scholarly journals Costimulation by B7-1 and LFA-3 targets distinct nuclear factors that bind to the interleukin-2 promoter: B7-1 negatively regulates LFA-3-induced NF-AT DNA binding.

1997 ◽  
Vol 17 (3) ◽  
pp. 1314-1323 ◽  
Author(s):  
E Parra ◽  
M Varga ◽  
G Hedlund ◽  
T Kalland ◽  
M Dohlsten
Keyword(s):  
T Cells ◽  
Dna Binding ◽  
Transcriptional Activity ◽  
Interleukin 2 ◽  
Luciferase Reporter ◽  
Jurkat T Cells ◽  
Response Elements ◽  
Nuclear Factors ◽  
Hla Dr ◽  
Nf Kappab

We have characterized the regulation of nuclear factors involved in transcriptional control of the interleukin-2 (IL-2) promoter-enhancer activity in Jurkat T cells stimulated with superantigen presented on HLA-DR transfectants combined with the ligands LFA-3 (CD58) and B7-1 (CD80). Gel shift analyses showed that NF-AT was strongly induced in LFA-3-costimulated Jurkat T cells, suggesting that NF-AT is a key target nuclear factor for the CD2-LFA-3 pathway. Studies using HLA-DR-B7-1-LFA-3 triple transfectants showed that the LFA-3-induced NF-AT DNA binding activity was negatively regulated by B7-1 costimulation. In contrast, induction of a CD28 response complex containing only c-Rel proteins was seen after B7-1 costimulation. Both LFA-3 costimulation and B7-1 costimulation induced the AP-1 and NF-kappaB nuclear factors. Distinct compositions of the NF-AT complexes were seen in B7-1- and LFA-3-costimulated cells. LFA-3 induced primarily Jun-D, Fra-1, and Fra-2, while B7-1 induced June-D-Fos complexes. In contrast, AP-1 and NF-kappaB complexes induced in B7-1- and LFA-3-costimulated T cells showed similar contents. Transient transfection of Jurkat T cells with a construct encoding the IL-2 enhancer-promoter region (position -500 to +60) linked to a luciferase reporter gene revealed that B7-1 costimulation was required to induce strong transcriptional activity. Combined B7-1-LFA-3 costimulation resulted in a synergistic increase in IL-2 transcriptional activity. Multimers of the AP-1, NF-AT, NF-kappaB, and CD28 response elements showed distinct kinetics and activity after LFA-3 and B7-1 costimulation and revealed that B7-1 and LFA-3 converge to superinduce transcriptional activity of the AP-1, NF-AT, and CD28 response elements. Transcriptional studies with an IL-2 enhancer-promoter carrying a mutation in the CD28 response element site revealed that the activity was reduced by 80% after B7-1 and B7-1-LFA-3 costimulation whereas the transcriptional activity induced by LFA-3 was unaffected. Our data strongly suggest a selectivity in induction of nuclear factors by the CD2-LFA-3 and CD28-B7-1 pathways. This selectivity may contribute to regulation of the levels of IL-2 induced by LFA-3 and B7-1 costimulation and favor autocrine and paracrine T-cell responses, respectively.

scholarly journals Detection of hyperreactive T cells in multiple myeloma by multivalent cross-linking of the CD3/TCR complex [see comments]

Blood ◽  
1991 ◽  
Vol 78 (7) ◽  
pp. 1770-1780 ◽  
Author(s):  
M Massaia ◽  
A Bianchi ◽  
C Attisano ◽  
S Peola ◽  
V Redoglia ◽  
...  
Keyword(s):  
Multiple Myeloma ◽  
T Cells ◽  
T Cell ◽  
Mononuclear Cells ◽  
Interleukin 2 ◽  
Normal Subjects ◽  
Cross Linking ◽  
Peripheral Blood Mononuclear ◽  
Hla Dr ◽  
Lower Ratio

