Journal: Nature Communications

Article Title: A tissue-intrinsic mechanism sensitizes HIV-1 particles for TLR-triggered innate immune responses

doi: 10.1038/s41467-026-72586-3

Figure Lengend Snippet: a Schematic representation of PRRs and their cognate ligands. PRRs: green, adapter proteins: pink, PAMPs: red, transcription factors: blue. b PRR inhibitor screening. MDMs were challenged with S or DC HIV-1 NL4.3 R5 virions in presence or absence of PRR inhibitors. IL-6 release was measured by ELISA from MDM supernatants. Data is normalized to mock infected condition (gray dotted line). c Measurement of IL-6 release from the supernatants of MDMs challenged with S or DC HIV-1 virions in presence or absence of the indicated inhibitors. d Volcano plot illustrating the differential gene expression between MDMs challenged with DC virus and mock challenged cells for 6 h. The number of down- and upregulated genes are indicated. e Volcano plot as in d. with highlighted gene signatures. Genes induced by TLR2 alone (blue), TLR8 alone (brown), co-regulated (light green) or associated with “Innate Immune Response” genes (red). f Volcano plot illustrating the differential gene expression between MDMs challenged with suspension or collagen primed HIV-1 virus for 6 h. The number of down- and upregulated genes are indicated. g Volcano plot as in f. with highlighted gene signatures. Genes induced by TLR2 alone (blue), TLR8 alone (brown), co-regulated (light green) or associated with Innate Immune Response genes (red). Top deregulated genes belonging to different gene ontology terms are highlighted. h Gene ontology analysis of biological processes repressed or induced in MDMs challenged with collagen primed virions as compared to cells challenged with suspension virus for 6 h. FDR is indicated (only upregulated pathways have a q -value < 0.05). Top deregulated genes for each pathway are highlighted. Results represent the mean ± SD of 3 independent donors. Symbols indicate individual donors ( b , c ) or individual genes ( d–g ). Significance is indicated by p-values, and was calculated by two-tailed paired t-tests or Wilcoxon matched-pairs signed rank test ( b , c ), differential gene expression analysis was performed using DESeq2: genes with adjusted p-value < 0.05 were considered significant ( d–g ), for GSEA analysis, p -values were corrected using Benjamini-Hochberg FDR ( h ). Source data are provided as a file.

Article Snippet: 5 μM of cGAS inhibitor (G140, Invivogen), 8 μM of TLR 1/2 inhibitor (Cu-CPT22, Selleckchem), 49 μM of TLRs 4& 2/6 inhibitor (GIT27, Tocris), 2 μM of TLR4 inhibitor (CLI-095, Invivogen), 100 μM of TLR2 inhibitor (TL2-C29, Invivogen) or 10 μM of TLR8 inhibitor (Cu-CPT9a, Invivogen) were incubated with MDMs 3 h prior to infection.

Techniques: Enzyme-linked Immunosorbent Assay, Infection, Gene Expression, Virus, Suspension, Two Tailed Test

Journal: Nature Communications

Article Title: A tissue-intrinsic mechanism sensitizes HIV-1 particles for TLR-triggered innate immune responses

doi: 10.1038/s41467-026-72586-3

Figure Lengend Snippet: a Representative structure of an HIV-1 Env trimer. Epitopes targeted by different bNAbs or nNAbs as well as sCD4 are highlighted (based upon a prediction of NL4.3 Env trimer structure using AlphaFold 3). b Experimental workflow. HIV-1 NL4.3 R5 Vpr.Int.GFP virions harvested from suspension or collagen cultures were sequentially incubated with primary, epitope-specific anti-gp120 antibodies, then with AF568 coupled secondary antibodies prior to processing for microscopy analysis. c . Representative micrographs of stained virions. Vpr.Int.GFP positive spots (green) can be seen bound by different antibodies or sCD4-PE (red). Scale bar: 0.5 µm. d Spider plot representing the binding frequency of antibody binding to HIV-1 NL4.3 R5 as in (c). e . sTLR2 is retained at the surface of NL4.3 R5 but not NL4.3 ΔEnv virions after collagen priming. One representative Western Blot analysis showing the ratio from HIV-1 NL4.3 virions cultured in S, DC or LC. The TLR2/p24 ratios are indicated below the blots after correction, after subtraction of the TLR2 background signals from the last two lanes. f Quantification of western blot analyses from three independent experiments as in e . g . Representative micrographs. MDMs challenged with HIV1 NL4.3 R5 Vpr.Int.GFP virions after culture in suspension or in DC. Yellow arrows indicate Vpr.Int.GFP/TLR8 colocalization. Scale bar: 5 µm. h Quantification of Virus/TLR8 colocalization from micrographs as in d. Results represent the mean ± SD of three independent experiments. Significance is indicated by p -values, and was calculated by two-tailed paired t-tests ( d , f , h ). Source data are provided as a file.

