Interaction of metargidin (ADAM-15) with alphavbeta3 and alpha5beta1 integrins on different haemopoietic cells

1999 ◽  
Vol 112 (4) ◽  
pp. 579-587 ◽  
Author(s):  
D. Nath ◽  
P.M. Slocombe ◽  
P.E. Stephens ◽  
A. Warn ◽  
G.R. Hutchinson ◽  
...  
Keyword(s):  
Cell Adhesion ◽  
Cell Line ◽  
Cell Lines ◽  
Solid Phase ◽  
Phase Cell ◽  
Cell Binding ◽  
Amino Terminal ◽  
Disintegrin Domain ◽  
Haemopoietic Cells ◽  
Adhesion Assays

Metargidin (ADAM-15) is a type I transmembrane glycoprotein belonging to the ADAM (A Disintegrin and Metalloprotease Domain) family of proteins and is widely expressed in different tissues and cell types. Members of this family contain an amino-terminal metalloprotease domain followed by a disintegrin domain, a cysteine-rich region and a membrane proximal EGF-like domain. The disintegrin domain of metargidin contains an RGD tripeptide sequence, suggesting that it may potentially interact with the integrin family of proteins. Here we identify integrin ligands for metargidin on haemopoietic cells, by using a chimeric protein containing the extracellular domain of metargidin fused to the Fc portion of human IgG. Binding activity to a panel of human cell lines was analysed by solid-phase cell-adhesion assays. Metargidin bound to a monocytic cell line, U937, and a T cell line, MOLT-4, in a specific manner. Adhesion was divalent cation- and temperature- dependent and strongly enhanced by Mn2+, all features of integrin-mediated binding. Using a panel of anti-integrin antibodies we show that alphavbeta3 is a ligand for metargidin on U937 cells. In contrast, for MOLT-4 cells, the integrin alpha5beta1 contributes to cell binding. Adhesion was mediated by the disintegrin domain of metargidin as RGD-based peptides inhibited cell binding to both cell lines. The specificity of the interaction between both alphavbeta3 and alpha5beta1 and metargidin was further confirmed by solid-phase adhesion assays using purified recombinant integrins. These results together indicate that metargidin can function as a cell adhesion molecule via interactions with alphavbeta3 and alpha5beta1 integrins.

Effects of polystyrene surface chemistry on the biological activity of solid phase fibronectin and vitronectin, analysed with monoclonal antibodies

1993 ◽  
Vol 104 (3) ◽  
pp. 793-803 ◽  
Author(s):  
P.A. Underwood ◽  
J.G. Steele ◽  
B.A. Dalton
Keyword(s):  
Monoclonal Antibodies ◽  
Cell Adhesion ◽  
Solid Phase ◽  
Divalent Cations ◽  
Biological Activities ◽  
Optimal Growth ◽  
Cell Types ◽  
Cell Binding ◽  
Urea Treatment ◽  
Bhk Cells

The conformation and biological activities of fibronectin (FN) and vitronectin (VN) coated on different plastic surfaces were investigated using cell adhesion and a panel of domain-specific monoclonal antibodies (mAbs). The adhesion of BHK fibroblasts was markedly better on FN coated on hydrophilic tissue culture polystyrene (TCPS) than on hydrophobic, untreated polystyrene (PS). mAbs A17 and 3E3, which inhibit the binding of BHK cells to the RGD-containing sequence within the cell binding region of FN, also bound preferentially to FN on TCPS. In contrast, two anti-FN mAbs, which have no effect on cell adhesion (A35 and A3), bound preferentially to the conformation of FN on the more hydrophobic PS. Mouse melanoma cells utilise an additional cell-binding site in the Hep II domain of FN and their preference for FN coated on TCPS was less marked than that of BHK cells. This reduced preference was again mimicked by the binding of a mAb, A32, which inhibits the binding of B16 cells to the Hep II domain of FN. In contrast, BHK cell adhesion to VN did not display a preference for TCPS over PS. The cell-binding activity of adsorbed VN was matched by the binding of a cell adhesion-inhibitory mAb, A18, which, unlike mAbs A17 and A32, displayed slightly increased binding to VN coated on PS, rather than TCPS. When the denaturating effect of coating FN and VN to PS in the presence of urea was investigated, similar correlations between BHK cell adhesion and the binding of inhibitory mAbs were observed. Urea treatment of FN significantly reduced both BHK cell adhesion and the binding of both cell adhesion-inhibitory mAbs, whereas the binding of A35 and A3 was unaffected. There was no significant effect of urea treatment of VN upon either BHK cell adhesion or mAb binding. A larger panel of anti-FN mAbs was used, together with the anti-VN mAbs, to determine whether there were differences in mAb recognition of FN and VN adsorbed on three different brands of TCPS. The mAbs segregated into four reactivity patterns, of which A17, A32, A35 and A18 respectively were representative. Significant differences were observed in mAb recognition of FN and VN adsorbed to different brands of TCPS. These may reflect differences in the ability of these surfaces to support optimal growth of different cell types. The effect of divalent cations upon adsorbed FN and VN was also investigated.(ABSTRACT TRUNCATED AT 400 WORDS)

