Journal: Pharmaceuticals

Article Title: CB2 Receptor Activation Attenuates IL-1β-Induced Inflammatory Transcriptomic Networks in Human Gingival Fibroblasts

doi: 10.3390/ph19091406

Figure Lengend Snippet: HU-308 attenuates IL-1β-induced interleukin expression in human gingival fibroblasts. Human gingival fibroblasts (HGFs) were divided into three experimental groups: untreated Control (black bars), IL-1β (10 ng/mL)-stimulated cells (red bars), and cells treated with IL-1β followed by the selective CB2 receptor agonist HU-308 (10 μM; blue bars). Twenty-four hours after IL-1β stimulation, transcript levels of proinflammatory and anti-inflammatory interleukins were quantified. IL-1β stimulation significantly increased the expression of the pro-inflammatory cytokines IL1B, IL6, IL23A, IL33, IL11, IL1α, and IL24, and treatment with HU-308 attenuated the IL-1β-induced expression of these cytokines, indicating suppression of the inflammatory response ( A – F ). In contrast, IL10 exhibited reduced expression following IL-1β stimulation, and HU-308 treatment partially restored its expression, supporting a shift toward a less inflammatory phenotype following CB2 receptor activation ( G ). Data are presented as mean ± SEM from three independent biological replicates ( n = 3 per group). Statistical significance was determined by one-way ANOVA followed by Tukey’s multiple-comparison test. * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001 was considered statistically significant.

Article Snippet: Recombinant human IL-1β was obtained from (InvivoGen, San Diego, CA, USA), reconstituted in water according to the manufacturer’s instructions, and used at a final concentration of 10 ng/mL.

Techniques: Expressing, Control, Activation Assay, Comparison

Journal: Pharmaceuticals

Article Title: CB2 Receptor Activation Attenuates IL-1β-Induced Inflammatory Transcriptomic Networks in Human Gingival Fibroblasts

doi: 10.3390/ph19091406

Figure Lengend Snippet: HU-308 modulates IL-1β-induced chemokine expression in human gingival fibroblasts. Human gingival fibroblasts (HGFs) were divided into three experimental groups: untreated Control (black bars), IL-1β (10 ng/mL)-stimulated cells (red bars), and cells treated with IL-1β followed by the selective CB2 receptor agonist HU-308 (10 μM; blue bars). Twenty-four hours after IL-1β stimulation, transcript levels of inflammatory chemokines were quantified. IL-1β stimulation significantly increased the expression of CXCL5, CXCL8, CXCL16, CCL20, and CCL28, consistent with activation of chemokine-mediated inflammatory signaling ( A – E ). Treatment with HU-308 attenuated the IL-1β-induced expression of these chemokines, indicating suppression of inflammatory chemokine responses and reduced potential for immune-cell recruitment ( A – E ). Data are presented as mean ± SEM from three independent biological replicates ( n = 3 per group). Statistical significance was determined by one-way ANOVA followed by Tukey’s multiple-comparison test. * p < 0.05, *** p < 0.001, and **** p < 0.0001 was considered statistically significant.

Article Snippet: Recombinant human IL-1β was obtained from (InvivoGen, San Diego, CA, USA), reconstituted in water according to the manufacturer’s instructions, and used at a final concentration of 10 ng/mL.

Techniques: Expressing, Control, Activation Assay, Immune Cell, Comparison

Journal: Pharmaceuticals

Article Title: CB2 Receptor Activation Attenuates IL-1β-Induced Inflammatory Transcriptomic Networks in Human Gingival Fibroblasts

doi: 10.3390/ph19091406

Figure Lengend Snippet: HU-308 attenuates IL-1β-induced matrix metalloproteinase expression in human gingival fibroblasts. Human gingival fibroblasts (HGFs) were divided into three experimental groups: untreated Control (black bars), IL-1β (10 ng/mL)-stimulated cells (red bars), and cells treated with IL-1β followed by the selective CB2 receptor agonist HU-308 (10 μM; blue bars). Twenty-four hours after IL-1β stimulation, transcript levels of MMP1, MMP3, and MMP10 were quantified. IL-1β stimulation significantly increased the expression of all three matrix metalloproteinases, consistent with enhanced extracellular matrix degradation and tissue remodeling associated with periodontal inflammation ( A – C ). Treatment with HU-308 significantly attenuated the IL-1β-induced expression of MMP1, MMP3, and MMP10, suggesting that CB2 receptor modulation suppresses matrix-degrading pathways and may contribute to the preservation of periodontal tissue integrity ( A – C ). Data are presented as mean ± SEM from three independent biological replicates ( n = 3 per group). Statistical significance was determined by one-way ANOVA followed by Tukey’s multiple-comparison test. * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001 was considered statistically significant.

