Journal: Oncology Reports

Article Title: Platelets and MMP?9 contribute to esophageal cancer invasion via CD40?CD154 interactions

doi: 10.3892/or.2025.8912

Figure Lengend Snippet: Functional analysis of cells stimulated with CD154. (A) Viability assay. No significant changes were observed in all four cell lines. (B) Migration assay. (C) Invasion assay. Migration and invasion were enhanced in TE-5 and TE-10 cells, which highly expressed CD40, after rsCD154 stimulation. No significant changes were observed in low CD40 expression cells (TE-4 and −11 cells). The Mann-Whitney U test was performed using GraphPad Prism 9 software. ns, not significant; OD, optical density; rsCD154, recombinant soluble CD154.

Article Snippet: CD154 , REA238 , FC , 1:50 , Miltenyi Biotec GmbH.

Techniques: Functional Assay, Viability Assay, Migration, Invasion Assay, Expressing, MANN-WHITNEY, Software, Recombinant

Journal: Oncology Reports

Article Title: Platelets and MMP?9 contribute to esophageal cancer invasion via CD40?CD154 interactions

doi: 10.3892/or.2025.8912

Figure Lengend Snippet: Changes in gene expression caused by the CD40-CD154 interaction in TE-10 cells. Screening of 40 genes involved in extracellular matrix remodeling. The expression ratios of target genes relative to GAPDH are shown on the left. On the right, CD154-induced changes in gene expression are displayed in descending order. Upregulation of gene expression in the order MMP-9 > PLG > LAMC2 was confirmed following rsCD154 stimulation.

Article Snippet: CD154 , REA238 , FC , 1:50 , Miltenyi Biotec GmbH.

Techniques: Gene Expression, Expressing

Journal: Oncology Reports

Article Title: Platelets and MMP?9 contribute to esophageal cancer invasion via CD40?CD154 interactions

doi: 10.3892/or.2025.8912

Figure Lengend Snippet: Changes in MMP-9 expression caused by the CD40-CD154 interaction in TE cells. (A) Qualitative analysis of MMP-9 secretion from TE cells using gelatin zymography. Increased MMP-9 secretion was observed in high CD40 expression cells (TE-5 and −10), regardless of the baseline MMP-9 secretion level without CD154 stimulation. (B) Quantitative analysis of MMP-9 secretion from TE cells using an ELISA. Increased MMP-9 secretion was observed in TE-5 and −10 cells. (C) Quantitative analysis of MMP-9 mRNA expression in TE cells via reverse transcription-quantitative PCR. MMP-9 mRNA expression upregulation was observed in high CD40 expression cells (TE-5 and −10). The Mann-Whitney U test was performed using GraphPad Prism 9 software. n.d., not detected; rsCD154, recombinant soluble CD154.

Article Snippet: CD154 , REA238 , FC , 1:50 , Miltenyi Biotec GmbH.

Techniques: Expressing, Zymography, Enzyme-linked Immunosorbent Assay, Reverse Transcription, Real-time Polymerase Chain Reaction, MANN-WHITNEY, Software, Recombinant

Journal: Oncology Reports

Article Title: Platelets and MMP?9 contribute to esophageal cancer invasion via CD40?CD154 interactions

doi: 10.3892/or.2025.8912

Figure Lengend Snippet: Experiments using CD40-knockdown TE-10 cells using siRNA. (A) Transfection with CD40 siRNA resulted in a reduction in CD40 mRNA expression. The average relative CD40 mRNA expression was compared using GraphPad Prism 9 software. After one-way ANOVA, Bonferroni post hoc analysis was performed. (B) CD154 stimulation of CD40 knockdown TE-10 cells was performed, and MMP-9 upregulation was suppressed. (C) Migration assay. CD40 knockdown reduced cell migration, and no increase in migration was observed following CD154 stimulation. The images on the right present a visualization of migrating cells under a 1.25× optical microscope, with the cellular regions indicated by green labelling. (D) Invasion assay. Similar to the migration assay, a decrease in invasion was observed following CD40 knockdown. The images on the right present a visualization of migrating cells under a 1.25× optical microscope, with the cellular regions indicated by green labelling. siRNA, small interfering RNA.

Article Snippet: CD154 , REA238 , FC , 1:50 , Miltenyi Biotec GmbH.

Techniques: Knockdown, Transfection, Expressing, Software, Migration, Microscopy, Invasion Assay, Small Interfering RNA

Journal: Oncology Reports

Article Title: Platelets and MMP?9 contribute to esophageal cancer invasion via CD40?CD154 interactions

doi: 10.3892/or.2025.8912

Figure Lengend Snippet: Survival analysis of patients who underwent esophagectomy was performed using the Kaplan-Meier method. The log-rank (Mantel-Cox) test was conducted using GraphPad Prism 9 software. (A) Association between DFS and MMP-9 levels. (B) Association between DFS and CD154 levels. (C) Association between OS and MMP-9 levels. (D) Association between OS and CD154 levels. DFS, disease-free survival; MST, median survival time; OS, overall survival.