Article Snippet: 5 μM of cGAS inhibitor (G140, Invivogen), 8 μM of TLR 1/2 inhibitor (Cu-CPT22, Selleckchem), 49 μM of TLRs 4& 2/6 inhibitor (GIT27, Tocris), 2 μM of TLR4 inhibitor (CLI-095, Invivogen), 100 μM of TLR2 inhibitor (TL2-C29, Invivogen) or 10 μM of TLR8 inhibitor (Cu-CPT9a, Invivogen) were incubated with MDMs 3 h prior to infection.

Techniques: Suspension, Incubation, Microscopy, Staining, Binding Assay, Western Blot, Cell Culture, Virus, Two Tailed Test

Journal: Frontiers in Immunology

Article Title: Human neutrophils recognize group B streptococci via formylated peptide receptors and toll-like receptor 8

doi: 10.3389/fimmu.2026.1828994

Figure Lengend Snippet: TLR8 is required for GBS-induced cytokine production. Neutrophils were treated with the TLR8 inhibitor CU-CPT9a (0.75, 1.50 and 3.00μM) or the TLR2 inhibitor TL2-C29 (5, 10 and 25μM) before stimulation with live [MOI of 5, (A, B) ] or heat-killed GBS [25 μg/ml, (C, D) ]. IL-8 (A, C) and TNF-α (B, D) were measured in 24h culture supernatants. Escherichia coli lipopolysaccharide (LPS; 10 ng/mL) was included as a positive control. (E) Effect of pretreatment with the TLR8 CU-CPT9a inhibitor (3μM) on the release of reactive oxygen species after stimulation with live GBS (MOI 100). Data are expressed as means ± standard deviations from five independent experiments, each performed in duplicate. *p < 0.05 and **p < 0.01, as determined by the Mann-Whitney test; ns, not significant.

Article Snippet: The contribution of FPRs and TLRs to IL-8 and ROS production induced by live bacteria was evaluated by pretreating neutrophils for 1 hour at 37 °C in 5% CO 2 with the selective FPR2 antagonist WRW4 (Abcam), the pan-FPR inhibitor Boc-2 (GenScript), the selective TLR8 inhibitor CU-CPT9a (InvivoGen) or the selective TLR2 inhibitor TL2-C29 (InvivoGen) at the indicated concentrations prior to stimulation with heat-killed (HK) or live bacteria.

Techniques: Positive Control, MANN-WHITNEY

Journal: Frontiers in Immunology

Article Title: Human neutrophils recognize group B streptococci via formylated peptide receptors and toll-like receptor 8

doi: 10.3389/fimmu.2026.1828994

Figure Lengend Snippet: Cytokine responses to GBS by nucleic acids. IL-8 (A) and TNF-α (B) responses to stimulation with graded doses of GBS nucleic acids (0.01, 0.1, 1 and 10 µg/ml). In experiments involving pretreatment with the TLR8 inhibitor CU-CPT9a (3 μΜ), neutrophils were then stimulated with 10 µg/ml of nucleic acids (RNA or DNA). Data are expressed as means ± standard deviations from three independent experiments, each performed in duplicate. *p < 0.05, as determined by the Mann-Whitney test.

Article Snippet: The contribution of FPRs and TLRs to IL-8 and ROS production induced by live bacteria was evaluated by pretreating neutrophils for 1 hour at 37 °C in 5% CO 2 with the selective FPR2 antagonist WRW4 (Abcam), the pan-FPR inhibitor Boc-2 (GenScript), the selective TLR8 inhibitor CU-CPT9a (InvivoGen) or the selective TLR2 inhibitor TL2-C29 (InvivoGen) at the indicated concentrations prior to stimulation with heat-killed (HK) or live bacteria.

