The subjects were exposed to cold air ( 4 degrees C ) twice/d , 30 min/exposure , for a total of 80 exposures .


The T3-subjects received placebo [ n = 8 ] and the T3 + subjects received T3 ( 30 micrograms/d ) [ n = 8 ] in a double-blind fashion .


Mononuclear leukocytes were isolated from peripheral blood before the cold exposure and drug regimen began , and then after every 20 exposures .

<cell_type>Mononuclear leukocytes</cell_type> were isolated from peripheral blood before the cold exposure and drug regimen began , and then after every 20 exposures .


The dissociation constant ( Kd ) and maximum binding capacity ( MBC ) of the NT3R values were log transformed to minimize between-subject variability .

The dissociation constant ( Kd ) and maximum binding capacity ( MBC ) of the <protein>NT3R</protein> values were log transformed to minimize between-subject variability .


In the T3+ group , serum total thyroxine ( TT4 ) , free T4 ( FT4 ) , and TSH were approx 50 % lower than both basal and T3-values .


The log10Kd increased 0.304 +/- 0.139 ( p < 0.04 ) and the log10MBC increased 0.49 +/- 0.10 ( p < 0.001 ) in the T3+ subjects compared to baseline .


This change in MBC represents a 311 % increase in the MBC over baseline and a fivefold increase over placebo-treated subjects .


The T3 -group showed no change in MBC over the study .


These results describe for the first time the rapid modulation of the NT3R in response to the combined influence of cold exposure and reduced circulating T4 and TSH .

These results describe for the first time the rapid modulation of the <protein>NT3R</protein> in response to the combined influence of cold exposure and reduced circulating T4 and TSH .


HIV-1 LTR activity in human CD40-activated B lymphocytes is dependent on NF-kappaB .

<dna>HIV-1 LTR</dna> activity in <cell_line>human CD40-activated B lymphocytes</cell_line> is dependent on <protein>NF-kappaB</protein> .


CD40-stimulated human B lymphocytes are highly permissive to a productive infection by the human immunodeficiency virus type 1 .

<cell_line>CD40-stimulated human B lymphocytes</cell_line> are highly permissive to a productive infection by the human immunodeficiency virus type 1 .


In these cells , nuclear factors involved in activation of the HIV-1 LTR , which contains the transcriptional control elements of the virus , are unknown .

In these cells , <protein>nuclear factors</protein> involved in activation of the <dna>HIV-1 LTR</dna> , which contains the <dna>transcriptional control elements</dna> of the virus , are unknown .


Transient expression assays with plasmids containing deleted parts of the LTR region linked to a reporter gene showed that the NF-kappaB binding site was essential for HIV-1 LTR activity in CD40-stimulated B lymphocytes .

Transient expression assays with plasmids containing deleted parts of the <dna>LTR region</dna> linked to a reporter gene showed that the <dna>NF-kappaB binding site</dna> was essential for <dna>HIV-1 LTR</dna> activity in <cell_line>CD40-stimulated B lymphocytes</cell_line> .


In addition , electrophoretic mobility shift and supershift assays revealed that important NF-kappaB binding activity composed of at least p50 , p65 , and c-Rel NF-kappaB subunits was present in nuclei of CD40-stimulated B cells .

In addition , electrophoretic mobility shift and supershift assays revealed that important <protein>NF-kappaB</protein> binding activity composed of at least <protein>p50</protein> , <protein>p65</protein> , and <protein>c-Rel</protein> <protein>NF-kappaB subunits</protein> was present in nuclei of <cell_line>CD40-stimulated B cells</cell_line> .


These results confirm at a molecular level the ability of HIV-1 to replicate in B cells and that this activity is strongly associated with NF-kappaB .

These results confirm at a molecular level the ability of HIV-1 to replicate in <cell_type>B cells</cell_type> and that this activity is strongly associated with <protein>NF-kappaB</protein> .


Sequence analysis and expression in cultured lymphocytes of the human FOSB gene ( G0S3 ) .

Sequence analysis and expression in <cell_type>cultured lymphocytes</cell_type> of the human <dna>FOSB gene</dna> ( <dna>G0S3</dna> ) .


G0S3 is a member of a set of putative G0/G1 switch regulatory genes ( G0S genes ) selected by screening cDNA libraries prepared from human blood mononuclear cells cultured for 2 hr with lectin and cycloheximide .

<dna>G0S3</dna> is a member of a set of <dna>putative G0/G1 switch regulatory genes</dna> ( <dna>G0S genes</dna> ) selected by screening <dna>cDNA libraries</dna> prepared from <cell_type>human blood mononuclear cells</cell_type> cultured for 2 hr with <protein>lectin</protein> and cycloheximide .


The sequence shows high homology with the murine FOSB gene , which encodes a component of the AP1 transcriptional regulator .

The sequence shows high homology with the <dna>murine FOSB gene</dna> , which encodes a component of the <protein>AP1 transcriptional regulator</protein> .


Comparison of cDNA and genomic sequences reveals a 4-exon structure characteristic of the FOS family of genes .

Comparison of <dna>cDNA</dna> and genomic sequences reveals a <dna>4-exon structure</dna> characteristic of the <dna>FOS family of genes</dna> .


Freshly isolated cells show high levels of FOSB/G0S3 and FOS/G0S7 mRNAs , which decline rapidly during incubation in culture medium .

Freshly isolated cells show high levels of <rna>FOSB/G0S3 and FOS/G0S7 mRNAs</rna> , which decline rapidly during incubation in culture medium .


The kinetics of expression suggest that the high initial levels are caused by the isolation procedure , and do not reflect constitutive expression .


In cells preincubated for a day , levels of FOS mRNA reach a maximum 20 min after the addition of lectin and decline to control levels over the next 3 hr .

In cells preincubated for a day , levels of <rna>FOS mRNA</rna> reach a maximum 20 min after the addition of <protein>lectin</protein> and decline to control levels over the next 3 hr .


Levels of FOSB mRNA reach a maximum 40 min after the addition of lectin and decline to control levels over the next 6 hr .

Levels of <rna>FOSB mRNA</rna> reach a maximum 40 min after the addition of <protein>lectin</protein> and decline to control levels over the next 6 hr .


In freshly isolated cells , both FOS and FOSB mRNAs increase dramatically in response to the protein synthesis inhibitor cycloheximide .

In freshly isolated cells , both <rna>FOS and FOSB mRNAs</rna> increase dramatically in response to the protein synthesis inhibitor cycloheximide .


In preincubated cells , the cycloheximide response is decreased , especially in the case of FOSB .

In preincubated cells , the cycloheximide response is decreased , especially in the case of <dna>FOSB</dna> .


These differences in expression of FOS and FOSB suggest different roles and regulation .

These differences in expression of <dna>FOS</dna> and <dna>FOSB</dna> suggest different roles and regulation .


Regions of low base order-dependent stem-loop potential in the region of the gene are defined .


These indicate where base order has been adapted for purposes other than stem-loop stability ( e.g. , encoding proteins or gene regulation ) .


Regions of low potential in a 68.5-kb genomic segment containing the FOSB gene suggest that the potential may help locate genes in uncharted DNA sequences .

Regions of low potential in a 68.5-kb genomic segment containing the <dna>FOSB gene</dna> suggest that the potential may help locate genes in <dna>uncharted DNA sequences</dna> .


Susceptibility to natural killer cells and down regulation of MHC class I expression in adenovirus 12 transformed cells are regulated by different E1A domains .

Susceptibility to <cell_type>natural killer cells</cell_type> and down regulation of <protein>MHC</protein> class I expression in <cell_line>adenovirus 12 transformed cells</cell_line> are regulated by different <dna>E1A domains</dna> .


All human adenoviruses transform rodent cells in vitro , but only cells transformed by serotypes belonging to subgroups A ( Ad12 ) and B ( Ad3 ) are tumorigenic for immunocompetent animals .

All human adenoviruses transform <cell_type>rodent cells</cell_type> in vitro , but only cells transformed by serotypes belonging to subgroups A ( Ad12 ) and B ( Ad3 ) are tumorigenic for immunocompetent animals .


In these cells , the expression of MHC-class I antigens is repressed and might allow them to escape from recognition by cytotoxic T lymphocytes ( CTL ) and to develop in tumor .

In these cells , the expression of <protein>MHC-class I antigens</protein> is repressed and might allow them to escape from recognition by <cell_type>cytotoxic T lymphocytes</cell_type> ( <cell_type>CTL</cell_type> ) and to develop in tumor .


Furthermore , these cell lines appear resistant to lysis by natural killer ( NK ) cells .

Furthermore , these <cell_line>cell lines</cell_line> appear resistant to lysis by <cell_type>natural killer ( NK ) cells</cell_type> .


To determine the E1A domain ( s ) responsible for these properties several cell lines were created by transforming baby rat kidney ( BRK ) cells with a set of plasmids expressing different Ad2/Ad12 hybrid E1A gene products .

To determine the <dna>E1A domain</dna> ( s ) responsible for these properties several <cell_line>cell lines</cell_line> were created by transforming <cell_line>baby rat kidney ( BRK ) cells</cell_line> with a set of <dna>plasmids</dna> expressing different <protein>Ad2/Ad12 hybrid E1A gene products</protein> .


The MHC class 1 gene expression was inhibited in cells expressing the Ad12 13S mRNA product and in cells transformed with Ad2/Ad12 hybrid E1A gene product harboring the C-terminal part of the conserved region ( CR ) 3 of Ad12 .

The <dna>MHC class 1 gene</dna> expression was inhibited in cells expressing the <protein>Ad12 13S mRNA product</protein> and in cells transformed with <protein>Ad2/Ad12 hybrid E1A gene product</protein> harboring the <protein>C-terminal part</protein> of the <dna>conserved region ( CR ) 3</dna> of Ad12 .


Susceptibility of these transformed cell lines to NK cells was determined by cytolytic assays .

Susceptibility of these transformed <cell_line>cell lines</cell_line> to <cell_type>NK cells</cell_type> was determined by cytolytic assays .


The results obtained suggest that two Ad12 E1A domains are required to induce resistance of the cell lines to NK cells .

The results obtained suggest that two <dna>Ad12 E1A domains</dna> are required to induce resistance of the <cell_line>cell lines</cell_line> to <cell_type>NK cells</cell_type> .


Cell specific expression of human Bruton 's agammaglobulinemia tyrosine kinase gene ( Btk ) is regulated by Sp1- and Spi-1/PU.1-family members .