Article Snippet: Recombinant human IL-1β was obtained from (InvivoGen, San Diego, CA, USA), reconstituted in water according to the manufacturer’s instructions, and used at a final concentration of 10 ng/mL.

Techniques: Expressing, Control, Tissue, Preserving, Comparison

Journal: Pharmaceuticals

Article Title: CB2 Receptor Activation Attenuates IL-1β-Induced Inflammatory Transcriptomic Networks in Human Gingival Fibroblasts

doi: 10.3390/ph19091406

Figure Lengend Snippet: HU-308 partially restores IL-1β-induced alterations in collagen gene expression in human gingival fibroblasts. Human gingival fibroblasts (HGFs) were divided into three experimental groups: untreated Control (black bars), IL-1β (10 ng/mL)-stimulated cells (red bars), and cells treated with IL-1β followed by the selective CB2 receptor agonist HU-308 (10 μM; blue bars). Twenty-four hours after IL-1β stimulation, transcript levels of COL1A1, COL3A1, COL11A1, and COL12A1 were quantified. IL-1β stimulation significantly reduced the expression of collagen-associated genes, consistent with impaired extracellular matrix synthesis and tissue remodeling during periodontal inflammation ( A – D ). Treatment with HU-308 partially restored the expression of these collagen transcripts toward control levels, suggesting that CB2 receptor activation may preserve extracellular matrix homeostasis and limit IL-1β-induced connective tissue damage ( A – D ). Data are presented as mean ± SEM from three independent biological replicates ( n = 3 per group). Statistical significance was determined by one-way ANOVA followed by Tukey’s multiple-comparison test. * p < 0.05, ** p < 0.01 and *** p < 0.001 was considered statistically significant.

Article Snippet: Recombinant human IL-1β was obtained from (InvivoGen, San Diego, CA, USA), reconstituted in water according to the manufacturer’s instructions, and used at a final concentration of 10 ng/mL.

Techniques: Gene Expression, Control, Expressing, Tissue, Activation Assay, Comparison

Journal: Pharmaceuticals

Article Title: CB2 Receptor Activation Attenuates IL-1β-Induced Inflammatory Transcriptomic Networks in Human Gingival Fibroblasts

doi: 10.3390/ph19091406

Figure Lengend Snippet: HU-308 modulates IL-1β-induced nitric oxide and oxidative stress-related gene expression in human gingival fibroblasts. Human gingival fibroblasts (HGFs) were divided into three experimental groups: untreated Control (black bars), IL-1β (10 ng/mL)-stimulated cells (red bars), and cells treated with IL-1β followed by the selective CB2 receptor agonist HU-308 (10 μM; blue bars). Twenty-four hours after IL-1β stimulation, transcript levels of NOS3, PRKG1 and PDE5A were quantified. IL-1β stimulation significantly altered the expression of genes involved in nitric oxide signaling, reflecting activation of redox-associated inflammatory pathways ( A – C ). Treatment with HU-308 modulated the IL-1β-induced expression of these genes, partially restoring their expression toward control levels and suggesting that CB2 receptor activation attenuates redox imbalance and nitric oxide signaling dysregulation associated with inflammatory activation ( A – C ). Data are presented as mean ± SEM from three independent biological replicates ( n = 3 per group). Statistical significance was determined by one-way ANOVA followed by Tukey’s multiple-comparison test. * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001 was considered statistically significant.

Article Snippet: Recombinant human IL-1β was obtained from (InvivoGen, San Diego, CA, USA), reconstituted in water according to the manufacturer’s instructions, and used at a final concentration of 10 ng/mL.

Techniques: Gene Expression, Control, Expressing, Activation Assay, Comparison

Journal: Pharmaceuticals

Article Title: CB2 Receptor Activation Attenuates IL-1β-Induced Inflammatory Transcriptomic Networks in Human Gingival Fibroblasts

doi: 10.3390/ph19091406

Figure Lengend Snippet: HU-308 modulates IL-1β-associated changes in solute carrier transporter transcript expression in human gingival fibroblasts. Human gingival fibroblasts (HGFs) were divided into three experimental groups: untreated Control (black bars), IL-1β (10 ng/mL)-stimulated cells (red bars), and cells treated with IL-1β followed by the selective CB2 receptor agonist HU-308 (10 μM; blue bars). Twenty-four hours after IL-1β stimulation, transcript levels of SLC2A1, SLC2A3, SLC2A5, SLC5A13, and SLC2A14 were quantified. IL-1β stimulation significantly altered the expression of SLC2A1, SLC2A3, SLC2A5, SLC2A13, and SLC2A14, and HU-308 treatment significantly modified several of these IL-1β-associated transcriptional responses ( A – E ). These findings demonstrate changes in transporter-associated transcript abundance but do not establish corresponding alterations in substrate uptake, glucose utilization, glycolytic activity, or cellular metabolic flux. Data are presented as mean ± SEM from three independent biological replicates ( n = 3 per group). Statistical significance was determined by one-way ANOVA followed by Tukey’s multiple-comparison test. * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001 was considered statistically significant.

