scholarly journals Novel Human Tenascin-C Function-Blocking Camel Single Domain Nanobodies

2021 ◽  
Vol 12 ◽  
Author(s):  
Sayda Dhaouadi ◽  
Rahma Ben Abderrazek ◽  
Thomas Loustau ◽  
Chérine Abou-Faycal ◽  
Ayoub Ksouri ◽  
...  
Keyword(s):  
Ulcerative Colitis ◽  
Dendritic Cells ◽  
Mesangial Cells ◽  
Single Domain ◽  
Myeloid Dendritic Cells ◽  
Electron Microscopic ◽  
Microscopic Imaging ◽  
Tenascin C ◽  
Fibronectin Type Iii ◽  
Fibronectin Type

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.

scholarly journals Tenascin-C contains distinct adhesive, anti-adhesive, and neurite outgrowth promoting sites for neurons.

1996 ◽  
Vol 132 (4) ◽  
pp. 681-699 ◽  
Author(s):  
B Götz ◽  
A Scholze ◽  
A Clement ◽  
A Joester ◽  
K Schütte ◽  
...  
Keyword(s):  
Neurite Outgrowth ◽  
Hippocampal Neurons ◽  
Neural Cells ◽  
Cell Binding ◽  
Tenascin C ◽  
Functional Studies ◽  
Distinct Cell ◽  
Fibronectin Type Iii ◽  
Alternatively Spliced ◽  
Fibronectin Type

The glia-derived extracellular matrix glycoprotein tenascin-C (TN-C) is transiently expressed in the developing CNS and may mediate neuron-glia interactions. Perturbation experiments with specific monoclonal antibodies suggested that TN-C functions for neural cells are encoded by distinct sites of the glycoprotein (Faissner, A., A. Scholze, and B. Götz. 1994. Tenascin glycoproteins in developing neural tissues--only decoration? Persp. Dev. Neurobiol. 2:53-66). To characterize these further, bacterially expressed recombinant domains were generated and used for functional studies. Several short-term-binding sites for mouse CNS neurons could be assigned to the fibronectin type III (FNIII) domains. Of these, the alternatively spliced insert TNfnA1,2,4,B,D supported initial attachment for both embryonic day 18 (E18) rat and postnatal day 6 (P6) mouse neurons. Only TNfn1-3 supported binding and growth of P6 mouse cerebellar neurons after 24 h, whereas attachment to the other domains proved reversible and resulted in cell detachment or aggregation. In choice assays on patterned substrates, repulsive properties could be attributed to the EGF-type repeats TNegf, and to TNfnA1,2,4. Finally, neurite outgrowth promoting properties for E18 rat hippocampal neurons and P0 mouse DRG explants could be assigned to TNfnB,D, TNfnD,6, and TNfn6. The epitope of mAb J1/tn2 which abolishes the neurite outgrowth inducing effect of intact TN-C could be allocated to TNfnD. These observations suggest that TN-C harbors distinct cell-binding, repulsive, and neurite outgrowth promoting sites for neurons. Furthermore, the properties of isoform-specific TN-C domains suggest functional significance of the alternative splicing of TN-C glycoproteins.

scholarly journals Anti-Inflammatory Effects of Saccharomyces Boulardii on Myeloid Dendritic Cells From Patients With Crohn's Disease and Ulcerative Colitis

2011 ◽  
Vol 140 (5) ◽  
pp. S-849
Author(s):  
Saskia Thomas ◽  
Diana Metzke ◽  
Yvonne Dörffel ◽  
Jürgen Schmitz ◽  
Andreas Radbruch ◽  
...  
Keyword(s):  
Ulcerative Colitis ◽  
Dendritic Cells ◽  
Crohn’S Disease ◽  
Crohn's Disease ◽  
Saccharomyces Boulardii ◽  
Myeloid Dendritic Cells ◽  
Anti Inflammatory

Concerted action of tenascin-C domains in cell adhesion, anti-adhesion and promotion of neurite outgrowth

1997 ◽  
Vol 110 (13) ◽  
pp. 1513-1522 ◽  
Author(s):  
D. Fischer ◽  
M. Brown-Ludi ◽  
T. Schulthess ◽  
R. Chiquet-Ehrismann
Keyword(s):  
Cell Adhesion ◽  
Neurite Outgrowth ◽  
Molecular Characteristics ◽  
Concerted Action ◽  
New Approach ◽  
Tenascin C ◽  
Type Iii ◽  
Starting Point ◽  
Fibronectin Type Iii ◽  
Fibronectin Type

We used a new approach to identify domains of chicken tenascin-C required for interaction with cells. Instead of expressing the parts of interest, we deleted them from an otherwise intact tenascin-C molecule and scored for the concomitant change in activity. As a starting point for all mutant constructs we expressed the smallest naturally occurring tenascin-C splice variant in vertebrate cells. The tenascin-C mutants had either deletions of all EGF-like repeats, all fibronectin type III repeats or of the fibrinogen globe. In double mutants the fibronectin type III repeats were deleted together with either the EGF-like repeats or the fibrinogen globe, respectively. All tenascin-C variants assembled correctly to hexameric molecules of the expected molecular characteristics. Intact tenascin-C and the mutant missing the fibrinogen globe did not promote adhesion of chick embryo fibroblasts, whereas both, the hexamers containing solely the fibrinogen globe or the EGF-like repeats were adhesive substrates and even supported cell spreading. When tenascin-C was added to the medium of fibroblasts plated on fibronectin-coated wells, cell adhesion was blocked by intact tenascin-C, but not by mutants missing the fibrinogen globe. In neurite outgrowth assays using dorsal root ganglia, processes formed on all substrates except on the mutant missing only the fibrinogen globe, where the ganglia failed to adhere. The mutants missing the fibronectin type III repeats allowed more rapid neurite outgrowth than all other tenascin-C variants and the mutant consisting essentially of oligomerized EGF-like repeats was as active a substrate for neurite outgrowth as laminin. From the combined data, it is concluded that the activities of intact tenascin-C cannot be mimicked by investigating domain by domain, but the concerted action of several domains leads to the diverse cellular responses.

