lps  (Novus Biologicals)

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

Journal: Cell Reports

Article Title: Ex vivo model of functioning human lymph node reveals role for innate lymphocytes and stroma in response to vaccine adjuvant

doi: 10.1016/j.celrep.2025.115938

Figure Lengend Snippet: Adjuvant directly activates TLR4 and the NLRP3 inflammasome in tissue monocytes/macrophages (A) Dot plot showing expression of TLR4 signaling components for each cell cluster within whole human LNs. Color indicates relative log-normalized level of expression across clusters and dot size the proportion of each cluster expressing each gene. (B) Volcano plot indicating DEG between LMQ-stimulated (right: upregulated) and control (left: upregulated) conditions in monocytes/macrophages. Genes with log 2 fold-change >0.25 are indicated in green, those with p value < 10 −5 in blue and those fulfilling both criteria in red. (C) Enriched canonical pathways among significantly ( p < 0.05) DEGs, which are up- (orange) and down- (purple) regulated in monocytes/macrophages by adjuvant (LMQ) stimulation. (D) Gene signature scoring (UCell) of the Canonical Inflammasome Complex Gene Ontology gene set according to cell type and sample type (magenta = control, lavender = LMQ-treated). The median UCell scores for each individual donor are indicated. ∗∗ p < 0.05 and ∗ p < 0.5 by two-way ANOVA with Wilcox test and Benjamini Hochberg correction. (E) Concentrations of IL-1β and IL-18 detected in slice culture supernatants after 20 h without (Ctrl) or with LMQ adjuvant (LMQ) from six donors, where each point is the average of three to four slices, ∗ p < 0.05 by Wilcoxon matched-pairs signed rank test. (F) Relative concentration of IL-1β and IL-18 in slice culture supernatants following 20 h stimulation with adjuvant LMQ in the absence or presence of inhibitors TAK242 (TLR4 inhibitor), MCC950 (NLRP3 inhibitor), or a combination of both. Proportion of each secreted cytokine was calculated relative to LMQ stimulation for each donor (indicated by dotted line at 1.0). Data are mean and SEM ( n = 3, each point is the average of three to four slices from one donor). ∗∗ p < 0.01 by mixed-effects analysis with Dunnett’s multiple comparisons test to LMQ alone. See also and and .

Article Snippet: Briefly, liposomes composed of 1,2-dioleoyl- sn -glycero-3-phosphocholine (DOPC, Merck-Avanti, USA) and cholesterol were combined with QS-21 (Desert King International, USA) and synthetic TLR4 ligand 3D-6-acyl-PHAD (3D6AP) (Merck-Avanti, USA).

Techniques: Adjuvant, Expressing, Control, Concentration Assay

Journal: Cell Reports

Article Title: Ex vivo model of functioning human lymph node reveals role for innate lymphocytes and stroma in response to vaccine adjuvant

doi: 10.1016/j.celrep.2025.115938

Figure Lengend Snippet: Indirect activation of ILCs by adjuvant results in downstream signaling to B cells in spatially preserved LN tissue (A) Proportion of each cell type in unstimulated control (magenta) and LMQ-stimulated (lavender) LN slices, each point represents an individual donor. ∗∗ p < 0.005, ∗∗∗ p < 0.0005, ANOVA using a linear model. (B) DEGs in IL2RA + IL22+ ILC3 between LMQ-stimulated (right: upregulated) and control (left: upregulated) conditions. Genes with log 2 fold-change >0.25 and p value < 10 −5 are indicated in red. (C) IL-22 concentration in slice culture supernatants after 20 h culture without (Ctrl) or with LMQ adjuvant (LMQ) from paired donor conditions ( n = 4, each point is the average of two to four slices from one donor), ∗ p < 0.05 by Wilcoxon matched-pairs signed rank test. (D) Proportion of IL2RA + IL22 + ILC3 cells per donor and per condition against IL-22 concentration detected in paired culture supernatants. r 2 value was calculated by simple linear regression. (E) DEGs in CD16 low NK cells as in (B). (F) IFN-γ concentration in slice culture supernatants as in (C). (G) Relative IFN-γ concentration in slice culture supernatants following 20 h stimulation with adjuvant LMQ in the absence or presence of inhibitors TAK242 (TLR4 inhibitor), MCC950 (NLRP3 inhibitor), or a combination of both. Proportion of each secreted cytokine was calculated relative to LMQ stimulation for each donor (indicated by dotted line at 1.0) ( n = 3, each point is the average of two to four slices from one donor). Data are mean and SEM. ∗∗ p < 0.01, ∗ p < 0.05 by mixed-effects analysis with Dunnett’s multiple comparisons test to LMQ alone. (H) Multiplex stained image of whole human LN for NK cells (CD3 − CD56 + , green triangles), macrophages (CD68, magenta triangles), and B cells (CD20, yellow triangles). Middle image scale bar indicates 400 μm. Inset images are zoomed in regions of boxed areas, where scale bars are 50 μm. Images are representative of three individuals. (I) DEGs in B cells as in (B). (J) Enriched canonical pathways among significant ( p < 0.05) DEGs which are up- (orange) and down- (purple) regulated in B cells in response to LMQ stimulation. See also and and .

