Journal: Molecular Therapy. Nucleic Acids

Article Title: Hemoglobin inhibits fibroblast-to-cardiomyocyte reprogramming via TLR2/TLR4-dependent chromatin compaction

doi: 10.1016/j.omtn.2026.102900

Figure Lengend Snippet: Hemoglobin activates TLR signaling in cardiac fibroblasts (A) Cardiac fibroblasts were incubated with Hb (5 mg/mL) for the indicated times. Protein extracts were analyzed by immunoblotting for p-NFκB and p-MAPK. Gapdh was used as a loading control. Representative immunoblots are shown on the left-hand side. Quantification was performed by normalizing p-NFκB and p-MAPK band densities with those of the loading control. N = 4. One-sample t tests were used to compare groups to the control group (∗∗∗ p < 0.001, ∗∗ p < 0.01, ∗ p < 0.05; ns, not significant). (B) Cardiac fibroblasts were incubated either vehicle or with varying concentrations of a TLR4 pharmacological inhibitor for 3 h. Hb (5 mg/mL) was then added. After 1 h, protein extracts were isolated and subsequently analyzed by immunoblotting for p-NFκB. Gapdh was used as a loading control. Representative immunoblots are shown on the left-hand side. Quantification was performed by normalizing p-NFκB band densities with those of the loading control. N = 4. One-sample t tests were used to compare groups to the control group (∗ p < 0.05; ns, not significant). (C) Cardiac fibroblasts were incubated with either LPS or Hb for 24 h, after which expression of the indicated pro-inflammatory cytokines was determined by qPCR. Expression values are shown relative to the housekeeping gene Gapdh. N = 6. ANOVA with Tukey post-hoc tests were used to determine significance (∗∗∗ p < 0.001, ∗∗ p < 0.01, ∗ p < 0.05; ns, not significant).

Article Snippet: Hb (Millipore Sigma, H2625), TLR2 signaling inhibitor-TL2-C29 (InvivoGen, catalog no. inh-c29), and TLR4 inhibitor-CLI-095 (InvivoGen, catalog no. tlrl-cli95-4) were used.

Techniques: Incubation, Western Blot, Control, Isolation, Expressing

Journal: Molecular Therapy. Nucleic Acids

Article Title: Hemoglobin inhibits fibroblast-to-cardiomyocyte reprogramming via TLR2/TLR4-dependent chromatin compaction

doi: 10.1016/j.omtn.2026.102900

Figure Lengend Snippet: Hemoglobin mediates gene repression through TLR2 and TLR4 (A and B) A study was conducted to determine the effect of hemoglobin (Hb) on (A) fibroblast-to-cardiomyocyte reprogramming and (B) fibroblast gene expression. With respect to fibroblast-to-cardiomyocyte reprogramming, cardiac fibroblasts were transfected with either miR combo or a non-targeting control miR. 24 h later, the cells were incubated with vehicle, a TLR2 pharmacological inhibitor, a TLR4 pharmacological inhibitor, or a combination of both pharmacological inhibitors for 3 h. After incubation with the indicated pharmacological inhibitors, Hb was added (5 mg/mL) to the media. All media was replaced the next day. Fourteen days after miR transfection, cells were analyzed for expression of the indicated cardiomyocyte specific genes by qPCR. Expression values were normalized to the housekeeping gene Gapdh. N = 6–10. One-sample t tests were used to compare groups to the control group (∗∗∗ p < 0.001, ∗∗ p < 0.01, ∗ p < 0.05; ns, not significant). t test was used to determine the significance between the miR combo groups (## p < 0.01, # p < 0.05; ns, not significant). With respect to fibroblast gene expression, cardiac fibroblasts were incubated with vehicle, a TLR2 pharmacological inhibitor, a TLR4 pharmacological inhibitor, or a combination of both pharmacological inhibitors for 3 h. After incubation with the indicated pharmacological inhibitors, Hb was added (5 mg/mL) to the media. All media was replaced the next day. Expression of the indicated fibroblast-specific genes was determined by qPCR and normalized to the housekeeping gene Gapdh. N = 10. One-sample t tests were used to compare groups to the control group (∗∗∗ p < 0.001, ∗∗ p < 0.01, ∗ p < 0.05; ns, not significant).

