CD44 is a transmembrane glycoprotein that binds to hyaluronic acid, performs roles in various mobile processes and is expressed in quite a lot of cell sorts. It’s up-regulated in stem cells and most cancers. Anti-CD44 monoclonal antibodies have an effect on cell motility and aggregation, and repress tumorigenesis and metastasis.
Right here we describe 4 new anti-CD44 monoclonal antibodies originating from B cells of a mouse injected with a plasmid expressing CD44 isoform 12. The 4 monoclonal antibodies bind to the terminal, extracellular, conserved area of CD44 isoforms. Primarily based on variations in western blot patterns of most cancers cell lysates, the 4 anti-CD44 mAbs separated into three distinct classes that embrace P4G9, P3D2, and P3A7, and P3G4.
Spot assay evaluation with peptides generated in Escherichia coli assist the conclusion that the monoclonal antibodies acknowledge unglycosylated sequences within the N-terminal conserved area between amino acid 21-220, and analyses with a peptide generated in human embryonic kidney 293 cells, exhibit that these monoclonal antibodies bind to those peptides solely after deglycosylation.
Western blots with lysates from three most cancers cell traces exhibit that a number of CD44 isoforms are unglycosylated within the anti-CD44 goal areas. The potential utility of the monoclonal antibodies in blocking tumorigenesis was examined by co-injection of cells of the breast cancer-derived tumorigenic cell line MDA-MB-231 with the anti-CD44 monoclonal antibody P3D2 into the mammary fats pads of mice.
All 5 management mice injected with MDA-MB-231 cells plus anti-IgG shaped palpable tumors, whereas solely one of many six check mice injected with MDA-MB-231 cells plus P3D2 shaped a tiny tumor, whereas the remaining 5 had been tumor-free, indicating that the 4 anti-CD44 mAbs could also be helpful therapeutically.
Anti-inflammatory exercise of CD44 antibodies in murine immune thrombocytopenia is mediated by Fcγ receptor inhibition
Monoclonal IgG antibodies to CD44 (anti-CD44) are anti-inflammatory in quite a few murine autoimmune fashions however the mechanisms are poorly understood. Anti-CD44 anti-inflammatory exercise reveals full therapeutic concordance with intravenous immunoglobulin (IVIg) in treating autoimmune illness fashions, making anti-CD44 a possible IVIg different.
In murine immune thrombocytopenia (ITP), there may be at the moment no mechanistic rationalization for anti-CD44 exercise though anti-CD44 ameliorates illness equally to IVIg. Right here we exhibit a novel anti-inflammatory mechanism of anti-CD44 that explains illness amelioration by anti-CD44 in murine ITP.
Macrophages handled with anti-CD44 in vitro had dramatically suppressed phagocytosis by means of FcγRs in two separate methods of IgG-opsonized platelets and erythrocytes. Phagocytosis inhibition by anti-CD44 was mediated by blockade of the FcγR IgG binding web site with out altering floor FcγR expression.
Anti-CD44 of various subclasses revealed that FcγR blockade was particular to receptors that may very well be engaged by the respective anti-CD44 subclass, and Fc-deactivated anti-CD44 variants misplaced all FcγR-inhibiting exercise. In vivo, anti-CD44 functioned analogously within the murine passive ITP mannequin and guarded mice from ITP when thrombocytopenia was induced by means of an FcγR that may very well be engaged by the CD44 antibody’s subclass.
In keeping with FcγR blockade, Fc-deactivated variants of anti-CD44 had been fully unable to ameliorate ITP. Collectively, anti-CD44 inhibits macrophage FcγR operate and ameliorates ITP according to an FcγR blockade mechanism. Anti-CD44 is a possible IVIg different and could also be of specific profit in ITP because of the vital function of FcγRs in human ITP pathophysiology.
Enhanced concentrating on of prostate cancer-initiating cells by salinomycin-encapsulated lipid-PLGA nanoparticles linked with CD44 antibodies.
Prostate most cancers is the fifth most typical explanation for cancer-associated mortality in males worldwide. The survival of prostate cancer-initiating cells (CICs) is a vital issue behind the metastasis and recurrence of prostate most cancers. The cluster of differentiation (CD) 44 antigen is taken into account an vital marker for prostate CICs.
Salinomycin is a potent therapeutic drug in opposition to CICs. The current research demonstrated that salinomycin exerts potent exercise in opposition to CD44+ prostate CICs. To additional improve this anticancer impact, salinomycin-encapsulated lipid-poly(lactic-co-glycolic acid) nanoparticles linked with CD44 antibodies (SM-LPN-CD44) had been generated.
The anticancer impact of the nanoparticles was investigated in a collection of assays, together with a cytotoxicity assay, stream cytometry and anticancer assay in prostate cancer-bearing mice in vivo. The outcomes revealed that SM-LPN-CD44 might effectively and particularly promote the supply of salinomycin to CD44+ prostate CICs, and there by obtain better inhibition of the cells in contrast with that achieved by salinomycin and non-targeted nanoparticles.
