Frequent use of IGHV3-30-3 in SARS-CoV-2 neutralizing antibody responses

Figures

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  Figure 1. Spike HCs frequently use IGHV3-30 group of genes. (A) A schematic representation of the SARS-CoV-2 cohort consisting of 14 individuals. PBMCs, sera and RNA samples were isolated from each of these individuals. The genotyping of 14 study participants was done using the RNA sample. Two of the 14 individuals’ PBMCs were used to isolated S-specific HCs and subsequent antibody expression. (B) A bar plot showing the IGHV gene usage of S-specific HCs with IGHV3-30 group highlighted in olive green. (C) Titration curves showing percent neutralization of CAB-F52 and REGN10987 against VoCs. (D) IC50 neutralization values of CAB-F52 and REGN10987 against the VoCs. (E) A table showing the genetic properties, specificity, and neutralization activity of CAB-F52 and several IGHV3-30-3-using mAbs from the CoVAbDab.

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  Figure 2. CAB-F52 targets RBD and retains neutralization activity upon germline reversion (A) Epitope mapping of CAB-F52 using HDX-MS. (A) A heat map showing the most probable (blue) and least probable (red) interaction sites of CAB-F52 on the RBD (B) A three-dimensional image of the targeted epitope (C) Amino acid sequence alignment of CAB-F52 VH region and germline allele IGHV3-30-3*01 allele. (D) Cartoon showing the gLCAB-F52 HC design (E) Titration curves showing percent neutralization of CAB-F52 and gLCAB-F52 against VoCs. (F) IC50 neutralization values of CAB-F52 and gLCAB-F52 against several VoCs.

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  Figure 3. Haplotype and genotype analysis reveals variation and IGHV3-30-3 deletion (A) Illustration of IGHV3-30, IGHV3-30-3, IGHV3-33 alleles presence on each chromosome of the haplotypable study participants. (B) A pie chart showing frequencies of individuals with the IGHV3-30-3 gene present (green) or absent (gray). (C) Combined IGHV3-30/IGHV3-30-3 frequencies in IgM libraries of individuals with the IGHV3-30-3 present (green) or absent (gray). ** refers to the significant difference between the groups indicated by p value ≤ 0.01.

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  Figure 4. IGHV3-30 group of genes are highly variable and can impact CAB-F52 function. (A) Alignment of all the amino acid sequences of IGHV3-30, IGHV3-30-3 and IGHV3-33 alleles present in the extended group of 46 cases. (B) Allelic content in the 46 cases: IGHV3-30 (top graph), IGHV3-30-3 (middle graph) and IGHV3-33 (lower graph). The IGHV3-30*04/IGHV3-30-3*03 (dark green) is shown in both the IGHV3-30-3 and IGHV3-30 plots since they cannot be distinguished (C) Cartoons showing the design of the germline-reverted CAB-F52 HC variants. (D) Titration curves showing percentage neutralization of the allele-swapped CAB-F52 mAb variants. (E) IC50 neutralization values of the allele-swapped CAB-F52 mAb variants.