Journal: Arthritis Research & Therapy

Article Title: Omentin-1 reduced synovial M1 macrophages through SIRT6 signaling pathway and alleviated knee osteoarthritis response in mice

doi: 10.1186/s13075-025-03680-y

Figure Lengend Snippet: Omentin-1 alleviated pain in ACLT induced SIRT6 −/− OA mice. A Timeline of animal experiments. We performed ACLT surgery to construct wild-type and SIRT6 −/− osteoarthritis (OA) mouse model, and injected 50µL Omentin-1 (300ng/joint/mouse) into the joint cavity of OA mice once a week for 5 consecutive weeks. Behavioral assessment was also performed through pain detection (PWTL) and gait analysis. Knee joint tissue was collected from mice at week 6, with a portion fixed in 4% paraformaldehyde for pathological staining and the remaining portion stored at −80℃ for Western blot analysis. The experimental groups were divided into three groups: Sham group, ACLT group, and Omentin-1 group, with 8 mice in each group. B Pain in OA mice was assessed before surgery and at 1, 2, 3, 4, 5, and 6 weeks after ACLT surgery. (C-E) At the 6th week after surgery, we conducted gait analysis and recorded the swing time, stride length, and paw area of the right hind limb. * p < 0.05, ** p < 0.01, *** p < 0.001

Article Snippet: 24 h after ACLT surgery, two types of mice were injected with Omentin-1 (300ng/50μL, HY-P701099, MedChemExpress) [ ] twice a week into the joint cavity for six consecutive weeks.

Techniques: Construct, Injection, Staining, Western Blot

Journal: Science Advances

Article Title: Targeted inhibition of hepatic de novo ceramide synthesis ameliorates MASH

doi: 10.1126/sciadv.adx2681

Figure Lengend Snippet: ( A ) Whole-body IVIS imaging of mice injected with nonlabeled LNP-siRNA (control group) or LNPs loaded Cy5-labeled siRNA. ( B ) Representative IVIS image illustrating the distribution of LNPs across various organs. ( C ) Flow cytometry analysis showing cellular uptake of Cy5-siRNA in hepatocytes. ASGR1 was used as a hepatocyte specific marker. ( D ) Distribution of Cy5-labeled siRNA among different liver cell populations in healthy mice and MASH mice fed an MCD diet for 8 weeks. Data were presented as the percentage of Cy5-positive cells within each cell type based on FACS analysis. EC, endothelial cells; HSCs, hepatic stellate cells; DCs, dendritic cells. ( E to G ) Time-course analysis of hepatic Sptlc2 mRNA levels and SPTLC2 protein levels following a single intravenous dose of LNP-siRNA (0.3 mg/kg) in MASH mice fed an MCD diet for 4 weeks. Glyceraldehyde-3-phosphate dehydrogenase was used as a loading control for Western blot analysis, and protein expression levels were normalized to that on day 2 postdosing of LNP-Scrambled siRNA (control). ( H ) Time-course analysis of hepatic Sptlc2 mRNA levels in healthy mice following siRNA treatment at 0.3 mg/kg per dose, administered either once or twice weekly. The once-weekly group received doses on days 0 and 7, while the twice-weekly group received doses on days 0, 3, 7, and 10. N = 3 mice per group. * P < 0.05, ** P < 0.01, and *** P < 0.001; one-way ANOVA.

Article Snippet: Cells were stained with an ASGR1 antibody (Proteintech, catalog no. CL488-11739, Illinois, USA) for 30 min at 4°C in the dark.

Techniques: Imaging, Injection, Control, Labeling, Flow Cytometry, Marker, Western Blot, Expressing

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Prototype Alzheimer's disease vaccine using the immunodominant B cell epitope from beta-amyloid and promiscuous T cell epitope pan HLA DR-binding peptide.

doi: 10.4049/jimmunol.174.3.1580

Figure Lengend Snippet: FIGURE 4. T cell proliferation in mice immunized with the epitope vaccine PADRE-A1–15-MAP and other control Ags. Splenocytes from individual mice were restimulated in vitro with the indicated peptides. Only the epitope vaccine PADRE- A1–15-MAP (A) induced the activa- tion of PADRE-specific, but not A-specific, T cells. In contrast, A1–33-MAP (B), fibrillar A42 (C), and A1–33 (D) Ags stimulated anti- A T cell responses.

