anti integrin beta 1  (Cell Signaling Technology Inc)

Journal: Bioactive Materials

Article Title: Mechanically sensitized hydrogel microspheres trigger membrane receptor switch for cartilage repair

doi: 10.1016/j.bioactmat.2026.03.017

Figure Lengend Snippet: Integrin receptor activation induced by membrane receptor switch. A) The fluorescence microscopy of MSCs loaded on HAMA or OBNC hydrogel after different treatments. B) Relative fluorescence intensity in the whole field of view for each group. C) Relative fluorescence intensity per cell for each group (∗ symbol represents comparison with HAMA group, # symbol represents comparison with OBNC group). D) Flow cytometry was used to detect integrin αvβ1 and α5β1 positive cells. E) Relative fluorescence intensity of each group. F) Flow cytometry was used to detect 12G10 positive cells and within integrin αvβ1 and α5β1 positive cells. G) Relative fluorescence intensity of each group, and relative proportions of 12G10 to integrins αvβ1 and α5β1. (ns: non-significant, ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001, ## P < 0.01, ### P < 0.001).

Article Snippet: TRPC1 inhibitor (0.3 nM, Pico145, CAS No. 1628287-16-0), TRPM7 inhibitor (1.0 μM, VPC4, CAS No. 945604-76-2), TRPV2 inhibitor (5.0 μM, compound IV2-1, CAS No. 2242724-49-6), TRPM4 inhibitor (1.5 μM, CBA, CAS No. 351424-20-9), PIEZO1 inhibitor (2.5 μM, GsMTx4, CAS No. 1209500-46-8), integrin αvβ5 inhibitor (8.0 nM, Compound 12, CAS No.: 2615912-33-7), integrin αvβ1 inhibitor (0.3 nM, Compound C8, CAS No. 1689540-62-2), integrin α5β1 inhibitor (10 μM, ATN-161, 904763-27-5), and CDK5 inhibitor (5 nM, CDK5-IN-1, 2,639,540-19-3) were purchased from MCE Biotechnology Co., LTD. After the MSCs were treated, the cRGD solution was added at a concentration of 1:200 and incubated in the dark for 15 min, and the results were observed by fluorescence microscopy.

Techniques: Activation Assay, Membrane, Fluorescence, Microscopy, Comparison, Flow Cytometry

Journal: Blood Advances

Article Title: VLA-4 agonist promotes engraftment and immune reconstitution of allogeneic hematopoietic stem cells ∗

doi: 10.1182/bloodadvances.2025017456

Figure Lengend Snippet: VLA-4 is a key target for regulating HSPC engraftment after allo-HSCT. (A-D) Data from a patient after allo-HSCT. (A) Schematic illustration for generation of scRNA-seq data from a patient after allo-HSCT. We focused on patients undergoing allo-HSCT who clinically presented with MDC and FDC statuses before and after manipulation of immunosuppression and/or DLI intervention. We collected clinical samples of BMMCs from patients undergoing allo-HSCT over a long period of time and finally enrolled a patient with acute myeloid leukemia undergoing haploidentical donor transplantation. (B) UMAP plots revealing 8 cell subtypes and the proportions of each cell subtype of BMMCs from the patient at MDC and FDC statuses. (C) Ligand-receptor pairs between 7 cell populations. Differentially expressed genes between samples of FDC and MDC were first identified, and ligand-receptor pairs were subsequently screened from these genes using the built-in ligand-receptor database in iTALK. NK cell population was not found due to no eligible gene between ligand-receptor. Solid lines linked ligand-receptor pairs with arrow. (D) Violin plots illustrating the smoothed expression distribution of receptor and ligand genes in HSPCs from the patient’s BM samples. (E-I) Data from MDC mouse model. (E) Schematic illustration of MDC mouse model protocol and the generation of scRNA-seq data. Two batches of samples (MDC-1 vs FDC-1 and MDC-2 vs FDC-2) were delivered for scRNA-seq analysis, and each sample consisted of a mixture of BMMCs from 3 individual mice per group. (F) UMAP plots revealing 13 cell subtypes and the proportions of each cell subtype in BMMCs from 2 batches of mouse model sample. (G) Ligand-receptor pairs between 12 cell populations in mouse BMMCs. Differentially expressed genes between samples of FDC and MDC were first identified, and ligand-receptor pairs were subsequently screened from these genes using the built-in ligand-receptor database in iTALK. Eosinophil cell population was not found due to no eligible gene between ligand-receptor. Solid lines linked ligand-receptor pairs with arrow. (H) Violin plots illustrating the smoothed expression distribution of marker genes in HSPCs from mouse BMMCs. (I) Relative mRNA expression levels of Igf1r , B2m , Itga4 , and Itgb1 genes in BM lineage-negative cells at MDC and FDC statuses after allo-BMT measured by real-time PCR analysis (n = 3). ns P > .05; ∗ P < .05; ∗∗ P < .01. B, B (CD19 + ) cell; DC, dendritic cell; Eosino, eosinophil; Neu, neutrophil; NK, natural killer cell; ns, not statistically significant; PCR, polymerase chain reaction; T, T (CD3 + ) cell; UMAP, uniform manifold approximation and projection.