Techniques: MANN-WHITNEY

Journal: FASEB BioAdvances

Article Title: A TLR8 Variant Identified From Whole Exome Sequencing as a Sepsis‐Prone Mutation

doi: 10.1096/fba.2026-00049

Figure Lengend Snippet: Correlation heatmap of TLR8 SNP rs3764880 with demographic and clinical parameters.

Article Snippet: TLR8 knockout cells were generated using CRISPR‐Cas9 targeting the sg_1 guide RNA (Figure ), followed by bulk RNA sequencing after ssRNA stimulation with ssRNA40/LyoVec, a known TLR8 agonist [ ], that has been shown to be a weak TLR7 and strong TLR8 agonist when tested using cell lines expressing either human or mouse TLR7 or TLR8 ( https://www.invivogen.com/ssrna40‐lv ).

Techniques:

Journal: FASEB BioAdvances

Article Title: A TLR8 Variant Identified From Whole Exome Sequencing as a Sepsis‐Prone Mutation

doi: 10.1096/fba.2026-00049

Figure Lengend Snippet: Single‐cell RNA‐seq analysis of PBMCs in sepsis patients and healthy donors. (a) Schematic of the experimental design for single‐cell RNA sequencing of PBMCs. (b) UMAP visualization of all immune cell types within PBMCs. (c) UMAP plot showing monocyte subclusters across study conditions (severe sepsis, mild sepsis, and healthy donors). (d) Dot plot of canonical marker genes defining the three monocyte subtypes. (e) Dot plot showing TLR8 expression across monocyte subtypes. (f) Dot plot showing TLR8 expression across the three study conditions. (g) Violin plot illustrating type I interferon signaling scores across monocyte subtypes. One‐way ANOVA with Bonferroni post hoc analysis was used. **** p < 0.0001. (h) Table summarizing the clinical demographics of patients with the TLR8 rs3764880 polymorphism.

Article Snippet: TLR8 knockout cells were generated using CRISPR‐Cas9 targeting the sg_1 guide RNA (Figure ), followed by bulk RNA sequencing after ssRNA stimulation with ssRNA40/LyoVec, a known TLR8 agonist [ ], that has been shown to be a weak TLR7 and strong TLR8 agonist when tested using cell lines expressing either human or mouse TLR7 or TLR8 ( https://www.invivogen.com/ssrna40‐lv ).

Techniques: Single Cell, RNA Sequencing, Marker, Expressing

Journal: FASEB BioAdvances

Article Title: A TLR8 Variant Identified From Whole Exome Sequencing as a Sepsis‐Prone Mutation

doi: 10.1096/fba.2026-00049

Figure Lengend Snippet: Enrichment of inflammatory signaling pathways in TLR8 high and TLR8 low monocytes. Violin plots show enrichment scores for interferon, IL‐1, IL‐4, IL‐6, IL‐10, and IL‐17 signaling pathways. Student t test was performed. * p < 0.05.

Article Snippet: TLR8 knockout cells were generated using CRISPR‐Cas9 targeting the sg_1 guide RNA (Figure ), followed by bulk RNA sequencing after ssRNA stimulation with ssRNA40/LyoVec, a known TLR8 agonist [ ], that has been shown to be a weak TLR7 and strong TLR8 agonist when tested using cell lines expressing either human or mouse TLR7 or TLR8 ( https://www.invivogen.com/ssrna40‐lv ).

Techniques: Protein-Protein interactions

Journal: FASEB BioAdvances

Article Title: A TLR8 Variant Identified From Whole Exome Sequencing as a Sepsis‐Prone Mutation

doi: 10.1096/fba.2026-00049

Figure Lengend Snippet: The role of the TLR8 SNP variant (rs3764880) in IFN‐β production. (a) Western blot showing TLR8 knockout in THP‐1 monocyte cells using CRISPR‐Cas9. (b) Dot plot showing expression patterns of IRF7 and CREBBP, key regulators of the TLR8–IFN‐β axis, across TLR8high and TLR8low monocyte subsets (P value < 0.001). (c) IFN‐β production by THP‐1 cells stimulated with the TLR8 agonist ssRNA40 (10 μg/mL); n = 4 per group. Student t test was performed. *** p < 0.001. (d) PCR‐RFLP assay detecting the presence of the TLR8 SNP variant (rs3764880) in whole blood from healthy volunteers. (e) IFN‐β production upon ssRNA40 (10 μg/mL) stimulation in carriers vs. non‐carriers of the rs3764880 variant; n = 3 per group. Student t test was performed. * p < 0.05.