Article Snippet: Hb (Millipore Sigma, H2625), TLR2 signaling inhibitor-TL2-C29 (InvivoGen, catalog no. inh-c29), and TLR4 inhibitor-CLI-095 (InvivoGen, catalog no. tlrl-cli95-4) were used.

Techniques: Gene Expression, Transfection, Control, Incubation, Expressing

Journal: Journal of Translational Autoimmunity

Article Title: Type I interferon-dependent and -independent signaling underlie autoantibody production in a murine lupus model

doi: 10.1016/j.jtauto.2026.100351

Figure Lengend Snippet: Induction of SLE autoantibodies occurs in the presence of LPS or Poly I:C, but not other TLR ligands. Wild type (WT) C57BL/6 mice received five immunizations with β2GPI in combination with one of the following TLR ligands: TLR2 : PAM2, FSL-1 (lipoprotein); TLR3 : Poly I:C (dsRNA); TLR4 : LPS; TLR7: IMQ (imiquimod); or TLR9 : CpG-ODN (ssDNA). Serum autoantibodies were detected by ELISA. Bars indicate the mean value +/standard error (S.E.) per group (n = 5 mice/group), and dots indicate values for individual mice. Autoantibody levels for control mice immunized with PBS and TLR ligand were OD 405 < 0.1. The OD 405 + 3 S.D. for unimmunized mice was ≤0.1 for all autoantibodies assayed. (A) Anti-β2GPI and anti-CL antibodies, and (B) hallmark SLE autoantibodies (anti-DNA, anti-Ro/SS-A, anti-La/SS-B, and anti-Sm) were analyzed statistically in separate groupings. For each of the two autoantibody categories, each different TLR ligand (immunized with β2GPI) was compared to the other TLR ligands by 2-way ANOVA, followed by a Tukey's multiple comparison test. ns = not significant. ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001, and ∗∗∗∗P < 0.0001.

Article Snippet: TLR2 −/− , TLR4 −/− , and MyD88 −/− mice, which originated from Dr. Shizuo Akira (Osaka University, Japan) and were obtained from Dr. Michel Chignard (Pasteur Institute, France), were provided by SQ.

Techniques: Enzyme-linked Immunosorbent Assay, Control, Comparison

Journal: The Cell Surface

Article Title: Pap1 is an adhesin involved in the interaction of Sporothrix schenckii and Sporothrix brasiliensis with the host

doi: 10.1016/j.tcsw.2025.100164

Figure Lengend Snippet: Cytokine stimulation by human peripheral blood mononuclear cells. Yeast-like cells and human peripheral blood mononuclear cells were coincubated for 24 h, and the secreted cytokines were quantified by ELISA. Panels A , C , and E are results generated with S. schenckii yeast-like cells (WT, 1099–18 ATCC MYA 4821 strain), while panels B , D , and F are results obtained with S. brasiliensis strains (WT, 5110 ATCC MYA 4823 strain). None, human cells preincubated with 5 μg mL −1 polymyxin B; Anti-MR, human cells preincubated with 5 μg mL −1 polymyxin B and 10 μg mL −1 anti-mannose receptor antibody. Anti-TLR4, human cells preincubated with 5 μg mL −1 polymyxin B and 10 μg mL −1 anti-TLR4. Data are means ± SD obtained with samples from eight donors, each assayed in duplicate wells. Results were analyzed with Dunnett's test and then the Mann-Whitney U test. * P < 0.05 when compared to strains WT, HSS67, or HSS68 (panels A , C , and E ). * P < 0.05 when compared to strains WT, HSB28, or HSB29 (panels B , D , and F ). † P < 0.05 when compared to the “None” group from the same strain.