Cell specific expression of <dna>human Bruton 's agammaglobulinemia tyrosine kinase gene</dna> ( <protein>Btk</protein> ) is regulated by <protein>Sp1-</protein> and <protein>Spi-1/PU.1-family members</protein> .


Bruton 's agammaglobulinemia tyrosine kinase ( Btk ) is a cytoplasmic tyrosine kinase involved in the human disease X-linked agammaglobulinemia ( XLA ) .

<protein>Bruton 's agammaglobulinemia tyrosine kinase</protein> ( <protein>Btk</protein> ) is a <protein>cytoplasmic tyrosine kinase</protein> involved in the human disease X-linked agammaglobulinemia ( XLA ) .


The gene is expressed in all hematopoietic cells with the exception of T-cells and plasma cells .

The gene is expressed in all <cell_type>hematopoietic cells</cell_type> with the exception of <cell_type>T-cells</cell_type> and <cell_type>plasma cells</cell_type> .


For this expression pattern the first 280 bp upstream of the major transcriptional start site seems to be sufficient .

For this expression pattern the first <dna>280 bp upstream</dna> of the <dna>major transcriptional start site</dna> seems to be sufficient .


In vitro footprinting analysis within this part of the promoter revealed two Sp1 binding sites as well as a PU-box .

In vitro footprinting analysis within this part of the <dna>promoter</dna> revealed two <dna>Sp1 binding sites</dna> as well as a <dna>PU-box</dna> .


The transcription factor Spi-1/PU.1 as well as the closely related factor Spi-B bound to the PU-box in B-cells .

The <protein>transcription factor</protein> <protein>Spi-1/PU.1</protein> as well as the closely related factor <protein>Spi-B</protein> bound to the <dna>PU-box</dna> in B-cells .


In the erythroleukemia cell line K562 , due to the absence of Spi-B , only PU.1 bound to the Btk promoter .

In the <cell_line>erythroleukemia cell line K562</cell_line> , due to the absence of <protein>Spi-B</protein> , only <protein>PU.1</protein> bound to the <dna>Btk promoter</dna> .


Mutation of either site reduced the expression in transient transfection experiments .


However , mutation of the PU box had no effect in the T-cell line Jurkat , where none of the Spi-1 family members is expressed .

However , mutation of the <dna>PU box</dna> had no effect in the <cell_line>T-cell line Jurkat</cell_line> , where none of the <protein>Spi-1 family members</protein> is expressed .


In addition Spi-B as well as PU.1 were able to transactivate Btk expression .

In addition <protein>Spi-B</protein> as well as <protein>PU.1</protein> were able to transactivate <protein>Btk</protein> expression .


In fetal liver of PU.1-/- mice , which lack lymphoid and myeloid cells , expression of Btk was reduced two- to threefold but not abolished .

In fetal liver of PU.1-/- mice , which lack <cell_type>lymphoid and myeloid cells</cell_type> , expression of <protein>Btk</protein> was reduced two- to threefold but not abolished .


Collectively this study shows that expression of the Btk gene is regulated by the combined action of Sp1- and PU.1-family members .

Collectively this study shows that expression of the <dna>Btk gene</dna> is regulated by the combined action of <protein>Sp1- and PU.1-family members</protein> .


T cell response to Epstein-Barr virus transactivators in chronic rheumatoid arthritis .

T cell response to <protein>Epstein-Barr virus transactivators</protein> in chronic rheumatoid arthritis .


Rheumatoid arthritis is a multistep disorder associated with autoimmune features of yet unknown etiology .


Implication of viruses such as Epstein-Barr virus ( EBV ) in rheumatoid arthritis pathogenesis has been suspected on the basis of several indirect observations , but thus far , a direct link between EBV and rheumatoid arthritis has not been provided .


Here we show that a large fraction of T cells infiltrating affected joints from a patient with chronic rheumatoid arthritis recognizes two EBV transactivators ( BZLF1 and BMLF1 ) in a major histocompatibility complex -restricted fashion .

Here we show that a large fraction of <cell_type>T cells</cell_type> infiltrating affected joints from a patient with chronic rheumatoid arthritis recognizes two <protein>EBV transactivators</protein> ( <protein>BZLF1</protein> and <protein>BMLF1</protein> ) in a <protein>major histocompatibility complex</protein> -restricted fashion .


Responses to these EBV antigens by synovial lymphocytes from several other chronic rheumatoid arthritis patients were readily detectable .

Responses to these <protein>EBV antigens</protein> by synovial lymphocytes from several other chronic rheumatoid arthritis patients were readily detectable .


Thus these results suggest a direct contribution of EBV to chronic rheumatoid arthritis pathogenesis .


They also demonstrate for the first time the occurrence of T cell responses against EBV transactivating factors , which might be central in the control of virus reactivation .

They also demonstrate for the first time the occurrence of T cell responses against <protein>EBV transactivating factors</protein> , which might be central in the control of virus reactivation .


CD14 -mediated signal pathway of Porphyromonas gingivalis lipopolysaccharide in human gingival fibroblasts .

<protein>CD14</protein> -mediated signal pathway of Porphyromonas gingivalis lipopolysaccharide in <cell_type>human gingival fibroblasts</cell_type> .


Lipopolysaccharide ( LPS ) induces expression of inflammatory cytokines in monocytes/macrophages via CD14 , one of the LPS receptors , which is expressed predominantly in these cells .

Lipopolysaccharide ( LPS ) induces expression of <protein>inflammatory cytokines</protein> in <cell_type>monocytes/macrophages</cell_type> via <protein>CD14</protein> , one of the <protein>LPS receptors</protein> , which is expressed predominantly in these cells .


It has been demonstrated that Porphyromonas gingivalis LPS ( P-LPS ) also is able to induce inflammatory cytokines in human gingival fibroblasts .

It has been demonstrated that Porphyromonas gingivalis LPS ( P-LPS ) also is able to induce <protein>inflammatory cytokines</protein> in <cell_type>human gingival fibroblasts</cell_type> .


Therefore , it is important to determine whether CD14 is expressed in gingival fibroblasts and to define the P-LPS-mediated signal-transducing mechanism in the cells .

Therefore , it is important to determine whether <protein>CD14</protein> is expressed in <cell_type>gingival fibroblasts</cell_type> and to define the P-LPS-mediated <protein>signal-transducing</protein> mechanism in the cells .


In this study , we observed unexpectedly by immunohistochemical , Western blotting ( immunoblotting ) , and Northern ( RNA ) blotting assays that CD14 is expressed at high density in human gingival fibroblasts .

In this study , we observed unexpectedly by immunohistochemical , Western blotting ( immunoblotting ) , and Northern ( RNA ) blotting assays that <protein>CD14</protein> is expressed at high density in <cell_type>human gingival fibroblasts</cell_type> .


P-LPS-induced expression of the monocyte chemoattractant protein 1 ( MCP-1 ) gene in the cells was inhibited markedly by treatment with anti-human CD14 antibody and was completely inhibited by herbimycin A , a potent inhibitor of tyrosine kinase .

P-LPS-induced expression of the <dna>monocyte chemoattractant protein 1 ( MCP-1 ) gene</dna> in the cells was inhibited markedly by treatment with anti-human <protein>CD14</protein> antibody and was completely inhibited by herbimycin A , a potent inhibitor of <protein>tyrosine kinase</protein> .


The inhibitor also dramatically inhibited monocyte chemotactic activity of and MCP-1 production by the cells .

The inhibitor also dramatically inhibited monocyte chemotactic activity of and <protein>MCP-1</protein> production by the cells .


Furthermore , P-LPS-induced expression of the MCP-1 gene in the cells also was blocked by inhibitors of two transcription factors , i.e. , curcumin , an inhibitor of AP-1 , and pyrolidine dithiocarbamate , an inhibitor of NF-kappaB .

Furthermore , P-LPS-induced expression of the <dna>MCP-1 gene</dna> in the cells also was blocked by inhibitors of two <protein>transcription factors</protein> , i.e. , curcumin , an inhibitor of <protein>AP-1</protein> , and pyrolidine dithiocarbamate , an inhibitor of <protein>NF-kappaB</protein> .


Both inhibitors inhibited monocyte chemotactic activity in the culture supernatant of P-LPS-treated cells .


Gel shift mobility assay showed stimulation of the AP-1 and NF-kappaB contents in P-LPS-treated cells .

Gel shift mobility assay showed stimulation of the <protein>AP-1</protein> and <protein>NF-kappaB</protein> contents in P-LPS-treated cells .


This study is the first to demonstrate the expression of CD14 in human gingival fibroblasts and to show that the signal-transducing pathway of P-LPS in the cells is mediated by CD14 .

This study is the first to demonstrate the expression of <protein>CD14</protein> in <cell_type>human gingival fibroblasts</cell_type> and to show that the signal-transducing pathway of P-LPS in the cells is mediated by <protein>CD14</protein> .


rel Is rapidly tyrosine-phosphorylated following granulocyte-colony stimulating factor treatment of human neutrophils .

<protein>rel</protein> Is rapidly tyrosine-phosphorylated following <protein>granulocyte-colony stimulating factor</protein> treatment of human neutrophils .


Stimulation of neutrophils with granulocyte-colony stimulating factor ( G-CSF ) results in an enhanced respiratory burst , prolonged survival , and increased tumor cell killing .

Stimulation of <cell_type>neutrophils</cell_type> with <protein>granulocyte-colony stimulating factor</protein> ( <protein>G-CSF</protein> ) results in an enhanced respiratory burst , prolonged survival , and increased tumor cell killing .


The effects of G-CSF are mediated by binding to specific , high affinity receptors .

The effects of <protein>G-CSF</protein> are mediated by binding to specific , high <protein>affinity receptors</protein> .


G-CSF receptors lack intrinsic tyrosine kinase activity , but activation of the receptor results in the rapid induction of tyrosine kinase activity .

<protein>G-CSF receptors</protein> lack intrinsic <protein>tyrosine kinase</protein> activity , but activation of the receptor results in the rapid induction of tyrosine kinase activity .


Antiphosphotyrosine immunoblots of whole cell lysates prepared from neutrophils show that the G-CSF rapidly induces prominent tyrosine phosphorylation of a protein of a relative molecular mass of 80 kDa .