Article Snippet: CD154 , REA238 , FC , 1:50 , Miltenyi Biotec GmbH.

Techniques: Software

Journal: Oncology Reports

Article Title: Platelets and MMP?9 contribute to esophageal cancer invasion via CD40?CD154 interactions

doi: 10.3892/or.2025.8912

Figure Lengend Snippet: Survival analysis of patients with esophageal cancer stratified by disease stage. (A) Association between DFS and MMP-9 levels in patients with pathological stage I. (B) Association between DFS and CD154 levels in patients with pathological stage I. (C) Association between OS and MMP-9 levels in patients with pathological stage I. (D) Association between OS and CD154 levels in patients with pathological stage I. (E) Association between DFS and MMP-9 levels in patients with pathological stage II–IV. (F) Association between DFS and CD154 levels in patients with pathological stage II–IV. (G) Association between OS and MMP-9 levels in patients with pathological stage II–IV. (H) Association between OS and CD154 levels in patients with pathological stage II–IV. Survival analysis was performed using the Kaplan-Meier method, and the log-rank (Mantel-Cox) test was conducted using GraphPad Prism 9 software. DFS, disease-free survival; MST, median survival time; OS, overall survival.

Article Snippet: CD154 , REA238 , FC , 1:50 , Miltenyi Biotec GmbH.

Techniques: Software

Journal: Oncology Reports

Article Title: Platelets and MMP?9 contribute to esophageal cancer invasion via CD40?CD154 interactions

doi: 10.3892/or.2025.8912

Figure Lengend Snippet: Schema of the CD40-CD154 interaction in the tumor microenvironment of esophageal cancer. In tumor cells, the CD40-CD154 interaction induces the secretion of molecules involved in ECM remodeling, including MMP-9, contributing to cell invasion, EMT and metastasis. In immune cells, the CD40-CD154 interaction is involved in antitumor immunity, leading to cancer cell death. ECM, extracellular matrix; EMT, epithelial-mesenchymal transition.

Article Snippet: CD154 , REA238 , FC , 1:50 , Miltenyi Biotec GmbH.

Techniques:

Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

Article Title: Associations Between Mean Platelet Volume and Various Factors in Type 2 Diabetes Patients: A Single-Center Study from Poland

doi: 10.12659/MSM.941109

Figure Lengend Snippet: ( A ) The median TPO concentration in patients with type 2 diabetes did not differ from that of the control group ( P =0.7210). ( B ) The median IL-6 concentration in patients with type 2 diabetes was significantly higher than that of the control group ( P <0.0001). ( C ) The median sP-selectin concentration in patients with type 2 diabetes was significantly higher than that of the control group ( P =0.0008). ( D ) The median sCD40L concentration in patients with type 2 diabetes was significantly higher than that of the control group ( P =0.0010). IL-6 – interleukin 6; sCD40L – soluble form of CD40L; sP-selectin – soluble form of selectin P; TPO – thrombopoietin. Statistical significance: *** P ≤0.001, **** P ≤0.0001. The figure was created with the use of the GraphPad Prism 8.0 software (GraphPad Software, San Diego, CA, USA).

Article Snippet: IL-6 concentration was measured using ELISA Quantikine Human IL-6 Immunoassay kit (Cat#: D6050; R&D Systems Europe Ltd.). sP-selectin concentration was measured using ELISA Quantikine Human P-Selectin Immunoassay kit (Cat#: DPSE00; R&D Systems Europe Ltd.). sCD40L concentration was measured using a human sCD40L ELISA kit (Cat#: 650 120 096; Diaclone SAS, Besançon, France).

Techniques: Concentration Assay, Control, Software

Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

Article Title: Associations Between Mean Platelet Volume and Various Factors in Type 2 Diabetes Patients: A Single-Center Study from Poland

doi: 10.12659/MSM.941109

Figure Lengend Snippet: Univariate and multivariate linear regression analysis results for PLT logarithm in patients with type 2 diabetes.

Article Snippet: IL-6 concentration was measured using ELISA Quantikine Human IL-6 Immunoassay kit (Cat#: D6050; R&D Systems Europe Ltd.). sP-selectin concentration was measured using ELISA Quantikine Human P-Selectin Immunoassay kit (Cat#: DPSE00; R&D Systems Europe Ltd.). sCD40L concentration was measured using a human sCD40L ELISA kit (Cat#: 650 120 096; Diaclone SAS, Besançon, France).

