Novel Human Tenascin-C Function-Blocking Camel Single Domain Nanobodies.
Animals
Antibodies, Neutralizing
/ immunology
Antibody Specificity
Binding Sites, Antibody
Camelus
/ immunology
Cell Adhesion
/ drug effects
Cell Line, Tumor
Colitis, Ulcerative
/ immunology
Colon
/ immunology
Enzyme-Linked Immunosorbent Assay
HEK293 Cells
Humans
Immunohistochemistry
Liver Neoplasms
/ immunology
Protein Binding
Single-Domain Antibodies
/ immunology
Tenascin
/ administration & dosage
diagnostic tool
extracellular matrix
fibronectin type III repeat
interaction modeling
nanobody
tenascin-c
therapeutic tool
tumor biomarker
Journal
Frontiers in immunology
ISSN: 1664-3224
Titre abrégé: Front Immunol
Pays: Switzerland
ID NLM: 101560960
Informations de publication
Date de publication:
2021
2021
Historique:
received:
29
11
2020
accepted:
19
02
2021
entrez:
1
4
2021
pubmed:
2
4
2021
medline:
22
9
2021
Statut:
epublish
Résumé
The extracellular matrix (ECM) molecule Tenascin-C (TNC) is well-known to promote tumor progression by multiple mechanisms. However, reliable TNC detection in tissues of tumor banks remains limited. Therefore, we generated dromedary single-domain nanobodies Nb3 and Nb4 highly specific for human TNC (hTNC) and characterized the interaction with TNC by several approaches including ELISA, western blot, isothermal fluorescence titration and negative electron microscopic imaging. Our results revealed binding of both nanobodies to distinct sequences within fibronectin type III repeats of hTNC. By immunofluroescence and immunohistochemical imaging we observed that both nanobodies detected TNC expression in PFA and paraffin embedded human tissue from ulcerative colitis, solid tumors and liver metastasis. As TNC impairs cell adhesion to fibronectin we determined whether the nanobodies abolished this TNC function. Indeed, Nb3 and Nb4 restored adhesion of tumor and mesangial cells on a fibronectin/TNC substratum. We recently showed that TNC orchestrates the immune-suppressive tumor microenvironment involving chemoretention, causing tethering of CD11c+ myeloid/dendritic cells in the stroma. Here, we document that immobilization of DC2.4 dendritic cells by a CCL21 adsorbed TNC substratum was blocked by both nanobodies. Altogether, our novel TNC specific nanobodies could offer valuable tools for detection of TNC in the clinical practice and may be useful to inhibit the immune-suppressive and other functions of TNC in cancer and other diseases.
Identifiants
pubmed: 33790905
doi: 10.3389/fimmu.2021.635166
pmc: PMC8006918
doi:
Substances chimiques
Antibodies, Neutralizing
0
Single-Domain Antibodies
0
TNC protein, human
0
Tenascin
0
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
635166Informations de copyright
Copyright © 2021 Dhaouadi, Ben Abderrazek, Loustau, Abou-Faycal, Ksouri, Erne, Murdamoothoo, Mörgelin, Kungl, Jung, Ledrappier, Benlasfar, Bichet, Chiquet-Ehrismann, Hendaoui, Orend and Bouhaouala-Zahar.
Déclaration de conflit d'intérêts
MM was employed by Colzyx AB. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.
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