Outside-In Signaling of Soluble and Solid-Phase Fibrinogen Through Integrin ΙΙbβ3 Is Different and Cooperative With Each Other in a Megakaryoblastic Leukemia Cell Line, CMK

Blood ◽  
1998 ◽  
Vol 92 (4) ◽  
pp. 1277-1286 ◽  
Author(s):  
Yumi Tohyama ◽  
Kaoru Tohyama ◽  
Misao Tsubokawa ◽  
Momoyo Asahi ◽  
Yataro Yoshida ◽  
...  
Keyword(s):  
Cell Adhesion ◽  
Signaling Pathways ◽  
Cell Line ◽  
Tyrosine Phosphorylation ◽  
Solid Phase ◽  
Leukemia Cell ◽  
Leukemia Cell Line ◽  
Megakaryoblastic Leukemia ◽  
Integrin Β3 ◽  
Coated Plate

Abstract The function and the outside-in signaling pathways of IIbβ3 were examined in relation to cell adhesion using a megakaryoblastic leukemia cell line, CMK. After 12-O-tetradecanoylphorbol-13-acetate (TPA) treatment, the cells adhered to the culture plate and underwent megakaryocytic differentiation with expression of IIbβ3. Binding of soluble fibrinogen to the cells via IIbβ3 was dependent on cell adhesion. Cell detaching reduced the affinity of this integrin for soluble fibrinogen, although its surface expression was almost unchanged. In contrast, detached cells became tightly adherent to the fibrinogen-coated plate (solid-phase fibrinogen). The same ligand, fibrinogen, present either in soluble or solid-phase form, triggered differential signaling pathways mediated by IIbβ3. By the stimulation with soluble fibrinogen, Syk was tyrosine-phosphorylated but FAK was dephosphorylated, whereas solid-phase fibrinogen promptly caused tyrosine phosphorylation of FAK followed by delayed phosphorylation of Syk. In addition, the binding of soluble fibrinogen to the cells adherent to fibrinogen-coated plate resulted in tyrosine phosphorylation of integrin β3 and a complex formation of integrin β3 with Syk. This implies the cooperation of both soluble and solid-phase fibrinogen-mediated signaling pathways. © 1998 by The American Society of Hematology.

scholarly journals A colony-stimulating factor 1 (CSF-1) receptor/platelet-derived growth factor-beta receptor gene fusion confers CSF-1 independence and tumorigenicity on a c-myc-immortalized monocyte cell line.

1992 ◽  
Vol 12 (1) ◽  
pp. 386-393
Author(s):  
M R Eccles ◽  
F J King ◽  
M D Cole
Keyword(s):  
Growth Factor ◽  
Tumor Cell ◽  
Cell Line ◽  
Cell Lines ◽  
Tumor Cell Line ◽  
Platelet Derived Growth Factor ◽  
Colony Stimulating Factor ◽  
Amino Terminal ◽  
Monocyte Cell ◽  
Stimulating Factor

Monocytes and macrophages express the receptor for the hematopoietic growth factor colony-stimulating factor 1 (CSF-1) and require this factor for growth in culture. A murine monocyte tumor cell line that lacks the usual requirement for CSF-1 was isolated. On the basis of the similarity of the structures of the CSF-1 and platelet-derived growth factor (PDGF) receptors and because monocytes normally secrete PDGF, we analyzed the tumor cell line for anomalous expression of the PDGF-R beta gene. Two different cDNAs that each contain sequences corresponding to the complete coding sequence of PDGF-R beta fused (in frame) to the amino-terminal half of the CSF-1 receptor were isolated. Introduction of these PDGF-R beta-related cDNAs into two partially transformed, CSF-1-dependent monocyte cell lines resulted in autonomous growth and cell transformation. These monocyte cell lines exhibit a novel form of growth factor receptor activation that can lead to oncogenic growth in collaboration with the c-myc oncogene.

scholarly journals Adhesive properties of carcinoembryonic antigen glycoforms expressed in glycosylation-deficient Chinese hamster ovary cell lines.

