Journal: Advanced Science

Article Title: Nanoparticle Immunoadjuvant Complexes Augment Germinal Center Responses to Vaccination

doi: 10.1002/advs.202517556

Figure Lengend Snippet: Development and characterization of GT8‐IL‐21‐NIC. (A) Model depicting GT8‐IL‐21‐NIC generated using Alphafold and Pymol; GT8 (light blue), IL‐21 (red), lumazine synthase scaffold (gray). IgE denotes signal sequence; GS denotes glycine‐serine linker. (B) Size exclusion chromatography (SEC) trace of GT8‐IL‐21‐NIC run at 215 nm. (C) Negative stain electron microscopy (nsEM) of the GT8‐60mer and GT8‐IL‐21‐NIC. (D) Binding ELISA of recombinant GT8‐60mer or GT8‐IL‐21‐NIC to IL‐21R probed with VRC01. (E) Representative contour plot of follicular B (FO B) cells labelled with Cell Trace Yellow from a Blimp‐1 reporter mouse cultured in vitro with recombinant GT8‐60mer, GT8‐IL‐21‐NIC, or co‐cultured with IL‐4/5. Plots show Blimp‐1+ plasma cells at 72 h post co‐culture. (F) Frequency of Blimp‐1+ plasma cells. (G,H) CD86 and CD80 geometric MFI (gMFI) of cultured follicular B cells with indicated stimuli. (I) Histogram showing pSTAT3 expression in purified FO B cells after culture with 10 µg/mL of the GT8‐60mer or GT8‐IL‐21‐NIC. (J) pSTAT3 gMFI in purified FO B cells with varying concentrations of the GT8‐60mer or GT8‐IL‐21‐NIC. Bars show mean with SD. Each dot represents one technical replicate (F–H); n = 3 technical replicates representative from one or two independent experiments. One‐way ANOVA (F‐H) or two‐way ANOVA (J) adjusted for multiple comparisons with Bonferroni correction used to compare groups; *** p < 0.001; **** p < 0.0001.

Article Snippet: 96‐well half area plates (Cat# 3690, Corning) were coated with 1 μg/mL mouse IL‐21R (Cat# 51184‐M08H, Sino Biologicals) overnight at 4°C.

Techniques: Generated, Sequencing, Size-exclusion Chromatography, Staining, Electron Microscopy, Binding Assay, Enzyme-linked Immunosorbent Assay, Recombinant, Cell Culture, In Vitro, Clinical Proteomics, Co-Culture Assay, Expressing, Purification

Journal: Nature Immunology

Article Title: Flt3L-mediated tumor cDC1 expansion enhances immunotherapy by priming stem-like CD8 + T cells in lymph nodes

doi: 10.1038/s41590-026-02419-4

Figure Lengend Snippet: AT-3 OVA or AT-3 OVA FL tumor cells (5 × 10 5 ) were injected subcutaneously into mice and treated, at days 10, 14 and 18 after tumor inoculation, with anti-CTLA-4 (150 µg per mouse). a – n , At day 9 post-treatment, CD8 + T cells from ( a – c ) tumor dLNs and ( d – n ) tumors of mice were analyzed by scRNA-seq with paired TCR-seq. Data were analyzed from n = 3 (dLNs) or n = 6 (tumors) mice per group from one experiment. a , UMAP of CD8 + T cells for all samples (left) and per treatment group (right). b , Percentage of CD8 + T cells in each cluster that had undergone clonal expansion (teal). c , Proportion of CD8 + T cells in the more clonally expanded L05 and L08 clusters in each treatment group. d , UMAP analysis of CD8 + T cells in all treatment groups (left), frequency of each cluster per treatment group (middle) and frequency of the more stem-like T04 and T06 clusters per individual sample (right). Statistical analysis: two-tailed unpaired t -test. e , f , GSEA of T04 cluster cells versus other clusters ( e ) and f ) UMAP of the CD62L + Tpex gene signature identified by Tsui et al. ( f ). g , Frequency of identified clusters. h , Violin plot depicting the T EX 2 (exhausted CD8 + ) gene signature score in each cluster. i , Prognostic value of T04 gene signature in melanoma patients before treatment with anti-PD-1 monotherapy ( n = 41) or combined anti-PD-1 and anti-CTLA-4 ( n = 31) . T PEX signature scores using the top 25 differentially expressed genes from T04 were calculated for each patient using single sample GSEA. Signature scores were benchmarked against RECIST ( https://recist.eortc.org/ ) and survival outcomes. j , GSEA analysis evaluating enrichment of genes associated with response to ICB in the T04 cluster. k , Volcano plot depicting genes upregulated in T13 cluster versus other clusters. l , Trajectory analysis and plotting of T cell gene expression across pseudotime. m , Chord plot depicting the proportion of TCRs shared between T13 and other clusters. n , Violin plot depicting expression of Il-21r in each cluster. o , Additional groups of tumor-bearing mice treated with anti-IL-21R alone (250 µg per mouse) or in combination with anti-CTLA-4 were as indicated. Tumor growth is presented as the mean ± s.e.m. of six mice per group from one experiment. p , Kaplan–Meier plots representing OS between patients receiving anti-CTLA-4 treatment stratified by IL-21 gene expression from Kovacs et al. . Statistical significance determined by two-way ANOVA ( o ). * P < 0.05. The scRNA-seq experiment was performed only once.

Article Snippet: Anti-mouse CTLA-4 (clone 9H10, cat. no. BE0131), PD-L1 (clone 10 F.9G2, cat. no. BE0101) and IL-21R (clone 4A9, cat. no. BE0258) antibodies were purchased from BioXCell.

Techniques: Injection, Two Tailed Test, Gene Expression, Expressing