Article Snippet: When required, the human cells were pre-incubated for 1 h at 37 °C and 5 % (v/v) CO 2 with one of the following immune receptor antagonists: 10 μg mL −1 of anti-mannose receptor (MR) (Thermo-Fisher Scientific, MA5–44033), or 10 μg mL −1 anti-TLR4 antibody (Santa Cruz Biotechnology, Dallas, TX, USA sc-293,072).

Techniques: Enzyme-linked Immunosorbent Assay, Generated, MANN-WHITNEY

Journal: The Cell Surface

Article Title: Pap1 is an adhesin involved in the interaction of Sporothrix schenckii and Sporothrix brasiliensis with the host

doi: 10.1016/j.tcsw.2025.100164

Figure Lengend Snippet: Phagocytosis of Sporothrix schenckii and Sporothrix brasiliensis PAP1 -silenced strains. Yeast-like cells were labeled with Acridine Orange and used to interact with human monocyte-derived macrophages for 2 h at 37 °C and 5 % (v/v) CO 2 . Then, macrophages were collected and analyzed by flow cytometry. Macrophages that were interacting with at least one red fluorescent yeast-like cell were included in the analysis. None, human cells preincubated with 5 μg mL-1 polymyxin B. Anti-MR, human cells preincubated with 5 μg mL −1 polymyxin B and 10 μg mL −1 anti-mannose receptor antibody. Anti-TLR4, human cells preincubated with 5 μg mL −1 polymyxin B and 10 μg mL −1 anti-TLR4. Panel A , results generated with S. schenckii yeast-like cells (WT, 1099–18 ATCC MYA 4821 strain); while in panel B are results obtained with S. brasiliensis strains (WT, 5110 ATCC MYA 4823 strain). Data are means ± SD obtained with samples from eight donors, each assayed in duplicate wells. Results were analyzed with Dunnett's test and then the Mann-Whitney U test. * P < 0.05 when compared to strains WT, HSS67, or HSS68 (panel A ). * P < 0.05 when compared to strains WT, HSB28, or HSB29 (panel B ). † P < 0.05 when compared to the “None” group from the same strain. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: When required, the human cells were pre-incubated for 1 h at 37 °C and 5 % (v/v) CO 2 with one of the following immune receptor antagonists: 10 μg mL −1 of anti-mannose receptor (MR) (Thermo-Fisher Scientific, MA5–44033), or 10 μg mL −1 anti-TLR4 antibody (Santa Cruz Biotechnology, Dallas, TX, USA sc-293,072).

Techniques: Labeling, Derivative Assay, Flow Cytometry, Generated, MANN-WHITNEY

Journal: The Cell Surface

Article Title: Pap1 is an adhesin involved in the interaction of Sporothrix schenckii and Sporothrix brasiliensis with the host

doi: 10.1016/j.tcsw.2025.100164

Figure Lengend Snippet: Cytokine stimulation by human monocyte-derived macrophages. Yeast-like cells and monocyte-derived macrophages were coincubated for 24 h, and the secreted cytokines were quantified by ELISA. Panels A and C are results generated with S. schenckii yeast-like cells (WT, 1099–18 ATCC MYA 4821 strain); while panels B and D are results obtained with S. brasiliensis strains (WT, 5110 ATCC MYA 4823 strain). None, human cells preincubated with 5 μg mL −1 polymyxin B. Anti-MR, human cells preincubated with 5 μg mL −1 polymyxin B and 10 μg mL −1 anti-mannose receptor antibody. Anti-TLR4, human cells preincubated with 5 μg mL −1 polymyxin B and 10 μg mL −1 anti-TLR4. Data are means ± SD obtained with samples from eight donors, each assayed in duplicate wells. Results were analyzed with Dunnett's test and then the Mann-Whitney U test. * P < 0.05 when compared to strains WT, HSS67, or HSS68 (panels A and C ). * P < 0.05 when compared to strains WT, HSB28, or HSB29 (panels B and D ). cells. † P < 0.05 when compared to the “None” group from the same strain.