2002 ◽  
Vol 49 (1) ◽  
pp. 273-283 ◽  
Author(s):  
Anna Krop-Watorek ◽  
Arkadiusz G Klopocki ◽  
Marcin Czerwinski ◽  
Elwira Lisowska
Keyword(s):  
Cell Adhesion ◽  
Carcinoembryonic Antigen ◽  
Chinese Hamster Ovary ◽  
Solid Phase ◽  
Chinese Hamster ◽  
Ovary Cell ◽  
Phase Cell ◽  
Adhesion Assay ◽  
Cell Surface Glycoprotein ◽  
Polypeptide Structure

Carcinoembryonic antigen (CEA) is an oncofoetal cell surface glycoprotein that serves as an important tumour marker for colorectal and some other carcinomas. Its immunoglobulin-like structure places CEA within the immunoglobulin superfamily. CEA functions in several biological roles including homotypic and heterotypic (with other CEA family members) cell adhesion. Cell-cell interaction can be modulated by different factors, e.g., post-translational modifications such as glycosylation. The purpose of this study was to examine whether changes in carbohydrate composition of CEA oligosaccharides can influence homotypic (CEA-CEA) interactions. In order to modulate glycosylation of CEA we used two different glycosylation mutants of Chinese hamster ovary (CHO) cells, Lec2 and Lec8. Lec2 cells should produce CEA with nonsialylated N-glycans, while Lec8 cells should yield more truncated sugar structures than Lec2. Parental CHO (Pro5) cells and the glycosylation deficient mutants were stably transfected with CEA cDNA. All three CEA glycoforms, tested in a solid-phase cell adhesion assay, showed an ability to mediate CEA-dependent cell adhesion, and no qualitative differences in the adhesion between the glycoforms were observed. Thus, it may be assumed that carbohydrates do not play a role in homotypic adhesion, and the interactions between CEA molecules depend solely on the polypeptide structure.

A colony-stimulating factor 1 (CSF-1) receptor/platelet-derived growth factor-beta receptor gene fusion confers CSF-1 independence and tumorigenicity on a c-myc-immortalized monocyte cell line

1992 ◽  
Vol 12 (1) ◽  
pp. 386-393
Author(s):  
M R Eccles ◽  
F J King ◽  
M D Cole
Keyword(s):  
Growth Factor ◽  
Tumor Cell ◽  
Cell Line ◽  
Cell Lines ◽  
Tumor Cell Line ◽  
Platelet Derived Growth Factor ◽  
Colony Stimulating Factor ◽  
Amino Terminal ◽  
Monocyte Cell ◽  
Stimulating Factor

Monocytes and macrophages express the receptor for the hematopoietic growth factor colony-stimulating factor 1 (CSF-1) and require this factor for growth in culture. A murine monocyte tumor cell line that lacks the usual requirement for CSF-1 was isolated. On the basis of the similarity of the structures of the CSF-1 and platelet-derived growth factor (PDGF) receptors and because monocytes normally secrete PDGF, we analyzed the tumor cell line for anomalous expression of the PDGF-R beta gene. Two different cDNAs that each contain sequences corresponding to the complete coding sequence of PDGF-R beta fused (in frame) to the amino-terminal half of the CSF-1 receptor were isolated. Introduction of these PDGF-R beta-related cDNAs into two partially transformed, CSF-1-dependent monocyte cell lines resulted in autonomous growth and cell transformation. These monocyte cell lines exhibit a novel form of growth factor receptor activation that can lead to oncogenic growth in collaboration with the c-myc oncogene.

scholarly journals Influence of Zika Virus in mechanisms related of cytotoxicity, cell adhesion, apoptosis and inflammatory markers on glioblastoma cells

2021 ◽  
Author(s):  
Daniel Marinowic ◽  
Fabiana Spillari Viola ◽  
Fernanda Majolo ◽  
Gabriele Goulart Zanirati ◽  
Pamella Nunes Azevedo ◽  
...  
Keyword(s):  
Cell Migration ◽  
Cell Adhesion ◽  
Cell Viability ◽  
Cell Line ◽  
Cell Lines ◽  
Zika Virus ◽  
Adhesion Behavior ◽  
Induction Of Apoptosis ◽  
Cd73 Expression ◽  
Potential Use

