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. Author manuscript; available in PMC: 2023 Jan 1.
Published in final edited form as: Sci Transl Med. 2022 Mar 16;14(636):eabl5399. doi: 10.1126/scitranslmed.abl5399

Fig. 1: Isolation and characterization of anti-Gn/Gc mAbs from PUUV-experienced donors.

Fig. 1:

(A) Phylogenetic tree of the M segment-encoded GPC sequences from Old World and New World hantaviruses (OWH/NWH) based on amino acid (aa) sequences shown in Table S1. ANDV, Andes virus; BAYV, Bayou virus; BCCV, Black Creek Canal virus; CHOV, Choclo virus; DOBV, Dobrava-Belgrade virus; HTNV, Hantaan virus; LGNV, Laguna Negra virus; MPRLV, Maporal virus; PHV, Prospect Hill virus; PUUV, Puumala virus; SEOV, Seoul virus; SNV, Sin Nombre virus; TULV, Tula virus. Scale bar represents 0.07 aa substitutions per site. (B) Frequency of hantavirus Gn/Gc-reactive B cells isolated with rVSV-mNG particles bearing PUUV or ANDV Gn/Gc from PUUV-naïve and PUUV-experienced donors. Fluorescence-activated cell sorting plots are gated on CD19+/CD20+/IgD−/IgM−/virus+ B cells. (C) Somatic hypermutation (SHM) load as determined by the number of VH and VL nucleotide substitutions away from the predicted VH and VL germline. Bar indicates median. (D) Binding reactivity of 180 isolated mAbs (25 nM) to rVSV-PUUV-Gn/Gc, as determined by ELISA. A450, absorbance at 450 nm. The red dashed line indicates the threshold for designating binders (A450=0.3). Bar indicates median. Averages for PUUV Gn/Gc binders (n=4) from two experiments. (E) Binding cross-reactivity of 135 isolated mAbs (25 nM) to rVSVs bearing Gn/Gc from the indicated viruses, as determined by ELISA. The red dashed line indicates the threshold for designating binders (A450=0.3). ANDV, SNV: averages (n=2) from one experiment. Other viruses, averages (n=4) from two experiments.