Abstract Cellular immunity was investigated in 43 patients with multiple myeloma (MM) by assessing 3HTdR uptake induced by monocyte-dependent [CD3 monoclonal antibodies (MoAbs), phytohemagglutinin (PHA)] and monocyte- independent (CD2 MoAbs, ionomycin + phorbolester) stimulations. The former were evaluated in peripheral blood mononuclear cells (PBMNC) and purified T cells; the latter were evaluated in purified T-cell preparations only. MM showed significantly lower PBMNC responses to PHA (P less than .001), soluble OKT3 (CD3) (P = .01), and immobilized OKT3 MoAbs (P = .01). On purification of T cells, MM responses were still defective to soluble T11(2) + T11(3) (CD2) MoAbs (P = .004), phorbol myristate acetate (PMA) plus ionomycin (P less than .001), but significantly higher to plastic-immobilized OKT3 (P = .004). In some MM, 3HTdR uptake, interleukin-2 (IL-2) receptor (CD25) expression, and IL-2 production were as high on stimulation with plastic-immobilized OKT3 as that observed in normal subjects under optimal conditions (ie, plastic-immobilized OKT3 plus accessory signals). CD3 hyperreactivity correlated with the number of CD8+ HLA-DR+ cells in MM T-cell preparations. MM patients with more than 10% CD8+ HLA-DR+ cells had significantly higher responses to immobilized OKT3 (P less than .001), but lower responses to T11(2) plus T11(3) (P = .01), and PMA plus ionomycin (P = .03) than patients with less than 10% CD8+ HLA-DR+ cells. Phenotyping of CD45RA (naive) and CD45R0 (memory) expressions in resting MM T cells showed a lower ratio of CD45RA to CD45R0 in both CD4 (P less than .05) and CD8 (P less than .001) subpopulations. These data indicate that (a) some MM T cells require significantly fewer accessory signals (if any) to express the IL-2 receptor fully, secrete IL-2, and proliferate on multivalent cross-linking of the CD3/TCR complex; and (b) this peculiar state of activation is associated with high HLA-DR expression in CD8+ lymphocytes.

scholarly journals Antigen presentation in an HLA-DR-restricted fashion by B-cell chronic lymphocytic leukemia cells

Blood ◽  
1988 ◽  
Vol 72 (1) ◽  
pp. 102-108 ◽  
Author(s):  
M Yasukawa ◽  
T Shiroguchi ◽  
A Inatsuki ◽  
Y Kobayashi
Keyword(s):  
T Cells ◽  
Chronic Lymphocytic Leukemia ◽  
B Cell ◽  
Antigen Presentation ◽  
Interleukin 2 ◽  
Class Ii ◽  
Hla Class Ii ◽  
Lymphocytic Leukemia ◽  
Hla Dr ◽  
Cell Chronic Lymphocytic Leukemia

The ability of B-cell chronic lymphocytic leukemia (B-CLL) cells to present antigen to antigen-specific T cells was investigated. B-CLL cells present herpes simplex virus (HSV) antigen and purified protein derivative (PPD) to HSV- and PPD-specific, interleukin-2-dependent T- cell lines in an antigen-specific manner. Treatment of B-CLL cells with the phorbol ester 12-O-tetradecanoylphorbol-13-acetate (TPA) induced markedly increased levels of HLA-DR expression. TPA-treated B-CLL cells showed substantially more effective presentation, especially at low antigen concentrations, than did untreated B-CLL cells. By coculturing different allogeneic combinations of B-CLL cells and T cells and by adding anti-HLA-DR monoclonal antibody to cultures, it was found that antigen presentation by B-CLL cells was restricted by HLA-DR in the same way as for macrophages. We concluded from these experiments that B- CLL cells have a capacity to serve as antigen-presenting cells in an HLA class II-restricted fashion and that increasing the amount of HLA class II antigen and activation of B-CLL cells resulted in effective antigen presentation.

scholarly journals Clausmarin A, Potential Immunosuppressant Revealed by Yeast-Based Assay and Interleukin-2 Production Assay in Jurkat T Cells

PLoS ONE ◽  
2015 ◽  
Vol 10 (8) ◽  
pp. e0136804 ◽  
Author(s):  
Pitipreya Suauam ◽  
Boon-ek Yingyongnarongkul ◽  
Tanapat Palaga ◽  
Tokichi Miyakawa ◽  
Chulee Yompakdee
Keyword(s):  
T Cells ◽  
Interleukin 2 ◽  
Jurkat T Cells

Manganese promotes phorbol ester-induced interleukin-2 production via AP-1 activation in Jurkat T-cells

2012 ◽  
Vol 211 (3) ◽  
pp. 312-318 ◽  
Author(s):  
Susumu Tanaka ◽  
Yasunori Masuda ◽  
Chihiro Honma ◽  
Kohei Hosaka ◽  
Katsunori Takahashi ◽  
...  
Keyword(s):  
T Cells ◽  
Phorbol Ester ◽  
Interleukin 2 ◽  
Jurkat T Cells