Article Snippet: When required, the human cells were pre-incubated for 1 h at 37 °C and 5 % (v/v) CO 2 with one of the following immune receptor antagonists: 10 μg mL −1 of anti-mannose receptor (MR) (Thermo-Fisher Scientific, MA5–44033), or 10 μg mL −1 anti-TLR4 antibody (Santa Cruz Biotechnology, Dallas, TX, USA sc-293,072).

Techniques: Derivative Assay, Enzyme-linked Immunosorbent Assay, Generated, MANN-WHITNEY

Journal: Cell Reports Medicine

Article Title: Taxane chemotherapy promotes response to TIM-3 checkpoint blockade via STING-mediated ER stress and HMGB1 secretion

doi: 10.1016/j.xcrm.2026.102788

Figure Lengend Snippet: Active secretion of HMGB1 during taxane treatment is dependent upon TLR4 (A) Flow cytometric detection of intracellular HMGB1 within live PyMT-B6 cells 24 h post-chemotherapy treatment. Data reflect mean ± SD of three biological replicates (right), with significance determined by one-way ANOVA with Sidak’s test for multiple comparisons, shown as ∗ p < 0.1 and ∗∗∗ p < 0.001. One of three representative experiments shown. (B) Confocal microscopy images (left) displaying HMGB1 (green) and DNA (DAPI, blue) in PyMT-B6 cells pretreated with leptomycin B or cytochalasin B for 1 h prior to DTX treatment. Scale bars, 25 μM. Quantification of nuclear HMGB1 is shown on the right. (C) Confocal microscopy images (left) displaying extracellular staining of LAMP1 (CD107a, orange) and DNA (DAPI, blue) in PyMT-B6 cells pretreated with cytochalasin B for 1 h prior to DTX treatment. Scale bars, 25 μM. Quantification of extracellular LAMP1 is shown on the right. (D) Confocal microscopy images (left) displaying HMGB1 (green) and DNA (DAPI, blue) in PyMT-B6 cells pretreated with TLR4 inhibitor ( M62812 ) for 1 h prior to DTX treatment. Scale bars, 10 μM. Quantification of nuclear HMGB1 is shown on the right. For (B–D), quantification reflects the MFI ± SD from individual cells in 2–3 images, with significance determined by one-way ANOVA with Sidak’s test for multiple comparisons, shown as ∗∗∗ p < 0.001. One of three representative experiments shown. (E) Extracellular flow cytometric detection of LAMP1 in PyMT-B6 cells pretreated with a TLR4 inhibitor ( M62812 ) for 1 h prior to DTX treatment. One of three independent experiments shown. Quantification reflects the MFI ±SD, with significance determined by one-way ANOVA with Sidak’s test for multiple comparisons, shown as ∗ p < 0.05. (F) Flow cytometric analysis of TLR4 expression in non-targeted control (NTC) and Tlr4 -deficient PyMT-B6 cells (top). Flow cytometric detection of nuclear HMGB1 in TLR4-proficient versus TLR4-deficient PyMT-B6 cells following PTX treatment (bottom). One of three independent experiments shown. Quantification reflects the MFI ± SD, with significance determined by one-way ANOVA with Sidak’s test for multiple comparisons, shown as ∗∗∗ p < 0.001. (G) Percentage change in tumor volume in mice bearing orthotopic Tlr4 -deficient PyMT-B6 tumors treated with PTX and either IgG 2a isotype control or αTIM-3. Treatment was initiated when tumors reached ∼50–100 mm 3 (day 0). One of two representative experiments shown with n = 5 mice per group. Data displayed mean ± SEM with significance determined by two-way ANOVA, shown as ∗∗ p < 0.01.

