A, JRFL-N141Q). The bars denote the mean with SEM from a single experiment performed in triplicate. *P < 0.05 (Kruskal–Wallis test with multiple comparisons)

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