Journal: Cancer Immunology, Immunotherapy : CII

Article Title: TLR8 agonists remodel the tumor immune microenvironment through PF4-dependent T cell recruitment and ancillary mechanisms

doi: 10.1007/s00262-026-04329-8

Figure Lengend Snippet: TLR8 agonist activates PF4-CXCR3 pathway to shape the TIME. A t-SNE plot showing the distribution of all cells from the control and agonist-treated groups combined ( n = 15,745 cells). B t-SNE plot depicting the distribution of distinct immune cell types. C Dot plot displaying the marker genes used to define each immune cell population. D Density plot showing the distribution of Tlr8-expressing cells across the t-SNE map. E Violin plot illustrating Tlr8 expression levels across different immune cell types. F Circle plot depicting the strength of Pf4–Cxcr3 receptor–ligand interactions between distinct cell types in the control and motolimod-treated groups. G Differential gene expression profiles of neutrophils, macrophages, cDC2, and CD8 + T cells. H Bubble plot comparing changes in specific receptor–ligand interactions between the control and motolimod-treated groups, focusing on macrophage-to-cCD4 + T signaling, cDC2-to-cCD4 + T signaling, and CD8 + T-to-cCD4 + T signaling. I Violin plot showing differential expression of Cxcr3 in cCD4 + T and Tregs, analyzed using the Wilcoxon rank-sum test. ns, not significant; ** P < 0.01

Article Snippet: Pf4 levels in culture supernatants were quantified using Mouse PF4 ELISA Kit (Elabscience, #E-EL-M3080) according to the manufacturer’s instructions.

Techniques: Control, Marker, Expressing, Gene Expression, Quantitative Proteomics

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: TLR8 agonists remodel the tumor immune microenvironment through PF4-dependent T cell recruitment and ancillary mechanisms

doi: 10.1007/s00262-026-04329-8

Figure Lengend Snippet: TLR8 agonist induces PF4 expression through NF-κB signaling pathway. A , B qRT-PCR analysis of Pf4 expression in mouse and human macrophages in the motolimod-treated and control groups ( n = 3). C , D Luminescence assay showing NF-κB pathway activation in mouse and human macrophages following motolimod stimulation ( n = 3). E , F qRT-PCR analysis of Rela expression in Rela -knockdown and control mouse and human macrophages with or without motolimod treatment ( n = 3). G , H Western blotting showing p65 protein levels in RELA -knockdown and control mouse and human macrophages with or without motolimod treatment. I , J qRT-PCR analysis of Pf4 / PF4 expression in Rela / RELA -knockdown and control mouse and human macrophages with or without motolimod treatment ( n = 3). K ELISA quantification of Pf4 levels in the culture supernatant from Rela -knockdown and control mouse macrophages with or without motolimod treatment ( n = 3). Data represent the mean ± SEM. Statistical comparisons were performed using Student’s t-tests. * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001

Article Snippet: Pf4 levels in culture supernatants were quantified using Mouse PF4 ELISA Kit (Elabscience, #E-EL-M3080) according to the manufacturer’s instructions.

Techniques: Expressing, Quantitative RT-PCR, Control, Luminescence Assay, Activation Assay, Knockdown, Western Blot, Enzyme-linked Immunosorbent Assay

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: TLR8 agonists remodel the tumor immune microenvironment through PF4-dependent T cell recruitment and ancillary mechanisms

doi: 10.1007/s00262-026-04329-8

Figure Lengend Snippet: PF4 remodels the TIME and suppresses tumor growth. A , B Tumor growth curves and terminal tumor weights in C57BL/6 mice bearing AKR or MC38 tumors with or without Pf4 overexpression, treated with or without motolimod administered every three days ( n = 6). C Percentage of CD45 + cells among live cells in AKR and MC38 tumor models ( n = 6). D Percentage of cCD4 + T cells within the CD45 + population in AKR and MC38 tumor models ( n = 6). E Percentage of CD8 + T cells within the CD45 + population in AKR and MC38 tumor models ( n = 6). F Percentage of Tregs within the CD45 + population in AKR and MC38 tumor models ( n = 6). G Ratios of cCD4 + T cells to Tregs in AKR and MC38 tumor models ( n = 6). H Ratios of CD8 + T cells to Tregs in AKR and MC38 tumor models ( n = 6). I Representative immunofluorescence staining of CD8α (red) and DAPI (blue) in MC38-Vector or MC38-Pf4-OE tumor sections with or without motolimod treatment. Scale bar: 50 μm. J Quantification of CD8α + positive cell ratio (%) relative to total DAPI + nucleated cells per section ( n = 6). Data represent the mean ± SEM. Statistical comparisons were performed using Student’s t-tests. * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001

Article Snippet: Pf4 levels in culture supernatants were quantified using Mouse PF4 ELISA Kit (Elabscience, #E-EL-M3080) according to the manufacturer’s instructions.

Techniques: Over Expression, Immunofluorescence, Staining, Plasmid Preparation