Article Snippet: Detection of CD4 T cells expressing IL-18R (Th1) or T1/ST2 (Th2) molecules Spleens from mice immunized with different immunogens or control animals were depleted of CD8 cells using MACS depletion kit (Miltenyi Biotec), and the remaining splenocytes were restimulated in HL-1 medium with the same peptide that was used for in vivo immunization.

Techniques: Control, In Vitro

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Prototype Alzheimer's disease vaccine using the immunodominant B cell epitope from beta-amyloid and promiscuous T cell epitope pan HLA DR-binding peptide.

doi: 10.4049/jimmunol.174.3.1580

Figure Lengend Snippet: FIGURE 6. Expression of IL-18R (Th1) and T1/ST2 (Th2) molecules on the surface of CD4 T cells. Spleno- cytes from all groups were depleted by CD8 T cells and restimulated with the same peptides that were used for in vivo injections, except splenocytes from mice immunized with A42 were activated in vitro with A40. Splenocytes were analyzed before in vitro activation (0 day) and after 7 days of activation (7 day). For details, see Materials and Methods.

Article Snippet: Detection of CD4 T cells expressing IL-18R (Th1) or T1/ST2 (Th2) molecules Spleens from mice immunized with different immunogens or control animals were depleted of CD8 cells using MACS depletion kit (Miltenyi Biotec), and the remaining splenocytes were restimulated in HL-1 medium with the same peptide that was used for in vivo immunization.

Techniques: Expressing, In Vivo, In Vitro, Activation Assay

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Prototype Alzheimer's disease vaccine using the immunodominant B cell epitope from beta-amyloid and promiscuous T cell epitope pan HLA DR-binding peptide.

doi: 10.4049/jimmunol.174.3.1580

Figure Lengend Snippet: FIGURE 5. Detection of splenocytes producing IFN- ((Th1), IL-4 (Th2), and TNF- in mice immunized with A1–33-MAP, PADRE-A1–15 MAP, fibrillar A42, or linear A1–15. The ELISPOT technique was used, as described in Materials and Methods.

Article Snippet: Detection of CD4 T cells expressing IL-18R (Th1) or T1/ST2 (Th2) molecules Spleens from mice immunized with different immunogens or control animals were depleted of CD8 cells using MACS depletion kit (Miltenyi Biotec), and the remaining splenocytes were restimulated in HL-1 medium with the same peptide that was used for in vivo immunization.

Techniques: Enzyme-linked Immunospot

Journal: Frontiers in Immunology

Article Title: Sensing Acute Cellular Rejection in Liver Transplant Patients Using Liver-Derived Extracellular Particles: A Prospective, Observational Study

doi: 10.3389/fimmu.2021.647900

Figure Lengend Snippet: EP gating strategy and in vitro results. (A) Transmission electron microscopy images of EP. (B) The EP gate was defined using 0.5 µm and 1.53 µm calibration beads. EP population shift was observed after staining with ASGR1 antibodies. (C) In vitro , TNF-α induced human hepatocyte–derived EP. Primary human hepatocytes following liver resection were isolated, cultured overnight, and stimulated with 10 and 20 ng/ml TNF-α. EP surface antigens were stained, and absolute AnnV + , CD130 + , Cx43 + , ASGR1 + EP were analyzed. The ordinary one-way ANOVA with Tukey’s post hoc test was used. The plots are indicated by the mean, and all error bars indicate the SD.

Article Snippet: Each sample containing 50 μl supernatant EP and 5 μL 10x AnnV binding buffer was subsequently incubated with antibodies: APC Alexa 700-conjugated Cx43 (clone: FAB7737N, R&D Systems, Minneapolis, MN, USA), BV421-conjugated CD130 (clone: AM64, BD Biosciences, San Jose, CA, USA), and PE-conjugated AnnV (Cat. No. 640908, BioLegend, San Diego, CA, USA), and FITC-conjugated ASGR1 (clone: REA608, Miltenyi Biotec, Auburn, CA, USA) or FITC-conjugated MDR3 (Cat. No. LS-C694886, LifeSpan BioSciences, Seattle, WA, USA) or FITC-conjugated CD31 (Cat. No. 303104, BioLegend).