Abstract Glioblastoma (GBM) is one of the most common brain tumors in adults. Despite the presence of available treatments, it remains one of the most lethal and difficult tumors to treat such that most patients die within two years. Studies reported that infection with Zika virus (ZIKV) causes inhibition of cell proliferation as well as induction of apoptosis; moreover, these manifestations show a predilection for developing neuronal cells. In the present study, two GBM cell lines U-138 and U-251 were infected with ZIKV at multiplicities of infection (MOI) 0.1, 0,01 and 0.001 and tested for cell viability, cell migration, cell adhesion, induction of apoptosis, interleukin levels, and cell surface markers (CD14 and CD73). Our study demonstrated that the ZIKV infection promotes loss of cell viability and increased apoptosis potential. It was not evidenced changes in cell migration, however, the two glioblastoma cell lines displayed increased the cell adhesion behavior. There was small increase in the IL-4 level in the U-251 cell line after exposure to ZIKV, with no change in relation to INF-γ levels. Furthermore, we observed an increase in the percentage of cells expressing the CD14 surface marker in both cell lines and increased CD73 expression in the U-251 cell line. Our results suggest that ZIKV may be associated with decrease of cell viability and increased CD73 expression, enhanced adherence, as well as increased apoptosis rates. Further investigations are required to explore the potential use of ZIKV in the treatment of GBM.

scholarly journals Chemical characterization (LC–MS–ESI), cytotoxic activity and intracellular localization of PAMAM G4 in leukemia cells

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
R. Flores-Mejía ◽  
M. J. Fragoso-Vázquez ◽  
L. G. Pérez-Blas ◽  
A. Parra-Barrera ◽  
S. S. Hernández-Castro ◽  
...  
Keyword(s):  
Tumor Cell ◽  
Cell Line ◽  
Cell Lines ◽  
Biological Effects ◽  
Intracellular Localization ◽  
Chemical Characterization ◽  
Tumor Cell Line ◽  
Exact Mass ◽  
Globular Structure ◽  
Amino Terminal

AbstractGeneration 4 of polyamidoamine dendrimer (G4-PAMAM) has several biological effects due to its tridimensional globular structure, repetitive branched amides, tertiary amines, and amino-terminal subunit groups liked to a common core. G4-PAMAM is cytotoxic due to its positive charges. However, its cytotoxicity could increase in cancer cells due to the excessive intracellular negative charges in these cells. Furthermore, this work reports G4-PAMAM chemical structural characterization using UHPLC-QTOF-MS/MS (LC–MS) by electrospray ionization to measure its population according to its positive charges. Additionally, the antiproliferative effects and intracellular localization were explored in the HMC-1 and K-562 cell lines by confocal microscopy. The LC–MS results show that G4-PAMAM generated multivalent mass spectrum values, and its protonated terminal amino groups produced numerous positive charges, which allowed us to determine its exact mass despite having a high molecular weight. Additionally, G4-PAMAM showed antiproliferative activity in the HMC-1 tumor cell line after 24 h (IC50 = 16.97 µM), 48 h (IC50 = 7.02 µM) and 72 h (IC50 = 5.98 µM) and in the K-562 cell line after 24 h (IC50 = 15.14 µM), 48 h (IC50 = 14.18 µM) and 72 h (IC50 = 9.91 µM). Finally, our results showed that the G4-PAMAM dendrimers were located in the cytoplasm and nucleus in both tumor cell lines studied.

scholarly journals ADAM disintegrin-like domain recognition by the lymphocyte integrins α4β1 and α4β7

2005 ◽  
Vol 387 (1) ◽  
pp. 101-108 ◽  
Author(s):  
Lance C. BRIDGES ◽  
Dean SHEPPARD ◽  
Ron D. BOWDITCH
Keyword(s):  
Monoclonal Antibody ◽  
Cell Adhesion ◽  
Jurkat Cell ◽  
Cell Attachment ◽  
Human Lymphocytes ◽  
Specific Region ◽  
Binding Properties ◽  
Disintegrin Domain ◽  
Dependent Cell ◽  
Adhesion Assays

