antimouse human cd44 neutralizing antibody  (Bio X Cell)

Journal: Journal of translational medicine

Article Title: ITIH5-mediated fibroblast/macrophage crosstalk exacerbates cardiac remodelling after myocardial infarction.

doi: 10.1186/s12967-025-06244-5

Figure Lengend Snippet: Fig. 8 Schematic illustration of ITIH5-mediated fibroblast–macrophage crosstalk exacerbates cardiac remodelling after MI. In myocardial tissue, ITIH5 expression is upregulated after MI and is localised to the ECM, regulating ECM-macrophage-fibroblast signalling network interaction thereby promoting macrophage profibrotic phenotype transformation, CFs activation and cardiac remodelling. In addition, highly expressed ITIH5 can be secreted into the peripheral blood and act as a biomarker in patients with ICM. In terms of mechanism of action, ITIH5 activates the CD44 receptors on the surface of the macrophage cell membrane in a hyaluronan-dependent manner and activates the macrophage STAT3 signalling pathway, promoting profibrotic phenotype transformation

Article Snippet: The antimouse/human CD44 neutralizing antibody (BE0039) and IgG2b isotype control (BE0090) were purchased from BioXcell (West Lebanon, USA).

Techniques: Expressing, Transformation Assay, Activation Assay, Biomarker Discovery, Membrane

Journal: Cell Communication and Signaling : CCS

Article Title: All-trans retinoic acid pretreatment of mesenchymal stem cells enhances the therapeutic effect on acute kidney injury

doi: 10.1186/s12964-024-01671-1

Figure Lengend Snippet: HA activates the downstream PI3K/AKT signaling pathway by binding to CD44. a, b Representative images and quantification of immunofluorescence staining of HK-2 cells with CD44 (red) and DAPI (blue) ( n = 3). Scale bars, 20 μm. c, d Representative immunofluorescence staining of CD44 in renal tissue ( n = 6). Scale bars, 20 μm. e, f Representative immunohistochemical staining of CD44 in renal tissue ( n = 6). Scale bars, 40 μm. g-j Representative western blot images and quantitative analysis of p-AKT and total AKT levels in kidney tissues ( n = 3). k-n Representative western blot images and quantitative analysis of p-AKT and total AKT levels in HK-2 cells ( n = 3). Data are presented as the means ± SD. ** P < 0.01, * P < 0.05

Article Snippet: To block HA interactions to CD44, anti-CD44 neutralizing antibody (10 μg/mL, 9400-01, Southern Biotech, USA)) was added to HK-2 cells culture medium before hypoxia treatment.

Techniques: Binding Assay, Immunofluorescence, Staining, Immunohistochemical staining, Western Blot

Journal: Cell Communication and Signaling : CCS

Article Title: All-trans retinoic acid pretreatment of mesenchymal stem cells enhances the therapeutic effect on acute kidney injury

doi: 10.1186/s12964-024-01671-1

Figure Lengend Snippet: Inhibition of the HA/CD44 axis reverses the anti-inflammatory, anti-apoptotic and pro-proliferative effects of ATRA-MSCs. a, c-i Western blot images and quantitative densitometry analysis of TNF-a, IL-6, Bax, Bcl-2 and cleaved caspase-3 in HK-2 cells ( n = 3). b, j-l Western blot images and quantitative densitometry analysis of p-AKT and total AKT in HK-2 cells ( n = 3). Data are presented as the means ± SD. **** P < 0.0001, *** P < 0.001, ** P < 0.01, * P < 0.05

Article Snippet: To block HA interactions to CD44, anti-CD44 neutralizing antibody (10 μg/mL, 9400-01, Southern Biotech, USA)) was added to HK-2 cells culture medium before hypoxia treatment.

