Journal: Materials Today Bio

Article Title: Tomatine as a versatile adjuvant boosts mRNA vaccine responses

doi: 10.1016/j.mtbio.2025.102360

Figure Lengend Snippet: Illustration of tomatine-containing mRNA vaccines against tumors and infectious diseases. Tomatine-based mRNA nano-vaccines were developed by simultaneously encapsulating tomatine and mRNA into two different types of nanocarriers including liposomes (T-LPX) and lipid nanoparticles (T-LNP). T-LPX and T-LNP exhibited superior efficacy owing to the significant enhancement of mRNA-encoded antigen expression and immunogenicity. 1) Tomatine regulated the entry of nano-vaccines into dendritic cells (DCs) mainly through the caveolin-mediated endocytosis. This bypassed lysosome-related degradation of mRNA, facilitating mRNA to reach the cytoplasm for translation. Moreover, tomatine remarkably enhanced mRNA translation by upregulating the mRNA translation initiation regulator (DExH-box helicase 29, DHX29), thereby promoting protein expression and antigen presentation. 2) Tomatine activated the nucleotide-binding oligomerization domain (NOD)-like receptor pathway to trigger the innate immune response, thereby promoting IFN- γ secretion and DCs maturation. Collectively, these processes significantly boosted the vaccine’s immunogenicity, hence achieving powerful immune responses against tumors and infectious diseases.

Article Snippet: IFN- γ ELISpotPLUS kits were purchased from Mabtech.

Techniques: Vaccines, Liposomes, Expressing, Immunopeptidomics, Binding Assay

Journal: Materials Today Bio

Article Title: Tomatine as a versatile adjuvant boosts mRNA vaccine responses

doi: 10.1016/j.mtbio.2025.102360

Figure Lengend Snippet: Tomatine amplified the immunogenicity of mRNA vaccines via the NOD-like receptor pathway. a-b Inflammatory cytokines secretion in the supernatant of BMDCs treated with PBS (Control), LPX and T-LPX was detected by ELISA. a IFN- β ; b IL-12; c TNF- α ; and d IFN- γ . e-j Tomatine stimulated the innate immune response through activation of the NOD-like receptor pathway: e Kyoto encyclopedia of genes and genomes (KEGG) pathway analysis of RNA sequencing (RNA-seq) of BMDCs treated with LPX and T-LPX indicated that tomatine incorporation affected the NOD-like receptor signaling pathway. f Quantitative polymerase chain reaction (qPCR) analysis was conducted to evaluate gene expression related to the NOD-like receptor signaling pathway in BMDCs treated with PBS (Control), LPX and T-LPX loaded with OVA-mRNA. g Western blotting was used to measure the expression levels of NOD1, phosphorylated TANK-binding kinase (p-TBK), and phosphorylated interferon regulatory factor 7 (p-IRF7) in extracts from BMDCs treated with PBS (Control), LPX and T-LPX. h Molecular docking between tomatine and NOD1 was carried out. i The docking scores for each molecular docking interaction between tomatine and NOD1 were calculated. j The per-residue confidence score (pLDDT) of the NOD1 structure was determined to be 84.2925, suggesting high credibility in the predicted results. Error bars represent the mean ± SEM for n = 3 experimental replicates. P values were determined by one-way analysis of variance (ANOVA).

Article Snippet: IFN- γ ELISpotPLUS kits were purchased from Mabtech.

Techniques: Amplification, Immunopeptidomics, Vaccines, Control, Enzyme-linked Immunosorbent Assay, Activation Assay, RNA Sequencing, Real-time Polymerase Chain Reaction, Gene Expression, Western Blot, Expressing, Binding Assay, Residue

Journal: Materials Today Bio

Article Title: Tomatine as a versatile adjuvant boosts mRNA vaccine responses

doi: 10.1016/j.mtbio.2025.102360

Figure Lengend Snippet: Tomatine enhanced mRNA expression and vaccine immunogenicity based on a lipid nanoparticle delivery system. a A schematic representation of the ionizable lipid nanoparticle co-loaded with mRNA and tomatine (T-LNP). b TEM images of T-LNP. The scale bar represents 200 nm. c The particle size, zeta potential and mRNA encapsulation efficiency of LNP (without tomatine) and T-LNP. d Transfection experiments involved the delivery of GFP-mRNA using LNP, a physical mixture of LNP and tomatine (LNP + T), and T-LNP into DC2.4 cells. Naked GFP-mRNA treated cells were used as control. The scale bar represents 100 μm. e The expression levels of LNP and T-LNP, each loaded with Luc-mRNA, were assessed at 6 h and 24 h post-intramuscular injection in BALB/c mice. f-g Tomatine also improved the immunogenicity of T-LNP: f Flow cytometric evaluation of BMDC maturation (CD11c + CD80 + CD86 + ) was conducted 24 h post-incubation with LNP and T-LNP, each loaded with OVA-mRNA. g The secretion of IFN- γ in the supernatants of BMDCs treated with LNP and T-LNP was detected by ELISA. PBS treated cells were used as control. Error bars represent the mean ± SEM for n = 3 experimental replicates. P values were determined by two-tailed student’s t-tests ( e ) and one-way analysis of variance (ANOVA) ( f and g ).

Article Snippet: IFN- γ ELISpotPLUS kits were purchased from Mabtech.

Techniques: Expressing, Immunopeptidomics, Zeta Potential Analyzer, Encapsulation, Transfection, Control, Injection, Incubation, Enzyme-linked Immunosorbent Assay, Two Tailed Test

Journal: Materials Today Bio

Article Title: Tomatine as a versatile adjuvant boosts mRNA vaccine responses

doi: 10.1016/j.mtbio.2025.102360

Figure Lengend Snippet: T-LNP elicited strong immune responses against SARS-CoV-2. a A schematic illustration details the immunization protocol and sample collection timeline: BALB/c mice were intramuscularly immunized with LNP and T-LNP, both carrying mRNA encoding the SARS-CoV-2 delta variant spike protein, at weeks 0 and 2. The control group received PBS. Serum samples were collected biweekly from immunized mice, and RBD-specific IgG titers were measured at weeks 2, 4, 6, 8, and 10 using ELISA. b IgG titers against the wild-type (WT), delta, and omicron RBD variants in the immunized mice were determined by ELISA ( n = 6). P value represents comparisons between LNP and T-LNP. c The T-cell immune response induced by LNP and T-LNP was evaluated: representative scatter plots and quantitative data from flow cytometric analyses of CD4 + and CD8 + T cells in the spleen and lymph nodes were obtained on week 10. d Splenocytes isolated from mice at week 10 post-immunization were stimulated with SARS-CoV-2 protein peptide pools for 36 h, and the quantity of IFN- γ -producing T cells was determined through ELISpot assay. e Safety evaluation of T-LNP. H&E analysis of heart, liver, spleen, lungs and kidney in PBS (Control), LNP and T-LPN treated mice. Scale bar, 200 μm. PBS treated mice were used as control. Error bars represent the mean ± SEM. P values were determined by one-way analysis of variance (ANOVA).

Article Snippet: IFN- γ ELISpotPLUS kits were purchased from Mabtech.

Techniques: Variant Assay, Control, Enzyme-linked Immunosorbent Assay, Isolation, Enzyme-linked Immunospot