Article Snippet: Recombinant human IL-1β was obtained from (InvivoGen, San Diego, CA, USA), reconstituted in water according to the manufacturer’s instructions, and used at a final concentration of 10 ng/mL.

Techniques: Expressing, Control, Modification, Activity Assay, Comparison

Journal: Pharmaceuticals

Article Title: CB2 Receptor Activation Attenuates IL-1β-Induced Inflammatory Transcriptomic Networks in Human Gingival Fibroblasts

doi: 10.3390/ph19091406

Figure Lengend Snippet: GPCR-related transcript modulation by IL-1β and HU-308 treatment in human gingival fibroblasts. Human gingival fibroblasts (HGFs) were divided into three experimental groups: untreated Control (black bars), IL-1β (10 ng/mL)-stimulated cells (red bars), and cells treated with IL-1β followed by the selective CB2 receptor agonist HU-308 (10 μM; blue bars). Twenty-four hours after IL-1β stimulation, transcript levels of GPR160, GPR180, GPR157, GPR61, GPR173, GPR155, GPR107, GPR75, GPR78, and GPR4 were quantified. IL-1β stimulation significantly altered the expression of multiple GPCR-related genes, including increased expression of GPR160, GPR180, GPR157, GPR61, GPR173, GPR155, GPR107, and GPR75, while reducing the expression of GPR78 and GPR4, indicating inflammatory remodeling of GPCR-associated signaling pathways in human gingival fibroblasts ( A – J ). Treatment with HU-308 modified several IL-1β-associated changes in GPCR-related transcript expression ( A – J ). These changes represent transcriptional associations and should not be interpreted as evidence of altered receptor activity or direct functional cross-talk with CB2. Data are presented as mean ± SEM from three independent biological replicates ( n = 3 per group). Statistical significance was determined by one-way ANOVA followed by Tukey’s multiple-comparison test. * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001 was considered statistically significant.

Article Snippet: Recombinant human IL-1β was obtained from (InvivoGen, San Diego, CA, USA), reconstituted in water according to the manufacturer’s instructions, and used at a final concentration of 10 ng/mL.

Techniques: Control, Expressing, Protein-Protein interactions, Modification, Activity Assay, Functional Assay, Comparison

Journal: Pharmaceuticals

Article Title: CB2 Receptor Activation Attenuates IL-1β-Induced Inflammatory Transcriptomic Networks in Human Gingival Fibroblasts

doi: 10.3390/ph19091406

Figure Lengend Snippet: Schematic summary of the coordinated anti-inflammatory effects of CB2 receptor activation in IL-1β-stimulated human gingival fibroblasts. IL-1β stimulation establishes a pro-inflammatory transcriptional landscape characterized by coordinated upregulation of inflammatory cytokines, chemokines, matrix metalloproteinases (MMPs), glucose transporter (SLC2/GLUT) genes, and multiple G protein-coupled receptor (GPCR)-related transcripts, together with suppression of collagen-associated genes and nitric oxide (NO)/cGMP signaling components. Collectively, these changes promote inflammatory amplification, extracellular matrix degradation, metabolic reprogramming, impaired tissue repair, and disruption of periodontal homeostasis ( left panel ). Pharmacological modulation of the cannabinoid receptor type 2 (CB2) with the selective agonist HU-308 shifts this inflammatory landscape toward a more homeostatic phenotype (center panel). HU-308 broadly attenuates IL-1β-induced inflammatory responses by reducing the expression of pro-inflammatory cytokines, chemokines, MMPs, glucose transporter-related genes, and several GPR-associated transcripts, while restoring collagen gene expression, NO/cGMP signaling, and selected receptor-associated pathways ( right panel ). Upward and downward arrows beneath each biological domain summarize the direction of transcript changes for individual genes following IL-1β stimulation (red arrows) and subsequent HU-308 treatment (blue arrows). Overall, the integrated pathway analysis demonstrates that CB2 modulation functions as an upstream regulator of multiple interconnected biological networks, including inflammatory signaling, extracellular matrix remodeling, redox homeostasis, metabolic adaptation, and GPR-associated signaling, rather than acting on a single molecular pathway, thereby supporting a systems-level mechanism through which CB2 modulation may preserve periodontal tissue homeostasis during inflammation. This figure represents a conceptual summary of selected transcriptional patterns and does not constitute a statistically derived gene-network, pathway-enrichment, or genome-wide differential-expression analysis.

