Journal: Cell Reports Medicine
Article Title: Engineering lipid nanoparticle-stabilized emulsions for spatiotemporal mRNA delivery and enhanced T cell immunity
doi: 10.1016/j.xcrm.2026.102667
Figure Lengend Snippet: Coordinating mRNA expression kinetics with immune dynamics for Th1-biased cellular immunity (A) Immunization regimen and timeline for immune kinetics analysis. C57BL/6J mice ( n = 6) received intramuscular injections of LNP or LSE formulations containing 5 μg Spike mRNA on days 0 and 28. GC B cell and Tfh cell responses in the dLNs were analyzed by flow cytometry on days 7, 14, 21, 28, 29, and 35 (blue). Serum antigen-specific IgG titers were measured by ELISA on days 7, 14, 21, 28, 35, 42, 49, 56, 63, 77, 91, 188, and 268 (green). T cell dynamics in splenocytes were evaluated by intracellular cytokine staining (ICS) on days 21, 28, 29, 35, and 56 (red). (B) Recruitment of DCs ( n = 6) at the injection site via flow cytometry at the indicated time points. (C–F) GC B cell (B220 + GL-7 + FAS + , C and E) and Tfh cell (CD4 + CXCR5 + PD-1 + , D and F) responses in dLNs. Gating strategy is shown in B. (G and H) Humoral immune responses. (G) Serum Spike-specific IgG titers. (H) IgG2a/IgG1 ratios at days 56 as an indicator of Th1/Th2 polarization. (I) Frequencies of Spike-specific CD4 + T cells producing TNF-α, IL-2, or IFN-γ in splenocytes, with LSE shown as red solid lines and LNP as blue dashed lines. Sample sizes were n = 6 for days 21–35 and n = 8 for day 56. Gating strategy is shown in C. (J and K) ELISpot analysis of cytokine-producing splenocytes at day 56 ( n = 8). IFN-γ-producing (G) and IL-4-producing (H) spot-forming cells were quantified among antigen-specific peptide-restimulated splenocytes, reported as spot-forming units per 10 6 cells. Representative ELISpot images (left) and corresponding statistical analyses (right) are shown. In (J and K), data were presented as mean ± SEM. Statistical significance was determined by one-way ANOVA compared to LSE-injected mice with post hoc Dunnett’s multiple comparisons tests. ∗∗∗∗ p < 0.0001.
Article Snippet: ELISpot Plus: Mouse IL-4 (ALP) , Mabtech , Cat# 3311-4APW-2.
Techniques: Expressing, Flow Cytometry, Enzyme-linked Immunosorbent Assay, Staining, Injection, Enzyme-linked Immunospot
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