scholarly journals Distinction between gamma c detection and function in YT lymphoid cells and in the granulocyte-macrophage colony-stimulating factor-responsive human myeloid cell line, Tf-1

Blood ◽  
1995 ◽  
Vol 86 (12) ◽  
pp. 4568-4578 ◽  
Author(s):  
NL Farner ◽  
SD Voss ◽  
TP Leary ◽  
J Gan ◽  
J Hakimi ◽  
...  
Keyword(s):  
Cell Line ◽  
Interleukin 2 ◽  
Cell Lineage ◽  
Myeloid Cell ◽  
Lymphoid Cells ◽  
Blood Monocytes ◽  
Cellular Environment ◽  
Myeloid Development ◽  
C Subunit ◽  
And Function

Peripheral blood monocytes respond to interleukin-2 (lL-2) and express the gamma common (gamma c) subunit of the lL-2 receptor (lL-2R) complex. However, the role of lL-2 in myeloid development has recently become of interest for several reasons, including the effect gamma c mutations may or may not have on myeloid development in patients with XSCID. Many studies of lL-2 function in the myeloid cell lineage have been performed on a murine background. To study gamma c expression and function in human myeloid precursors, we introduced the human myelomonocytic cell line, Tf-1, with a retroviral vector containing the human lL-2R beta subunit to create functional human intermediate lL-2R consisting of beta gamma c dimers. We have characterized this transfected variant of Tf-1 (Tf-1 beta) with regard to its response to lL-2. Unlike the parental Tf-1 cell line that is deficient in both lL- 2R alpha and lL-2R beta expression, the Tf-1 beta transfectant binds and responds to lL-2 through intermediate-affinity lL-2Rs. Scatchard analyses indicate the number of intermediate-affinity receptors on Tf-1 beta is similar to the number found on the well-characterized YT cell line. However, detection of gamma c on Tf-1 beta cells is dramatically less than on YT cells by Western blot analysis and is undetectable by flow cytometric studies and surface iodinations. The gamma c component on YT cells is readily detected by all three methods. We conclude from these studies that the intermediate-affinity lL-2Rs on the Tf-1 cell line behave differently than those on YT cells with respect to gamma c detection. Either the gamma c molecule itself is different, or the cellular environment in which it functions is altered. Elucidation of gamma c function on this cell line will allow for its use as a model in which other cytokines using gamma c (including lL-2, lL-4, and lL-15) can be studied on the same cellular background.

CD300f Triggering Modifies Myeloid Cell Function

Blood ◽  
2008 ◽  
Vol 112 (11) ◽  
pp. 1255-1255
Author(s):  
Derek Hart ◽  
Courtney Modra ◽  
Georgina Clark
Keyword(s):  
Cell Line ◽  
Cell Lines ◽  
Peripheral Blood ◽  
Cell Function ◽  
Myeloid Cell ◽  
Significant Inhibition ◽  
Blood Monocytes ◽  
Peripheral Blood Monocytes ◽  
Monocyte Migration ◽  
Phosphoinositide 3 Kinase

Abstract CD300f, a member of the CD300 family of immunoregulatory molecules, is capable of signalling through association with both SHP-1 phosphatase and the p85α subunit of phosphoinositide 3-kinase. On normal cells, CD300f is expressed on monocytes and dendritic cells in the blood and bone marrow. CD300f is expressed on myeloid derived cell lines and Acute Myeloid Leukaemias (AMLs) and is an acknowledged candidate for antibody targeting of AML (Modra CJ et al. Blood2006;108:225B-225B and Zhao X et al Blood2007;110:531a–532a). We have generated a monoclonal antibody, MMRI- 23, specific for the extracellular domain of CD300f. MMRI-23 immunoprecipitates a 57kd protein from the myeloid derived cell lines HEL, THP-1 and U937 and this protein binds to a polyclonal antibody to CD300f (LMIR3) in Western blot analysis. Binding of MMRI-23 to myeloid cells was blocked by the CD300f recombinant proteins or the polyclonal CD300f antibody. The MMRI-23 mAb was used as a surrogate ligand to study the functional consequences of crosslinking CD300f on normal monocytes and myeloid derived cell lines. Purified peripheral blood monocytes were cultured for 18 hours in the presence of immobilised MMRI-23 or control mAb. MMRI-23 binding was not altered by activation of peripheral blood monocytes but crosslinking monocytes with MMRI- 23 induced downregulation of CD14 and CD33. There was significant inhibition of IL-6 but not IL-1β, IL-8 or TNFα secretion. Crosslinking monocytes or the U937 cell line with MMRI-23 increased specific chemotaxis towards CXCL12 (SDF-1). This increased migration index following MMRI-23 crosslinking was reversed in the presence of wortmannin indicating that MMRI-23 induces signalling through phosphoinositide 3-kinase. The effect of crosslinking did not enhance CXCR4 upregulation but did induce localization of CD300f and CXCR4 to the lipid rafts in myeloid cell line, U937. Thus CD300f plays a significant role in the regulation of monocyte migration to CXCR4. As CD300f is upregulated on around 70% of AMLs, this regulation of homing has major implications for the treatment of AML.

Production of laminin B2 chain protein and messenger RNA by a murine neutrophil precursor cell line, 32Dc13 [see comments]

Blood ◽  
1990 ◽  
Vol 76 (7) ◽  
pp. 1398-1404 ◽  
Author(s):  
DJ Tweardy ◽  
M Sasaki ◽  
JJ Jr Cardamone ◽  
JP Jr McCoy ◽  
MJ Bonidie ◽  
...  
Keyword(s):  
Cell Surface ◽  
Cell Line ◽  
Messenger Rna ◽  
Precursor Cell ◽  
Laminin Receptors ◽  
Precursor Cell Line ◽  
Myeloid Development ◽  
Exogenous Origin ◽  
And Function ◽  
Murine Neutrophil

Laminin is a heterotrimeric glycoprotein that plays a central role in promoting neutrophil chemotaxis, motility, and attachment to basement membrane. Rabbit peritoneal exudate neutrophils stain positively for laminin, which is presumed to be of exogenous origin and bound to laminin receptors on the cell surface. We examined 32Dc13 cells, a murine neutrophil precursor cell line, by immunoprecipitation. Northern blot analysis, flow cytometry, and electron microscopy for the endogenous production of laminin. Our results demonstrate that 32Dc13 cells endogenously produce a laminin B2 chain protein and messenger RNA (mRNA) without producing any detectable A or B1 chain protein or mRNA. The B2 chain protein was not secreted by the cells; rather it could be detected on the cell surface after treatment of cells with neuraminidase. These findings suggest the possibility of a novel role for the laminin B2 chain in myeloid development and function.

scholarly journals Establishment and characterization of a human mixed-lineage, T- lymphoid/myeloid cell line (USP-91)

Blood ◽  
1993 ◽  
Vol 82 (6) ◽  
pp. 1829-1837 ◽  
Author(s):  
T Umiel ◽  
CW Rettenmier ◽  
S Siegel ◽  
S Ota ◽  
H Shimada ◽  
...  
Keyword(s):  
T Cell ◽  
Cell Line ◽  
Phorbol Esters ◽  
Myeloid Cell ◽  
Mononuclear Phagocytes ◽  
Lymphoid Cells ◽  
Cell Surface Receptor ◽  
Initial Growth ◽  
Monocytic Differentiation ◽  
Biopsy Material

Abstract We report the establishment of a novel cell line from a pediatric patient with recurrent non-Hodgkin's lymphoma. This cell line, termed USP-91, showed both T-lymphoid cell as well as myeloid (ie, nonlymphoid) cell characteristics using a comprehensive multiparameter approach. The initial growth of this cell line was dependent on the presence of the murine stromal cell line, 14F1.1. Subsequently, a phenotypically stable, stroma-independent cell line was established. Although the recurrent biopsy material and the derivative cell line, USP-91, were clonally-derived from T-lineage lymphoid cells, as evidenced by the same rearrangement of the T-cell receptor-beta locus, USP-91 coexpressed both the T-cell antigens CD7, CD3, and CD4, and the myeloid antigens CD13, CD33, CD11b, and CD34. The myeloid features of USP-91 were most consistent with monocytic differentiation as these cells expressed alpha-napthol acetate esterase, lysozyme, alpha-1- antitrypsin, alpha-1-antichymotrypsin, as well as the cell surface receptor for macrophage colony-stimulating factor. In addition, incubation in the presence of phorbol esters induced USP-91 to exhibit morphologic and functional properties of mature mononuclear phagocytes. The expression of this bilineage phenotype suggests that USP-91 represents the malignant transformation of a progenitor cell capable of either myelomonocytic or T-lymphoid differentiation.

Abelson virus transformation of an interleukin 2-dependent antigen-specific T-cell line

1987 ◽  
Vol 7 (7) ◽  
pp. 2631-2635
Author(s):  
W D Cook ◽  
B Fazekas de St Groth ◽  
J F Miller ◽  
H R MacDonald ◽  
R Gabathuler
Keyword(s):  
Tyrosine Kinase ◽  
Cell Line ◽  
Short Circuit ◽  
Interleukin 2 ◽  
Lymphoid Cells ◽  
Specific Cell ◽  
Colony Stimulating Factor ◽  
Macrophage Colony Stimulating Factor ◽  
Macrophage Colony ◽  
Stimulating Factor

Abelson murine leukemia virus (A-MuLV) carries the gene v-abl, one of a group of oncogenes with structural and functional (tyrosine kinase) homology to three growth factor receptors. Work in this and other laboratories has shown that A-MuLV infection can render myeloid and lymphoid cells independent of the growth factors interleukin 3 and granulocyte-macrophage colony-stimulating factor. We have now shown that v-abl can also relieve interleukin 2 (IL-2) dependence in T cells. We infected a cloned IL-2-dependent antigen-specific cell line. Transformed cells were generated which were factor independent and tumorigenic. The transformants each bore unique v-abl DNA inserts and expressed v-abl mRNA. No elevation of expression of either IL-2 or its receptor could be detected in these cells. Thus, A-MuLV can short-circuit the dependence of hematopoietic cells on IL-2, IL-3, and possibly granulocyte-macrophage colony-stimulating factor, none of whose receptors are known to be of the tyrosine kinase type.

Induction of allopeptide-specific human CD4+CD25+ regulatory T cells ex vivo

Blood ◽  
2003 ◽  
Vol 102 (6) ◽  
pp. 2180-2186 ◽  
Author(s):  
Shuiping Jiang ◽  
Niels Camara ◽  
Giovanna Lombardi ◽  
Robert I. Lechler
Keyword(s):  
T Cells ◽  
T Cell ◽  
Regulatory T Cells ◽  
Cell Line ◽  
Ex Vivo ◽  
Interleukin 2 ◽  
Cell Lineage ◽  
Human Leukocyte ◽  
Regulatory Cells ◽  
Antigen Specificity

Abstract Although CD4+CD25+ regulatory T cells are pivotal in the prevention of autoimmunity and appear to mediate transplantation tolerance, little is known concerning their antigen specificity. Here we describe the induction of a human CD4+CD25+ regulatory T-cell line specific for a defined peptide alloantigen (human leukocyte antigen A2 [HLA-A2] 138-170) by priming purified CD4+CD25+ cells ex vivo. The regulatory cells were anergic and retained their ability to suppress antigen-driven responses of CD4+CD25– cells. They inhibited not only interleukin 2 (IL-2) secretion by CD4+CD25– T cells specific for the same peptide but also direct alloresponse of naive CD4+CD25– T cells stimulated by semiallogeneic dendritic cells (DCs) in the presence of the peptide (“linked suppression”). They also suppressed the response of CD4+ T cells specific for viral and bacterial antigens. The suppressive T-cell line showed sustained high CD25 expression. These findings suggest that peripheral CD4+CD25+ regulatory cells are a precommitted cell lineage from which cells with specificity for non–self-peptides can be selected. This may pave the way for inducing and expanding peptide antigen-specific regulatory T cells ex vivo for cell therapy in transplantation, allergy, and autoimmune disease.

BCR/ABL alters the function of NK cells and the acquisition of killer immunoglobulin-like receptors (KIRs)

Blood ◽  
2003 ◽  
Vol 101 (9) ◽  
pp. 3527-3533 ◽  
Author(s):  
Elena G. Chiorean ◽  
Scott J. Dylla ◽  
Krista Olsen ◽  
Todd Lenvik ◽  
Yvette Soignier ◽  
...  
Keyword(s):  
Nk Cells ◽  
Cell Line ◽  
Kinase Inhibitor ◽  
Blast Crisis ◽  
Interleukin 2 ◽  
Chronic Phase ◽  
Lymphoid Cells ◽  
Myelogenous Leukemia ◽  
Parental Cell Line ◽  
Natural Cytotoxicity

Natural killer (NK) cells decrease in function during chronic myelogenous leukemia (CML) progression from chronic phase to blast crisis, and they can becomeBCR/ABL+ late in the disease course. To study this altered function, NK92 cells were transduced with the BCR/ABL oncogene. In contrast to the parental cells, which died when deprived of interleukin 2 (IL-2), p210+ NK92 cells proliferated and survived indefinitely in the absence of IL-2. BCR/ABL also decreased the natural cytotoxicity of NK92 cells against K562 targets, without affecting IL-2, interferon γ (IFN-γ), or tumor necrosis factor α (TNF-α) production. Although the ABL-specific tyrosine kinase inhibitor imatinib mesylate (STI-571) had no effect on parental NK92 cells, it markedly decreased the growth and survival of IL-2–independent p210+ NK92 cells. In contrast to the parental cell line, serial analysis of p210+ NK92 cells detected small populations that clonally expressed one or more killer immunoglobulin-like receptors (KIRs). Unlike the decreased natural cytotoxicity, the function of the activating CD158j receptor remained intact. Southern blotting and hybridization with an enhanced green fluorescence protein (eGFP) probe showed that KIR− and KIR+ NK92 cells were all derived from the same clone, suggesting that KIR acquisition remains dynamic at the maturational stage represented by the NK92 cell line. When tested in primary CD56+bright NK cells, p210 induced partial IL-2–independent growth and increased KIR expression similar to findings in NK92 cells. This is the first study to show thatBCR/ABL, well known for its effects on the myeloid lineage, can alter the function of lymphoid cells, which may be associated with the defect in innate immunity associated with CML progression.

scholarly journals Production of laminin B2 chain protein and messenger RNA by a murine neutrophil precursor cell line, 32Dc13 [see comments]

Blood ◽  
1990 ◽  
Vol 76 (7) ◽  
pp. 1398-1404
Author(s):  
DJ Tweardy ◽  
M Sasaki ◽  
JJ Jr Cardamone ◽  
JP Jr McCoy ◽  
MJ Bonidie ◽  
...  
Keyword(s):  
Cell Surface ◽  
Cell Line ◽  
Messenger Rna ◽  
Precursor Cell ◽  
Laminin Receptors ◽  
Precursor Cell Line ◽  
Myeloid Development ◽  
Exogenous Origin ◽  
And Function ◽  
Murine Neutrophil

Abstract Laminin is a heterotrimeric glycoprotein that plays a central role in promoting neutrophil chemotaxis, motility, and attachment to basement membrane. Rabbit peritoneal exudate neutrophils stain positively for laminin, which is presumed to be of exogenous origin and bound to laminin receptors on the cell surface. We examined 32Dc13 cells, a murine neutrophil precursor cell line, by immunoprecipitation. Northern blot analysis, flow cytometry, and electron microscopy for the endogenous production of laminin. Our results demonstrate that 32Dc13 cells endogenously produce a laminin B2 chain protein and messenger RNA (mRNA) without producing any detectable A or B1 chain protein or mRNA. The B2 chain protein was not secreted by the cells; rather it could be detected on the cell surface after treatment of cells with neuraminidase. These findings suggest the possibility of a novel role for the laminin B2 chain in myeloid development and function.

scholarly journals Regulated expression and function of CD122 (interleukin-2/interleukin- 15R-beta) during lymphoid development

Blood ◽  
1996 ◽  
Vol 87 (1) ◽  
pp. 190-201 ◽  
Author(s):  
T Reya ◽  
JA Yang-Snyder ◽  
EV Rothenberg ◽  
SR Carding
Keyword(s):  
B Cells ◽  
Fetal Liver ◽  
Interleukin 2 ◽  
Lymphoid Cells ◽  
Lymphocyte Development ◽  
Hematopoietic Stem ◽  
Embryo Sections ◽  
And Function ◽  
Receptor Beta

To determine whether signaling via CD122 (interleukin-2 [IL-2]/IL-15 receptor beta-chain) plays a role in regulating the expansion and differentiation of lymphocyte precursors, we have characterized its expression and evaluated its ability to influence the activity of developing lymphoid cells. A significant fraction of Sca1+Lin- hematopoietic stem cells in day 12 fetal liver were found to be CD122+. CD122-mRNA+ and IL-2-mRNA+ cells were also localized in embryo sections within pharyngeal blood vessels adjacent to and surrounding the thymic analgen. This distribution is consistent with the migration of CD122+ progenitor cells from the liver to the developing thymus where a majority of Sca1+ intrathymic T-cell progenitors were CD122+. Analysis of CD122 expression in the day 12 fetal liver revealed that the majority of B220+ cells were CD122+. Furthermore, CD122 expression was restricted to the earliest B220+ cells (CD43+CD24-; prepro B cells; fraction A) that proliferate vigorously to IL-2 in the absence of any stromal cells, but not to IL-15. Consistent with a role for the IL-2/IL- 2R pathway in lymphocyte development is the progressive loss of B cells seen in IL-2-deficient mice. Together, these observations suggest that CD122 plays a role in regulating normal lymphocyte development in vivo.

scholarly journals Bipotential cell differentiation of KU-812: evidence of a hybrid cell line that differentiates into basophils and macrophage-like cells

Blood ◽  
1987 ◽  
Vol 70 (3) ◽  
pp. 612-619
Author(s):  
T Fukuda ◽  
K Kishi ◽  
Y Ohnishi ◽  
A Shibata
Keyword(s):  
Cell Line ◽  
Hybrid Cell ◽  
Philadelphia Chromosome ◽  
Cell Lineage ◽  
Myeloid Cell ◽  
Culture Conditions ◽  
Cloning Efficiency ◽  
Hla Dr ◽  
F Cells ◽  
Cytological Features

KU-812-F is a subclone of KU-812, which has cytological features similar to the parent clone, and which carries the Philadelphia chromosome. We studied the effects of various chemical agents and serum- free culture (SF-C) condition on its differentiation. The KU-812-F subclone differentiated into macrophage-like cells with phorbol myristate acetate (PMA), but not with other agents. Unexpectedly, under SF-C, KU-812-F cells differentiated into mature basophil-like cells, and the histamine content of cell lysates increased in proportion to maturation. The addition of the condensed supernatant of SF-C promoted the cloning efficiency of KU-812-F cells in semisolid SF-C, whereas the cloning efficiency was reduced in SF-C alone. KU-812-F cells expressed myelomonocytic antigens. Additionally, My4 was induced with PMA and SF- C, but less often. HLA-DR was not expressed in any culture conditions, despite blastic morphology of KU-812-F cells or macrophage-like cells induced with PMA. The IgE receptor could not be demonstrated under any conditions. Correspondingly, immature and mature KU-812-F cells did not respond to anti-IgE or concanavalin A (con A), while histamine release was induced with PMA. In conclusion, KU-812-F belongs to the myeloid cell lineage and is at least a bipotential cell that can differentiate into basophils and macrophage-like cells. Although the functions appear to be dissimilar to those of normal basophils, the KU-812-F cell line may be a good model for basophil differentiation. Moreover, KU-812-F cells may provide new insights because they grow in semisolid culture by an autostimulating mechanism.

scholarly journals Phenotypic changes induced by interleukin-2 (IL-2) and IL-3 in an immature T-lymphocytic leukemia are associated with regulated expression of IL-2 receptor beta chain and of protein tyrosine kinases LCK and LYN

Blood ◽  
1992 ◽  
Vol 80 (4) ◽  
pp. 1017-1025
Author(s):  
R O'Connor ◽  
T Torigoe ◽  
JC Reed ◽  
D Santoli
Keyword(s):  
Cell Line ◽  
Cell Lines ◽  
Tyrosine Kinases ◽  
Lymphoblastic Leukemia ◽  
Interleukin 2 ◽  
Myeloid Cell ◽  
Lymphocytic Leukemia ◽  
Beta Chain ◽  
Protein Tyrosine ◽  
Phenotypic Changes

We have previously reported the establishment of an interleukin-3 (IL- 3)-dependent and phenotypically myeloid cell line (TALL-103/3), obtained by culturing cells from an immature T-lymphoblastic leukemia in the presence of IL-3. These cells differentiated into a T-lymphoid cell line (TALL-103/2) upon removal of IL-3 and incubation in IL-2. Despite the different phenotype, the two cell lines remained karyotypically and genotypically identical. Here, we have analyzed the phenotypic changes and the signaling events induced by these two lymphokines in TALL-103/3 cells by switching them to temporary growth in IL-2 and returning them to IL-3. All four sublines obtained (the myeloid in IL-3 and the lymphoid in IL-2) expressed RNA for CD3, IL-2 receptor (R) alpha, and T-cell receptor (TCR)-gamma and -delta chains. However, cells cultured in IL-3 failed to express detectable levels of the IL-2R beta chain at both the protein and RNA levels, whereas cells exposed to IL-2 always expressed IL-2R beta. In parallel with the changes in IL-2R beta expression, the SRC-like protein tyrosine kinase (PTK) p56 LCK could not be detected in IL-3-dependent cells, but was abundant in the IL-2-dependent cells and underwent markedly increased autophosphorylation in response to IL-2. In contrast, p53/p56 LYN was highly expressed in IL-3-dependent cells, and greatly decreased when these cells were switched to growth in IL-2. LYN kinase autophosphorylation modestly increased in response to IL-3. None of the other kinases in the SRC family that were tested underwent increased autophosphorylation after lymphokine stimulation, indicating the specificity of IL-2 for LCK and of IL-3 for LYN. The TALL-103 cell lines provide a unique system to study the interaction between lymphokines and SRC-family PTKs in signal transduction pathways leading to hematopoietic cell differentiation.

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