Microtubule depolymerization inhibits clathrin coated-pit internalization in non-adherent cell lines while interleukin 2 endocytosis is not affected

1997 ◽  
Vol 110 (19) ◽  
pp. 2441-2447 ◽  
Author(s):  
A. Subtil ◽  
A. Dautry-Varsat
Keyword(s):  
Cell Lines ◽  
Interleukin 2 ◽  
Lymphoid Cells ◽  
Membrane Receptors ◽  
Adherent Cell ◽  
Transferrin Receptors ◽  
Hemopoietic Cell ◽  
Microtubule Depolymerization ◽  
Microtubule Cytoskeleton ◽  
Microtubule Disruption

The microtubule cytoskeleton is generally not considered to be essential for the first steps of clathrin-mediated endocytosis of membrane receptors. Its role in clathrin-independent endocytosis has not been investigated. We have previously shown that the cytokine interleukin 2 (IL2) is internalized in lymphoid cells expressing its receptors when clathrin-dependent endocytosis is inhibited. Here we compare the internalization of IL2 and of transferrin, a marker of clathrin-dependent endocytosis, after microtubule disruption. In hemopoietic cell lines, which express IL2 receptors, transferrin receptor entry was inhibited by about 40%. However, in adherent cell lines, transferrin entry was unaffected by microtubule disruption, as previously reported. Unlike the case for transferrin, internalization of IL2 receptors was not affected by depolymerization of the microtubule cytoskeleton in hemopoietic cell lines. These results show that IL2 and transferrin receptors do not have the same endocytic properties and support our previous conclusion that these receptors follow different pathways of endocytosis.

scholarly journals Specific adsorption of HTLV-I to various target human and animal cells

Blood ◽  
1987 ◽  
Vol 70 (5) ◽  
pp. 1303-1311 ◽  
Author(s):  
K Krichbaum-Stenger ◽  
BJ Poiesz ◽  
P Keller ◽  
G Ehrlich ◽  
J Gavalchin ◽  
...  
Keyword(s):  
Cell Lines ◽  
Mononuclear Cells ◽  
Cultured Cells ◽  
Interleukin 2 ◽  
Flow Cytometric Analysis ◽  
Lymphoid Cells ◽  
Core Antigen ◽  
Target Cells ◽  
Animal Cells ◽  
Homologous Virus

In this report, we describe a flow cytometric analysis of HTLV-I specific binding to fresh and cultured cells on a single cell basis. This assay uses rhodamine hydrocarbon tagged, purified HTLV-I virions according to the procedure originally described for avian retroviruses. Successful HTLV-I transmission was detected by analysis of integrated HTLV-I DNA, virion-associated reverse transcriptase, and/or intracellular HTLV-I core antigen p19 expression. Only a specific virus- cell interaction was detected because nonrhodamine-tagged homologous virus or related HTLV-II interfered with tagged HTLV-I binding. In contrast, an unrelated, nonlabeled animal retrovirus was unable to block tagged HTLV binding. Of the cell lines tested, 2 nonlymphoid mammalian and 3 human lymphoid bound significantly high to moderate levels of HTLV-I-tagged virions. The other three human lymphocyte cell lines were insensitive to HTLV-I adsorption. A direct correlation was observed between HTLV-I binding sites and infectivity of human lymphoid cells alone and not other nonlymphoid animal cells. Fresh normal human mononuclear cells bound low levels of HTLV-I virions. As expected, T lymphocytes demonstrated more binding than did the non-T cell population. Enhancement of HTLV-I cell binding in a subpopulation of mononuclear target cells was achieved with phytohemagglutinin (PHA) activation and interleukin 2 (IL2) stimulation, which correlates well with previously published infectivity studies.

scholarly journals CD2 Rescues T Cells From T-Cell Receptor/CD3 Apoptosis: A Role for the Fas/Fas-L System

Blood ◽  
1997 ◽  
Vol 89 (10) ◽  
pp. 3717-3726 ◽  
Author(s):  
Emira Ayroldi ◽  
Graziella Migliorati ◽  
Lorenza Cannarile ◽  
Rosalba Moraca ◽  
Domenico V. Delfino ◽  
...  
Keyword(s):  
T Cell ◽  
Fas Ligand ◽  
Cell Clone ◽  
Costimulatory Molecule ◽  
Interleukin 2 ◽  
Experimental System ◽  
Lymphoid Cells ◽  
Membrane Receptors ◽  
Peripheral Tolerance ◽  
Fas L

