Fig. 4: Somatic hypermutation is similarly induced by mRNA and Rabipur and is not required for neutralization. | Nature Communications

Fig. 4: Somatic hypermutation is similarly induced by mRNA and Rabipur and is not required for neutralization.

From: Unmodified rabies mRNA vaccine elicits high cross-neutralizing antibody titers and diverse B cell memory responses

Fig. 4

a Violin plots showing the mean level of SHM of antigen-specific Sanger sequences per animal across different groups and timepoints. Lines represent group mean. Statistical significance was calculated using Mann–Whitney test for non-paired data and Wilcoxon signed-rank test for paired data. b Maximum likelihood tree inferred by the multiple sequence alignment of VDJH amino acid antigen-specific sequences for all groups. The tree is drawn to scale, with branch lengths in the same units as those of the evolutionary distances used to infer the phylogenetic tree. The scale bar indicates the distance of 0.2 substitutions per sequence position. This analysis involved 334 amino acid sequences. The reference rabies antibodies binding to Site I and to Site III are indicated with magenta and cyan bars respectively. Twelve sequences were selected for cloning monoclonal antibodies based on lowest, medium and highest SHM (named LowRab1-4, MedRab1-4 and HighRab1-4) as well as based on proximity with the reference antibodies when possible. c Top and side view of a trimeric RABV-G model with antigenic sites I and III highlighted in magenta and cyan respectively. PMDB ID: PM0079619. d Binding potency (EC50) of the cloned mAbs is shown together with that of the reference rabies antibodies and negative control. Light gray, dark gray and black bars indicate the mAbs with the lowest, medium and highest SHM (LowRab, MedRab and HighRab) respectively. The magenta and cyan bars refer to the reference rabies antibodies binding to Site I and to Site III respectively. e Summary of competition between the cloned mAbs with the reference rabies mAbs. The color gradient from red to blue indicates high to no competition. f Spearman correlation between % of SHM and neutralization (IC50) in the different cloned mAbs. The dashed line indicates the limit of detection (LOD) for neutralization which corresponds to the highest concentration of mAb used for the assay (5 μg/mL). All statistical tests comparing the study groups were two-tailed tests.

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