Article Snippet: VLA-4 agonists (1-[anilinocarbonyl] proline [activator-1, A1], integrin modulator 1 [activator-2, A2], and THI0019 [activator-3, A3]) and VLA-4 inhibitor (BIO5192 ) were purchased from MedChemExpress.

Techniques: Transplantation Assay, Expressing, Marker, Real-time Polymerase Chain Reaction, Polymerase Chain Reaction

Journal: Blood Advances

Article Title: VLA-4 agonist promotes engraftment and immune reconstitution of allogeneic hematopoietic stem cells ∗

doi: 10.1182/bloodadvances.2025017456

Figure Lengend Snippet: VLA-4 agonist promotes long-term engraftment and leads to skewed lymphocyte output of HSCs. (A) Schematic illustration of the experimental design testing VLA-4 A2 in the MDC mouse model of allo-BMT.VLA-4 A2 was injected intraperitoneally with a dose of 5 mg/kg per day from day 8 (d8) to day 56 (d56) after transplantation (n = 17). An equal amount of DMSO was used as vehicle (n = 10). (B) Chimerism levels of donor cells in PB from 2 weeks (2Ws) to 8 weeks (8Ws) after allo-BMT for the following defined populations: total leukocytes (PB-Total), Myeloid cells (PB-Mye cells), B cells (PB-B cells), T cells (PB-T cells), mature Erythrocytes (PB-Erys), and nucleated Erythrocytes (PB-nErys). (C) Lineage compositions in donor-derived PB cells from 2Ws to 8Ws after transplantation. (D) Dynamics of myeloid and lymphoid lineage compositions in donor-derived PB cells from 2Ws to 8Ws after transplantation. (E-F) Representative flow cytometry plots and statistical analyses of HPCs, MPPs, and SLAM HSCs within the donor-derived LSK cell population in the BM at 2 weeks (E) or 4 weeks (F) after transplantation. VLA-4 A2 was injected intraperitoneally from d8 to d14 or from d8 to d28, with DMSO used as the vehicle (n = 7-9 per group). (G-H) Quantification of SLAM + HSCs in the BM of vehicle- and VLA-4 A2-treated MDC mice at 2 weeks (G) or 4 weeks (H) after transplantation. (I) Chimerism levels of donor cells in BM at 8Ws after transplantation for the following populations: LSKs, HPCs, MPPs, and SLAM HSCs. VLA-4 A2 was injected intraperitoneally from d8 to d56 (n = 17). DMSO was used as a vehicle (n = 10). (J) Statistical plots of burst-forming-unit-erythroid (BFU-E), colony-forming-unit-granulocyte and -macrophage (CFU-GM) and CFU-GEMM for BM cells from recipient mice after VLA-2 A2 and vehicle (DMSO)-treated from d8 to d56 after transplantation. A total of 20 000 whole BM cells were incubated in methylcellulose medium supplemented with growth factors for 7 to 12 days in triplicate. Each dot indicates an individual recipient mouse. ns P > .05; ∗ P < .05; ∗∗ P < .01; ∗∗∗ P < .001. DMSO, dimethyl sulfoxide; ns, not statistically significant.