Antiphosphotyrosine immunoblots of whole cell lysates prepared from <cell_type>neutrophils</cell_type> show that the <protein>G-CSF</protein> rapidly induces prominent tyrosine phosphorylation of a protein of a relative molecular mass of 80 kDa .


Using monospecific antibodies , the 80-kDa tyrosine-phosphorylated protein has been shown to be p80c-rel , a proto-oncogene belonging to a family of transcriptional regulators which include NF-kB .

Using <protein>monospecific antibodies</protein> , the 80-kDa <protein>tyrosine-phosphorylated protein</protein> has been shown to be <protein>p80c-rel</protein> , a <dna>proto-oncogene</dna> belonging to a family of <protein>transcriptional regulators</protein> which include <protein>NF-kB</protein> .


The induction of tyrosine phosphorylation of p80c-rel was unique to G-CSF in that granulocyte-macrophage colony stimulating factor which also stimulates neutrophils and induces tyrosine phosphorylation does not result in tyrosine phosphorylation of p80c-rel .

The induction of tyrosine phosphorylation of <protein>p80c-rel</protein> was unique to <protein>G-CSF</protein> in that <protein>granulocyte-macrophage colony stimulating factor</protein> which also stimulates <cell_type>neutrophils</cell_type> and induces tyrosine phosphorylation does not result in tyrosine phosphorylation of <protein>p80c-rel</protein> .


The consequences of p80c-rel tyrosine phosphorylation are not yet known ; however , tyrosine-phosphorylated p80c-rel is capable of binding to DNA , and G-CSF stimulation results in an increase in the amount of p80c-rel which binds to DNA .

The consequences of <protein>p80c-rel</protein> tyrosine phosphorylation are not yet known ; however , tyrosine-phosphorylated <protein>p80c-rel</protein> is capable of binding to DNA , and <protein>G-CSF</protein> stimulation results in an increase in the amount of <protein>p80c-rel</protein> which binds to DNA .


These results demonstrate that one of the first biochemical events which occurs in neutrophils following G-CSF stimulation , activation of a tyrosine kinase , leads directly to the tyrosine phosphorylation of p80c-rel .

These results demonstrate that one of the first biochemical events which occurs in <cell_type>neutrophils</cell_type> following <protein>G-CSF</protein> stimulation , activation of a <protein>tyrosine kinase</protein> , leads directly to the tyrosine phosphorylation of <protein>p80c-rel</protein> .


Thus , the tyrosine kinase activated by G-CSF appears to directly transduce a signal to a protein which functions as a transcriptional regulator .

Thus , the <protein>tyrosine kinase</protein> activated by <protein>G-CSF</protein> appears to directly transduce a signal to a protein which functions as a <protein>transcriptional regulator</protein> .


Patterns of Pan expression and role of Pan proteins in endocrine cell type-specific complex formation .

Patterns of <protein>Pan</protein> expression and role of <protein>Pan proteins</protein> in endocrine cell type-specific complex formation .


The Pan gene encodes at least two distinct transcripts , Pan-1 and Pan-2 ( also known as E47 and E12 , respectively ) , by the mechanism of alternative RNA splicing .

The <dna>Pan gene</dna> encodes at least two distinct transcripts , <protein>Pan-1</protein> and <protein>Pan-2</protein> ( also known as <protein>E47</protein> and <protein>E12</protein> , respectively ) , by the mechanism of alternative RNA splicing .


Northern blot analyses performed on rat and mouse tissues have detected ubiquitously expressed Pan transcripts , but the abundance , distribution , and form of Pan proteins have not been clearly defined .

Northern blot analyses performed on rat and mouse tissues have detected ubiquitously expressed Pan transcripts , but the abundance , distribution , and form of <protein>Pan proteins</protein> have not been clearly defined .


Studies of cell lines representing endocrine , fibroblast , and lymphoid lineages using polyclonal antisera to detect E2A proteins have suggested that significant E2A protein expression is restricted to B-lymphocytes .

Studies of <cell_line>cell lines</cell_line> representing <cell_type>endocrine , fibroblast , and lymphoid lineages</cell_type> using polyclonal antisera to detect <protein>E2A proteins</protein> have suggested that significant <protein>E2A protein</protein> expression is restricted to <cell_type>B-lymphocytes</cell_type> .


We have developed a monoclonal antibody , Yae , which is specific for Pan/E2A proteins , and have used the Yae antibody to examine a variety of endocrine and nonendocrine cell lineages for differences in Pan/ E2A protein expression , subcellular localization , and heteromeric complex formation .

We have developed a <protein>monoclonal antibody</protein> , <protein>Yae</protein> , which is specific for <protein>Pan/E2A proteins</protein> , and have used the <protein>Yae antibody</protein> to examine a variety of <cell_type>endocrine and nonendocrine cell lineages</cell_type> for differences in Pan/ <protein>E2A protein</protein> expression , subcellular localization , and heteromeric complex formation .


In contrast to previous results obtained using polyclonal antiseras to detect Pan/ E2A proteins , we report comparable levels of Pan proteins in GH/PRL- and insulin-producing , B- and T-lymphocyte cells .

In contrast to previous results obtained using polyclonal antiseras to detect Pan/ <protein>E2A proteins</protein> , we report comparable levels of <protein>Pan proteins</protein> in <cell_line>GH/PRL- and insulin-producing , B- and T-lymphocyte cells</cell_line> .


IEF-1 , a pancreatic beta-cell type-specific complex believed to regulate insulin expression , is demonstrated to consist of at least two distinct species , one of which does not contain Pan molecules .

<protein>IEF-1</protein> , a pancreatic beta-cell type-specific complex believed to regulate insulin expression , is demonstrated to consist of at least two distinct species , one of which does not contain <protein>Pan molecules</protein> .


Although it has been postulated that pituitary endocrine cells and pancreatic endocrine beta-cells share identical Pan/E2A complexes , native-Western analyses of pituitary and endocrine beta-cells detect Pan proteins in distinct cell type-specific complexes .

Although it has been postulated that <cell_type>pituitary endocrine cells</cell_type> and <cell_type>pancreatic endocrine beta-cells</cell_type> share identical <protein>Pan/E2A complexes</protein> , native-Western analyses of pituitary and <cell_type>endocrine beta-cells</cell_type> detect <protein>Pan proteins</protein> in distinct <protein>cell type-specific complexes</protein> .


Visualization of the endogenous NF-kappa B p50 subunit in the nucleus of follicular dendritic cells in germinal centers .

Visualization of the <protein>endogenous NF-kappa B p50 subunit</protein> in the nucleus of <cell_type>follicular dendritic cells</cell_type> in germinal centers .


NF-kappa B , a 50 kDa/65 kDa ( p50/p65 ) heterodimer , is a ubiquitous transcription factor involved in the positive regulation of various immune genes .

<protein>NF-kappa B</protein> , a <protein>50 kDa/65 kDa ( p50/p65 ) heterodimer</protein> , is a <protein>ubiquitous transcription factor</protein> involved in the positive regulation of various <dna>immune genes</dna> .


The aim of this study was to determine whether NF-kappa B is related to a particular cell type and/or differentiation step during immunopoiesis .

The aim of this study was to determine whether <protein>NF-kappa B</protein> is related to a particular cell type and/or differentiation step during immunopoiesis .


Using in situ hybridization on sections from non HIV hyperplastic lymph nodes , we found that the gene of the 105 kDa precursor of p50 was overexpressed in the light zone of germinal centers , with a network aspect , which suggested the involvement of follicular dendritic cells ( FDC ) .

Using in situ hybridization on sections from non HIV hyperplastic lymph nodes , we found that the gene of the <protein>105 kDa precursor</protein> of <protein>p50</protein> was overexpressed in the light zone of germinal centers , with a network aspect , which suggested the involvement of <cell_type>follicular dendritic cells</cell_type> ( <cell_type>FDC</cell_type> ) .


By immunohistochemistry , p50 protein was detected in the cytoplasm and nucleus of FDC , confirming the involvement of FDC .

By immunohistochemistry , <protein>p50</protein> protein was detected in the cytoplasm and nucleus of <cell_type>FDC</cell_type> , confirming the involvement of <cell_type>FDC</cell_type> .


Furthermore , p50 protein was detected in the cytoplasm of all lymphocytes .

Furthermore , <protein>p50</protein> protein was detected in the cytoplasm of all <cell_type>lymphocytes</cell_type> .


Thus , we focused our study on isolated FDC clusters from normal tonsils .

Thus , we focused our study on isolated <cell_type>FDC</cell_type> clusters from normal tonsils .


As showed on tissue sections , we detected the p50 in both cytoplasm and nucleus of FDC .

As showed on tissue sections , we detected the <protein>p50</protein> in both cytoplasm and nucleus of <cell_type>FDC</cell_type> .


Nuclei of lymphocytes from FDC clusters were negative .

Nuclei of <cell_type>lymphocytes</cell_type> from <cell_type>FDC</cell_type> clusters were negative .


We next studied p65 and c-Rel protein expression in FDC clusters .

We next studied p65 and c-Rel protein expression in <cell_type>FDC</cell_type> clusters .


p65 was detected in the cytoplasm of FDC , whereas nuclei were negative .

<protein>p65</protein> was detected in the cytoplasm of <cell_type>FDC</cell_type> , whereas nuclei were negative .


Furthermore , p65 was detected in the nuclei of some lymphocytes .

Furthermore , <protein>p65</protein> was detected in the nuclei of some <cell_type>lymphocytes</cell_type> .


c-Rel protein was detected only in the cytoplasm of lymphocytes and not in the nucleus and cytoplasm of FDC .

<protein>c-Rel protein</protein> was detected only in the cytoplasm of <cell_type>lymphocytes</cell_type> and not in the nucleus and cytoplasm of <cell_type>FDC</cell_type> .


Our results indicated that , in the context of T cell-dependent B cell immunopoiesis occurring in FDC clusters , p50 is mainly related to FDC with a massive overexpression in the nuclei , whereas p65 is expressed in a scattered manner in the nuclei of lymphocytes and c-Rel protein exclusively in the cytoplasm of lymphocytes from FDC clusters .

Our results indicated that , in the context of T cell-dependent B cell immunopoiesis occurring in <cell_type>FDC</cell_type> clusters , <protein>p50</protein> is mainly related to <cell_type>FDC</cell_type> with a massive overexpression in the nuclei , whereas <protein>p65</protein> is expressed in a scattered manner in the nuclei of <cell_type>lymphocytes</cell_type> and c-Rel protein exclusively in the cytoplasm of <cell_type>lymphocytes</cell_type> from <cell_type>FDC</cell_type> clusters .