Article Snippet: Briefly, liposomes composed of 1,2-dioleoyl- sn -glycero-3-phosphocholine (DOPC, Merck-Avanti, USA) and cholesterol were combined with QS-21 (Desert King International, USA) and synthetic TLR4 ligand 3D-6-acyl-PHAD (3D6AP) (Merck-Avanti, USA).

Techniques: Activation Assay, Adjuvant, Control, Concentration Assay, Multiplex Assay, Staining

Journal: Cell Reports

Article Title: Ex vivo model of functioning human lymph node reveals role for innate lymphocytes and stroma in response to vaccine adjuvant

doi: 10.1016/j.celrep.2025.115938

Figure Lengend Snippet: LN non-hematopoietic stromal cells (NHSCs) are primed both directly and indirectly by vaccine adjuvant to mediate inflammatory cell recruitment (A) Proportion of adjuvant-positive cell populations within CD45 − populations of human LN slices after 20 h of culture ( n = 3). Stromal cells were defined as CD45 − CD31 − Pdpn + and endothelial cells as CD45 − CD31 + . Data are mean and SD. (B) DEGs in ECs between LMQ-stimulated (right: upregulated) and control (left: upregulated) LN slices. Genes with log 2 fold-change >0.25 are indicated in green, those with p value < 10 −5 in blue, and those fulfilling both criteria in red. (C) Enriched canonical gene sets among significant ( p < 0.05) DEGs that are up- (orange) and down- (purple) regulated in ECs following adjuvant (LMQ) stimulation. (D) DEGs in stromal cells as in (C). (E) Enriched gene ontology biological processes gene sets in stromal cells as in (C). (F) CXCL8 concentration in slice culture supernatants after 20 h without (Ctrl) or with adjuvant LMQ (LMQ) ( n = 6, each point is the average of three to four slices from one donor), ∗ p < 0.05 by Wilcoxon matched-pairs signed rank test. (G) UMAP plot of sub-clustered NHSCs from human LN slices colored by cluster and predicted stromal cell type. (H) Dot plot showing expression of stromal/endothelial marker genes and TLR4 receptor components for each NHSC cluster. Color indicates relative log-normalized level of expression across clusters and dot size the proportion of each cluster expressing each gene. (I) Left, relative expression of CXCL2 (top) and CXCL8 (bottom) across NHSC clusters, and right, median transcript expression across NHSC cell types in human LN slices without (Ctrl; magenta) or with LMQ adjuvant (LMQ; lavender), each dot represents an individual donor. See also and and .

Article Snippet: Briefly, liposomes composed of 1,2-dioleoyl- sn -glycero-3-phosphocholine (DOPC, Merck-Avanti, USA) and cholesterol were combined with QS-21 (Desert King International, USA) and synthetic TLR4 ligand 3D-6-acyl-PHAD (3D6AP) (Merck-Avanti, USA).

Techniques: Adjuvant, Control, Concentration Assay, Expressing, Marker