Techniques: Inhibition, Western Blot

Journal: Cell Communication and Signaling : CCS

Article Title: All-trans retinoic acid pretreatment of mesenchymal stem cells enhances the therapeutic effect on acute kidney injury

doi: 10.1186/s12964-024-01671-1

Figure Lengend Snippet: Schematic illustration of ATRA promoted HA production by MSCs and activated the PI3K/AKT pathway by binding to CD44 in renal tubular epithelial cells, thereby improving renal repair after AKI

Article Snippet: To block HA interactions to CD44, anti-CD44 neutralizing antibody (10 μg/mL, 9400-01, Southern Biotech, USA)) was added to HK-2 cells culture medium before hypoxia treatment.

Techniques: Binding Assay

Journal: Scientific reports

Article Title: Melanocortin-4 receptor in macrophages attenuated angiotensin II-induced abdominal aortic aneurysm in mice.

doi: 10.1038/s41598-023-46831-4

Figure Lengend Snippet: Figure 4. An osteopontin receptor CD44 mediates vascular vulnerability of MC4RTB/TB mice. (A) Leptin (100 nM)-induced Spp1 gene expression in primary vascular smooth muscle cells from WT male mice pretreated with or without the PI3K inhibitor LY294002 (LY, 10 nM; n = 4). (B) Representative pictures and (C) quantification of osteopontin (OPN) immunostaining in the aorta of WD-fed MC4R+/+ and MC4RTB/TB mice (n = 6, scale bar is 100 μm). (D) Representative pictures of DAPI, α-smooth muscle cells, and merged images in the aorta of WD-fed MC4RTB/TB mice (scale bars represent 20 μm). (E) Changes in the sBP of 14–18-weeks- old MC4RTB/TB mice during Ang II (500 ng/kg/min) infusion concomitantly administered with anti-CD44 or control antibodies (Ab; n = 6–12). (F) Representative pictures of the aorta and (G) AAA incidence and (H) diameter after Ang II infusion into WD-fed MC4R+/+ and MC4RTB/TB mice administered with anti-CD44 or control Abs (n = 6–12). *P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: In the experiment of α-MSH (M4135, Sigma-Aldrich, 1 mg/kg/day) or anti-CD44 neutralizing antibody (BE0039, BioXCell, 200 μg/day) treatment, they were injected intraperitoneally into ApoE KO or MC4RTB/TB mice for 4 weeks starting on the day the minipumps were implanted.

Techniques: Gene Expression, Immunostaining, Control

Journal: Scientific Reports

Article Title: Melanocortin-4 receptor in macrophages attenuated angiotensin II-induced abdominal aortic aneurysm in mice

doi: 10.1038/s41598-023-46831-4

Figure Lengend Snippet: An osteopontin receptor CD44 mediates vascular vulnerability of MC4R TB/TB mice. ( A ) Leptin (100 nM)-induced Spp1 gene expression in primary vascular smooth muscle cells from WT male mice pretreated with or without the PI3K inhibitor LY294002 (LY, 10 nM; n = 4). ( B ) Representative pictures and ( C ) quantification of osteopontin (OPN) immunostaining in the aorta of WD-fed MC4R + / + and MC4R TB/TB mice (n = 6, scale bar is 100 μm). ( D ) Representative pictures of DAPI, α-smooth muscle cells, and merged images in the aorta of WD-fed MC4R TB/TB mice (scale bars represent 20 μm). ( E ) Changes in the sBP of 14–18-weeks-old MC4R TB/TB mice during Ang II (500 ng/kg/min) infusion concomitantly administered with anti-CD44 or control antibodies (Ab; n = 6–12). ( F ) Representative pictures of the aorta and ( G ) AAA incidence and ( H ) diameter after Ang II infusion into WD-fed MC4R + / + and MC4R TB/TB mice administered with anti-CD44 or control Abs (n = 6–12). * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: In the experiment of α-MSH (M4135, Sigma-Aldrich, 1 mg/kg/day) or anti-CD44 neutralizing antibody (BE0039, BioXCell, 200 μg/day) treatment, they were injected intraperitoneally into ApoE KO or MC4R TB/TB mice for 4 weeks starting on the day the minipumps were implanted.