Article Snippet: M62812 TLR4 inhibitor , Fisher Scientific , Cat# 570510.

Techniques: Confocal Microscopy, Staining, Expressing, Control

Journal: Cell Reports Medicine

Article Title: Taxane chemotherapy promotes response to TIM-3 checkpoint blockade via STING-mediated ER stress and HMGB1 secretion

doi: 10.1016/j.xcrm.2026.102788

Figure Lengend Snippet: Taxanes induce ROS-dependent DNA damage and PARP activation (A) Flow cytometric detection of lipid peroxidation in PyMT-B6 cells pretreated with a TLR4 inhibitor ( M62812 ) or Nox2/4 inhibitor (GLX481304) for 1 h prior to DTX treatment. Quantification on the right reflects the MFI ± SD in live cells from three biological replicates, with significance determined by one-way ANOVA with Sidak’s test for multiple comparisons, shown as ∗∗∗ p < 0.001. One of three independent experiments shown. (B) Confocal microscopy images (left) displaying intracellular staining of HMGB1 (green) and DNA (DAPI, blue) in PyMT-B6 cells pretreated with Nox2/4 inhibitor GLX481304 or N-acetyl cysteine (NAC) for 1 h prior to DTX treatment, with quantification of nuclear HMGB1 (right). Scale bars, 25 μM. (C) Confocal microscopy images (left) displaying interaction of poly-ADP ribosylated chains (PAR) and HMGB1 by proximity ligation assay, with a positive signal indicating proximity of both signals less than 40 nm apart (red), along with DNA staining (DAPI, blue) in PyMT-B6 cells pretreated with a TLR4 inhibitor ( M62812 ), NAC, or Nox2/4 inhibitor (GLX481304) for 1 h prior to DTX treatment for 12 h, with quantification of PARylated HMGB1 (right). Scale bars, 25 μM. (D) Confocal microscopy images (left) displaying HMGB1 (green) and DNA (DAPI, blue) in PyMT-B6 cells pretreated with PARP1/2 inhibitors (niraparib, olaparib, or rucaparib) for 1 h prior to DTX treatment for 24 h, with quantification of nuclear HMGB1 (right). Scale bars, 25 μM. For (B–D), quantification reflects the MFI ± SD in individual cells from 2 to 3 images, with significance determined by one-way ANOVA with Sidak’s test for multiple comparisons, shown as ∗∗∗ p < 0.001. One of three independent experiments shown. (E) Intracellular flow cytometric detection (left) of phosphorylated H2AX (γH2AX) in PyMT cells pretreated with a TLR4 inhibitor ( M62812 ) or NAC for 1 h prior to DTX treatment for 24 h. H 2 O 2 was added as a positive control for staining. Data reflect the mean ± SD of three biological replicates (right), with one representative experiment of three shown. Significance determined by one-way ANOVA with Sidak’s test for multiple comparisons, shown as ∗∗ p < 0.01 and ∗∗∗ p < 0.001. (F) Confocal microscopy images (left) displaying cytoplasmic dsDNA (green) and DNA (DAPI, blue) in PyMT-B6 cells treated with DTX for 24 h. Scale bars are 25 μM. Quantification reflects dsDNA MFI ± SD in individual cells from 2 to 3 images, with significance determined by one-way ANOVA with Sidak’s test for multiple comparisons, shown as ∗∗∗ p < 0.001. One of three independent experiments shown.

Article Snippet: M62812 TLR4 inhibitor , Fisher Scientific , Cat# 570510.