The ADAM (adisintegrin and metalloprotease) family of proteins possess both proteolytic and adhesive domains. We have established previously that the disintegrin domain of ADAM28, an ADAM expressed by human lymphocytes, is recognized by the integrin α4β1. The present study characterizes the integrin binding properties of the disintegrin-like domains of human ADAM7, ADAM28 and ADAM33 with the integrins α4β1, α4β7 and α9β1. Cell-adhesion assays demonstrated that, similar to ADAM28, the ADAM7 disintegrin domain supported α4β1-dependent Jurkat cell adhesion, whereas the ADAM33 disintegrin domain did not. The lymphocyte integrin α4β7 was also found to recognize both disintegrin domains of ADAM7 and ADAM28, but not of ADAM33. This is the first demonstration that mammalian disintegrins are capable of interacting with α4β7. All three disintegrin domains supported α9β1-dependent cell adhesion. Recognition by both α4β1 and α4β7 of ADAM7 and ADAM28 was activation-dependent, requiring either the presence of Mn2+ or an activating monoclonal antibody for cell attachment. Charge-to-alanine mutagenesis experiments revealed that the same residues within an individual ADAM disintegrin domain function in recognizing multiple integrins. However, the residues within a specific region of each ADAM disintegrin-like domain required for integrin binding were distinct. These results establish that ADAM7 and ADAM28 are recognized by the leucocyte integrins α4β1, α4β7 and α9β1. ADAM33 exclusively supported only α9β1-dependent adhesion.

Expression of the cell–cell adhesion molecule E-cadherin in tongue carcinoma cell lines

1994 ◽  
Vol 108 (11) ◽  
pp. 957-961 ◽  
Author(s):  
A. R. Kinsella ◽  
G. L. Bowie ◽  
J. K. Fields ◽  
A. S. Jones
Keyword(s):  
Cell Adhesion ◽  
Cell Line ◽  
Cell Lines ◽  
Adhesion Molecule ◽  
Cell Adhesion Molecule ◽  
Carcinoma Cell ◽  
Tumour Progression ◽  
Tongue Carcinoma ◽  
Carcinoma Cell Lines ◽  
E Cadherin

AbstractA reduction in cell adhesiveness and cell invasion are essential steps in tumour progression. In the present study six tongue carcinoma cell lines were compared with regard to their invasive potential in two in vitro invasion assay systems and for their patterns of expression of the cell–cell adhesion molecule E-cadherin. The three cell lines negative for E-cadherin expression were invasive in both assays. One cell line with strong E-cadherin expression was strongly invasive and one weakly invasive. One cell line with reduced E-cadherin expression was weakly invasive. There was no significant pattern to these findings (x2 = 0.375; p = 0.54). This supports previous studies from this group that suggest that E-cadherin is only one of the presumably many molecules involved in tumour progression in squamous cell carcinoma of the tongue.

Hypoxia induces lymphocyte adhesion to human mesenchymal cells via an LFA-1-dependent mechanism

1993 ◽  
Vol 264 (3) ◽  
pp. C617-C624 ◽  
Author(s):  
I. Ginis ◽  
S. J. Mentzer ◽  
D. V. Faller
Keyword(s):  
Endothelial Cells ◽  
Cell Adhesion ◽  
Cell Line ◽  
Cell Lines ◽  
Muscle Cell ◽  
Mesenchymal Cells ◽  
Peripheral Blood Lymphocytes ◽  
Lymphoid Cells ◽  
Lymphocyte Cell ◽  
Lymphocyte Cell Line

We and others have previously reported that mesenchymal cells, including endothelial and muscle cells, sense oxygen tension and respond in a specific way during exposure to hypoxic environment. We have examined the interactions of human muscle and endothelial cells, which have been exposed to hypoxic environments, with T and B lymphoid cell lines and peripheral blood lymphocytes (PBL), not subjected to hypoxia. The adhesion of B lymphocyte cell line (JY) and the adhesion of T lymphocyte cell line (Jurkat) to muscle cell monolayers that had been incubated at PO2 of 50 Torr for 3 h increased more than four- and twofold, respectively. Hypoxia appears to upregulate a saturable muscle cell-associated adhesion mechanism, which is capable of withstanding distraction forces greater than 45 g, and is inhibitable by LFA-1-specific monoclonal antibodies (MAbs). Hypoxia also induced a reciprocal decrease in lymphocyte-muscle cell adhesion mechanisms inhibitable by VCAM-1- or VLA-4-specific MAbs. Cultured human endothelial cells when subjected to hypoxic conditions also increased their adhesion for lymphoid cells and cell lines. This induction of adhesion could again be attenuated by anti-LFA-1, but not by anti-ICAM-1 MAb, suggesting that hypoxia activates an adhesion molecule on human mesenchymal cells that is likely to be a new ligand for LFA-1. This report is the first demonstration of a direct induction of cell adhesion mechanisms by hypoxic environments.

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