Techniques: In Vitro, Transmission Assay, Electron Microscopy, Staining, Derivative Assay, Isolation, Cell Culture

Journal: Frontiers in Immunology

Article Title: Sensing Acute Cellular Rejection in Liver Transplant Patients Using Liver-Derived Extracellular Particles: A Prospective, Observational Study

doi: 10.3389/fimmu.2021.647900

Figure Lengend Snippet: Time course of EP after LT (n = 11). (A) Total EP (/µl) (B) AnnV + , CD130 + , CD31 + EP (/µl) (C) Cx43 + , ASGR1 + , MDR3 + EP (/µl) were stained and analyzed. The Wilcoxon matched-pairs signed ranked test was used. The plots are indicated by the median, and all error bars indicate the interquartile range (IQR). A single asterisk indicates significance at p < 0.05 (D) Patients at risk for acute rejection. EP dynamics from preoperative state to POD 1 of the total, AnnV + , CD130 + , Cx43 + , ASGR1 + , MDR3 + , CD31 + EP (/µl) and their fold change were analyzed. Absolute EP were non-normally distributed; the two-tailed Mann-Whitney U test was used. The plots are indicated by the mean, and all error bars indicate the SD.

Article Snippet: Each sample containing 50 μl supernatant EP and 5 μL 10x AnnV binding buffer was subsequently incubated with antibodies: APC Alexa 700-conjugated Cx43 (clone: FAB7737N, R&D Systems, Minneapolis, MN, USA), BV421-conjugated CD130 (clone: AM64, BD Biosciences, San Jose, CA, USA), and PE-conjugated AnnV (Cat. No. 640908, BioLegend, San Diego, CA, USA), and FITC-conjugated ASGR1 (clone: REA608, Miltenyi Biotec, Auburn, CA, USA) or FITC-conjugated MDR3 (Cat. No. LS-C694886, LifeSpan BioSciences, Seattle, WA, USA) or FITC-conjugated CD31 (Cat. No. 303104, BioLegend).

Techniques: Staining, Two Tailed Test, MANN-WHITNEY

Journal: Frontiers in Immunology

Article Title: Sensing Acute Cellular Rejection in Liver Transplant Patients Using Liver-Derived Extracellular Particles: A Prospective, Observational Study

doi: 10.3389/fimmu.2021.647900

Figure Lengend Snippet: EP profiles during histology-proven ACR. EP were determined in the blood plasma of patients with concomitant LBs. The patients were then classified as histology-proven ACR or non-ACR. EP surface antigens (A) AnnV + , (B) CD130 + , (C) Cx43 + , (D) ASGR1 + , (E) MDR3 + , (F) CD31 + EP were stained, and relative EP (%) were analyzed. (G) Example FSC-A (forward scatter) histograms of MDR3 + EP (%) (H) demonstrating the highest receiver operating characteristic (ROC) of all single antigens. One-way analysis of variance followed by Tukey’s post hoc test was used. The plots are indicated by the median, and all error bars indicate the IQR.

Article Snippet: Each sample containing 50 μl supernatant EP and 5 μL 10x AnnV binding buffer was subsequently incubated with antibodies: APC Alexa 700-conjugated Cx43 (clone: FAB7737N, R&D Systems, Minneapolis, MN, USA), BV421-conjugated CD130 (clone: AM64, BD Biosciences, San Jose, CA, USA), and PE-conjugated AnnV (Cat. No. 640908, BioLegend, San Diego, CA, USA), and FITC-conjugated ASGR1 (clone: REA608, Miltenyi Biotec, Auburn, CA, USA) or FITC-conjugated MDR3 (Cat. No. LS-C694886, LifeSpan BioSciences, Seattle, WA, USA) or FITC-conjugated CD31 (Cat. No. 303104, BioLegend).