These results suggested that the two subunits of NF-kappa B and the c-Rel protein have different roles in different cell types during B cell immunopoiesis .

These results suggested that the two subunits of <protein>NF-kappa B</protein> and the <protein>c-Rel protein</protein> have different roles in different cell types during B cell immunopoiesis .


Cloning and characterization of NF-ATc and NF-ATp : the cytoplasmic components of NF-AT .

Cloning and characterization of <protein>NF-ATc</protein> and <protein>NF-ATp</protein> : the cytoplasmic components of <protein>NF-AT</protein> .


Present evidence indicates a pathway of signal transmission in T cells that is outlined in figure 1 .

Present evidence indicates a pathway of signal transmission in <cell_type>T cells</cell_type> that is outlined in figure 1 .


The elevation in intracellular calcium that is induced by interactions at the antigen receptor leads to the activation of the calcium-dependent phosphatase calcineurin .

The elevation in intracellular calcium that is induced by interactions at the <protein>antigen receptor</protein> leads to the activation of the <protein>calcium-dependent phosphatase</protein> <protein>calcineurin</protein> .


This in turn leads to the nuclear association of the cytosolic component of NF-ATc .

This in turn leads to the nuclear association of the <protein>cytosolic component</protein> of <protein>NF-ATc</protein> .


The activation of calcineurin and the nuclear import of NF-ATc can both be blocked by cyclosporin A or FK506 in complex with their respective immunophilins .

The activation of <protein>calcineurin</protein> and the nuclear import of <protein>NF-ATc</protein> can both be blocked by cyclosporin A or FK506 in complex with their respective <protein>immunophilins</protein> .


Once in the nucleus , NF-ATc interacts with NF-ATn to form an active transcriptional complex .

Once in the nucleus , <protein>NF-ATc</protein> interacts with <protein>NF-ATn</protein> to form an <protein>active transcriptional complex</protein> .


NF-ATn is a ubiquitous protein , can be synthesized in response to PMA , and has many similarities to AP-1 .

<protein>NF-ATn</protein> is a ubiquitous protein , can be synthesized in response to PMA , and has many similarities to <protein>AP-1</protein> .


The mechanism by which NF-ATc enters the nucleus is unknown , and although it appears to require calcineurin , NF-ATc has not yet been shown to be an in vivo substrate of calcineurin .

The mechanism by which <protein>NF-ATc</protein> enters the nucleus is unknown , and although it appears to require <protein>calcineurin</protein> , <protein>NF-ATc</protein> has not yet been shown to be an in vivo substrate of <protein>calcineurin</protein> .


Alternative mechanisms include the possibility that NF-ATc operates on some cytoplasmic anchor or that other proteins that are controlled by calcineurin carry out the nuclear import of NF-ATc .

Alternative mechanisms include the possibility that <protein>NF-ATc</protein> operates on some <protein>cytoplasmic anchor</protein> or that other proteins that are controlled by <protein>calcineurin</protein> carry out the nuclear import of <protein>NF-ATc</protein> .


Although NF-ATp copurifies with NF-ATc , there is as yet no understanding of how NF-ATp is functioning in vivo .

Although <protein>NF-ATp</protein> copurifies with <protein>NF-ATc</protein> , there is as yet no understanding of how <protein>NF-ATp</protein> is functioning in vivo .


Now that these proteins are purified and cloned , the major goals will be to understand their role and the roles of other family members in thymic development .


Induction of vascular cell adhesion molecule-1 by low-density lipoprotein .

Induction of <protein>vascular cell adhesion molecule-1</protein> by <protein>low-density lipoprotein</protein> .


Low-density lipoprotein ( LDL ) is a well-established risk factor for atherosclerosis .

<protein>Low-density lipoprotein</protein> ( <protein>LDL</protein> ) is a well-established risk factor for atherosclerosis .


When endothelial cells are incubated with this lipoprotein in pathophysiologic amounts , the cells are activated .


Among the documented cellular responses to LDL is increased recruitment of monocytes , which are believed to play a major role in promoting intimal plaque formation .

Among the documented cellular responses to <protein>LDL</protein> is increased recruitment of <cell_type>monocytes</cell_type> , which are believed to play a major role in promoting intimal plaque formation .


The findings presented here link an atheogenic lipoprotein , LDL , with the induction of an adhesion molecule important in atherogenesis Human LDL induces the vascular cell adhesion molecule-1 ( VCAM-1 ) transcriptionally with an increase in mRNA levels through activation of the VCAM promoter .

The findings presented here link an <protein>atheogenic lipoprotein</protein> , <protein>LDL</protein> , with the induction of an adhesion molecule important in atherogenesis Human <protein>LDL</protein> induces the <protein>vascular cell adhesion molecule-1</protein> ( <protein>VCAM-1</protein> ) transcriptionally with an increase in mRNA levels through activation of the <dna>VCAM promoter</dna> .


This effect is blocked by anti-VCAM antibodies .

This effect is blocked by <protein>anti-VCAM antibodies</protein> .


After a 2-day incubation in LDL , the binding of NF-kappa B , which is believed to be a key oxidative-stress sensor for VCAM regulation , remains at basal level .

After a 2-day incubation in <protein>LDL</protein> , the binding of <protein>NF-kappa B</protein> , which is believed to be a key oxidative-stress sensor for <protein>VCAM</protein> regulation , remains at basal level .


In contrast , the binding activities of AP-1 and GATA , on the other hand , are increased by LDL .

In contrast , the binding activities of <protein>AP-1</protein> and <protein>GATA</protein> , on the other hand , are increased by <protein>LDL</protein> .


Thus , a component of LDL -enhanced endothelial recruitment of monocytes is attributed to VCAM-1 expression , which appears to be mediated through AP-1 and GATA .

Thus , a component of <protein>LDL</protein> -enhanced endothelial recruitment of <cell_type>monocytes</cell_type> is attributed to <protein>VCAM-1</protein> expression , which appears to be mediated through <protein>AP-1</protein> and <protein>GATA</protein> .


These data identify LDL as a VCAM -inducer possibly distinct from cytokines and endotoxin .

These data identify <protein>LDL</protein> as a <protein>VCAM</protein> -inducer possibly distinct from <protein>cytokines</protein> and <protein>endotoxin</protein> .


Involvement of nuclear factor-kappa B activation in IgE synthesis in human B cells .

Involvement of <protein>nuclear factor-kappa B</protein> activation in <protein>IgE</protein> synthesis in <cell_type>human B cells</cell_type> .


Nuclear factor-kappa B ( NF-kappa B ) is a transcription factor that binds to the consensus DNA sequence in the cis-acting elements of various genes .

<protein>Nuclear factor-kappa B</protein> ( <protein>NF-kappa B</protein> ) is a transcription factor that binds to the <dna>consensus DNA sequence</dna> in the <dna>cis-acting elements</dna> of <dna>various genes</dna> .


Although NF-kappa B activates the expression of many genes involved in immune and inflammatory responses , little is known about the role of NF-kappa B activation in the induction of IgE synthesis in human B cells .

Although <protein>NF-kappa B</protein> activates the expression of many genes involved in immune and inflammatory responses , little is known about the role of <protein>NF-kappa B</protein> activation in the induction of IgE synthesis in <cell_type>human B cells</cell_type> .


Therefore we first examined the participation of NF-kappa B in germline C epsilon transcription in a human Burkitt lymphoma B cell line , DND39 .

Therefore we first examined the participation of <protein>NF-kappa B</protein> in <cell_line>germline C epsilon</cell_line> transcription in a <cell_line>human Burkitt lymphoma B cell line</cell_line> , <cell_line>DND39</cell_line> .


Stimulation of DND39 cells with IL-4 or anti-CD40 monoclonal antibody ( mAb ) activated phosphatidylinositol 3-kinase and subsequently induced nuclear expression of NF-kappa B , which was identified by electrophoretic mobility shift assays .

Stimulation of <cell_line>DND39 cells</cell_line> with <protein>IL-4</protein> or <protein>anti-CD40 monoclonal antibody</protein> ( <protein>mAb</protein> ) activated <protein>phosphatidylinositol 3-kinase</protein> and subsequently induced nuclear expression of <protein>NF-kappa B</protein> , which was identified by electrophoretic mobility shift assays .


n-Acetyl-L-cysteine ( NAC ) , a potent antioxidant , blocked NF-kappa B activation caused by IL-4 and by anti-CD40 mAb .

n-Acetyl-L-cysteine ( NAC ) , a potent antioxidant , blocked <protein>NF-kappa B</protein> activation caused by <protein>IL-4</protein> and by <protein>anti-CD40 mAb</protein> .


Although inhibition of IL-4-driven germline C epsilon transcription by NAC was not sufficient , the agent remarkably diminished anti-CD40 mAb -mediated up-regulation of germline C epsilon transcription .

Although inhibition of <cell_line>IL-4-driven germline C epsilon</cell_line> transcription by NAC was not sufficient , the agent remarkably diminished <protein>anti-CD40 mAb</protein> -mediated up-regulation of <dna>germline</dna> C epsilon transcription .


Second , we studied the effect of NAC on IgE synthesis in human normal B cells costimulated with IL-4 and anti-CD40 mAb .

Second , we studied the effect of NAC on <protein>IgE</protein> synthesis in human normal B cells costimulated with <protein>IL-4</protein> and <protein>anti-CD40 mAb</protein> .


NAC was effective in inhibiting mature C epsilon transcription and IgE synthesis in the T cell-independent culture system .

NAC was effective in inhibiting mature C epsilon transcription and <protein>IgE</protein> synthesis in the T cell-independent culture system .


However , NAC did not significantly affect the spontaneous production of IgE by atopic B cells .

However , NAC did not significantly affect the spontaneous production of <protein>IgE</protein> by <cell_type>atopic B cells</cell_type> .


These results indicate that NF-kappa B activity is commonly inducible in DND39 cells by IL-4 and anti-CD40 mAb and suggest that NF-kappa B sensitive to NAC may play a role in regulating IgE synthesis in B cells .

These results indicate that <protein>NF-kappa B</protein> activity is commonly inducible in <cell_line>DND39 cells</cell_line> by <protein>IL-4</protein> and <protein>anti-CD40 mAb</protein> and suggest that <protein>NF-kappa B</protein> sensitive to NAC may play a role in regulating <protein>IgE</protein> synthesis in <cell_type>B cells</cell_type> .