Abstract Anti-CD3 monoclonal antibodies (MoAbs) and glucocorticoid hormones induce apoptosis in immature thymocytes and peripheral T lymphocytes. This process is inhibited by a number of growth factors, including interleukin-2 (IL-2), IL-3, and IL-4, as well as by triggering of the adhesion molecule CD44, which would indicate that signals generated by membrane receptors can modulate the survival of lymphoid cells. To investigate whether triggering of CD2 may also affect apoptosis in lymphoid cells, we analyzed the effect of stimu-lation with anti-CD2 MoAbs on T-cell apoptosis induced by two stimuli, anti-CD3 MoAbs and dexamethasone (DEX), using a hybridoma T-cell line and a T-helper cell clone. The results show that CD2 engagement decreased anti-CD3 MoAb-induced apoptosis, but did not influence DEX-induced cell death. Furthermore, the decrease appeared to be related to the expression of Fas/APO-1 (CD95) and Fas-ligand (Fas-L). In fact, we show that CD2 stimulation inhibits apoptosis by preventing the CD3-induced upregulation of Fas and Fas-L in a Fas-dependent experimental system. These data suggest that a costimulatory molecule may control a deletion pathway and may therefore contribute to the regulation of peripheral tolerance.

scholarly journals Fas transduces activation signals in normal human T lymphocytes.

1993 ◽  
Vol 178 (6) ◽  
pp. 2231-2235 ◽  
Author(s):  
M R Alderson ◽  
R J Armitage ◽  
E Maraskovsky ◽  
T W Tough ◽  
E Roux ◽  
...  
Keyword(s):  
T Cells ◽  
T Cell ◽  
Cell Lines ◽  
Interleukin 2 ◽  
Tumor Necrosis Factor Receptor ◽  
Lymphoid Cells ◽  
Culture Vessel ◽  
Cell Surface Molecule ◽  
Transformed Cell ◽  
Transformed Cell Lines

The Fas gene encodes a cell surface molecule that is a member of the the nerve growth factor/tumor necrosis factor receptor family of proteins and can mediate programmed cell death (apoptosis) in certain transformed cell lines. To characterize further the biological function of Fas, particularly with regard to its function in normal cells, a panel of monoclonal antibodies (mAbs) was generated against the extracellular portion of human Fas. Some of these mAbs induced apoptosis in transformed cell lines expressing Fas, but only when immobilized on the culture vessel. One of the new Fas mAbs (M38) was used for studies on normal lymphoid cells and found to stimulate the proliferation of purified human T cells and thymocytes when immobilized on culture wells along with CD3 antibody. T cell proliferation induced by Fas mAb was largely interleukin 2 independent and was demonstrated to be due to a direct effect on the precursor T cell. Thus, the data demonstrate that in addition to a role in the induction of apoptosis in certain transformed cell lines, the Fas protein may also play an important role in the activation and proliferation of normal T cells.

scholarly journals Specific adsorption of HTLV-I to various target human and animal cells

Blood ◽  
1987 ◽  
Vol 70 (5) ◽  
pp. 1303-1311 ◽  
Author(s):  
K Krichbaum-Stenger ◽  
BJ Poiesz ◽  
P Keller ◽  
G Ehrlich ◽  
J Gavalchin ◽  
...  
Keyword(s):  
Cell Lines ◽  
Mononuclear Cells ◽  
Cultured Cells ◽  
Interleukin 2 ◽  
Flow Cytometric Analysis ◽  
Lymphoid Cells ◽  
Core Antigen ◽  
Target Cells ◽  
Animal Cells ◽  
Homologous Virus

Abstract In this report, we describe a flow cytometric analysis of HTLV-I specific binding to fresh and cultured cells on a single cell basis. This assay uses rhodamine hydrocarbon tagged, purified HTLV-I virions according to the procedure originally described for avian retroviruses. Successful HTLV-I transmission was detected by analysis of integrated HTLV-I DNA, virion-associated reverse transcriptase, and/or intracellular HTLV-I core antigen p19 expression. Only a specific virus- cell interaction was detected because nonrhodamine-tagged homologous virus or related HTLV-II interfered with tagged HTLV-I binding. In contrast, an unrelated, nonlabeled animal retrovirus was unable to block tagged HTLV binding. Of the cell lines tested, 2 nonlymphoid mammalian and 3 human lymphoid bound significantly high to moderate levels of HTLV-I-tagged virions. The other three human lymphocyte cell lines were insensitive to HTLV-I adsorption. A direct correlation was observed between HTLV-I binding sites and infectivity of human lymphoid cells alone and not other nonlymphoid animal cells. Fresh normal human mononuclear cells bound low levels of HTLV-I virions. As expected, T lymphocytes demonstrated more binding than did the non-T cell population. Enhancement of HTLV-I cell binding in a subpopulation of mononuclear target cells was achieved with phytohemagglutinin (PHA) activation and interleukin 2 (IL2) stimulation, which correlates well with previously published infectivity studies.

