Development of a Novel Anti-CD44 Variant 8 Monoclonal Antibody C₄₄Mab-94 against Gastric Carcinomas

Published: May 15, 2023

Gastric cancer (GC) is the third leading cause of cancer-related deaths worldwide. GC with peritoneal metastasis exhibits a poor prognosis due to lack diagnostic biomarkers and effective therapy. A comprehensive analysis malignant ascites identified genomic alterations significant amplifications driver genes, including CD44. CD44 its splicing variants are overexpressed in tumors, play crucial roles acquisition invasiveness, stemness, resistance treatments. Therefore, development CD44-targeting monoclonal antibodies (mAbs) important for diagnosis In this study, we immunized mice CD44v3–10-overexpressed PANC-1 cells established several dozens clones that produce anti-CD44v3–10 mAbs. One (C44Mab-94; IgG1, kappa) recognized variant-8-encoded region peptide, indicating C44Mab-94 specific mAb CD44v8. Furthermore, could recognize CHO/CD44v3–10 cells, oral squamous cell carcinoma line (HSC-3), or lines (MKN45 NUGC-4) flow cytometric analyses. detect exogenous CD44v3–10 endogenous CD44v8 western blotting stained formalin-fixed paraffin-embedded gastric immunohistochemistry. These results indicate useful detecting various applications expected be application

Language: Английский

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A Novel Anti-CD44 Variant 10 Monoclonal Antibody is Useful for Immunohistochemical Analysis against Oral Squamous cell carcinomas

Published: May 26, 2023

CD44 is known as a cancer stem cell marker of head and neck squamous carcinoma (HNSCC) plays critical role in malignant progression. Splicing variant isoforms (CD44v) are overexpressed cancers considered promising target for therapy. Several monoclonal antibodies (mAbs) against have been developed by immunizing mice with CD44v3–10-overexpressed cells. In this study, we characterized novel clone, C44Mab-18 (IgM, kappa). reacted CHO/CD44v3–10, but not CHO/CD44s flow cytometry. Enzyme-linked immunosorbent assay revealed that the epitope determined to be border sequence between 10 constant exon 16-encoded sequence. Flow cytometry showed recognizes HSC-3, an oral carcinoma(OSCC) line. The apparent dissociation (KD) CHO/CD44v3–10 HSC-3 was 1.6 × 10−7 M 1.7 M, respectively. detected CD44v3–10, western blotting. Furthermore, endogenous CD44v10 immunohistochemistry using OSCC tissues. Taken together, useful detecting immunohistochemistry.

Language: Английский

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Epitope Mapping of an Anti-CD44v4 Monoclonal Antibody (C₄₄Mab-108) using Enzyme-Linked Immunosorbent Assay

Published: Nov. 3, 2023

CD44 is a type I transmembrane glycoprotein, and possesses various isoforms which are largely classified into standard variant (CD44v) isoforms. Some variant-encoded regions play critical roles in tumor progression. However, the function of 4 (CD44v4)-encoded region has not been fully understood. Using peptide immunization, we developed an anti-CD44v4 mAb, C44Mab-108, useful for flow cytometry, western blotting, immunohistochemistry. In this study, determined epitope C44Mab-108 by enzyme-linked immunosorbent assay (ELISA). We used alanine (or glycine)-substituted peptides CD44v4-encoded (amino acids 271-290 human CD44v3-10), found that did recognize alanine-substituted D280A W281A. Furthermore, these could inhibit recognition cytometry The results indicate binding includes Asp280 Trp281 CD44v3-10.