Prostaglandin E2 induction of binding activity to CRE and AP-2 elements in human T lymphocytes .

Prostaglandin E2 induction of binding activity to <dna>CRE</dna> and <dna>AP-2 elements</dna> in <cell_type>human T lymphocytes</cell_type> .


Prostaglandins of the E series are immunomodulatory agents which exert inhibitory as well as stimulatory effects on a variety of immune responses .


Since it is known that PGE2 is able to increase cAMP levels , we investigated whether it can affect gene expression through the activation of the transcription factors which bind enhancer elements in the promoter regions of cAMP-regulated genes .

Since it is known that PGE2 is able to increase cAMP levels , we investigated whether it can affect gene expression through the activation of the <protein>transcription factors</protein> which bind enhancer elements in the promoter regions of <dna>cAMP-regulated genes</dna> .


Using electrophoretic mobility shift assay , we demonstrated that a short treatment of human T lymphocytes with PGE2 induces specific binding activity to CRE and AP-2 , but not AP-1 , DNA elements .

Using electrophoretic mobility shift assay , we demonstrated that a short treatment of <cell_type>human T lymphocytes</cell_type> with PGE2 induces specific binding activity to <dna>CRE</dna> and <dna>AP-2</dna> , but not <dna>AP-1 , DNA elements</dna> .


Since the okadaic acid , a potent protein phosphatase inhibitor , prolongs the induction of the binding activity , phosphorylation events are likely to occur .


This activity seems to be due to increased cAMP levels because forskolin and IBMX mimic the effects of PGE2 .


More interestingly , transfection experiments with CRE-CAT plasmide show that PGE2 activates the transcription of a CRE-containing promoter .

More interestingly , transfection experiments with <dna>CRE-CAT plasmide</dna> show that PGE2 activates the transcription of a <dna>CRE-containing promoter</dna> .


These data support the positive role for PGE2 on some immune functions .


Nasal NK- and T-cell lymphomas share the same type of Epstein-Barr virus latency as nasopharyngeal carcinoma and Hodgkin 's disease .

<cell_type>Nasal NK- and T-cell lymphomas</cell_type> share the same type of Epstein-Barr virus latency as nasopharyngeal carcinoma and Hodgkin 's disease .


Nasal T/NK-cell lymphomas can be further separated into those of natural killer ( NK ) cell lineage or of T-cell lineage , with differences in cellular phenotype , T-cell receptor ( TcR ) gene rearrangement and TcR transcript expression .

<cell_type>Nasal T/NK-cell lymphomas</cell_type> can be further separated into those of <cell_type>natural killer ( NK ) cell lineage</cell_type> or of <cell_type>T-cell lineage</cell_type> , with differences in cellular phenotype , <dna>T-cell receptor ( TcR ) gene</dna> rearrangement and TcR transcript expression .


Both NK- and T-cell subtypes are closely associated with Epstein-Barr virus ( EBV ) .

Both <cell_type>NK- and T-cell subtypes</cell_type> are closely associated with Epstein-Barr virus ( EBV ) .


In this study , EBV gene expression was determined in 23 cases of nasal lymphoma ( NL ) by in situ hybridisation ( ISH ) , reverse transcriptase -polymerase chain reaction ( RT-PCR ) and immunohistochemistry ( IH ) .

In this study , <dna>EBV gene</dna> expression was determined in 23 cases of nasal lymphoma ( NL ) by in situ hybridisation ( ISH ) , <protein>reverse transcriptase</protein> -polymerase chain reaction ( RT-PCR ) and immunohistochemistry ( IH ) .


Of the 23 cases , 19 were classified as NK-cell and 4 as T-cell tumours .


ISH for EBV-encoded small non-polyadenylated RNAs showed that all cases , whether NK or T , harboured EBV in virtually all tumour cells .

ISH for <rna>EBV-encoded small non-polyadenylated RNAs</rna> showed that all cases , whether NK or T , harboured EBV in virtually all <cell_type>tumour cells</cell_type> .


RT-PCR demonstrated that NL of both subtypes expressed EBNAI of the QUK splice pattern , the latent membrane proteins , LMP1 and 2 and the BamHI A rightward transcripts in the absence of EBNA2 mRNAs , compatible with the latency type II pattern .

RT-PCR demonstrated that <cell_type>NL</cell_type> of both subtypes expressed <protein>EBNAI</protein> of the QUK splice pattern , the <protein>latent membrane proteins</protein> , <protein>LMP1 and 2</protein> and the <rna>BamHI A rightward transcripts</rna> in the absence of <rna>EBNA2 mRNAs</rna> , compatible with the latency type II pattern .


In addition , analysis of EBV protein expression by IH revealed a heterogeneous pattern of EBV gene expression at the single-cell level consisting of both LMP1+ and LMP1- tumour cells , suggesting a mixture of latency I and II .

In addition , analysis of EBV protein expression by IH revealed a heterogeneous pattern of <dna>EBV gene</dna> expression at the single-cell level consisting of both <cell_type>LMP1+ and LMP1- tumour cells</cell_type> , suggesting a mixture of latency I and II .


Although 2 early lytic transcripts , BZLF1 and BHRF1 , were also detected in 13 and 10 cases , respectively , the lack of ZEBRA staining in any case indicates that these lytic transcripts are most likely expressed by rare cells in the biopsies entering lytic cycle .

Although 2 <protein>early lytic transcripts</protein> , <protein>BZLF1</protein> and <protein>BHRF1</protein> , were also detected in 13 and 10 cases , respectively , the lack of <protein>ZEBRA</protein> staining in any case indicates that these lytic transcripts are most likely expressed by rare cells in the biopsies entering lytic cycle .


The viral transcriptional pattern similar to that of nasopharyngeal carcinoma and Hodgkin 's disease suggests that EBV can exploit common regulatory mechanisms for gene transcription in diverse host cell types .


Down-regulation of immunogenic proteins ( EBNA2-EBNA6 ) in nasal lymphoma may enable tumour cells to evade host cytotoxic T-cell surveillance .

Down-regulation of <protein>immunogenic proteins</protein> ( <protein>EBNA2-EBNA6</protein> ) in nasal lymphoma may enable <cell_type>tumour cells</cell_type> to evade host cytotoxic T-cell surveillance .


Potent gene regulatory and antiproliferative activities of 20-methyl analogues of 1 , 25 dihydroxyvitamin D3 .


The biological active form of vitamin D3 , 1 , 25-dihydroxyvitamin D3 ( VD ) , regulates cellular growth and differentiation .


This provides the hormone with an interesting therapeutic potential .


However , hypercalcemia is a side effect , which is caused by VD 's classical action , the regulation of calcium homeostasis .


This made the need for VD analogues with selectively increased cell regulatory properties .


Studies with 20-epi analogues pointed out the importance of the carbon-20 position and led to the development of 20-methyl derivatives of VD .


In this report the biological properties of the compounds ZK161422 and ZK157202 , which are 20-methyl- and 20-methyl-23-eneanalogues , respectively , have been analyzed in comparison with VD .


Both compounds show about 2-fold lower affinity to the VD receptor ( VDR ) than VD .


However , compared to VD , their antiproliferative effect is up to 30-fold higher on human peripheral blood mononuclear cells and even up to 300-fold higher on human breast cancer MCF-7 cells .

However , compared to VD , their antiproliferative effect is up to 30-fold higher on <cell_type>human peripheral blood mononuclear cells</cell_type> and even up to 300-fold higher on <cell_line>human breast cancer MCF-7 cells</cell_line> .


Whereas the hypercalcemic effect for ZK157202 is also increased 10-fold , ZK161422 has the same calcium-mobilizing potency as VD .


Moreover , ZK161422 , but not ZK157202 , showed preference for gene activation from a promoter carrying a VD response element with a palindromic arrangement of two hexameric receptor binding sites spaced by 9 nucleotides ( IP9 ) rather than for activation from a response element formed by a direct repeat spaced by 3 nucleotides ( DR3 ) .

Moreover , ZK161422 , but not ZK157202 , showed preference for gene activation from a <dna>promoter</dna> carrying a <dna>VD response element</dna> with a palindromic arrangement of two <dna>hexameric receptor binding sites</dna> spaced by 9 nucleotides ( IP9 ) rather than for activation from a <dna>response element</dna> formed by a direct repeat spaced by 3 nucleotides ( DR3 ) .


This observation supports a model , in which promoter selectivity reflects the selectively increased antiproliferative effect of VD analogues .


Calcium-dependent immediate-early gene induction in lymphocytes is negatively regulated by p21Ha-ras .

Calcium-dependent <dna>immediate-early gene</dna> induction in <cell_type>lymphocytes</cell_type> is negatively regulated by <protein>p21Ha-ras</protein> .


The induction of immediate-early ( IE ) response genes , such as egr-1 , c-fos , and c-jun , occurs rapidly after the activation of T lymphocytes .

The induction of <dna>immediate-early ( IE ) response genes</dna> , such as <dna>egr-1</dna> , <dna>c-fos</dna> , and <dna>c-jun</dna> , occurs rapidly after the activation of <cell_type>T lymphocytes</cell_type> .


The process of activation involves calcium mobilization , activation of protein kinase C ( PKC ) , and phosphorylation of tyrosine kinases .

The process of activation involves calcium mobilization , activation of <protein>protein kinase C</protein> ( <protein>PKC</protein> ) , and phosphorylation of <protein>tyrosine kinases</protein> .


p21 ( ras ) , a guanine nucleotide binding factor , mediates T-cell signal transduction through PKC -dependent and PKC -independent pathways .

<protein>p21 ( ras )</protein> , a <protein>guanine nucleotide binding factor</protein> , mediates T-cell signal transduction through <protein>PKC</protein> -dependent and <protein>PKC</protein> -independent pathways .


The involvement of p21 ( ras ) in the regulation of calcium-dependent signals has been suggested through analysis of its role in the activation of NF-AT .

The involvement of <protein>p21 ( ras )</protein> in the regulation of calcium-dependent signals has been suggested through analysis of its role in the activation of <protein>NF-AT</protein> .


We have investigated the inductions of the IE genes in response to calcium signals in Jurkat cells ( in the presence of activated p21 ( ras ) ) and their correlated consequences .

We have investigated the inductions of the <dna>IE genes</dna> in response to calcium signals in <cell_line>Jurkat cells</cell_line> ( in the presence of activated <protein>p21 ( ras )</protein> ) and their correlated consequences .