Jagged2 promotes the development of natural killer cells and the establishment of functional natural killer cell lines

Blood ◽  
2005 ◽  
Vol 105 (9) ◽  
pp. 3521-3527 ◽  
Author(s):  
Sarah L. DeHart ◽  
Marc J. Heikens ◽  
Schickwann Tsai
Keyword(s):  
Natural Killer Cells ◽  
Nk Cells ◽  
Natural Killer ◽  
Cell Lines ◽  
Nk Cell ◽  
Killer Cell ◽  
Interleukin 2 ◽  
Lymphoid Cells ◽  
Killer Cells ◽  
Hematopoietic Stem

AbstractEmerging evidence indicates that Notch receptors and their ligands play important roles in the development of T cells and B cells. However, little is known about their possible roles in the development of other lymphoid cells. Here we demonstrate that Jagged2, a Notch ligand, stimulates the development of natural killer (NK) cells from Lin- Sca-1+ c-kit+ hematopoietic stem cells. Our culture system supports NK cell development for 2 to 3 months, often leading to the establishment of continuous NK cell lines. The prototype of such cell lines is designated as KIL. KIL depends on interleukin-7 for survival and proliferation and is NK1.1+ CD3- TCRαβ- TCRδγ- CD4- CD8- CD19- CD25+ CD43+ CD45+ CD49b- CD51+ CD94+ NKG2D+ Mac-1-/low B220- c-kit+ perforin I+ granzyme B+ Notch-1+, and cytotoxic. Like normal natural killer cells, the T-cell receptor-β loci of KIL remain in the germ-line configuration. In response to interleukin-2, KIL proliferates extensively (increasing cell number by approximately 1010-fold) and terminally differentiates into adherent, hypergranular NK cells. Our findings indicate that Jagged2 stimulates the development of natural killer cells and the KIL cell line preserves most properties of the normal NK precursors. As such, KIL provides a valuable model system for NK cell research.

scholarly journals Both Sphingomyelin and Cholesterol in the Host Cell Membrane Are Essential for Rubella Virus Entry

2017 ◽  
Vol 92 (1) ◽  
Author(s):  
Noriyuki Otsuki ◽  
Masafumi Sakata ◽  
Kyoko Saito ◽  
Kiyoko Okamoto ◽  
Yoshio Mori ◽  
...  
Keyword(s):  
Cell Lines ◽  
Rubella Virus ◽  
Binding Sites ◽  
Lymphoid Cells ◽  
Congenital Rubella Syndrome ◽  
Adherent Cell ◽  
Independent Manner ◽  
Neutral Ph ◽  
Congenital Rubella ◽  
E1 Protein

ABSTRACTRubella virus (RuV) causes a systemic infection, and transplacental fetal infection causes congenital rubella syndrome. In this study, we showed that treatment of cells with sphingomyelinase inhibited RuV infection. Assays using inhibitors of serine palmitoyl transferase and ceramide transport protein demonstrated the contribution of sphingomyelin (SM) to RuV infection. Compelling evidence for direct binding of RuV to lipid membranes at neutral pH was obtained using liposome coflotation assays. The absence of either SM or cholesterol (Chol) abrogated the RuV-liposome interaction. SM and Chol (SM/Chol) were also critical for RuV binding to erythrocytes and lymphoid cells. Removal of Ca2+from the assay buffer or mutation of RuV envelope E1 protein Ca2+-binding sites abrogated RuV binding to liposomes, erythrocytes, and lymphoid cells. However, RuV bound to various nonlymphoid adherent cell lines independently of extracellular Ca2+or SM/Chol. Even in these adherent cell lines, both the E1 protein Ca2+-binding sites and cellular SM/Chol were essential for the early stage of RuV infection, possibly affecting envelope-membrane fusion in acidic compartments. Myelin oligodendrocyte glycoprotein (MOG) has recently been identified as a cellular receptor for RuV. However, RuV bound to MOG-negative cells in a Ca2+-independent manner. Collectively, our data demonstrate that RuV has two distinct binding mechanisms: one is Ca2+dependent and the other is Ca2+independent. Ca2+-dependent binding observed in lymphoid cells occurs by the direct interaction between E1 protein fusion loops and SM/Chol-enriched membranes. Clarification of the mechanism of Ca2+-independent RuV binding is an important next step in understanding the pathology of RuV infection.IMPORTANCERubella has a significant impact on public health as infection during early pregnancy can result in babies being born with congenital rubella syndrome. Even though effective rubella vaccines are available, rubella outbreaks still occur in many countries. We studied the entry mechanism of rubella virus (RuV) and found that RuV binds directly to the host plasma membrane in the presence of Ca2+at neutral pH. This Ca2+-dependent binding is specifically directed to membranes enriched in sphingomyelin and cholesterol and is critical for RuV infection. Importantly, RuV also binds to many cell lines in a Ca2+-independent manner. An unidentified RuV receptor(s) is involved in this Ca2+-independent binding. We believe that the data presented here may aid the development of the first anti-RuV drug.