Language: Английский

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CRISPR/Cas9-mediated silencing of CD44: unveiling the role of hyaluronic acid-mediated interactions in cancer drug resistance

Naunyn-Schmiedeberg s Archives of Pharmacology, Journal Year: 2023, Volume and Issue: 397(5), P. 2849 - 2876

Published: Nov. 22, 2023

Language: Английский

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Epitope Mapping of an Anti-Mouse CD39 Monoclonal Antibody using PA Scanning and RIEDL Scanning

Published: Dec. 6, 2023

A cell-surface ectonucleotidase CD39 mediates the conversion of extracellular ATP into immunosuppressive adenosine with another CD73. The elevated in tumor microenvironment (TME) attenuates antitumor immunity, which promotes cell immunologic escape and progression. Anti-CD39 monoclonal antibodies (mAbs), suppress enzymatic activity, can be applied to therapy. Therefore, an understanding relationship between inhibitory activity epitope mAbs is important. We previously established anti-mouse (mCD39) mAb, C39Mab-1 using Cell-Based Immunization Screening (CBIS) method. In this study, we determined critical flow cytometry. performed PA tag (12 amino acids)-substituted analysis (named scanning) RIEDL (5 determine By combination scanning scanning, identified conformational epitope, spanning three segments 275th 279th, 282nd 291st, 306th 323rd acids mCD39. These analyses would contribute identification membrane proteins.

Language: Английский

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Beta Cells Deficient forRenalaseCounteract Autoimmunity by Shaping Natural Killer Cell Activity

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2024, Volume and Issue: unknown

Published: March 4, 2024

ABSTRACT Type 1 diabetes (T1D) arises from autoimmune-mediated destruction of insulin-producing pancreatic beta cells. Recent advancements in the technology generating cells human pluripotent stem (SC-beta cells) have facilitated exploration cell replacement therapies for treating T1D. However, persistent threat autoimmunity poses a significant challenge to survival transplanted SC-beta Genetic engineering is promising approach enhance immune resistance as we previously showed by inactivating Renalase ( Rnls ) gene. Here demonstrate that loss-of-function shape mediating regulatory Natural Killer (NK) phenotype important induction tolerogenic antigen presenting -deficient mediate cell-cell-contact-independent hallmark anti-inflammatory cytokine Tgfβ1 NK In addition, surface expression key checkpoints CD47 and Ceacam1 are markedly elevated on deficient . Enhanced glucose metabolism mutant responsible upregulation expression. These findings crucial better understand how genetically engineered giving valuable insights future therapeutic treat cure Graphical summary

Language: Английский

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Epitope Mapping of an Anti-CD44v4 Monoclonal Antibody (C₄₄Mab-108) Using Enzyme-Linked Immunosorbent Assay

Monoclonal Antibodies in Immunodiagnosis and Immunotherapy, Journal Year: 2024, Volume and Issue: 43(3), P. 85 - 89

Published: March 20, 2024

CD44 is a type I transmembrane glycoprotein and possesses various isoforms which are largely classified into standard (CD44s) variant (CD44v) isoforms. Some variant-encoded regions play critical roles in tumor progression. However, the function of 4 (CD44v4)-encoded region has not been fully understood. Using peptide immunization, we developed an anti-CD44v4 monoclonal antibody, C

Language: Английский

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Anti-CD44 Variant 10 Monoclonal Antibody Exerts Antitumor Activity in Mouse Xenograft Models of Oral Squamous Cell Carcinomas

Published: Aug. 6, 2024

CD44 regulates cell adhesion, proliferation, survival, and stemness has been considered a tumor therapy target. possesses the shortest standard (CD44s) variety of variant (CD44v) isoforms. Since expression CD44v is restricted in epithelial cells carcinomas compared to CD44s, promising target for monoclonal antibody (mAb) therapy. We previously developed an anti‐CD44v10 mAb, C44Mab-18 (IgM, kappa) recognize exon 10-encoded region. In present study, mouse IgG2a version (C44Mab-18-mG2a) was generated evaluate antitumor activities against CD44-positive with established anti-pan C44Mab-46-mG2a. C44Mab-18-mG2a exhibited higher reactivity C44Mab-46-mG2a CD44v3–10-overexpressed CHO-K1 (CHO/CD44v3–10) oral squamous carcinoma lines (HSC-2 SAS) flow cytometry. exerted superior antibody‐dependent cellular cytotoxicity (ADCC) CD44v3–10. contrast, showed complement‐dependent (CDC) A similar tendency observed ADCC CDC HSC-2 SAS. Furthermore, administering or significantly suppressed CD44v3–10, HSC-2, SAS xenograft growth control IgG2a. These results indicate that could be therapeutic regimen CD44v10-positive tumors.

Language: Английский

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