Article Snippet: VLA-4 agonists (1-[anilinocarbonyl] proline [activator-1, A1], integrin modulator 1 [activator-2, A2], and THI0019 [activator-3, A3]) and VLA-4 inhibitor (BIO5192 ) were purchased from MedChemExpress.

Techniques: Injection, Transplantation Assay, Derivative Assay, Flow Cytometry, Incubation

Journal: Blood Advances

Article Title: VLA-4 agonist promotes engraftment and immune reconstitution of allogeneic hematopoietic stem cells ∗

doi: 10.1182/bloodadvances.2025017456

Figure Lengend Snippet: VLA-4 A2 promotes HSC self-renewal capacity and skewing lymphocyte output in vivo. (A-F) Data from competitive transplantation mouse model. (A) Schematic illustration of competitive transplantation assay with in vivo VLA-4 A2- and vehicle-treated BM cells. Primary (first) recipients received VLA-4 A2 (5 mg/kg per day, intraperitoneally) from day 8 to day 14 (orange line, 2-week group) or from day 8 to day 28 (purple line, 4-week group) after allo-BMT. BM from these primary recipients was harvested on day 14 or day 28 and transplanted, together with competitor BM cells into lethally irradiated secondary (second) recipients. (Total n = 20 per group of secondary recipients, sacrificed and detected at different time points). (B) Representative flow cytometry plots and statistical analyses of MPP4 (Lin − Sca-1 + c-Kit + Flk2 + CD150 − ) in BM LSK cells from primary recipients at 2Ws after transplantation. VLA-4 A2/vehicle was injected intraperitoneally from day 8 to day 14. (C) Representative flow cytometry graphs for the percentages of CD45.1 + CD45.2 − (donor) and CD45.1 + CD45.2 + (competitor) cells in total PB cells (PB-total), T cells (CD3 + ), spleen cells (splenocytes), and BM LSK cells (BM-LSKs) from secondary recipients at 2Ws after transplantation. (D) Relative percentages of CD45.1 + cells in secondary recipients at 2Ws after transplantation. (E) Relative percentages of CD45.1 + cells in secondary recipients at 24Ws after secondary transplantation. The secondary recipients receiving BM cells from primary recipient mice administered VLA-4 A2/vehicle from d5 to d28. (F) Relative percentages of CD45.1 + cells within the CD4 + and CD8 + cell populations in the spleens of secondary recipients at 24 weeks after transplantation. (G-N) Data from xenotransplantation NOG mouse model of human UCB. (G) Schematic illustration of assessing VLA-4 A2 on hematopoietic reconstitution of human UCB CD34 + cells in xenotransplantation model (n = 5 per group). (H) The percentages of human CD45 + cells in PB of the primary NOG recipient mice at 12Ws. (I) The percentages of human CD45 + cells, myeloid cells (CD33 + ), and B cells (CD19 + ) in BM at 12Ws (BM+12Ws). (J) The percentages of human CD45 + cells in spleen at 12Ws. (K) Quantification of human CD45 + cells in the BM of the primary NOG recipient mice at 12Ws after transplantation with 10 000 human UCB CD34 + cells. (L-M) The frequencies of human SRCs in UCB CD34 + cells treated with VLA-4 A2 or vehicle in vivo. (L) Poisson statistical analysis of data from . Shapes represent the percentages of negative mice for each dose of cells. Solid lines indicate the best-fit linear model for each data set. Dotted lines represent 95% confidence intervals. (M) HSC frequencies (line in the box) and 95% confidence intervals (box) presented as the number of SRCs in 1 × 10 6 CD34 + cells. (N) The percentages and quantification of human CD45 + cell in BM of secondary NOG recipient mice transplanted with 5 × 10 6 BM cells from primary NOG recipient mice (n = 5 per group). Each dot indicates an individual recipient mouse. ns P > .05; ∗ P < .05; ∗∗ P < .01; ∗∗∗ P < .001; ∗∗∗∗ P < .0001. ns, not statistically significant.