scholarly journals CD44 (Pgp-1) inhibits CD3 and dexamethasone-induced apoptosis

Blood ◽  
1995 ◽  
Vol 86 (7) ◽  
pp. 2672-2678 ◽  
Author(s):  
E Ayroldi ◽  
L Cannarile ◽  
G Migliorati ◽  
A Bartoli ◽  
I Nicoletti ◽  
...  
Keyword(s):  
T Cell ◽  
Interleukin 2 ◽  
Lymphoid Cells ◽  
Membrane Receptors ◽  
Receptor Stimulation ◽  
Similar Activity ◽  
T Cell Apoptosis ◽  
Induced Apoptosis ◽  
T Cell Survival ◽  
Immature Thymocytes

Abstract Anti-CD3 monoclonal antibodies (MoAbs) and glucocorticoid hormones (GCH) induce apoptosis in immature thymocytes and peripheral T lymphocytes. This process is inhibited by a number of growth factors, including interleukin-2 (IL-2), IL-3, and IL-4, indicating that signals generated by membrane receptors can modulate the survival of lymphoid cells. To investigate whether signals activated by adhesion receptors have a similar activity, we analyzed the effect of CD44 (Pgp-1) adhesion molecule receptor stimulation on T-cell apoptosis induced by three stimuli (anti-CD3 MoAbs, dexamethasone [DEX] treatment, and exposure to ultraviolet irradiation [UV]) on a 3DO T-cell line. The results show that CD44 engagement, either by hyaluronic acid (HA) or anti-CD44 MoAbs, inhibits DNA fragmentation and apoptosis induced by DEX and anti-CD3 MoAbs, whereas that induced by UV, a p53-dependent phenomenon, was not inhibited. Furthermore, the antiapoptotic effect exerted through CD44 activation does not seem related to overexpression of bcl-2 or to have appreciable effects on cell proliferation. Our results indicate that adhesion molecules modulate T-cell survival by counteracting apoptosis induced by DEX or anti-CD3 MoAbs.

scholarly journals CD44 (Pgp-1) inhibits CD3 and dexamethasone-induced apoptosis

Blood ◽  
1995 ◽  
Vol 86 (7) ◽  
pp. 2672-2678 ◽  
Author(s):  
E Ayroldi ◽  
L Cannarile ◽  
G Migliorati ◽  
A Bartoli ◽  
I Nicoletti ◽  
...  
Keyword(s):  
T Cell ◽  
Interleukin 2 ◽  
Lymphoid Cells ◽  
Membrane Receptors ◽  
Receptor Stimulation ◽  
Similar Activity ◽  
T Cell Apoptosis ◽  
Induced Apoptosis ◽  
T Cell Survival ◽  
Immature Thymocytes

Anti-CD3 monoclonal antibodies (MoAbs) and glucocorticoid hormones (GCH) induce apoptosis in immature thymocytes and peripheral T lymphocytes. This process is inhibited by a number of growth factors, including interleukin-2 (IL-2), IL-3, and IL-4, indicating that signals generated by membrane receptors can modulate the survival of lymphoid cells. To investigate whether signals activated by adhesion receptors have a similar activity, we analyzed the effect of CD44 (Pgp-1) adhesion molecule receptor stimulation on T-cell apoptosis induced by three stimuli (anti-CD3 MoAbs, dexamethasone [DEX] treatment, and exposure to ultraviolet irradiation [UV]) on a 3DO T-cell line. The results show that CD44 engagement, either by hyaluronic acid (HA) or anti-CD44 MoAbs, inhibits DNA fragmentation and apoptosis induced by DEX and anti-CD3 MoAbs, whereas that induced by UV, a p53-dependent phenomenon, was not inhibited. Furthermore, the antiapoptotic effect exerted through CD44 activation does not seem related to overexpression of bcl-2 or to have appreciable effects on cell proliferation. Our results indicate that adhesion molecules modulate T-cell survival by counteracting apoptosis induced by DEX or anti-CD3 MoAbs.