Techniques:

Journal: Inflammation and Regeneration

Article Title: Evaluation of immunosuppression protocols for MHC-matched allogeneic iPS cell-based transplantation using a mouse skin transplantation model

doi: 10.1186/s41232-021-00190-7

Figure Lengend Snippet: Blockade of leukocyte costimulatory molecules permits long-term engraftment of mouse skin grafts from MHC-matched allogeneic donors. ( A ) CB-based immunoregulation protocol. Immunosuppression was performed by using a protocol of administrating a combination of anti-CD40L mAb and CTLA4-Ig daily (CB) with or without rapamycin on every 3 days from POD 1 (CB + rapa). ( B ) Skin graft survival under immunosuppression with CB ( n = 9, C3129F1; n = 9, BALB/c; n = 9, C57BL/6; n = 9, CBA/N). ( C ) Skin graft survival under immunosuppression with CB + rapa ( n = 9, C3129F1; n = 8, BALB/c; n = 9, C57BL/6; n = 9, CBA/N). ( D ) Macroscopic observation (upper panels) and hematoxylin and eosin stained section (lower panels) of autologous (C3129F1), CBA/N, and C57BL/6 skin grafts on day 100. Scale bars: 100 μm. ( E ) Immunohistochemical staining for CD3 of C57BL/6 skin grafts harvested from CB or CB + rapa treated groups on 100 days post-transplantation. Upper and lower panels represent CD3 specific staining and isotype control stained section, respectively. Scale bars: 100 μm. ( F ) Recipient T cell response in MHC-matched but minor antigen-mismatched skin transplantation. The T cell proliferation rates in each treatment group were normalized to that of C3129F1 mice stimulated with autologous irradiated splenocytes. Error bars indicate standard error of technical triplicates. Similar results were obtained in two independent experiments. * p < 0.05, ** p < 0.01 (Tukey’s HSD test). ( G ) De novo anti-donor antibody production in the recipients. Error bars indicate standard error of biological replicates ( n = 8, naive; n = 4, non-treatment; n = 9, CB; n = 9, CB + rapa). tx, transplantation; mAb, monoclonal antibody; CB, co-stimulatory molecule blocking; rapa, rapamycin; POD, post-operative day

Article Snippet: For co-stimulatory molecule blockade (CB) therapy, anti-CD40 ligand (CD40L) (MR-1 for mouse and #BE0292 for human, BioXcell) and CTLA4Ig (Orencia®, Bristol Myers Squibb for mouse and #BE0099, BioXcell for human) were administered at a dose of 500 μg for anti-CD40L, 400 μg for CTLA4Ig on days 0, 2, 4, and 6 after transplantation.

Techniques: Staining, Immunohistochemical staining, Transplantation Assay, Control, Irradiation, Blocking Assay

Journal: Vaccines

Article Title: Chimeric Antigen by the Fusion of SARS-CoV-2 Receptor Binding Domain with the Extracellular Domain of Human CD154: A Promising Improved Vaccine Candidate

doi: 10.3390/vaccines10060897

Figure Lengend Snippet: Construction and expression of RBD-CD in HEK-293 cells. ( A ) Schematic representation of the construction encoding the protein RBD-CD. The diagram indicates in that order from the N - towards the C -terminal end: a fragment of the human cytomegalovirus enhancer/promoter (hCMV), the Igk-chain leader sequence, a segment of RBD, a six-histidine tail plus a linker, and the extracellular domain of human CD154. ( B ) Protein expression in cell pools obtained after lentiviral transduction and selection with blasticidin determined by ACE2 binding in ELISA. Data are represented as mean + STD of six replicates. A Kruskal–Wallis and Dunn’s multiple comparison test were performed. Asterisks represent statistical differences with controls. (** p < 0.01) ( C ) ELISA to compare individual clones obtained by limiting dilution. Data are represented as mean + STD of three replicates.

Article Snippet: In parallel, a sequence coding for 633 bp of the extracellular domain of Homo sapiens CD40 ligand (CD154) (NM_000074.2) was amplified by PCR from pCMV3-CD40LG-His (HG10239-CH, Sino Biological) using primers CD154-F and CD154-R ( ).

Techniques: Expressing, Sequencing, Transduction, Selection, Binding Assay, Enzyme-linked Immunosorbent Assay, Clone Assay

Journal: Vaccines

Article Title: Chimeric Antigen by the Fusion of SARS-CoV-2 Receptor Binding Domain with the Extracellular Domain of Human CD154: A Promising Improved Vaccine Candidate

doi: 10.3390/vaccines10060897

Figure Lengend Snippet: RBD-CD ESI-MS characterization ( A ) Amino acid sequence of the RBD-CD recombinant protein. The verified sequence by ESI-MS/MS analysis is underlined. The first residues correspond to additional 18 amino acids from the pDisplay subcloning vector. In black the receptor-binding domain (RBD) of SARS-CoV-2 S protein (residues Glu324-Gly535). The lines represent disulfide bonds that link C336-C361, C379-C432, C480-C488, and C391-C525 (for RBD) and C178-C218 for CD154. Residues indicated as N correspond to potential N-glycosylation sites. ( B ) ESI-MS spectrum of tryptic digestion of the RBD-CD protein in a buffer-free solution. The signals marked with diamonds correspond to the RBD protein peptides, the triangles to the CD154 fragment, and the ovals to the six His tails plus the spacer arm.

Article Snippet: In parallel, a sequence coding for 633 bp of the extracellular domain of Homo sapiens CD40 ligand (CD154) (NM_000074.2) was amplified by PCR from pCMV3-CD40LG-His (HG10239-CH, Sino Biological) using primers CD154-F and CD154-R ( ).

Techniques: Sequencing, Recombinant, Tandem Mass Spectroscopy, Subcloning, Plasmid Preparation, Binding Assay