Article Snippet: VLA-4 agonists (1-[anilinocarbonyl] proline [activator-1, A1], integrin modulator 1 [activator-2, A2], and THI0019 [activator-3, A3]) and VLA-4 inhibitor (BIO5192 ) were purchased from MedChemExpress.

Techniques: In Vivo, Transplantation Assay, Irradiation, Flow Cytometry, Injection

Journal: Blood Advances

Article Title: VLA-4 agonist promotes engraftment and immune reconstitution of allogeneic hematopoietic stem cells ∗

doi: 10.1182/bloodadvances.2025017456

Figure Lengend Snippet: Reconstituted cellular immunity induced by VLA-4 A2 exhibits potent antiviral capabilities without aggravating acute GVHD. (A) Schematic illustration of testing VLA-4 A2 on immune reconstitution and antiviral infection effects. Mice were infected with 50 000 PFU of MHV68-H2bYFP intranasally on day 22 after transplantation. Virus titers in the lungs were quantified on day 3 and day 6 after infection, and virus-infected cells were analyzed by multicolor flow cytometry on day 16 after infection (total n = 13-15 per group, sacrificed and detected at 3 time points). (B) The white blood cell (WBC) and lymphocyte counts in the PB at day 14 after BMT (n = 13-15). The counting was performed by Sysmex pocH-100i Automated Hematology Analyzer. (C) The counts of CD45.1 + CD3 + T cells and CD45.1 + B220 + B cells in the PB at day 14 after BMT (n = 13-15). (D) Lineage compositions of donor-derived (CD45.1 + ) cells in PB at 2 weeks (2Ws) after BMT. (E) Reconstitution of splenic donor-derived CD45.1 + CD4 + T cells, including Th1 cells (IFN-γ + ), Τh2 cells (ΙL-4 + ), Th17 cells (IL-17a + ), and naive/other CD4 + T cell subsets at day 28 after transplantation. WT C57BL/6J mice were used as control (n = 5 per group). (F) Thymic reconstitution at day 28 after transplantation revealing CD4 − CD8 − , CD4 + CD8 + , CD4 + CD8 − , and CD4 − CD8 + thymocyte subsets. WT C57BL/6J mice were used as control (n = 5 per group). (G) The percentages of YFP + cells in splenocytes on day 16 after infection (n = 10 per group). (H) Counts of B-cell subsets in spleens at day 16 after infection, including precursor B cells (pre-B, B220 + IgM − CD19 + CD43 − ), pre–progenitor B cells (pre–pro-B, B220 + IgM − CD19 − CD43 + ), progenitor B cells (pro-B, B220 + IgM − CD19 + CD43 + ), immature B cells (B220 low IgM + ), and mature B cells (B220 high IgM + ) (n = 5 per group). (I) Percentages of plasma cells (CD3 − B220 low CD138 + ) in splenocytes at day 16 after infection (n = 10 per group). (J) Representative flow cytometry graphs and statistical plots of the percentages of IFN-γ + cells in spleens CD45.1 + CD4 + cells at day 16 after infection (n = 10 per group). (K) The CD45.1 + CD4 + IFN-γ + cell counts in the spleen of mice at day 16 after infection (n = 10 per group). (L) Representative flow cytometry graphs and statistical plots of the percentages of IFN-γ + cells in spleens of CD45.1 + CD4 + cells at day 16 after infection (n = 10 per group). (M) The CD45.1 + CD4 + IFN-γ + cell counts in the spleen of mice at day 16 after infection (n = 10 per group). (N) Schematic illustration of testing VLA-4 A2 on a GVHD mouse model. (O) The GVHD scores of GVHD mice in all experimental groups. (P) The overall survival of GVHD mice in all experimental groups. Each dot indicates an individual recipient mouse. ns P > .05; ∗ P < .05; ∗∗ P < .01; ∗∗∗∗ P < .0001. ns, not statistically significant.