The expression of activated p21 ( ras ) negatively regulated the induction of IE genes by calcium ionophore .

The expression of activated <protein>p21 ( ras )</protein> negatively regulated the induction of <dna>IE genes</dna> by calcium ionophore .


This inhibition of calcium-activated IE gene induction was reversed by treatment with cyclosporin A , suggesting the involvement of calcineurin in this regulation .

This inhibition of calcium-activated <dna>IE gene</dna> induction was reversed by treatment with cyclosporin A , suggesting the involvement of <protein>calcineurin</protein> in this regulation .


A later result of inhibition of this activation pathway by p21 ( ras ) was down-regulation of the activity of the transcription factor AP-1 and subsequent coordinate reductions in IL-2 gene expression and protein production .

A later result of inhibition of this activation pathway by <protein>p21 ( ras )</protein> was down-regulation of the activity of the <protein>transcription factor</protein> <protein>AP-1</protein> and subsequent coordinate reductions in <dna>IL-2 gene</dna> expression and protein production .


These results suggest that p2l ( ras ) is an essential mediator in generating not only positive but also negative modulatory mechanisms controlling the competence of T cells in response to inductive stimulations .

These results suggest that <protein>p2l ( ras )</protein> is an essential mediator in generating not only positive but also negative modulatory mechanisms controlling the competence of <cell_type>T cells</cell_type> in response to inductive stimulations .


Calcineurin acts in synergy with PMA to inactivate I kappa B/MAD3 , an inhibitor of NF-kappa B .

<protein>Calcineurin</protein> acts in synergy with PMA to inactivate <protein>I kappa B/MAD3</protein> , an inhibitor of <protein>NF-kappa B</protein> .


The interleukin-2 ( IL-2 ) promoter consists of several independent T cell receptor ( TcR ) responsive elements .

The <dna>interleukin-2 ( IL-2 ) promoter</dna> consists of several independent <dna>T cell receptor ( TcR ) responsive elements</dna> .


The induction of promoters dependent on these elements is inhibitable by the immunosuppressants cyclosporin A ( CsA ) and tacrolimus ( FK-506 ) .

The induction of <dna>promoters</dna> dependent on these elements is inhibitable by the immunosuppressants cyclosporin A ( CsA ) and tacrolimus ( FK-506 ) .


Calcineurin , a Ca2+/calmodulin-dependent protein phosphatase , is the FK-506 -and CsA-sensitive enzyme required for TcR mediated activation of the IL-2 promoter .

<protein>Calcineurin</protein> , a <protein>Ca2+/calmodulin-dependent protein phosphatase</protein> , is the FK-506 -and CsA-sensitive enzyme required for TcR mediated activation of the <dna>IL-2 promoter</dna> .


We report that a constitutively active form of calcineurin partially substitutes for the Ca2+ co-stimulus required to activate the IL-2 promoter elements IL-2A ( which binds the factors OAP and Oct-1 ) and IL-2E ( which binds NF-AT ) , and completely substitutes for the Ca2+ co-stimulus required to stimulate an NF-kappa B -dependent element .

We report that a constitutively active form of <protein>calcineurin</protein> partially substitutes for the Ca2+ co-stimulus required to activate the <dna>IL-2 promoter elements</dna> <dna>IL-2A</dna> ( which binds the factors <protein>OAP</protein> and <protein>Oct-1</protein> ) and <dna>IL-2E</dna> ( which binds <protein>NF-AT</protein> ) , and completely substitutes for the Ca2+ co-stimulus required to stimulate an <protein>NF-kappa B</protein> -dependent element .


Calcineurin stimulates the NF-kappa B element by enhancing inactivation of I kappa B/MAD3 , an inhibitor of NF-kappa B , thereby increasing the amount of nuclear NF-kappa B DNA binding activity .

<protein>Calcineurin</protein> stimulates the <dna>NF-kappa B element</dna> by enhancing inactivation of <protein>I kappa B/MAD3</protein> , an inhibitor of <protein>NF-kappa B</protein> , thereby increasing the amount of nuclear <protein>NF-kappa B</protein> DNA binding activity .


These data provide the first demonstration in vivo that activation of a protein phosphatase can inactivate I kappa B , and suggest one possible explanation for mechanism-based toxicities associated with FK-506 and CsA by demonstrating that these drugs can inhibit the calcineurin -dependent activation of a virtually ubiquitous transcription factor .

These data provide the first demonstration in vivo that activation of a <protein>protein phosphatase</protein> can inactivate <protein>I kappa B</protein> , and suggest one possible explanation for mechanism-based toxicities associated with FK-506 and CsA by demonstrating that these drugs can inhibit the <protein>calcineurin</protein> -dependent activation of a virtually <protein>ubiquitous transcription factor</protein> .


Detection of minimal residual disease in a patient with acute promyelocytic leukemia by RT-PCR : necessity of chemotherapy following ATRA therapy .


The PML/RAR alpha fusion gene resulting from the t ( 15 ; 17 ) translocation is a specific marker for acute promyelocytic leukemia ( APL ) .

The <dna>PML/RAR alpha fusion gene</dna> resulting from the <dna>t ( 15 ; 17 ) translocation</dna> is a specific marker for acute promyelocytic leukemia ( APL ) .


We examined bone marrow cells by reverse transcriptase-polymerase chain reaction ( RT-PCR ) to detect residual PML/RAR alpha mRNA-containing cells following treatment with all-trans retinoic acid ( ATRA ) and cytotoxic chemotherapy in a patient with APL .

We examined bone marrow cells by reverse transcriptase-polymerase chain reaction ( RT-PCR ) to detect residual <cell_type>PML/RAR alpha mRNA-containing cells</cell_type> following treatment with all-trans retinoic acid ( ATRA ) and cytotoxic chemotherapy in a patient with APL .


This RT-PCR assay can detect one leukemic cell in 10 ( 2 ) normal cells in vitro .

This RT-PCR assay can detect one <cell_type>leukemic cell</cell_type> in 10 ( 2 ) <cell_type>normal cells</cell_type> in vitro .


We show that PML/RAR alpha mRNA was still detectable despite clinical remission following ATRA treatment , but undetectable following consolidation with chemotherapy .

We show that <rna>PML/RAR alpha mRNA</rna> was still detectable despite clinical remission following ATRA treatment , but undetectable following consolidation with chemotherapy .


These data show that this technique is useful for the identification of minimal residual disease in patients with APL and that cytotoxic chemotherapy following ATRA therapy is required for the elimination of APL cells .

These data show that this technique is useful for the identification of minimal residual disease in patients with APL and that cytotoxic chemotherapy following ATRA therapy is required for the elimination of <cell_type>APL cells</cell_type> .


Glucocorticoid-mediated inhibition of interleukin-2 receptor alpha and -beta subunit expression by human T cells .


To determine the mechanism of glucocorticoid ( GC ) -mediated inhibition of T cell functions , the effect of dexamethasone ( DM ) on T cell proliferation and interleukin-2 receptor ( IL-2R ) generation were studied .

To determine the mechanism of glucocorticoid ( GC ) -mediated inhibition of T cell functions , the effect of dexamethasone ( DM ) on <cell_type>T cell</cell_type> proliferation and <protein>interleukin-2 receptor</protein> ( <protein>IL-2R</protein> ) generation were studied .


Dexamethasone inhibited IL-2 -induced T cell proliferation by 30 % -88 % , relative to its concentration within the cultures .

Dexamethasone inhibited <protein>IL-2</protein> -induced <cell_type>T cell</cell_type> proliferation by 30 % -88 % , relative to its concentration within the cultures .


The effect of DM on expression of IL-2R alpha ( Tac , p55 , CD25 ) and beta ( p75 ) genes in activated T cells was examined next .

The effect of DM on expression of <protein>IL-2R alpha</protein> ( <protein>Tac</protein> , <protein>p55</protein> , <protein>CD25</protein> ) and <dna>beta ( p75 ) genes</dna> in <cell_type>activated T cells</cell_type> was examined next .


In T cells stimulated with purified phytohemagglutinin ( PHA-p ) and 4 beta-phorbol 12-myristate 13-acetate ( PMA ) addition of DM to the cultures resulted in a 60 % reduction in IL-2R alpha and a 30 % reduction in IL-2R beta membrane expression compared to T cells cultured in the absence of DM ( p < 0.01 ) .

In <cell_type>T cells</cell_type> stimulated with purified <protein>phytohemagglutinin</protein> ( <protein>PHA-p</protein> ) and 4 beta-phorbol 12-myristate 13-acetate ( PMA ) addition of DM to the cultures resulted in a 60 % reduction in <protein>IL-2R alpha</protein> and a 30 % reduction in <protein>IL-2R beta</protein> membrane expression compared to <cell_type>T cells</cell_type> cultured in the absence of DM ( p < 0.01 ) .


Inhibition of membrane IL-2R alpha and IL-2R beta expression by 10 ( -6 ) M DM was partially reversible by recombinant human IL-2 ( rhIL-2 ) .

Inhibition of membrane <protein>IL-2R alpha</protein> and <protein>IL-2R beta</protein> expression by 10 ( -6 ) M DM was partially reversible by <protein>recombinant human IL-2</protein> ( <protein>rhIL-2</protein> ) .


By Northern blot analysis , DM caused a comparable decrease in IL-2R alpha and in IL-2R beta mRNA levels to membrane receptor expression in mitogen-stimulated T cells .

By Northern blot analysis , DM caused a comparable decrease in <protein>IL-2R alpha</protein> and in <protein>IL-2R beta</protein> mRNA levels to membrane receptor expression in <cell_type>mitogen-stimulated T cells</cell_type> .


By in vitro transcription assays , DM regulated IL-2R alpha gene expression at a transcriptional level while transcription of IL-2R beta gene was unaffected by DM .

By in vitro transcription assays , DM regulated <protein>IL-2R alpha</protein> gene expression at a transcriptional level while transcription of <dna>IL-2R beta gene</dna> was unaffected by DM .


The mechanism of action of DM on IL-2R alpha transcription was examined by determining the mRNA levels of the p50 subunit of nuclear factor kappa B ( NF-kappa B ) , a transcription factor that stimulates IL-2R alpha gene expression .

The mechanism of action of DM on <protein>IL-2R alpha</protein> transcription was examined by determining the mRNA levels of the p50 subunit of nuclear factor kappa B ( <protein>NF-kappa B</protein> ) , a <protein>transcription factor</protein> that stimulates <protein>IL-2R alpha</protein> gene expression .