Article Snippet: TLR8 knockout cells were generated using CRISPR‐Cas9 targeting the sg_1 guide RNA (Figure ), followed by bulk RNA sequencing after ssRNA stimulation with ssRNA40/LyoVec, a known TLR8 agonist [ ], that has been shown to be a weak TLR7 and strong TLR8 agonist when tested using cell lines expressing either human or mouse TLR7 or TLR8 ( https://www.invivogen.com/ssrna40‐lv ).

Techniques: Variant Assay, Western Blot, Knock-Out, CRISPR, Expressing, RFLP Assay

Journal: FASEB BioAdvances

Article Title: A TLR8 Variant Identified From Whole Exome Sequencing as a Sepsis‐Prone Mutation

doi: 10.1096/fba.2026-00049

Figure Lengend Snippet: Bulk RNA‐seq analysis of ssRNA‐stimulated WT and TLR8‐knockout THP‐1 monocytes. (a) Volcano plot of differentially expressed genes (DEGs) in ssRNA‐stimulated THP‐1 monocytes compared with unstimulated controls. (b) Volcano plot of DEGs in ssRNA‐stimulated TLR8‐knockout THP‐1 monocytes relative to stimulated wild‐type controls. (c) Gene Ontology enrichment analysis of upregulated genes in ssRNA‐stimulated THP‐1 monocytes. (d) Gene Ontology enrichment analysis of downregulated genes in ssRNA‐stimulated THP‐1 monocytes. (e) Table of interferon‐stimulated genes (ISGs) in ssRNA‐stimulated TLR8‐knockout THP‐1 monocytes relative to stimulated wild‐type controls.

Article Snippet: TLR8 knockout cells were generated using CRISPR‐Cas9 targeting the sg_1 guide RNA (Figure ), followed by bulk RNA sequencing after ssRNA stimulation with ssRNA40/LyoVec, a known TLR8 agonist [ ], that has been shown to be a weak TLR7 and strong TLR8 agonist when tested using cell lines expressing either human or mouse TLR7 or TLR8 ( https://www.invivogen.com/ssrna40‐lv ).

Techniques: RNA Sequencing, Knock-Out

Journal: FASEB BioAdvances

Article Title: A TLR8 Variant Identified From Whole Exome Sequencing as a Sepsis‐Prone Mutation

doi: 10.1096/fba.2026-00049

Figure Lengend Snippet: Proposed mechanistic model linking the TLR8 rs3764880 variant to enhanced interferon responses in sepsis. (A) In the wild‐type allele (A), translation initiation occurs at the canonical AUG start codon in exon 1 of the TLR8 gene (exons 1–3). Under these conditions, the TLR8v2 isoform is predominantly produced. TLR8v2 lacks exon 1 in the mature transcript and contains exons 2–3. (B) The rs3764880 variant (A → G; Met1Val) alters the canonical translation initiation site in exon 1 (AUG → GUG), modifying translation initiation and favoring production of the TLR8v1 isoform. TLR8v1 contains exons 1–3, resulting in a shift in isoform balance toward increased TLR8v1 relative to TLR8v2. (C) Upon stimulation with bacterial single‐stranded RNA (ssRNA), TLR8 signaling is activated, particularly in monocyte populations enriched for TLR8 expression such as non‐classical monocytes. In this model, the rs3764880‐associated shift toward the TLR8v1 isoform is proposed to enhance downstream TLR8 signaling, leading to increased production of interferon‐β (IFN‐β) and amplified innate immune responses.

Article Snippet: TLR8 knockout cells were generated using CRISPR‐Cas9 targeting the sg_1 guide RNA (Figure ), followed by bulk RNA sequencing after ssRNA stimulation with ssRNA40/LyoVec, a known TLR8 agonist [ ], that has been shown to be a weak TLR7 and strong TLR8 agonist when tested using cell lines expressing either human or mouse TLR7 or TLR8 ( https://www.invivogen.com/ssrna40‐lv ).