Activation of the Human Immunodeficiency Virus-1 Long Terminal Repeat by Respiratory Burst Oxidants of Neutrophils

Blood ◽  
1999 ◽  
Vol 93 (1) ◽  
pp. 350-356
Author(s):  
Seymour J. Klebanoff ◽  
Catherine M. Headley
Keyword(s):  
Human Immunodeficiency Virus ◽  
T Cells ◽  
Long Terminal Repeat ◽  
Respiratory Burst ◽  
Terminal Repeat ◽  
Luciferase Reporter ◽  
Human Neutrophils ◽  
Jurkat T Cells ◽  
Immunodeficiency Virus ◽  
Hiv 1

The human immunodeficiency virus type 1 (HIV-1) long terminal repeat (LTR) introduced in association with the luciferase reporter gene into Jurkat T cells was strongly activated by a combination of human neutrophils and phorbol myristate acetate (PMA). Activation was not observed when normal neutrophils were replaced by neutrophils which lack a respiratory burst, ie, from a patient with chronic granulomatous disease (CGD), was strongly inhibited by catalase, was potentiated by vanadate, was stimulated by relatively low concentrations of azide, and was inhibited by selective inhibitors of protein kinase C (PKC). The PMA affected activation in three ways: (1) by directly activating the LTR in Jurkat LTRluc; (2) by inducing a respiratory burst in neutrophils with the formation of H2O2; and (3) by increasing the sensitivity of Jurkat LTRluc to the activating effect of H2O2. When PMA was replaced by opsonized zymosan as the neutrophil stimulus, activation of the LTR was low unless azide was added. Activation in the presence of azide was not seen when CGD neutrophils were used or when catalase was added, suggesting that azide acts by inhibiting the degradation of H2O2. These findings indicate that activation of the HIV-1 LTR in Jurkat T cells can be induced by H2O2 released by neutrophils, particularly when PKC is concomitantly activated.

scholarly journals Loureirin B Exerts its Immunosuppressive Effects by Inhibiting STIM1/Orai1 and KV1.3 Channels

2021 ◽  
Vol 12 ◽  
Author(s):  
Shujuan Shi ◽  
Qianru Zhao ◽  
Caihua Ke ◽  
Siru Long ◽  
Feng Zhang ◽  
...  
Keyword(s):  
T Cells ◽  
Molecular Mechanisms ◽  
Cell Model ◽  
Interleukin 2 ◽  
Immunosuppressive Effect ◽  
Dependent Manner ◽  
Jurkat T Cells ◽  
In The Wild ◽  
Resina Draconis ◽  
Loureirin B

Loureirin B (LrB) is a constituent extracted from traditional Chinese medicine Resina Draconis. It has broad biological functions and an impressive immunosuppressive effect that has been supported by numerous studies. However, the molecular mechanisms underlying Loureirin B-induced immune suppression are not fully understood. We previously reported that Loureirin B inhibited KV1.3 channel, calcium ion (Ca2+) influx, and interleukin-2 (IL-2) secretion in Jurkat T cells. In this study, we applied CRISPR/Cas9 to edit KV1.3 coding gene KCNA3 and successfully generated a KV1.3 knockout (KO) cell model to determine whether KV1.3 KO was sufficient to block the Loureirin B-induced immunosuppressive effect. Surprisingly, we showed that Loureirin B could still inhibit Ca2+ influx and IL-2 secretion in the Jurkat T cells in the absence of KV1.3 although KO KV1.3 reduced about 50% of Ca2+ influx and 90% IL-2 secretion compared with that in the wild type cells. Further experiments showed that Loureirin B directly inhibited STIM1/Orai1 channel in a dose-dependent manner. Our results suggest that Loureirin B inhibits Ca2+ influx and IL-2 secretion in Jurkat T cells by inhibiting both KV1.3 and STIM1/Orai1 channels. These studies also revealed an additional molecular target for Loureirin B-induced immunosuppressive effect, which makes it a promising leading compound for treating autoimmune diseases.

scholarly journals Further evidence for lymphokine overproduction in severe aplastic anemia [see comments]

Blood ◽  
1988 ◽  
Vol 72 (1) ◽  
pp. 266-272
Author(s):  
W Hinterberger ◽  
G Adolf ◽  
G Aichinger ◽  
R Dudczak ◽  
K Geissler ◽  
...  
Keyword(s):  
T Cells ◽  
Aplastic Anemia ◽  
Cd8 T Cells ◽  
Mononuclear Cells ◽  
Interleukin 2 ◽  
Serum Levels ◽  
Severe Aplastic Anemia ◽  
Hematopoietic Progenitor Cell ◽  
Ifn Gamma ◽  
Hla Dr