The data indicate that 10 ( -6 ) M DM increased T cell p50 NF-kappa B mRNA levels by four-fold compared to the levels obtained in the absence of DM .

The data indicate that 10 ( -6 ) M DM increased <cell_type>T cell</cell_type> p50 <rna>NF-kappa B mRNA</rna> levels by four-fold compared to the levels obtained in the absence of DM .


Further , the level of nuclear proteins capable of binding to the NF-kappa B sites in activated T cells increased in response to DM .

Further , the level of <protein>nuclear proteins</protein> capable of binding to the <dna>NF-kappa B sites</dna> in <cell_type>activated T cells</cell_type> increased in response to DM .


In sum , DM regulates T cell membrane expression of IL-2R by more than one molecular mechanism .

In sum , DM regulates <cell_type>T cell</cell_type> membrane expression of <protein>IL-2R</protein> by more than one molecular mechanism .


Alteration of structural order of human erythrocyte ghost membrane by glucocorticoids and the influence of the glucocorticoid receptor antagonist RU 486 .


High-dose pulse glucocorticoid therapy has been used successfully in the clinic in severe pathological conditions for about 20 years .


The mode of glucocorticoid action after administration of such megadoses is inexplicable up to now .


It is supposed that some effects may be due to membrane alterations .


In the present in-vitro experiments the effect of dexamethasone , of further glucocorticoids , and of the glucocorticoid receptor antagonist RU 486 , on structural order of human erythrocyte ghost membranes was investigated by determining the steady-state fluorescence anisotropy of diphenylhexatriene ( DPH ) .


Dexamethasone was found to induce a significant decrease in membrane structural order at concentrations of about 10 ( -6 ) M in a concentration-dependent manner .


We found a correlation between the uptake of dexamethasone by the ghost membranes and the decrease in the structural order .


The other glucocorticoids tested , methylprednisolone and corticosterone , were also effective at concentrations of 10 ( -5 ) M or greater .


We observed no change in membrane structural order with RU 486 up to a concentration of 10 ( -4 ) M .


However , simultaneous incubation of RU 486 with dexamethasone caused a distinct interference of RU 486 with dexamethasone .


Thus , the glucocorticoid-induced membrane perturbation , the possibility to inhibit it by RU 486 , and the inactivity of the structurally related progesterone , refer to relatively specific binding sites for the glucocorticoids in the membrane of erythrocyte ghosts .

Thus , the glucocorticoid-induced membrane perturbation , the possibility to inhibit it by RU 486 , and the inactivity of the structurally related progesterone , refer to relatively specific <dna>binding sites</dna> for the glucocorticoids in the membrane of <cell_type>erythrocyte ghosts</cell_type> .


Mutation of tyrosines 492/493 in the kinase domain of ZAP-70 affects multiple T-cell receptor signaling pathways .

Mutation of tyrosines 492/493 in the <protein>kinase domain</protein> of <protein>ZAP-70</protein> affects multiple <protein>T-cell receptor</protein> signaling pathways .


The protein-tyrosine kinase ZAP-70 is implicated , together with the Src kinase p56 ( lck ) , in controlling the early steps of the T-cell antigen receptor ( TCR ) signaling cascade .

The <protein>protein-tyrosine kinase</protein> <protein>ZAP-70</protein> is implicated , together with the <protein>Src kinase</protein> <protein>p56 ( lck )</protein> , in controlling the early steps of the T-cell antigen receptor ( <protein>TCR</protein> ) signaling cascade .


To help elucidate further the mechanism by which ZAP-70 regulates these initial events , we used a dominant-negative mutant approach .

To help elucidate further the mechanism by which <protein>ZAP-70</protein> regulates these initial events , we used a dominant-negative mutant approach .


We overexpressed in the Jurkat T-cell line ZAP-70 mutated on Tyr-492 and Tyr-493 in the putative regulatory loop of its kinase domain .

We overexpressed in the <cell_line>Jurkat T-cell line</cell_line> <protein>ZAP-70</protein> mutated on Tyr-492 and Tyr-493 in the putative regulatory loop of its <protein>kinase domain</protein> .


This mutant inhibited TCR -induced activation of nuclear factor of activated T cells by interfering with both intracellular calcium increase and Ras -regulated activation of extracellular signal-regulated kinases .

This mutant inhibited <protein>TCR</protein> -induced activation of <protein>nuclear factor of activated T cells</protein> by interfering with both intracellular calcium increase and <protein>Ras</protein> -regulated activation of <protein>extracellular signal-regulated kinases</protein> .


Moreover , TCR -induced phosphorylation of pp36-38 , thought to play a role upstream of these pathways , was found to be reduced .

Moreover , <protein>TCR</protein> -induced phosphorylation of <protein>pp36-38</protein> , thought to play a role upstream of these pathways , was found to be reduced .


In contrast , overexpression of wild-type ZAP-70 induced constitutive activation of nuclear factor of activated T cells .

In contrast , overexpression of <protein>wild-type ZAP-70</protein> induced constitutive activation of <protein>nuclear factor of activated T cells</protein> .


The ZAP-70 mutant studied here could be phosphorylated on tyrosine when associated to the TCR zeta chain and was able to bind p56 ( lck ) .

The <protein>ZAP-70</protein> mutant studied here could be phosphorylated on tyrosine when associated to the <protein>TCR zeta chain</protein> and was able to bind <protein>p56 ( lck )</protein> .


This result demonstrates that Tyr-492 and Tyr-493 are not responsible for the Src homology domain 2-mediated association of p56 ( lck ) with ZAP-70 .

This result demonstrates that Tyr-492 and Tyr-493 are not responsible for the Src homology domain 2-mediated association of <protein>p56 ( lck )</protein> with <protein>ZAP-70</protein> .


Our data are most consistent with a model in which recruitment to the TCR allows ZAP-70 autophosphorylation and binding to p56 ( lck ) , which in turn phosphorylates Tyr-492 and/or Tyr-493 with consequent up-regulation of the ZAP-70 kinase activity .

Our data are most consistent with a model in which recruitment to the <protein>TCR</protein> allows <protein>ZAP-70</protein> autophosphorylation and binding to <protein>p56 ( lck )</protein> , which in turn phosphorylates Tyr-492 and/or Tyr-493 with consequent up-regulation of the <protein>ZAP-70</protein> kinase activity .


ZAP-70 will then be able to effectively control phosphorylation of its substrates and lead to gene activation .

<protein>ZAP-70</protein> will then be able to effectively control phosphorylation of its substrates and lead to gene activation .


Effects of glucocorticoids on lymphocyte activation in patients with steroid-sensitive and steroid-resistant asthma .


BACKGROUND : Glucocorticoids are important medications used to control the airway inflammation associated with asthma .


Synthetic glucocorticoids vary in their binding affinity for the glucocorticoid receptor ( GCR ) .


METHODS : We compared hydrocortisone , beclomethasone dipropionate , triamcinolone acetonide , flunisolide , and budesonide with regard to their capacity to inhibit phytohemagglutinin -induced peripheral blood mononuclear cell proliferation from six patients with steroid-sensitive asthma and seven patients with steroid-resistant asthma .

METHODS : We compared hydrocortisone , beclomethasone dipropionate , triamcinolone acetonide , flunisolide , and budesonide with regard to their capacity to inhibit <protein>phytohemagglutinin</protein> -induced <cell_type>peripheral blood mononuclear cell</cell_type> proliferation from six patients with steroid-sensitive asthma and seven patients with steroid-resistant asthma .


Peripheral blood mononuclear cell GCR binding affinities for dexamethasone and budesonide were also determined for both patient groups by using a radioligand binding assay and Scatchard analysis .

Peripheral blood mononuclear cell <protein>GCR</protein> binding affinities for dexamethasone and budesonide were also determined for both patient groups by using a radioligand binding assay and Scatchard analysis .


RESULTS : Dose-dependent inhibition was demonstrated for all glucocorticoids in both patient groups , with the steroid-resistant group requiring approximately 2 log-fold more glucocorticoids for an equivalent degree of inhibition .


The mean concentrations necessary to cause 50 % inhibition of lymphocyte proliferation ( IC50s ) for the steroid-sensitive group ranged from 2 x 10 ( -10 ) mol/L for budesonide to 7 x 10 ( -8 ) mol/L for hydrocortisone , whereas the mean IC50s for the steroid-resistant group ranged from approximately 2 x 10 ( -8 ) mol/L for budesonide to greater than 10 ( -6 ) mol/L for hydrocortisone .


In addition , a significant correlation was noted between the degree of inhibition of lymphocyte proliferation ( IC50 ) and the binding affinity of dexamethasone to the GCR .

In addition , a significant correlation was noted between the degree of inhibition of lymphocyte proliferation ( IC50 ) and the binding affinity of dexamethasone to the <protein>GCR</protein> .


Patients with steroid-resistant asthma have been shown to have a reduced GCR binding affinity .

Patients with steroid-resistant asthma have been shown to have a reduced <protein>GCR</protein> binding affinity .


The GCR binding affinity for budesonide was significantly higher in both groups ( i.e. , lower dissociation constant ) than that obtained for dexamethasone .

The <protein>GCR</protein> binding affinity for budesonide was significantly higher in both groups ( i.e. , lower dissociation constant ) than that obtained for dexamethasone .


CONCLUSION : These data suggest that glucocorticoids such as budesonide , by virtue of their high GCR binding affinities and greater ability to suppress lymphocyte proliferation , may therefore be beneficial in the management of difficult-to-control asthma .

CONCLUSION : These data suggest that glucocorticoids such as budesonide , by virtue of their high <protein>GCR</protein> binding affinities and greater ability to suppress lymphocyte proliferation , may therefore be beneficial in the management of difficult-to-control asthma .


Characterization of the murine cyclin-dependent kinase inhibitor gene p27Kip1 .

Characterization of the <dna>murine cyclin-dependent kinase inhibitor gene</dna> <dna>p27Kip1</dna> .


The cyclin-dependent kinase inhibitor p27Kip1 plays an important role in regulating cell-cycle progression .

The <dna>cyclin-dependent kinase inhibitor</dna> <dna>p27Kip1</dna> plays an important role in regulating cell-cycle progression .


p27Kip1 directly inhibits the catalytic activity of cyclin/cdks ( cyclin-dependent kinase ) complexes and/or interferes physically with cyclin/cdks activation by CAK .