Article Snippet: Recombinant human IL-1β was obtained from (InvivoGen, San Diego, CA, USA), reconstituted in water according to the manufacturer’s instructions, and used at a final concentration of 10 ng/mL.

Techniques: Activation Assay, Amplification, Tissue, Disruption, Expressing, Gene Expression, Analysis, Derivative Assay, Genome Wide, Quantitative Proteomics

Journal: Cell Reports Medicine

Article Title: HIV tri-specific killer engagers directly enhance NK cell function and safely expand NK cells in vivo

doi: 10.1016/j.xcrm.2026.102847

Figure Lengend Snippet: TriKE binds CD16 + NK cells and induces NK cell proliferation and activation (A–C) Schematic of the human VRC (A) and CD4 (B) TriKEs and the rhesusized CD4 (C) TriKE. (D) Whole PBMCs from PWOH (black, n = 3) and PWH (red, n = 3) were incubated with or without TriKE (10 nM), and NK cells (live CD56 + CD3 − ) were assessed for CD16 expression and bound TriKE (anti-His) by flow cytometry. (E–I) Whole PBMCs from PWOH (black, n = 5) and PWH (red, n = 5) were CellTrace labeled and cultured with listed treatments for 7 days, and (E) CD56 + CD3 − NK cell proliferation (CTB dilution) and activation by CD38 (F), HLA-DR (G), CD69 (H), and CD57 (I) was assessed by flow cytometry. NT, no treatment, IL-15 (10 nM), TriKEs (10 nM). Statistical assessment was performed with a matched repeated measures two-way ANOVA with a Geisser-Greenhouse correction. Multiple comparisons were controlled for by a Dunnett’s multiple comparisons test. Each dot represents a unique donor, and error bars represent mean ± SEM.

Article Snippet: To quantify biological activity of the recombinant human (rH) and TriKE contained IL-15, HEK-Blue IL-2/IL-15 cells (InvivoGen, Cat# hkb-il2-2) were utilized as reporter cells.

Techniques: Activation Assay, Incubation, Expressing, Flow Cytometry, Labeling, Cell Culture

Journal: Cell Reports Medicine

Article Title: HIV tri-specific killer engagers directly enhance NK cell function and safely expand NK cells in vivo

doi: 10.1016/j.xcrm.2026.102847

Figure Lengend Snippet: HIV TriKEs mediate enhanced NK cell function and killing of HIV-infected cells Primary human PBMCs were co-cultured with CellTrace-labeled HIV-uninfected (A3, blue diamonds), productively infected (8E5, red circles), or latent HIV (ACH-2, green squares) or no targets (PB alone, black triangles) at an E:T of 2:1 with listed treatments. After 5 h, NK cells were evaluated for CD107a (A and B), IFN-γ (C and D), and TNF-α (E and F) expression and killing of target cells (G and H) by flow cytometry. NT, no treatment, IL-15 (10 nM), VRC01 bNab (10 μg/mL), TriKEs (10 nM). Statistical assessment was performed with a matched repeated measures two-way ANOVA with a Geisser-Greenhouse correction. Multiple comparisons were controlled for by a Dunnett’s multiple comparisons test. Data are represented as mean ± SEM. ∗ p < 0.05, ∗∗ p < 0.005, ∗∗∗ p < 0.0005. N = 5 for each PWOH and PWH.

Article Snippet: To quantify biological activity of the recombinant human (rH) and TriKE contained IL-15, HEK-Blue IL-2/IL-15 cells (InvivoGen, Cat# hkb-il2-2) were utilized as reporter cells.