Interferon-gamma (IFN-gamma) and tumor necrosis factor (TNF) are lymphokines with a potent hematopoietic progenitor cell suppressive capacity. In untreated and immunosuppressed patients with severe aplastic anemia (SAA) and in control individuals we measured (a) serum levels of IFN-gamma and TNF and its production by peripheral blood mononuclear cells (PBMNC); (b) serum levels of neopterin, a product that reflects endogenous IFN production; (c) resting and activated lymphocyte subpopulations; and (d) serum levels of soluble interleukin- 2 receptor (IL-2R). Serum levels of IFN and TNF did not differ significantly in untreated and treated SAA patients and control individuals. Spontaneous and phytohemagglutinin-induced production of IFN and TNF by PBMNC, however, were highly increased in both untreated and treated SAA patients. Increased and decreased neopterin serum levels in untreated and treated SAA patients, respectively, suggest modulation of endogenous lymphokine release subsequent to immunosuppression. HLA-DR+ antigen was mainly expressed by CD8 T cells. Circulating numbers of activated (CD4 and CD8) T cells and serum levels of IL-2R were not increased in both untreated and treated SAA patients. The proportion of HLA-DR+ T cells in the PBMNC of untreated SAA patients correlated with the extent of lectin-induced IFN production. Although we were unable to confirm previous reports in SAA on (a) detectable IFN in blood and bone marrow serum, (b) improvement of stem cell growth upon neutralization of endogenous IFN, (c) absolutely increased numbers of circulating activated T cells, and (d) normalization of these abnormalities subsequent to successful immunosuppression, our data clearly support previous reports on abnormal lymphokine production in severe aplastic anemia. Our failure to relate this phenomenon to the severity of disease states, however, further raises doubts on the pathogenetic significance of lymphokine overproduction in SAA.

Abstract 627: Regulation of Nuclear Factor of Activated T Cells by BMP-Binding Endothelial Regulator Signaling

2015 ◽  
Vol 35 (suppl_1) ◽  
Author(s):  
Pamela P Lockyer ◽  
Hua Mao ◽  
Xi Li ◽  
Xinchun Pi
Keyword(s):  
T Cells ◽  
Endothelial Cells ◽  
Transcriptional Activity ◽  
Ldl Receptor ◽  
Bmp Signaling ◽  
Luciferase Reporter ◽  
Nuclear Factor ◽  
Activated T Cells ◽  
Morphogenetic Protein ◽  
Endothelial Integrity

Dysfunction of the vascular endothelium results in various cardiovascular, circulatory and blood diseases and exemplifies the importance of endothelial integrity. BMP-binding endothelial regulator (BMPER), a well recognized extracellular modulator of Bone morphogenetic protein (BMP) signaling, has been identified as a vital component in the vascular response to stress. Microarray analysis revealed nuclear factor of activated T cells (NFAT) as one of the genes found to be most highly upregulated by BMPER treatment in mouse endothelial cells (MECs), as well as many genes with NFAT consensus binding sites. Therefore we hypothesize that BMPER is an important regulator of NFAT transcriptional activity. Initially we have investigated the effect of BMPER on NFATc1 activation with MECs and human primary endothelial cells. Our data show that the translocation of NFATc1 from the cytoplasm to the nucleus following BMPER treatment, determined by immunofluorescent analysis. By using the nuclear fractionation assays, we observed the similar result that the translocation of NFATc1 to the nucleus of HUVECs took place after 30 minutes of BMPER treatment. Next, we wanted to determine whether the increased NFATc1 protein level in nucleus results in the enhanced transcriptional activity. Indeed, when HUVECs are treated with BMPER and then analyzed with luciferase reporter assay, a 1.5-fold significant increase in NFAT activity over baseline was observed. Our previous data demonstrate that LDL receptor related protein (LRP1) interacts with BMPER and regulates BMPER’s activity through endocytosis in endothelial cells. Interesting, we observe that LRP1 also interacts with NF45, the 45-kDa subunit of NFAT protein. It strongly suggests that BMPER positively regulates NFAT activity through LRP1. This novel signaling pathway indicates that BMPER may acts as a new ligand and exhibits BMP-independent activity in endothelial cells and therefore contribute to the regulation of vascular homeostasis.

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