Techniques: Clinical Proteomics, Staining

Journal: Frontiers in Immunology

Article Title: Sensing Acute Cellular Rejection in Liver Transplant Patients Using Liver-Derived Extracellular Particles: A Prospective, Observational Study

doi: 10.3389/fimmu.2021.647900

Figure Lengend Snippet: Clustering and color coding with FlowSOM and viSNE for identifying EP subpopulations via density plots. (A) Qualitative analysis and population identification were performed using viSNE and FlowSOM on samples of patient 52, who had episodes of graft dysfunction caused by ACR and non-ACR. Several LBs were performed over time, and their concomitant blood samples presented the following data. t-SNE (t-distributed stochastic neighbor embedding), an unsupervised nonlinear dimensionality reduction algorithm, was used to fit six-dimensional data into two dimensions. All clusters were created via FlowSOM analysis. Clusters were formed based on FACS channels. The arrow indicates the novel ASGR1 + CD130 + AnnV + EP population. Coordinates for each t-SNE dimension (t-SNE1 and t-SNE2) were calculated for each microparticle after dimensionality reduction. (B) Color-coded t-SNE density plots showing different antigen expression levels by the channel of above-mentioned samples.

Article Snippet: Each sample containing 50 μl supernatant EP and 5 μL 10x AnnV binding buffer was subsequently incubated with antibodies: APC Alexa 700-conjugated Cx43 (clone: FAB7737N, R&D Systems, Minneapolis, MN, USA), BV421-conjugated CD130 (clone: AM64, BD Biosciences, San Jose, CA, USA), and PE-conjugated AnnV (Cat. No. 640908, BioLegend, San Diego, CA, USA), and FITC-conjugated ASGR1 (clone: REA608, Miltenyi Biotec, Auburn, CA, USA) or FITC-conjugated MDR3 (Cat. No. LS-C694886, LifeSpan BioSciences, Seattle, WA, USA) or FITC-conjugated CD31 (Cat. No. 303104, BioLegend).

Techniques: Expressing

Journal: Frontiers in Immunology

Article Title: Sensing Acute Cellular Rejection in Liver Transplant Patients Using Liver-Derived Extracellular Particles: A Prospective, Observational Study

doi: 10.3389/fimmu.2021.647900

Figure Lengend Snippet: Analysis of ASGR1 + CD130 + AnnV + EP (%) population discovered by viSNE maps and FlowSOM algorithm. (A) The particular EP subpopulation was identified in the plasma of patients with concomitant LBs. The patients were then classified as histology-proven ACR or non-ACR. ROC was constructed by comparing (B) ACR vs non-ACR and (C) ACR vs control. One-way analysis of variance followed by Tukey’s post hoc test was used. ROC curves and AUC were generated for relative EP. The plots are indicated by the median, and all error bars indicate the IQR.

Article Snippet: Each sample containing 50 μl supernatant EP and 5 μL 10x AnnV binding buffer was subsequently incubated with antibodies: APC Alexa 700-conjugated Cx43 (clone: FAB7737N, R&D Systems, Minneapolis, MN, USA), BV421-conjugated CD130 (clone: AM64, BD Biosciences, San Jose, CA, USA), and PE-conjugated AnnV (Cat. No. 640908, BioLegend, San Diego, CA, USA), and FITC-conjugated ASGR1 (clone: REA608, Miltenyi Biotec, Auburn, CA, USA) or FITC-conjugated MDR3 (Cat. No. LS-C694886, LifeSpan BioSciences, Seattle, WA, USA) or FITC-conjugated CD31 (Cat. No. 303104, BioLegend).

Techniques: Clinical Proteomics, Construct, Control, Generated

Journal: Frontiers in Immunology

Article Title: Sensing Acute Cellular Rejection in Liver Transplant Patients Using Liver-Derived Extracellular Particles: A Prospective, Observational Study

doi: 10.3389/fimmu.2021.647900

Figure Lengend Snippet: Diagnosis sensitivity and specificity of ASGR1 + Annexin V + CD130 + EP (%) for ACR in liver transplant.

Article Snippet: Each sample containing 50 μl supernatant EP and 5 μL 10x AnnV binding buffer was subsequently incubated with antibodies: APC Alexa 700-conjugated Cx43 (clone: FAB7737N, R&D Systems, Minneapolis, MN, USA), BV421-conjugated CD130 (clone: AM64, BD Biosciences, San Jose, CA, USA), and PE-conjugated AnnV (Cat. No. 640908, BioLegend, San Diego, CA, USA), and FITC-conjugated ASGR1 (clone: REA608, Miltenyi Biotec, Auburn, CA, USA) or FITC-conjugated MDR3 (Cat. No. LS-C694886, LifeSpan BioSciences, Seattle, WA, USA) or FITC-conjugated CD31 (Cat. No. 303104, BioLegend).

Techniques: Biomarker Discovery