Removal of both B-1–mediated anti-PC natural IgM and B-2–mediated anti-PEG IgM responses abrogates ABC in mice. Removal of both B-1–mediated anti-PC natural.

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Removal of both B-1–mediated anti-PC natural IgM and B-2–mediated anti-PEG IgM responses abrogates ABC in mice. Removal of both B-1–mediated anti-PC natural IgM and B-2–mediated anti-PEG IgM responses abrogates ABC in mice. (A–E) C57Bl6/J WT or sIgM−/− mice were injected i.v. with human Epo encoding mRNA formulated with LNP weekly for 5 wk (arrows on x-axis indicate the day of injection). Human Epo protein expression was assessed in the serum of WT (open symbol) or sIgM−/− (closed symbol) mice by ELISA 6 and 24 h postinjection. Significance was calculated using two-way ANOVA with Sidak posttest versus C57Bl6/J WT mice at each timepoint. (B) Anti-PEG IgM or (C) anti-PC IgM levels were quantified by flow cytometric bead assay 24 h postinjection. Significance was calculated using one-way ANOVA with Sidak posttest versus WT mice at each timepoint. (D) Anti-PEG IgG1 (triangles) or anti-PEG IgG3 (squares) levels in serum were quantified from C57Bl6/J WT (open symbols) or sIgM−/− (closed symbols) mice by flow cytometric bead assay 24 h postinjection. Significance was calculated using one-way ANOVA with Sidak posttest versus WT mice at each timepoint. (E) Human Epo mRNA distribution (brown) and nuclei (blue) in the liver 24 h after fifth injection was visualized by in situ hybridization and hematoxylin staining, respectively. Scale bar, 100 μm. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. n.s., not significant. Gilles Besin et al. ImmunoHorizons 2019;3:282-293 Copyright © 2019 The Authors