Techniques: Expressing, Immunostaining

Journal: Arteriosclerosis, Thrombosis, and Vascular Biology

Article Title: Single-Cell RNA Sequencing of Coronary Perivascular Adipose Tissue From End-Stage Heart Failure Patients Identifies SPP1 + Macrophage Subpopulation as a Target for Alleviating Fibrosis

doi: 10.1161/ATVBAHA.123.319828

Figure Lengend Snippet: Conditioned medium from perivascular adipose tissue (PVAT) of patients with atherosclerosis (AS) significantly promotes the fibrogenesis of human fibroadipogenic progenitors (FAPs) through CD44/integrin axis. A , Schematic representation of the experimental procedure. B , Representative images of transwell migration assay ( top ) and EdU (5-ethynyl-2’-deoxyuridine) proliferation assay ( bottom ) of human FAP cells. Scale bar, 100 µm. C and D , Quantitative analysis of transwell migration assay ( C ) and EdU proliferation assay ( D ) of human FAP cells. Four points in each group in C and D represent 4 biological replicates. Comparisons of the cell migration or proliferation between each 2 groups were performed using the Student t test; P values were adjusted for multiple hypothesis testing using the Benjamini-Hochberg method.

Article Snippet: FAP cells were pretreated with vehicle or anti-CD44 (cluster of differentiation 44) neutralizing monoclonal antibody (10 μg/mL, 15675-1-AP; Proteintech), RGD (arginylglycylaspartic acid) peptide to inhibit integrin receptors (10 μM, A8052; Sigma Aldrich), or both in combination for 30 minutes as described previously.

Techniques: Transwell Migration Assay, Proliferation Assay, Migration

Journal: Stem Cell Research & Therapy

Article Title: Hyaluronan supports the limbal stem cell phenotype during ex vivo culture

doi: 10.1186/s13287-022-03084-8

Figure Lengend Snippet: Effects of HA on TKE2 cells cultured on differently coated dishes or in the presence of HA as a supplement to the media. TKE2 cells were cultured on differently coated dishes ( A ) or on uncoated dishes with HA, HA + anti-CD44 or 4MU + Hyalase supplemented to the media ( B ). The number of ∆Np63 positive cells and the total number of cells (DAPI) were quantified using QuPath and the percentage of ∆Np63 + cells calculated ( C ). *Represents p ≤ 0.05, scale bar represents 50 μm

Article Snippet: In order to verify the mechanism by which HA supports LESCs, anti-CD44 neutralizing antibody (R&D systems, Minneapolis, USA#AF6127) was added to the culture media in order to block the binding of HA to CD44.

Techniques: Cell Culture

Journal: Stem Cell Research & Therapy

Article Title: Hyaluronan supports the limbal stem cell phenotype during ex vivo culture

doi: 10.1186/s13287-022-03084-8

Figure Lengend Snippet: Effects of HA on the differentiation of TKE2 cells cultured on differently coated dishes or in the presence of HA as a supplement to the media. TKE2 cells were cultured on differently coated dishes ( A ) or on uncoated dishes with HA of different molecular weights, HA + anti-CD44 or 4MU + Hyalase supplemented to the media ( B ) under differentiating conditions. The number of ∆Np63 positive cells and the total number of cells (DAPI) were quantified using Qupath and the percentage of ∆Np63 + cells calculated for TKE cells maintained on differently coated dishes ( C ) and with HA, HA + anti-CD44 and 4MU + Hyalase supplemented to the media ( D ). *Represents p ≤ 0.05, scale bar represents 50 μm

Article Snippet: In order to verify the mechanism by which HA supports LESCs, anti-CD44 neutralizing antibody (R&D systems, Minneapolis, USA#AF6127) was added to the culture media in order to block the binding of HA to CD44.

Techniques: Cell Culture