Journal: bioRxiv

Article Title: Coordinated Tuning of Ionizable Lipids and Formulation Redirects mRNA Vaccines Toward Lymphoid-Specific CD4 + T Cell Immunity

doi: 10.64898/2026.04.16.719092

Figure Lengend Snippet: (a) Experimental scheme for in vivo screening. C57BL/6N mice were intramuscularly administered fLuc mRNA-loaded LNPs (0.05 mg/kg) and bioluminescence was assessed at 6 h post-injection by IVIS imaging. (b) Representative in vivo IVIS bioluminescence images of nine LNP formulations. (c) Quantification of bioluminescence signals at the injection site. (d) Quantification of bioluminescence signals in the liver. (e) Injection site-to-liver bioluminescence ratio (n = 2 per group). N4Y and N4Z were selected as lead candidates based on high injection site expression and minimal hepatic off-target signal. (f, g) Time-dependent serum levels of MCP-1 (f) and IL-6 (g) following intramuscular administration of N4Y- or N4Z-based LNPs encapsulating SARS-CoV-2 spike mRNA (0.5 mg/kg) in BALB/c mice (n = 4 per group). Statistical significance was determined by unpaired two-tailed Student’s t-test at each time point. Data are presented as mean ± SEM. *p < 0.05, **p < 0.01, ***p < 0.001.

Article Snippet: Luciferase expression was assessed by whole-body bioluminescence imaging and ex vivo imaging of harvested organs using an IVIS system (PerkinElmer).

Techniques: In Vivo, Injection, Imaging, Expressing, Two Tailed Test

Journal: bioRxiv

Article Title: Coordinated Tuning of Ionizable Lipids and Formulation Redirects mRNA Vaccines Toward Lymphoid-Specific CD4 + T Cell Immunity

doi: 10.64898/2026.04.16.719092

Figure Lengend Snippet: (a) Schematic comparison of lipid compositions of N4Z and optimized N4Z (N4Z-opt) formulations. (b) mRNA encapsulation efficiency (n = 3). (c) Hydrodynamic diameter (n = 3). (d) Polydispersity index (PDI) (n = 3). (e) Representative flow cytometry contour plots showing GFP expression in RAW 264.7 macrophages following delivery of PBS, N4Z, and N4Z-opt. (f) Mean fluorescence intensity (MFI) of GFP expression in RAW 264.7 macrophages (n = 3). (g) Representative in vivo IVIS bioluminescence images following intramuscular administration of fLuc mRNA-loaded N4Z or N4Z-opt LNPs. (h) Injection site-to-body bioluminescence ratio (n = 3). (i) Representative ex vivo organ images showing bioluminescence in liver, spleen, and lymph node. (j) Spleen-to-liver bioluminescence ratio (n = 3). (k) Lymph node-to-liver bioluminescence ratio (n = 3). Statistical significance for (b–d) was determined by unpaired two-tailed Student’s t -test. Statistical significance for (f, h, j, k) was determined by one-way ANOVA followed by Tukey’s multiple comparisons test. Data are presented as mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Luciferase expression was assessed by whole-body bioluminescence imaging and ex vivo imaging of harvested organs using an IVIS system (PerkinElmer).

Techniques: Comparison, Encapsulation, Flow Cytometry, Expressing, Fluorescence, In Vivo, Injection, Ex Vivo, Two Tailed Test