Techniques: Cell Function Assay, Infection, Cell Culture, Labeling, Expressing, Flow Cytometry

Journal: Cell Reports Medicine

Article Title: HIV tri-specific killer engagers directly enhance NK cell function and safely expand NK cells in vivo

doi: 10.1016/j.xcrm.2026.102847

Figure Lengend Snippet: CD38 + CD57 + NK cells exhibit the greatest polyfunctional response mediated by HIV-targeting TriKEs against 8E5 HIV-infected cells Polyfunctional NK cell responses were assessed by co-expression of CD38, CD57, CD107a, IFN-γ, and TNF-α and analyzed using SPICE (Simplified Presentation of Incredibly Complex Evaluations). Whole PBMCs from HIV− (A–E) and HIV+ (F–J) donors were co-cultured with 8E5 target cells at a ratio of 2:1 and either left untreated or treated with 10 nM IL-15 (A and F), 10 μg/mL VRC01 bNAb (B and G), IL-15+bNAb (C and H), 10 nM VRC TriKE (D and I), or 10 nM CD4 TriKE (E and J). A Wilcoxon rank-sum test was run to assess statistical differences between each condition compared to the IL-15 control. (K–N) Bar plots showing polyfunctional NK cell responses (CD107a+IFN-γ+TNF-α+) within NK cell subsets that are CD38 + CD57 + (K), CD38 + CD57 − (L), CD38 − CD57 + (M), and CD38 − CD57 − (N). Each dot represents a unique donor, and error bars represent mean ± SEM. N = 5 for each PWOH and PWH.

Article Snippet: To quantify biological activity of the recombinant human (rH) and TriKE contained IL-15, HEK-Blue IL-2/IL-15 cells (InvivoGen, Cat# hkb-il2-2) were utilized as reporter cells.

Techniques: Infection, Expressing, Cell Culture, Control

Journal: bioRxiv

Article Title: Opposing immunomodulatory effects of the Alternaria mycotoxin tenuazonic acid in immune and intestinal epithelial cells

doi: 10.64898/2026.06.24.734282

Figure Lengend Snippet: Cytotoxic effects of the Alternaria mycotoxin tenuazonic acid (TeA) on Caco-2 (a) and HCEC-1CT (b) cells. Cell viability was assessed by applying the CellTiter-Blue ® assay. Cells were pre-incubated with TeA or 1 μM dexamethasone (Dexa; used as a negative control) for 2 h, followed by co-stimulation with 25 ng/mL IL-1β for an additional 18 h. Triton X-100 (0.1 %) was used as a positive control for cytotoxicity. Data are presented as mean + SD of at least three independent experiments and are expressed relative to the positive control (25 ng/mL IL-1β), as indicated by dotted lines. Statistical significance between treatments and the positive control was assessed using Student’s t -test (*p < 0.05, **p < 0.01, ***p < 0.001). One-way ANOVA followed by Fisher’s LSD post hoc test was applied to evaluate differences between different concentrations of TeA (a–e; p < 0.05)

Article Snippet: Lipopolysaccharide (LPS) from Escherichia coli and dexamethasone (Dexa) were acquired from Sigma Aldrich (St. Louis, USA), and the recombinant human IL-1β and the Quanti-LucTM reagent from Invivogen (San Diego, USA).

Techniques: CtB Assay, Incubation, Negative Control, Positive Control

Journal: bioRxiv

Article Title: Opposing immunomodulatory effects of the Alternaria mycotoxin tenuazonic acid in immune and intestinal epithelial cells

doi: 10.64898/2026.06.24.734282

Figure Lengend Snippet: Impact of the Alternaria mycotoxin tenuazonic acid (TeA) on IL-6, IL-8, IL-10, and TNF-α gene transcription and protein secretion in IL-1β-stimulated Caco-2 (a,d) and HCEC-1CT (b,c,e,f) cells. Cells were pre-incubated with non-cytotoxic concentrations of TeA or 1 μM dexamethasone (Dexa; used as a negative control) for 2 h, followed by co-stimulation with 25 ng/mL IL-1β for an additional 3 h (a,b,d,e) or 18 h (c,f). Panels a–c show relative gene expression determined by qRT-PCR using the 2 −ΔΔCT method and normalized to GAPDH. Panels d–f depict corresponding cytokine secretion levels measured by ELISA. Results were normalized to the positive control (25 ng/mL IL-1β), as indicated by dotted lines. Data are presented as mean + SD of at least three independent experiments. Statistical significance between treatments and the positive control was assessed using Student’s t -test (*p < 0.05, **p < 0.01, ***p < 0.001). One-way ANOVA followed by Fisher’s LSD post hoc test was applied to evaluate differences between TeA concentrations (a–e; p < 0.05)

Article Snippet: Lipopolysaccharide (LPS) from Escherichia coli and dexamethasone (Dexa) were acquired from Sigma Aldrich (St. Louis, USA), and the recombinant human IL-1β and the Quanti-LucTM reagent from Invivogen (San Diego, USA).

Techniques: Incubation, Negative Control, Gene Expression, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Positive Control