Techniques: Activation Assay, Confocal Microscopy, Staining, Proximity Ligation Assay, Positive Control

Journal: Cell Reports Medicine

Article Title: Taxane chemotherapy promotes response to TIM-3 checkpoint blockade via STING-mediated ER stress and HMGB1 secretion

doi: 10.1016/j.xcrm.2026.102788

Figure Lengend Snippet: The cGAS-STING pathway is required for HMGB1 release (A) Confocal microscopy images depicting HMGB1 (green) and DNA (DAPI, blue) in sgNTC, cGas- deficient, and Sting- deficient PyMT-B6 cells treated with DTX. Representative images of one of three independent experiments shown (left). Scale bars are 25 μM. Quantification reflects MFI ± SD in individual cells from 2 to 3 images, with significance determined by t test, shown as ∗∗∗ p < 0.001 (right). (B) Tumor volume in mice bearing orthotopic Cgas- or Sting -deficient PyMT-B6 tumors treated with PTX and either IgG 2a isotype control or αTIM-3. Treatment was initiated when tumors reached ∼50–100 mm 3 (day 0). One of two representative experiments shown with n = 5–10 mice per group as indicated. Data displayed as mean ± SEM, with significance determined by two-way ANOVA, shown as ∗∗ p < 0.01. (C) Relative mRNA levels for Cxcl9 , Ifnb1 , and Ifna1 in PyMT-B6 cells treated with DTX or DMXAA, along with blockade of IFNAR1, as determined by RT-PCR. (D) Release of IFN-β measured by ELISA following treatment with DTX, PTX, or DMXAA. For (C and D), data reflect the mean ± SD of three biological replicates from one of two independent experiments, with significance determined by one-way ANOVA and Sidak’s test for multiple comparisons, shown as ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001. (E) Western blot of phospho-IRF3 (pIRF3) and IRF3 protein present in PyMT-B6 cells exposed to PTX, Tunicamycin, or DMXAA or DTX ± TLR4 inhibitor as indicated. β-actin used as loading control. Molecular weights in kDa are shown to the left. Representative images from one of two independent experiments shown. (F) Confocal microscopy images (left) depicting the colocalization of the Golgi (red), STING (green), and DNA (DAPI, blue) in control, cGas- deficient, and Sting- deficient PyMT-B6 cells treated with DTX. Scale bars, 25 μM. Quantification reflects the colocalization of STING with the Golgi as determined by Mander’s overlap (top right) and the MFI of STING (bottom right). (G) Confocal microscopy images (left) depicting calreticulin (ER marker, red), STING (green), and DNA (DAPI, blue) in control, cGas- deficient, and Sting- deficient PyMT-B6 cells treated with DTX. Scale bars, 25 μM. Quantification reflects the colocalization of STING with the ER as determined by Mander’s overlap (top right) and the MFI of calreticulin (bottom right). For (F) and (G), data reflect the mean ± SD in individual cells from two to three images, with significance determined by t test and shown as ∗∗ p < 0.01, ∗∗∗ p < 0.001, and one of three independent experiments is shown.

Article Snippet: M62812 TLR4 inhibitor , Fisher Scientific , Cat# 570510.

Techniques: Confocal Microscopy, Control, Reverse Transcription Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Western Blot, Marker

Journal: Cell Reports Medicine

Article Title: Taxane chemotherapy promotes response to TIM-3 checkpoint blockade via STING-mediated ER stress and HMGB1 secretion