Techniques: Variant Assay, Produced, Expressing, Amplification

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: Dynamics of immune cell subsets in response to agonist stimulation. A Schematic of the experimental workflow. Immune profiling by flow cytometry was performed 48 h after a single intratumoral injection of PRR agonists. B Percentage of CD45 + leukocytes among live cells following intratumoral administration of different agonists: TLR3 agonist poly IC (25 µg/tumor), TLR7 agonist imiquimod hydrochloride (25 µg/tumor), TLR8 agonist TL8-506 (10 µg/tumor), TLR9 agonist CpG ODN 2395 (50 µg/tumor), STING agonist ADU-S100 ammonium salt (25 µg/tumor), NOD1 agonist Tri-DAP (10 µg/tumor), and NOD2 agonist murabutide (5 µg/tumor). C Proportion of cDCs within the CD45 + population. D Percentage of ZsGreen + cells among DCs. E Proportion of macrophages within the CD45 + population. F Percentage of ZsGreen + cells among macrophages. G Proportion of CD8 + T cells within the CD45 + compartment. H Proportion of cCD4 + T cells within the CD45 + compartment. I Proportion of Tregs within the CD45 + compartment, pooled from two independent experiments. J Ratio of cCD4 + T cells to Tregs. (K) Ratio of CD8 + T cells to Tregs. Data represent the mean ± SEM ( n = 8 mice per group). Statistical comparisons were performed using one-way ANOVA. * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001

Article Snippet: When tumors reached approximately 200 mm 3 , PRR agonists were administered via intratumoral injection at the following doses: TLR3 agonist poly(I:C) (Sigma-Aldrich, #42,424–50-0; 25 μg/tumor); TLR7 agonist imiquimod hydrochloride (MedChemExpress, #HY-B0180A; 25 μg/tumor); TLR8 agonist TL8-506 (InvivoGen, #tlrl-tl8506; 10 μg/tumor); TLR8 agonist motolimod (MedChemExpress, #HY-13773; 50 μg/tumor); TLR9 agonist CpG ODN 2395 (Class C) (InvivoGen, #tlrl-2395–1; 50 μg/tumor); STING agonist ADU-S100 ammonium salt (MedChemExpress, #HY-12885B; 25 μg/tumor); NOD1 agonist Tri-DAP (InvivoGen, #tlrl-tdap; 10 μg/tumor); and NOD2 agonist murabutide (InvivoGen, #tlrl-mbt; 5 μg/tumor).

Techniques: Flow Cytometry, Injection

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: Immune modulation and antitumor effects of TLR8 agonists. A Flow cytometric analysis of CD45 + leukocytes was performed 48 h after three intratumoral injections of TL8-506 (10 µg/tumor) or motolimod (50 µg/tumor) ( n = 6). B Percentage of cDCs within the CD45 + population and proportion of ZsGreen + cells among cDCs ( n = 6). C Percentage of macrophages within the CD45 + population and proportion of ZsGreen + cells among macrophages ( n = 6). D Frequency of cCD4 + T cells, CD8 + T cells, and Tregs within the CD45 + compartment ( n = 6). E Ratios of cCD4 + T cells to Tregs and CD8 T cells to Tregs ( n = 6). F , G Tumor growth curves and terminal tumor weights of AKR and MC38 tumors in C57BL/6 mice administered TL8-506 or motolimod every 3 days ( n = 5). H , I Tumor growth curves and terminal tumor weights of AKR and MC38 tumors in nude mice administered motolimod every three days ( n = 5). J , K Macrophage phagocytosis of CFSE-labeled tumor cells following motolimod stimulation ( n = 5). Data represent the mean ± SEM. Data in panels A–G were analyzed using one-way ANOVA, and data in panels H–K were analyzed using Student’s t-tests. * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001

Article Snippet: When tumors reached approximately 200 mm 3 , PRR agonists were administered via intratumoral injection at the following doses: TLR3 agonist poly(I:C) (Sigma-Aldrich, #42,424–50-0; 25 μg/tumor); TLR7 agonist imiquimod hydrochloride (MedChemExpress, #HY-B0180A; 25 μg/tumor); TLR8 agonist TL8-506 (InvivoGen, #tlrl-tl8506; 10 μg/tumor); TLR8 agonist motolimod (MedChemExpress, #HY-13773; 50 μg/tumor); TLR9 agonist CpG ODN 2395 (Class C) (InvivoGen, #tlrl-2395–1; 50 μg/tumor); STING agonist ADU-S100 ammonium salt (MedChemExpress, #HY-12885B; 25 μg/tumor); NOD1 agonist Tri-DAP (InvivoGen, #tlrl-tdap; 10 μg/tumor); and NOD2 agonist murabutide (InvivoGen, #tlrl-mbt; 5 μg/tumor).