Article Snippet: VLA-4 agonists (1-[anilinocarbonyl] proline [activator-1, A1], integrin modulator 1 [activator-2, A2], and THI0019 [activator-3, A3]) and VLA-4 inhibitor (BIO5192 ) were purchased from MedChemExpress.

Techniques: Infection, Transplantation Assay, Virus, Flow Cytometry, Derivative Assay, Control, Clinical Proteomics

Journal: Blood Advances

Article Title: VLA-4 agonist promotes engraftment and immune reconstitution of allogeneic hematopoietic stem cells ∗

doi: 10.1182/bloodadvances.2025017456

Figure Lengend Snippet: VLA-4 A2 regulates HSC function through ERK1/2 phosphorylation. (A) Schematic illustration of the investigation into the effect of VLA-4 A2 on HSC engraftment after allo-BMT and the generation of scRNA-seq data. (B) UMAP plots revealing 10 cell types and proportions of each cell type in BMMCs of VLA-4 A2- and vehicle-treated MDC mice at day 14 (d14) and day 35 (d35) after allo-BMT. VLA-4 A2 was administered intraperitoneally at 5 mg/kg per day from days 8 to 14 or 35 after transplantations. Each batch comprised pooled cells from 3 individual mice. (C) Violin plots revealing the smoothed expression distribution of intramodular hub genes of HSPCs in MDC mice at d14 and d35 after allo-BMT. (D) Histogram plots revealing the frequencies of CD34 + Flk2 + cells in HSPCs in MDC mice at d14 and d35 after allo-BMT. (E) Relative mRNA expression levels of Bcl11b , CD7 , Trdc , Notch1 , and Tcf1 genes of BM lineage–negative cells in MDC mice at d14 and d35 after allo-BMT (n = 3). (F) Representative flow cytometry graph (left) and statistical plot (right) for the effects of VLA-4 A2 on phosphorylation of ERK 1/2 (phosphor-ERK). Flow-sorted UCB CD34 + cells were serum starved for 16 hours and subsequently incubated with VLA-4 A2 (1μM), VLA-4 A2 (1μM) + U0126 (1μM), VCAM-1 (1 μg/mL), VCAM-1 (1 μg/mL) + U0126 (1μM), or vehicle for 2 hours. (G) Statistical plots of CFU-GEMM for mouse BM cells. CFU assays were performed using murine BM cells cultured under different conditions: control, VLA-4 agonist A2 (10nM), A2 plus MEK1/2 inhibitor U0126 (1μM), VCAM-1 (1 μg/mL), and VCAM-1 plus U0126. A total of 20 000 whole BM cells were incubated in methylcellulose medium supplemented with growth factors for 7 to 12 days in triplicate. (H) Representative flow cytometry graphs and statistical plots of the frequencies of early lymphoid progenitors (CD5 + CD7 + cells) from UCB CD34 + cells after incubation with VLA-4 A2 (10nM), VCAM-1 (1 μg/mL) plus U0126 (1μM) or not in StemSpan Lymphoid Progenitor Expansion Supplement medium for 14 days, respectively. (I) Frequencies of CD4 + CD8 + double-positive T cells differentiated from UCB CD34 + cells after 14 days in StemSpan Lymphoid Progenitor Expansion medium followed by 21 days in StemSpan T Cell Progenitor Maturation medium under the same treatment conditions. Each dot indicates an individual replicate well. StemSpan T Cell Generation Kit (StemCell Technologies, no. 09940). ns P > .05; ∗ P < .05; ∗∗∗∗ P < .0001. ns, not statistically significant; UMAP, uniform manifold approximation and projection.

Article Snippet: VLA-4 agonists (1-[anilinocarbonyl] proline [activator-1, A1], integrin modulator 1 [activator-2, A2], and THI0019 [activator-3, A3]) and VLA-4 inhibitor (BIO5192 ) were purchased from MedChemExpress.