Anti-inflammatory effects ofSaccharomyces boulardiimediated by myeloid dendritic cells from patients with Crohn's disease and ulcerative colitis

2011 ◽  
Vol 301 (6) ◽  
pp. G1083-G1092 ◽  
Author(s):  
Saskia Thomas ◽  
Diana Metzke ◽  
Jürgen Schmitz ◽  
Yvonne Dörffel ◽  
Daniel C. Baumgart
Keyword(s):  
Ulcerative Colitis ◽  
Dendritic Cells ◽  
Crohn’S Disease ◽  
T Cell ◽  
Crohn's Disease ◽  
Transforming Growth Factor ◽  
Mucosal Healing ◽  
Cell Phenotype ◽  
Myeloid Dendritic Cells ◽  
Tnf Α

Saccharomyces boulardii ( Sb) is a probiotic yeast that has demonstrated efficacy in pilot studies in patients with inflammatory bowel disease (IBD). Microbial antigen handling by dendritic cells (DC) is believed to be of critical importance for immunity and tolerance in IBD. The aim was to characterize the effects of Sb on DC from IBD patients. Highly purified (>95%), lipopolysaccharide-stimulated CD1c+CD11c+CD123−myeloid DC (mDC) from patients with ulcerative colitis (UC; n = 36), Crohn's disease (CD; n = 26), or infectious controls (IC; n = 4) were cultured in the presence or absence of fungal supernatant from Sb ( SbS). Phenotype and cytokine production and/or secretion of IBD mDC were measured by flow cytometry and cytometric bead arrays, respectively. T cell phenotype and proliferation were assessed in a mixed lymphocyte reaction (MLR) with allogenic CD4+CD45RA+naïve T cells from healthy donors. Mucosal healing was investigated in epithelial wounding and migration assays with IEC-6 cells. SbS significantly decreased the frequency of CD40-, CD80-, and CD197 (CCR7; chemokine receptor-7)-expressing IBD mDC and reduced their secretion of tumor necrosis factor (TNF)-α and interleukin (IL)-6 while increasing IL-8. In the MLR, SbS significantly inhibited T cell proliferation induced by IBD mDC. Moreover, SbS inhibited TH1 (TNF-α and interferon-γ) polarization induced by UC mDC and promoted IL-8 and transforming growth factor-β-dependent mucosal healing. In summary, we provide novel evidence of synergistic mechanisms how Sb controls inflammation (inhibition of T cell costimulation and inflammation-associated migration and mobilization of DC) and promotes epithelial restitution relevant in IBD.

Analysis of combinatorial variability reveals selective accumulation of the fibronectin type III domains B and D of tenascin-C in injured brain

2010 ◽  
Vol 225 (1) ◽  
pp. 60-73 ◽  
Author(s):  
Alexandre Dobbertin ◽  
Stefan Czvitkovich ◽  
Ursula Theocharidis ◽  
Jeremy Garwood ◽  
Melissa R. Andrews ◽  
...  
Keyword(s):  
Tenascin C ◽  
Type Iii ◽  
Injured Brain ◽  
Fibronectin Type Iii ◽  
Selective Accumulation ◽  
Fibronectin Type

The glia-derived extracellular matrix glycoprotein tenascin-C promotes embryonic and postnatal retina axon outgrowth via the alternatively spliced fibronectin type III domain TNfnD

2008 ◽  
Vol 4 (4) ◽  
pp. 271-283 ◽  
Author(s):  
Sonia Siddiqui ◽  
Andrea Horvat-Bröcker ◽  
Andreas Faissner
Keyword(s):  
Extracellular Matrix ◽  
Neurite Outgrowth ◽  
Ganglion Cells ◽  
Axon Growth ◽  
Fc Fragment ◽  
Tenascin C ◽  
Type Iii ◽  
Fibronectin Type Iii ◽  
Fibronectin Type Iii Domain ◽  
Fibronectin Type

Tenascin-C (Tnc) is an astrocytic multifunctional extracellular matrix (ECM) glycoprotein that potentially promotes or inhibits neurite outgrowth. To investigate its possible functions for retinal development, explants from embryonic day 18 (E18) rat retinas were cultivated on culture substrates composed of poly-d-lysine (PDL), or PDL additionally coated with Tnc or laminin (LN)-1, which significantly increased fiber length. When combined with LN, Tnc induced axon fasciculation that reduced the apparent number of outgrowing fibers. In order to circumscribe the stimulatory region, Tnc-derived fibronectin type III (TNfn) domains fused to the human Ig-Fc-fragment TNfnD6-Fc, TNfnBD-Fc, TNFnA1A2-Fc and TNfnA1D-Fc were studied. The fusion proteins TNfnBD-Fc and to a lesser degree TNfnA1D-Fc were stimulatory when compared with the Ig-Fc-fragment protein without insert. In contrast, the combination TNfnA1A2-Fc reduced fiber outgrowth beneath the values obtained for the Ig-Fc domain, indicating potential inhibitory properties. The monoclonal J1/tn2 antibody (clone 578) that is specific for domain TNfnD blocked the stimulatory properties of the TNfn-Fc fusions. When postnatal day 7 retinal ganglion cells were used rather that explants, Tnc and Tnc-derived proteins proved permissive for neurite outgrowth. The present study highlights a strong retinal axon growth-promoting activity of the Tnc domain TNfnD, which is modulated by neighboring domains.

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