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. Author manuscript; available in PMC: 2021 May 13.
Published in final edited form as: Cell Host Microbe. 2020 May 13;27(5):710–724.e7. doi: 10.1016/j.chom.2020.04.007

Figure 1. Fifteen hmAbs isolated from 3 children post DENV3 infection are DENV3-specific.

Figure 1.

Memory B cells were immortalized from children experiencing primary or secondary DENV3 infections in Nicaragua. TS hmAbs were isolated that bound and neutralized DENV3.

A) DENV Immune PBMCs from children in a Nicaraguan Cohort study experienced DENV3 as either a primary infection or secondary infection after a DENV2 primary infection.

B) Fifteen hmAbs were tested against DENV1–4 by Vero-81 focus reduction neutralization test (FRNT). Positive controls were as follows, DENV1-specific hmAbs 1F4 and 14c10 for DENV1, DENV2-specific hmAb 2D22 for DENV2, DENV3-specific hmAb 5J7 for DENV3 and crossreactive hmAb EDEI C8 for DENV4. All assays were performed twice in duplicate.

C) DENV 3 neutralization by hmAbs was evaluated by FRNT using wildtype and recombinant viruses. Comparison of DENV3-specific hmAbs to 5J7. Using wild-type DENV3, EC50 values from repeat experiments performed on different days were averaged to show reproducibility. EC50 value denotes amount of hmAb needed to neutralize 50% of the virus in a Vero-81 FRNT. Error bars indicate standard deviation.

D) DENV4/3 M16 ic-PyMOL representation of DENV3 residues (blue) that capture the hmAb 5J7 epitope transplanted into a DENV4 backbone.

E) DENV3-specific hmAbs do not use 5J7 epitope. DENV4/3 M16 ic is neutralized by hmAb 5J7 but not by the panel of 15 DENV3-specific hmAbs. EC50 values in a FRNT of hmAbs of DENV4/3 M16 ic, and parental DENV3 ic and DENV4ic.