Techniques: Labeling

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: When tumors reached approximately 200 mm 3 , PRR agonists were administered via intratumoral injection at the following doses: TLR3 agonist poly(I:C) (Sigma-Aldrich, #42,424–50-0; 25 μg/tumor); TLR7 agonist imiquimod hydrochloride (MedChemExpress, #HY-B0180A; 25 μg/tumor); TLR8 agonist TL8-506 (InvivoGen, #tlrl-tl8506; 10 μg/tumor); TLR8 agonist motolimod (MedChemExpress, #HY-13773; 50 μg/tumor); TLR9 agonist CpG ODN 2395 (Class C) (InvivoGen, #tlrl-2395–1; 50 μg/tumor); STING agonist ADU-S100 ammonium salt (MedChemExpress, #HY-12885B; 25 μg/tumor); NOD1 agonist Tri-DAP (InvivoGen, #tlrl-tdap; 10 μg/tumor); and NOD2 agonist murabutide (InvivoGen, #tlrl-mbt; 5 μg/tumor).

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: When tumors reached approximately 200 mm 3 , PRR agonists were administered via intratumoral injection at the following doses: TLR3 agonist poly(I:C) (Sigma-Aldrich, #42,424–50-0; 25 μg/tumor); TLR7 agonist imiquimod hydrochloride (MedChemExpress, #HY-B0180A; 25 μg/tumor); TLR8 agonist TL8-506 (InvivoGen, #tlrl-tl8506; 10 μg/tumor); TLR8 agonist motolimod (MedChemExpress, #HY-13773; 50 μg/tumor); TLR9 agonist CpG ODN 2395 (Class C) (InvivoGen, #tlrl-2395–1; 50 μg/tumor); STING agonist ADU-S100 ammonium salt (MedChemExpress, #HY-12885B; 25 μg/tumor); NOD1 agonist Tri-DAP (InvivoGen, #tlrl-tdap; 10 μg/tumor); and NOD2 agonist murabutide (InvivoGen, #tlrl-mbt; 5 μg/tumor).

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: TLR8 agonist enhances the efficacy of anti-PD-1 therapy. A Experimental workflow showing treatment schedule for AKR tumor-bearing C57BL/6 mice receiving intratumoral motolimod and intraperitoneal anti-PD-1 (αPD-1) antibody, alone or in combination. B , C Tumor growth curves and survival analysis of AKR tumor-bearing mice treated with the indicated regimens ( n = 7 mice per group). D Experimental workflow showing treatment schedule and details for MC38 tumors in C57BL/6 mice receiving intratumoral motolimod and intraperitoneal αPD-1 antibody, alone or in combination. E , F Tumor growth curves and survival analysis of MC38 tumor-bearing mice treated with the indicated regimens ( n = 7 mice per group). Data represent the mean ± SEM. Tumor growth curves were analyzed using one-way ANOVA, and survival curves were analyzed using log-rank testing. * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001

Article Snippet: When tumors reached approximately 200 mm 3 , PRR agonists were administered via intratumoral injection at the following doses: TLR3 agonist poly(I:C) (Sigma-Aldrich, #42,424–50-0; 25 μg/tumor); TLR7 agonist imiquimod hydrochloride (MedChemExpress, #HY-B0180A; 25 μg/tumor); TLR8 agonist TL8-506 (InvivoGen, #tlrl-tl8506; 10 μg/tumor); TLR8 agonist motolimod (MedChemExpress, #HY-13773; 50 μg/tumor); TLR9 agonist CpG ODN 2395 (Class C) (InvivoGen, #tlrl-2395–1; 50 μg/tumor); STING agonist ADU-S100 ammonium salt (MedChemExpress, #HY-12885B; 25 μg/tumor); NOD1 agonist Tri-DAP (InvivoGen, #tlrl-tdap; 10 μg/tumor); and NOD2 agonist murabutide (InvivoGen, #tlrl-mbt; 5 μg/tumor).

Techniques: