Evaluation of skin sensitization based on interleukin‑2 promoter activation in Jurkat cells

Purpose: Targeting programmed death-ligand 1 (PD-L1) may be an effective intervention for osteosarcoma and PD-L1 expression is controlled by diverse regulatory factors. Low-density lipoprotein receptor-related protein 8 (LRP8) regulates osteoblast differentiation and it is unclear whether and how LRP8 could contribute to osteosarcoma pathogenesis. In this study, we investigated the LRP8/signal transducer and activator of transcription 3 (STAT3)/PD-L1 network in osteosarcoma. Methods: The expression of LRP8, STAT3, and PD-L1 was measured in osteosarcoma tissues and paired normal tissues. The effects of LRP8 on STAT3 and PD-L1 expression were investigated in an osteosarcoma cell line. The effects on immunosuppression were investigated in an in vitro co-culture system with Jurkat cell line and osteosarcoma cell line. The effects of LRP8 were blocked by a LRP8 neutralizing antibody, dominant-negative STAT3, or STAT3 inhibitor. Results: LRP8 was overexpressed in osteosarcoma compared to normal tissues and its level was correlated with phospho-STAT3 (p-STAT3) level in osteosarcoma tissues. In osteosarcoma cell lines, LRP8 increased p-STAT3 level and promoted nuclear translocation of STAT3. STAT3 activation also increased PD-L1 mRNA, protein, and promoter activity. In addition, LRP8 enhanced PD-L1 expression via STAT3. In a co-culture system, LRP8 overexpression in an osteosarcoma cell line impaired viability and interleukin-2 secretion of Jurkat cells and induced apoptosis of Jurkat cells. The effects of LRP8 could be blocked by neutralizing LRP8 antibody or STAT3 inhibitor. Blocking LRP8 inhibits proliferation and induces apoptosis of osteosarcoma cells. Conclusions: Our results provide evidence for a novel regulation network of LRP8/STAT3/PD-L1 in osteosarcoma and LRP8 may be a potential therapeutic target in osteosarcoma.

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Stimulation of T cells through the CD3/T-cell receptor complex: role of cytoplasmic calcium, protein kinase C translocation, and phosphorylation of pp60c-src in the activation pathway.

Molecular and Cellular Biology ◽

10.1128/mcb.7.2.650 ◽

1987 ◽

Vol 7 (2) ◽

pp. 650-656 ◽

Cited By ~ 44

Author(s):

J A Ledbetter ◽

L E Gentry ◽

C H June ◽

P S Rabinovitch ◽

A F Purchio

Keyword(s):

T Cells ◽

T Cell ◽

T Cell Receptor ◽

Solid Phase ◽

Interleukin 2 ◽

Cell Receptor ◽

Jurkat Cells ◽

Receptor Complex ◽

Kinase C ◽

Stimulation Of

Stimulation of T cells or the Jurkat T-cell line with soluble antibodies to the CD3/T-cell receptor complex causes mobilization of cytoplasmic Ca2+, which is blocked by pertussis toxin but not by ethylene glycol-bis(beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid, and translocation of protein kinase C activity from the cytoplasm to the membrane. Such stimulation also causes phosphorylation of pp60c-src at an amino-terminal serine residue. These activities are consistent with induction of phosphatidylinositol metabolism after antibody binding. Anti-CD3 stimulation with antibody in solution, however, does not cause Jurkat cells to release interleukin 2 and blocks rather than induces proliferation of T cells. Induction of interleukin 2 production by Jurkat cells and proliferation by normal T cells requires anti-CD3 stimulation with antibody on a solid support, such as Sepharose beads or a plastic dish. Thus, we examined phosphorylation of pp60c-src after stimulation of Jurkat cells with anti-CD3 in solution or on solid phase. Both of these caused serine phosphorylation of pp60c-src that was indistinguishable even after 4 h of stimulation. These results indicate that the mode of anti-CD3 stimulation (in solution or on solid phase) controls a cellular function that modifies the consequences of signal transduction through phosphatidylinositol turnover.

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Signalling through CD28 T-cell activation pathway involves an inositol phospholipid-specific phospholipase C activity

Biochemical Journal ◽

10.1042/bj2930835 ◽

1993 ◽

Vol 293 (3) ◽

pp. 835-842 ◽

Cited By ~ 30

Author(s):

J Nunes ◽

S Klasen ◽

M D Franco ◽

C Lipcey ◽

C Mawas ◽

...

Keyword(s):

T Cell ◽

Phospholipase C ◽

T Cell Activation ◽

Cell Activation ◽

Interleukin 2 ◽

Jurkat Cells ◽

Cd28 Molecule ◽

Human T Cell ◽

Protein Kinase C Inhibitor ◽

Human T Cell Line

Stimulation of the human T-cell line, Jurkat, by a monoclonal antibody (mAb) directed against the CD28 molecule leads to sustained increases in intracellular levels of Ca2+ ([Ca2+]i); the initial rise in Ca2+ comes from internal stores, followed by Ca2+ entry into the cells. The CD28 molecule also appears to activate polyphosphoinositide (InsPL)-specific phospholipase C (PLC) activity in Jurkat cells, as demonstrated by PtdInsP2 breakdown, InsP3 and 1,2-diacylglycerol generation and PtdIns resynthesis. We also observed that interleukin-2 (IL2) production induced via CD28 triggering was sensitive to a selective protein kinase C inhibitor. Of the four other anti-CD28 mAbs (CD28.2, CD28.4, CD28.5, CD28.6) tested, only one (CD28.5) was unable to generate any InsPL-specific PLC or IL2 secretion. However, the cross-linking of cell-bound CD28.5 with anti-mouse Ig antibodies led to an increase in [Ca2+]i. CD28-molecule clustering in itself appears to be a sufficient signal for induction of PLC activity.

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Inhibition or activation of human T cell receptor transfectants is controlled by defined, soluble antigen arrays.

Journal of Experimental Medicine ◽

10.1084/jem.176.5.1421 ◽

1992 ◽

Vol 176 (5) ◽

pp. 1421-1430 ◽

Cited By ~ 34

Author(s):

D E Symer ◽

R Z Dintzis ◽

D J Diamond ◽

H M Dintzis

Keyword(s):

T Cells ◽

T Cell ◽

T Cell Receptor ◽

Interleukin 2 ◽

Cell Receptor ◽

Jurkat Cells ◽

Soluble Antigen ◽

Human T Cell ◽

Human T Cells ◽

Binding Avidity

We present evidence that direct T cell receptor (TCR) occupancy by antigen can either activate or inhibit T cells, depending upon whether or not a threshold number of local TCRs are crosslinked by multivalent arrays of the antigen. Variants of Jurkat cells were previously transfected with TCR alpha and beta chains that bind fluorescein, yielding FL-TCR+ human T cells. The transfectants are activated upon binding soluble multivalent antigen arrays at concentrations well below those required for monovalent interactions. This activation, measured by calcium fluxes and interleukin 2 (IL-2) production, indicates the superior binding avidity of multivalent ligands. Smaller, less multivalent arrays do not activate the cells, but antagonize larger arrays, demonstrating that antigen can bind TCR as either agonist or antagonist. The balance between activation and inhibition depends upon antigen array size, ligand valence, and concentration, indicating that a threshold extent of receptor crosslinking, and not individual perturbations of single TCR, is required for activation by antigen. Approximately 100 stimulatory arrays specifically bind per FL-TCR+ cell at concentrations where IL-2 production is half-maximal.

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Submicromolar La3+ concentrations block the calcium release-activated channel, and impair CD69 and CD25 expression in CD3- or thapsigargin-activated Jurkat cells

Biochemical Journal ◽

10.1042/bj3130909 ◽

1996 ◽

Vol 313 (3) ◽

pp. 909-913 ◽

Cited By ~ 56

Author(s):

Claude AUSSEL ◽

Rachid MARHABA ◽

Claudette PELASSY ◽

Jean-Philippe BREITTMAYER

Keyword(s):

T Cell ◽

T Cell Activation ◽

Cell Activation ◽

Calcium Release ◽

Cell Signalling ◽

Interleukin 2 ◽

Jurkat Cells ◽

T Cell Signalling ◽

A Chain ◽

20 Nm

The calcium release-activated channel (CRAC) opened in Jurkat cells activated either with CD3 monoclonal antibody or the endoplasmic reticulum Ca2+-ATPase blocker, thapsigargin, is blocked by La3+ with an IC50 of 20 nM. Similarly, the entry of Mn2+, used as a surrogate for Ca2+, is also blocked by submicromolar La3+ concentrations. La3+ seems to play its role simply by plugging the CRAC because this ion does not penetrate the cells, as demonstrated by chelation experiments with EGTA. Blocking the Ca2+ influx in activated Jurkat cells results in a lack of expression of CD25, a chain of the interleukin-2 receptor and of CD69, a marker of T-cell activation. By contrast, the very early steps of the T-cell signalling pathway such as the release of Ca2+ from intracellular stores and the subsequent inhibition of phosphatidylserine synthesis are not affected by La3+.

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Effects of modulators of cytosolic Ca2+ on phytohemagglutin-dependent Ca2+ response and interleukin-2 production in Jurkat cells

Journal of Leukocyte Biology ◽

10.1002/jlb.53.1.66 ◽

1993 ◽

Vol 53 (1) ◽

pp. 66-72 ◽

Cited By ~ 6

Author(s):

Gilles Dupuis ◽

Fawzi Aoudjit ◽

Isabelle Ricard ◽

Marcel D. Payet

Keyword(s):

Interleukin 2 ◽

Jurkat Cells

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A novel immunosuppressive factor in bovine colostrum blocks activation of the interleukin 2 gene enhancer at the NFAT site

Biochemistry and Cell Biology ◽

10.1139/o96-063 ◽

1996 ◽

Vol 74 (4) ◽

pp. 585-593 ◽

Cited By ~ 7

Author(s):

Damaraju Sambasivarao ◽

Jonathan Hooton ◽

Axel Dost ◽

Verner Paetkau

Keyword(s):

Interleukin 2 ◽

Leukemia Cell ◽

Calcium Ionophore ◽

Bovine Colostrum ◽

Jurkat Cells ◽

Leukemia Cell Line ◽

Luciferase Gene ◽

Gene Transcriptional Regulation ◽

Gene Enhancer ◽

Similar Mode

A factor in bovine colostrum (colostrum inhibitory factor, CIF) inhibits interleukin 2 (IL2) production in activated T helper cells by blocking the accumulation of IL2 mRNA. To determine whether CIF blocks at the level of IL2 transcription, we introduced reporter plasmids into the human T leukemia cell line Jurkat by transient transfection. These contained the luciferase gene under the control of either the human IL2 upstream enhancer region (segments −326 to +45) or three repeats of the NFAT element contained within it (segments −255 to −285). Expression of luciferase in these cells was induced by phorbol myristate acetate plus a calcium ionophore. CIF inhibited induction of either construct as did cyclosporine, which is known to block activation of the NFAT element. CIF failed to inhibit several other enhancer elements. The NFAT-controlled luciferase gene system distinguishes CIF from other T cell inhibitory activities present in colostrum, in particular, TGFβ1 and TGFβ2 and the glucocorticoids. Stably transfected Jurkat cells behaved similarly to the transiently transfected ones with respect to inhibition by CIF and cyclosporine. The NFAT-luc assay is a useful technique for the rapid, sensitive measurement of CIF or other immunosuppressants with a similar mode of action.Key words: immunosuppression, cyclosporine, NFAT, reporter gene, transcriptional regulation.

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Prolonged high glucose suppresses phorbol 12-myristate 13-acetate and ionomycin-induced interleukin-2 mRNA expression in Jurkat cells

Biochimica et Biophysica Acta (BBA) - General Subjects ◽

10.1016/j.bbagen.2008.10.002 ◽

2009 ◽

Vol 1790 (1) ◽

pp. 8-15 ◽

Cited By ~ 6

Author(s):

Koji Higai ◽

Masatoshi Tsukada ◽

Yumiko Moriya ◽

Yutaro Azuma ◽

Kojiro Matsumoto

Keyword(s):

Mrna Expression ◽

High Glucose ◽

Interleukin 2 ◽

Jurkat Cells

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A yeast-based screening system identified bakkenolide B contained in Petasites japonicus as an inhibitor of interleukin-2 production in a human T cell line

Bioscience Biotechnology and Biochemistry ◽

10.1093/bbb/zbab130 ◽

2021 ◽

Author(s):

Shota Uesugi ◽

Mayuka Hakozaki ◽

Yuko Kanno ◽

Honoka Takahashi ◽

Yui Kudo ◽

...

Keyword(s):

T Cell ◽

Cell Line ◽

Screening Method ◽

Interleukin 2 ◽

Jurkat Cells ◽

Yeast Cells ◽

Wild Plant ◽

Human T Cell ◽

Petasites Japonicus ◽

Human T Cell Line

Abstract Ca2+ signaling is related to various diseases such as allergies, diabetes, and cancer. We explored Ca2+ signaling inhibitors in natural resources using a yeast-based screening method, and found bakkenolide B from the flower buds of edible wild plant, Petasites japonicus, using the YNS17 strain (zds1Δ erg3Δ pdr1/3Δ). Bakkenolide B exhibited growth-restoring activity against the YNS17 strain and induced Li+ sensitivity of wild-type yeast cells, suggesting that it inhibits the calcineurin pathway. Additionally, bakkenolide B inhibited interleukin-2 production at gene and protein levels in Jurkat cells, a human T cell line, but not the in vitro phosphatase activity of human recombinant calcineurin, an upstream regulator of interleukin-2 production. Furthermore, bakkenolide A showed weak activity in YNS17 and Jurkat cells compared with bakkenolide B. These findings revealed new biological effects and the structure-activity relationships of bakkenolides contained in Petasites japonicus as inhibitors of interleukin-2 production in human T cells.

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