To the very best of our data, that is the primary research to report improved therapeutic results in opposition to prostate CICs achieved by the enhancement of focused drug supply by way of nanoparticles conjugated with CD44 antibodies. Due to this fact, SM-LPN-CD44 nanoparticle-based remedy represents a novel method to eradicate prostate CICs and is a promising potential therapy technique for prostate most cancers.
A novel monoclonal antibody KMP1 has potential antitumor exercise of bladder most cancers by blocking CD44 in vivo and in vitro.
Bladder most cancers turns into a critical medical and social concern attributable to its excessive recurrence and mortality charges. Thus, it’s pressing to go looking a novel prognostic biomarker and focused remedy with excessive sensitivity and specificity. On this research, we used the human bladder most cancers cell line EJ as an immunogen to generate a novel mouse monoclonal antibody KMP1 that particularly sure to bladder most cancers, after which, the antitumor impact of KMP1 in opposition to bladder most cancers was investigated each in vivo and in vitro.
The outcomes confirmed that expression of the KMP1 epitope is according to medical severity and prognosis of bladder most cancers. Moreover, KMP1 not solely considerably inhibited the proliferation, migration, and adhesion of EJ cells in vivo, but additionally suppressed the xenograft tumor progress in nude mice in contrast with the management group handled with mIgG.
Subsequently, the underlying mechanism of KMP1 in opposition to bladder most cancers was explored by way of antigen affinity chromatography and mass spectrometry. CD44 positioned on the cytomembrane was discovered because the antigen of KMP1. Utilizing RNA interference expertise to knock down CD44 expression, we additional recognized that KMP1 has the antitumor exercise by binding to CD44 and blocking its capabilities.
In conclusion, KMP1 may be precious for improvement as a promising particular diagnostic biomarker or focused agent for bladder most cancers.
Mouse and human HSPC immobilization in liquid tradition by CD43- or CD44–antibody coating.
Retaining observe of particular person cell identifications is crucial to the research of dynamic single-cell habits over time. Extremely motile hematopoietic stem and progenitor cells migrate rapidly and don’t adhere, and thus should be imaged very continuously to maintain cell identifications. Even worse, they’re additionally flushed away throughout medium change. To beat these limitations, we examined antibody coating for decreasing HSPC motility in vitro.
Anti-CD43- and anti-CD44-antibody coating diminished the cell motility of mouse and human HSPCs in a concentration-dependent method. This permits 2-dimensional colony formation with out cell mixing in liquid cultures, massively will increase time-lapse imaging throughput, and likewise maintains cell positions throughout media change. Anti-CD43 however not anti-CD44 coating reduces mouse HSPC proliferation with rising concentrations.
CD44 Antigen (CD44) Antibody |
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| abx139276-500l | Abbexa | 500 µl | Ask for price |
CD44 Antigen (CD44) Antibody |
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| abx139277-01mg | Abbexa | 0.1 mg | EUR 444 |
CD44 Antigen (CD44) Antibody |
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| abx139277-100l | Abbexa | 100 µl | EUR 287.5 |
CD44 Antigen (CD44) Antibody |
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| abx139277-500l | Abbexa | 500 µl | Ask for price |
CD44 Antigen (CD44) Antibody |
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| 20-abx134411 | Abbexa |
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CD44 Antigen (CD44) Antibody |
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| abx137093-01mg | Abbexa | 0.1 mg | EUR 487.5 |
CD44 Antigen (CD44) Antibody |
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| 20-abx137093 | Abbexa |
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CD44 Antigen (CD44) Antibody |
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| abx137102-01mg | Abbexa | 0.1 mg | EUR 487.5 |
CD44 Antigen (CD44) Antibody |
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| 20-abx137102 | Abbexa |
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CD44 Antigen (CD44) Antibody |
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| 20-abx137492 | Abbexa |
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CD44 Antigen (CD44) Antibody |
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| abx137492-50tests | Abbexa | 50 tests | EUR 225 |
CD44 Antigen (CD44) Antibody |
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| abx130862-100l | Abbexa | 100 µl | EUR 250 |
CD44 Antigen (CD44) Antibody |
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| abx130862-500l | Abbexa | 500 µl | EUR 312.5 |
CD44 Antigen (CD44) Antibody |
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| abx176874-1096tests | Abbexa | 10 × 96 tests | EUR 687.5 |
CD44 Antigen (CD44) Antibody |
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| abx176874-596tests | Abbexa | 5 × 96 tests | EUR 325 |
CD44 Antigen (CD44) Antibody |
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| abx176874-96tests | Abbexa | 96 tests | EUR 262.5 |
CD44 Antigen (CD44) Antibody |
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| abx176876-1096tests | Abbexa | 10 × 96 tests | EUR 725 |
No related results on cell survival or myeloid and megakaryocyte differentiation of hematopoietic stem cells and multipotent progenitors 1-5 had been detected. Human umbilical wire hematopoietic CD34+ cell survival, proliferation, and differentiation weren’t affected by both coating. This method each massively simplifies and accelerates steady evaluation of suspension cells, and allows the research of their habits in dynamic somewhat than static tradition situations over time.
