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. 2023 Nov 2;8:169. doi: 10.1038/s41541-023-00766-z

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© The Author(s) 2023

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 4 — a Enhanced anti-PEG IgM production induced by repeated LNP injection. ▲Anti-PEG IgM (Log₁₀ CONC) means Anti-PEG IgM (Log₁₀ CONC_{2nd injection}) (log₁₀-transformed concentration of anti-PEG IgM induced during the second injection cycle) subtracted corresponding Anti-PEG IgM (Log₁₀ CONC_{1st injection}) (log₁₀-transformed concentrations of anti-PEG IgM induced during the first injection cycle). Peak levels of ▲Anti-PEG IgM (Log₁₀ CONC) induced by different LNP doses were 0.654 ± 0.471 for L-LNP (▲Day 7), 1.574 ± 0.399 for M-LNP (▲Day 3) and 2.277 ± 0.410 for H-LNP (Day 3), respectively, with significant difference among three groups (P < 0.001 for M-LNP vs L-LNP; P < 0.0001 for H-LNP vs L-LNP; P < 0.0001 for H-LNP vs M-LNP). b Time-course of enhanced anti-PEG IgM induced by repeated injection of LNP. c Enhanced anti-PEG IgG production induced by repeated injection of LNP. ▲Anti-PEG IgG (Log₁₀ CONC) means Anti-PEG IgG (Log₁₀ CONC_{2nd injection}) (log10-transformed concentration of anti-PEG IgG induced during the second injection cycle) subtracted corresponding Anti-PEG IgG (Log₁₀ CONC_{1st injection}) (log₁₀-transformed concentrations of anti-PEG IgG induced during the first injection cycle). Peak levels of ▲Anti-PEG IgG (Log₁₀ CONC) induced by different LNP doses were 0.888 ± 0.459 for M-LNP (▲Day 5) and 1.354 ± 0.308 for H-LNP (▲Day 7), respectively, with significant difference between these two groups (P < 0.01). d Time-course of enhanced anti-PEG IgG induced by repeated injection of LNP. In (a) and (c), data were presented as “mean ± standard deviation”, with n = 8 for Control group and n = 15 for all LNP-treated groups. Differences in ▲Anti-PEG IgM (Log₁₀ CONC) or ▲Anti-PEG IgG (Log₁₀ CONC) among various groups were analyzed using Mann–Whitney U test, with P values adjusted for FDR (false discovery rate). *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001. In (b) and (d), changing curves of average level of ▲Anti-PEG IgM (Log₁₀ CONC) or ▲Anti-PEG IgG (Log₁₀ CONC) over time for various doses were fitted by the R package called “ggalt”.