scholarly journals CD2 Rescues T Cells From T-Cell Receptor/CD3 Apoptosis: A Role for the Fas/Fas-L System

Blood ◽  
1997 ◽  
Vol 89 (10) ◽  
pp. 3717-3726 ◽  
Author(s):  
Emira Ayroldi ◽  
Graziella Migliorati ◽  
Lorenza Cannarile ◽  
Rosalba Moraca ◽  
Domenico V. Delfino ◽  
...  
Keyword(s):  
T Cell ◽  
Fas Ligand ◽  
Cell Clone ◽  
Costimulatory Molecule ◽  
Interleukin 2 ◽  
Experimental System ◽  
Lymphoid Cells ◽  
Membrane Receptors ◽  
Peripheral Tolerance ◽  
Fas L

Anti-CD3 monoclonal antibodies (MoAbs) and glucocorticoid hormones induce apoptosis in immature thymocytes and peripheral T lymphocytes. This process is inhibited by a number of growth factors, including interleukin-2 (IL-2), IL-3, and IL-4, as well as by triggering of the adhesion molecule CD44, which would indicate that signals generated by membrane receptors can modulate the survival of lymphoid cells. To investigate whether triggering of CD2 may also affect apoptosis in lymphoid cells, we analyzed the effect of stimu-lation with anti-CD2 MoAbs on T-cell apoptosis induced by two stimuli, anti-CD3 MoAbs and dexamethasone (DEX), using a hybridoma T-cell line and a T-helper cell clone. The results show that CD2 engagement decreased anti-CD3 MoAb-induced apoptosis, but did not influence DEX-induced cell death. Furthermore, the decrease appeared to be related to the expression of Fas/APO-1 (CD95) and Fas-ligand (Fas-L). In fact, we show that CD2 stimulation inhibits apoptosis by preventing the CD3-induced upregulation of Fas and Fas-L in a Fas-dependent experimental system. These data suggest that a costimulatory molecule may control a deletion pathway and may therefore contribute to the regulation of peripheral tolerance.

scholarly journals Chitosan-Coated Gold Nanoparticles Induce Low Cytotoxicity and Low ROS Production in Primary Leucocytes, Independent of Their Proliferative Status

Pharmaceutics ◽  
2021 ◽  
Vol 13 (7) ◽  
pp. 942
Author(s):  
Helen Yarimet Lorenzo-Anota ◽  
Diana G. Zarate-Triviño ◽  
Jorge Alberto Uribe-Echeverría ◽  
Andrea Ávila-Ávila ◽  
José Raúl Rangel-López ◽  
...  
Keyword(s):  
Gold Nanoparticles ◽  
Cell Death ◽  
Cancer Cells ◽  
Cell Lines ◽  
Mononuclear Cells ◽  
Bone Marrow Cells ◽  
Lymphoid Cells ◽  
Ros Production ◽  
Biomedical Sciences ◽  
Peripheral Blood Mononuclear

(1) Background: Chitosan-coated gold nanoparticles (CH-AuNPs) have important theranostic applications in biomedical sciences, including cancer research. However, although cell cytotoxicity has been studied in cancerous cells, little is known about their effect in proliferating primary leukocytes. Here, we assessed the effect of CH-AuNPs and the implication of ROS on non-cancerous endothelial and fibroblast cell lines and in proliferative lymphoid cells. (2) Methods: The Turkevich method was used to synthetize gold nanoparticles. We tested cell viability, cell death, ROS production, and cell cycle in primary lymphoid cells, compared with non-cancer and cancer cell lines. Concanavalin A (ConA) or lipopolysaccharide (LPS) were used to induce proliferation on lymphoid cells. (3) Results: CH-AuNPs presented high cytotoxicity and ROS production against cancer cells compared to non-cancer cells; they also induced a different pattern of ROS production in peripheral blood mononuclear cells (PBMCs). No significant cell-death difference was found in PBMCs, splenic mononuclear cells, and bone marrow cells (BMC) with or without a proliferative stimuli. (4) Conclusions: Taken together, our results highlight the selectivity of CH-AuNPs to cancer cells, discarding a consistent cytotoxicity upon proliferative cells including endothelial, fibroblast, and lymphoid cells, and suggest their application in cancer treatment without affecting immune cells.

Sign in / Sign up

Export Citation Format

Share Document