Techniques: Phospho-proteomics, Expressing, Flow Cytometry, Incubation, Cell Culture, Control

Journal: Bioactive Materials

Article Title: A large puncture closer of aortic wall by multi-memory actions with thrombo-hemodynamic control

doi: 10.1016/j.bioactmat.2025.12.042

Figure Lengend Snippet: 18 Fr puncture of hemostasis in porcine aorta using VWP by validating the memory programming effect of each part. a, As a challenging model for application of large-diameter catheters, i) an 18 Fr (6 mm) puncture is created into the porcine thoracic aorta (diameter: 10 mm) so that the size-matched VWP is deployed, followed by measuring proximal and distal blood pressure. ii) The experimental groups are designed first to exam the memory programming effect of collaboration between Ring squeezing with Body expansion on self-locking (SL) to enable efficient hemostasis. Next, the effect of Wing shape recovery from curve to flat is examined on hemodynamic control (HC) in cooperation with the actions of Body and Ring to handle hemostasis. No recovery of Wing shape is expected to induce excessive thrombosis. iii) Four experimental groups are established using a total of 12 pigs (N = 12) with immediate sacrifice following deployment (N = 3 each). Group 1 [SL(−) HC(−)] represents no memory programming. Group 2 [SL(+) w/flat Wing] has the effects of Body and Ring actions except the hemostatic sealing by keeping the flat Wing. Group 3 [SL(+) HC(+)] possesses the complete memory effects of the three parts. Group 4 [SL(+) w/bump Wing] is expected to have excessive thrombosis because of no shape recovery from the curved Wing while maintaining the memory actions of Body and Wing. b, Each group is visually explained in the illustrations. c , In VWP actions, (left) the bleeding condition preserves the normal sinusoidal waveform of high proximal pressure (green) in contrast to the disturbed waveform of low distal pressure (red). (middle) Hemostatic closure results in similar high sinusoidal waveform at both pressure sites. (right) Excessive thrombosis does not disturb the waveform, but the distal pressure level becomes lower than the proximal one. d, When reperfusion starts by removing the clamp post-deployment (blue), only Group 3 [SL(+) HC(+)] reaches the hemostatic closure, as evidenced by flow stabilization (red) with a 5 s plateau at both pressure sites. Group 4 [SL(+) w/bump Wing] exhibits the pattern of over-thrombosis. e, H&E images show bleeding in Group 1 as an indication of incomplete closure in contrast to moderate, minimal, and dense thrombotic features observed in Group 2, 3, and 4 respectively as further supported by the signals of activated platelets (green, CD41-positive) and fibrinogen (red) [Scale bars = 0.5 mm (4 mm in box)]. f, Compared to Group 1 [SL(−) HC(−)] and 2 [SL(+) w/flat Wing], Group 3 [SL(+) HC(+)] shows the fastest i) hemostasis and ii) arterial pressure equilibration, indicating the most efficient hemostatic response. g, These outcomes in Group 3 include i) the smallest difference between the proximal and distal pressures with ii) the smallest thrombus area in contrast the largest area of Group 4 [SL(+) w/bump Wing] as an indication of excessive thrombosis. h , The marker gene expression of thrombotic feature (vWF, PF-4, and P-sel) significantly increases from Group 2 to Group 3 and further to Group 4 except the comparison of vWF expression between Group 2 and 3 (ns: no significance). Data are shown as mean ± SD, N = 3 biologically independent animals per group. Significance was determined using one-way ANOVA with Tukey's test between groups.

Article Snippet: Primary antibodies against CD41 (1:100, 24552-1-AP, proteintech), fibrinogen (1:100, ab232793, Abcam), CD31 (1:100, sc-376764, Santa Cruz Biotechnology), CD68 (1:100, ab125212, Abcam), and ARG-1 (1:200, LS-C447907, LSBio) were applied overnight at 4°C.

Techniques: Control, Marker, Gene Expression, Comparison, Expressing