Extended Data Fig. 5: Biosafety and immune responses investigation.
a–h, Liver functions assessment of the mice with different treatments: the levels of ALT (a), ALP (b), AST (c), γ-GT (d), TBIL (e), DBIL (f), ALB (g), and TBA (h). The area between the two gray dashed lines represented the normal range (NR). i,j, Detection of PEG-specific antibodies including IgM (i) and IgG (j) in mice serum. k–m, Liver tissues staining: ORO staining for lipid accumulation (k), DHE staining for intracellular reactive oxygen species (l), and H&E staining for pathological features (m). All scale bars = 100 μm. In a–m, C57BL/6 normal mice received three i.m. injections of EB-LNP or PEG-LNP on day 0, 3, and 5. The blood samples and livers were collected on day 6 to determine liver functions. PEG-specific antibodies in serum were continuously monitored until day 28. n–p, FCM analysis of inguinal LNs dissected from C57BL/6 normal mice 7 days after a single i.m. injection of EB-Vax or PEG-Vax: the percentages of CD11c+ DCs (n), CD11c+H2Kb+ DCs (o), and CD11c+CD80+CD86+ DCs (p). q, Determination of CD8+IFN-γ+ T cells in spleen. Abbreviations: EB-Vax for EB-LNP@mRNAOVA, and PEG-Vax for PEG-LNP@mRNAOVA. Data in a–j and n–q were presented as mean ± s.d. from n = 3 biologically independent samples. Statistical significance in a–h and n–q was determined by one-way ANOVA with Tukey’s correction: n.s., not significant, *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. Statistical significance in i and j was determined by one-way ANOVA with Dunnett’s multiple comparisons test: n.s., not significant, *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. For a–h and n–q, the center line represented the median, and the whiskers represented the maximum and minimum values.