doi: 10.1016/j.xcrm.2026.102788

Figure Lengend Snippet: STING-dependent ER stress and HMGB1 release in a model of TNBC (A) Confocal microscopy images (left) depicting HMGB1 (green) and DNA (DAPI, blue) in KP1 cells treated with PTX or DTX, with and without TLR4 inhibitor. Scale bars are 25 μM. Representative images of one of three independent experiments shown. Quantification reflects the MFI ± SD in individual cells from 2 to 3 images, with significance determined by t test and shown as ∗∗∗ p < 0.001 (right). (B) Quantification of MFI ± SD in individual cells from two to three confocal microscopy images assaying cytosolic dsDNA post-PTX or -DTX treatment, with significance determined by t test and shown as ∗∗ p < 0.01 and ∗∗∗ p < 0.001. (C) Live cell imaging with ER Tracker Blue-White depicting ER expansion in KP1 cells pretreated with NAC or TLR4 inhibitor for 1 h prior to treatment with DTX. Data reflect the MFI ± SD of three biological replicates. (D) Extracellular flow cytometric detection of LAMP1 on KP1 cells 24 h post-pretreatment with NAC or TLR4 inhibitor for 1 h prior to treatment with DTX. Data reflect the percent positivity ± SD of three biological replicates. For (C and D), significance was determined by one-way ANOVA with Sidak’s test for multiple comparisons, shown as ∗ p < 0.05 and ∗∗∗ p < 0.001. One of three independent experiments shown. (E) Tumor volume in mice bearing orthotopic sgNTC or sg Sting KP1 tumors treated with PTX and either IgG 2a isotype control or αTIM-3. Treatment was initiated when tumors reached ∼50–100 mm 3 (day 0). (F) Tumor volume in mice bearing orthotopic sgNTC or sg Sting KP1 tumors treated with PTX and either IgG 2a isotype control or αPD-1. Treatment was initiated when tumors reached ∼50–100 mm 3 (day 0). (G) Flow cytometric analysis of CD103 + cDC1s and Ly6G + neutrophils in the tumors from (F). Data from one of two independent experiments with n = 5 mice per group. Data shown as mean ± SEM, with significance determined by one-way ANOVA with Sidak’s test for multiple comparisons, shown as ∗ p < 0.05 and ∗∗ p < 0.01. (H) Tumor volume in mice bearing orthotopic sgNTC control or Ifnb1 -deficient KP1 tumors treated with PTX and IgG 2a isotype control or αPD-1. Treatment was initiated when tumors reached ∼50–100 mm 3 (day 0). For (E), (F), and (H), data merged from two independent experiments, with n = 9–10 mice per group. Data shown as mean ± SEM with significance determined by two-way ANOVA displayed with ∗ p < 0.05 and ∗∗ p < 0.01.

Article Snippet: M62812 TLR4 inhibitor , Fisher Scientific , Cat# 570510.

Techniques: Confocal Microscopy, Live Cell Imaging, Control

Journal: Frontiers in Immunology

Article Title: Pharmacological intervention of the HMGB1-pCTS-L axis to ameliorate inflammatory diseases

doi: 10.3389/fimmu.2026.1843251

Figure Lengend Snippet: Divergent modulation of HMGB1 functions through distinct molecular interactions. (A) Interfacial residues of human HMGB1 for interacting with TLR4, RAGE, or TN P2. The binding regions for TLR4 (residues 89–108) and RAGE (residues 150–183) are highlighted in italicized red and blue text, respectively. Key P2-interacting residues are highlighted in non-italicized purple text. (B) Divergent functional outcomes stemming from HMGB1’s interaction with various proteins. HMGB1 engages TLR4 and RAGE receptors, leading to the induction of cytokine/chemokine production and macrophage pyroptosis, respectively. Conversely, some endogenous proteins, such as HP, can bind to HMGB1 to induce anti-inflammatory IL-10. Similarly, TN can sequester HMGB1 to facilitate the endocytosis of HMGB1/TN complexes, and subsequently induce macrophage pyroptosis and immunosuppression. In contrast, a TN-derived P2–1 peptide binds HMGB1 to prevent its interaction with a pro-endocytic receptor, RAGE, thereby suppressing macrophage pyroptosis and HMGB1-mediated pCTS-L upregulation.

Article Snippet: Although clinical trials involving TLR4 ligand antagonists (e.g., LPS mimetics, eritoran) ( ) and early cytokines (e.g., TNF) neutralizing mAbs ( , ) failed to improve sepsis survival, it is crucial to distinguish between targeting general TLR4 activation or early cytokines, and specifically interfering with HMGB1/pCTS-L-TLR4 interactions.

Techniques: Binding Assay, Functional Assay, Derivative Assay