1. ABC effect in WT or T cell– or B cell–deficient mice.
ABC effect in WT or T cell– or B cell–deficient mice. (A–C) BALB/c (black, n = 3), nude (red, n = 3) or JHT mice (blue, n = 3) were injected i.v. with human Epo mRNA formulated with LNP weekly for 5 wk. (A) Human Epo protein expression in the serum was assessed by ELISA at day 1 (D1) and 24 h after the fourth injection on day 22 (D22). Significance was calculated using two-way ANOVA with Sidak posttest versus human Epo protein expression level preinjection. (B) Human Epo mRNA distribution (brown) and nuclei (blue) in the liver were visualized 24 h after the fourth injection by RNAscope in situ hybridization. Scale bar, 100 μm. (C) Anti-PEG IgM levels in serum were quantified by flow cytometric bead assay at D1 and 24 h after the fourth injection on D22. Significance was calculated using two-way ANOVA with Sidak posttest versus anti-PEG IgM level preinjection. (D) C57Bl6/J WT mice (n = 8) were injected i.v. with firefly luciferase mRNA formulated with LNP weekly (white bars) or every 2 wk (black bars). Firefly luciferase expression was measured by bioluminescence 24 h after each injection. Significance was calculated using two-way ANOVA with Sidak posttest versus luciferase activity after the first injection. (E) C57BL/6 WT mice (n = 8) were injected i.v. with firefly luciferase mRNA formulated with LNP weekly for 3 wk, followed by a fourth injection 2 wk after the third injection. Firefly luciferase expression was measured by bioluminescence 24 h after each injection. Significance was calculated using two-way ANOVA with Sidak posttest versus luciferase activity after the first injection. *p < 0.05, **p < 0.01, ***p < n.s., not significant.
Gilles Besin et al. ImmunoHorizons 2019;3:
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