<dna>p27Kip1</dna> directly inhibits the catalytic activity of <protein>cyclin/cdks ( cyclin-dependent kinase ) complexes</protein> and/or interferes physically with <protein>cyclin/cdks</protein> activation by <protein>CAK</protein> .


Interestingly , the expression level of p27Kip1 mRNA was maximal in resting Go T-cells and rapidly declined following anti-CD3 activation .

Interestingly , the expression level of <rna>p27Kip1 mRNA</rna> was maximal in <cell_type>resting Go T-cells</cell_type> and rapidly declined following <protein>anti-CD3</protein> activation .


We report here the cloning of p27Kip1 gene from murine genomic DNA and the functional analysis of the promoter of the p27Kip1 gene .

We report here the cloning of <dna>p27Kip1 gene</dna> from <dna>murine genomic DNA</dna> and the functional analysis of the promoter of the <dna>p27Kip1</dna> gene .


The gene consists of at least three exons and spans more than 5.6 kb of DNA .

The gene consists of at least three exons and spans more than <dna>5.6 kb of DNA</dna> .


Primer extension and nuclease S1 protection analysis revealed two major transcription initiation sites .

Primer extension and nuclease S1 protection analysis revealed two major <dna>transcription initiation sites</dna> .


The promoter region lacked a TATA box but contained potential binding sites for the transcriptional factors including two Sp1 , CRE , Myb and NFkB located at positions -153 , -178 , -286 , -875 , and -1011 , respectively .

The <dna>promoter region</dna> lacked a <dna>TATA box</dna> but contained potential binding sites for the <protein>transcriptional factors</protein> including two <protein>Sp1</protein> , <protein>CRE</protein> , <protein>Myb</protein> and <protein>NFkB</protein> located at <dna>positions -153 , -178 , -286 , -875 , and -1011</dna> , respectively .


To analyze the regulatory mechanisms controlling p27Kip1 gene expression , we characterized the 5'-flanking region from nt -1609 to +178 .

To analyze the regulatory mechanisms controlling <dna>p27Kip1</dna> gene expression , we characterized the <dna>5'-flanking region</dna> from <dna>nt -1609 to +178</dna> .


The -326 to -615 region contained positive regulatory elements .

The <dna>-326 to -615 region</dna> contained <dna>positive regulatory elements</dna> .


Nuclear NF-ATp is a hallmark of unstimulated B cells from B-CLL patients .

<protein>Nuclear NF-ATp</protein> is a hallmark of <cell_type>unstimulated B cells</cell_type> from B-CLL patients .


B lymphocytes from the peripheral blood of patients with chronic lymphocytic leukaemia ( CLL ) were analysed for the nuclear presence and DNA binding of a panel of transcription factors which are involved in the gene control of lymphoid cells .

<cell_type>B lymphocytes</cell_type> from the peripheral blood of patients with chronic lymphocytic leukaemia ( CLL ) were analysed for the nuclear presence and DNA binding of a panel of <protein>transcription factors</protein> which are involved in the gene control of <cell_type>lymphoid cells</cell_type> .


The following transcription factors were studied : the Octamer factors Oct-1 and Oct-2 , members of the AP-1 factor family , NF-AT factors , in particular NF-ATp , and members of the Rel/NF-kB family .

The following <protein>transcription factors</protein> were studied : the <protein>Octamer factors</protein> <protein>Oct-1</protein> and <protein>Oct-2</protein> , members of the <protein>AP-1 factor family</protein> , <protein>NF-AT factors</protein> , in particular <protein>NF-ATp</protein> , and members of the <protein>Rel/NF-kB family</protein> .


We show that the constitutive nuclear translocation of NF-ATp , a member of the growing family of NF-AT factors , is a hallmark of nonstimulated B cells from CLL patients that distinguishes B-CLL cells from 'normal ' B lymphocytes .

We show that the constitutive nuclear translocation of <protein>NF-ATp</protein> , a member of the growing family of <protein>NF-AT factors</protein> , is a hallmark of <cell_type>nonstimulated B cells</cell_type> from CLL patients that distinguishes <cell_type>B-CLL cells</cell_type> from <cell_type>'normal ' B lymphocytes</cell_type> .


Constitutive nuclear appearance was also observed for NF-kB2/p52 .


Constitutive binding of further factor proteins to DNA , such as JunD , c-Fos and FosB , was detected in several patients whereas the localisation and DNA binding of other factors such as c-Jun , RelA/p65 and c-Rel was unaltered .

Constitutive binding of further <protein>factor proteins</protein> to DNA , such as <protein>JunD</protein> , <protein>c-Fos</protein> and <protein>FosB</protein> , was detected in several patients whereas the localisation and DNA binding of other factors such as <protein>c-Jun</protein> , <protein>RelA/p65</protein> and <protein>c-Rel</protein> was unaltered .


It is remarkable that in B-CLL cells the nuclear appearance and DNA binding of specific transcription factors is dramatically affected whereas other members of the same factor family remained unaltered in these leukemic cells .

It is remarkable that in <cell_type>B-CLL cells</cell_type> the nuclear appearance and DNA binding of specific <protein>transcription factors</protein> is dramatically affected whereas other members of the same <protein>factor family</protein> remained unaltered in these leukemic cells .


It remains to be shown which molecular events lead to the specific 'pre-activation ' , i.e .


constitutive nuclear translocation and DNA binding , of these members of NF-AT , NF-kB and AP-1 factor families .

constitutive nuclear translocation and DNA binding , of these members of <protein>NF-AT</protein> , <protein>NF-kB</protein> and <protein>AP-1 factor families</protein> .


Naive ( CD45RA+ ) T lymphocytes are more sensitive to oxidative stress-induced signals than memory ( CD45RO+ ) cells .

<cell_type>Naive ( CD45RA+ ) T lymphocytes</cell_type> are more sensitive to oxidative stress-induced signals than <cell_type>memory ( CD45RO+ ) cells</cell_type> .


Formation of reactive oxygen intermediates ( ROI ) after oxidative stress has been shown to be an activation signal for T lymphocytes , e.g. , expression of IL-2 and its receptor are induced .

Formation of reactive oxygen intermediates ( ROI ) after oxidative stress has been shown to be an activation signal for <cell_type>T lymphocytes</cell_type> , e.g. , expression of <protein>IL-2</protein> and its <protein>receptor</protein> are induced .


These ROI-induced effects can , to a large extent , be attributed to the activation of the transcription factor NF-kappaB .

These ROI-induced effects can , to a large extent , be attributed to the activation of the <protein>transcription factor</protein> <protein>NF-kappaB</protein> .


Now we have examined whether naive and memory T lymphocytes differ in their sensitivity to ROI-mediated signals .

Now we have examined whether <cell_type>naive and memory T lymphocytes</cell_type> differ in their sensitivity to ROI-mediated signals .


When CD45RA+ ( naive ) and CD45RO+ ( memory ) T lymphocytes were directly stimulated with H2O2 , NF-kappaB nuclear translocation was stronger in naive cells than in memory cells and it could be induced with lower doses .

When <cell_type>CD45RA+ ( naive )</cell_type> and <cell_type>CD45RO+ ( memory ) T lymphocytes</cell_type> were directly stimulated with H2O2 , <protein>NF-kappaB</protein> nuclear translocation was stronger in <cell_type>naive cells</cell_type> than in <cell_type>memory cells</cell_type> and it could be induced with lower doses .


The composition of the induced nuclear NF-kappaB ( levels of p50 and RelA proteins ) was similar in these cell types .

The composition of the induced <protein>nuclear NF-kappaB</protein> ( levels of <protein>p50</protein> and <protein>RelA proteins</protein> ) was similar in these cell types .


The magnitude and kinetics of intracellular ROI were similar , suggesting that there were no differences in ROI-forming mechanisms or antioxidative capacities .


The probable regulatory point was the cytoplasmic IkappaB inhibitor : in CD45RA+ cells , H2O2 caused a more profound depression in the levels of IkappaB alpha .

The probable regulatory point was the cytoplasmic <protein>IkappaB</protein> inhibitor : in CD45RA+ cells , H2O2 caused a more profound depression in the levels of <protein>IkappaB alpha</protein> .


These findings indicate that T cells representing different activation and/or differentiation stages can be differentially responsive to ROI-mediated signals .

These findings indicate that <cell_type>T cells</cell_type> representing different activation and/or differentiation stages can be differentially responsive to ROI-mediated signals .


Rapid shuttling of NF-AT in discrimination of Ca2+ signals and immunosuppression .

Rapid shuttling of <protein>NF-AT</protein> in discrimination of Ca2+ signals and immunosuppression .


Cells need to distinguish between transient Ca2+ signals that induce events such as muscle contraction , secretion , adhesion and synaptic transmission , and sustained Ca2+ signals that are involved in cell proliferation and differentiation .


The latter class of events is blocked in lymphocytes by the immunosuppressive drugs cyclosporin A and FK506 , which inhibit calcineurin , a Ca2+-activated serine/threonine phosphatase necessary for the nuclear import of NF-AT transcription factors .

The latter class of events is blocked in <cell_type>lymphocytes</cell_type> by the immunosuppressive drugs cyclosporin A and FK506 , which inhibit <protein>calcineurin</protein> , a <protein>Ca2+-activated serine/threonine phosphatase</protein> necessary for the nuclear import of <protein>NF-AT</protein> <protein>transcription factors</protein> .


Here we report that sustained high concentrations of Ca2+ , but not transient pulses , are required to maintain NF-AT transcription factors in the nucleus , where they participate in Ca2+-dependent induction of genes required for lymphocyte activation and proliferation .

Here we report that sustained high concentrations of Ca2+ , but not transient pulses , are required to maintain <protein>NF-AT</protein> <protein>transcription factors</protein> in the nucleus , where they participate in Ca2+-dependent induction of <dna>genes</dna> required for <cell_type>lymphocyte</cell_type> activation and proliferation .


Furthermore , overexpression and constitutive nuclear localization of NF-AT , but not Jun , Fos , NF-kappaB , Oct or Ets family members , renders the interleukin-2 enhancer in Jurkat T lymphocytes resistant to FK506 and cyclosporin A .

Furthermore , overexpression and constitutive nuclear localization of <protein>NF-AT</protein> , but not <protein>Jun , Fos , NF-kappaB , Oct or Ets family members</protein> , renders the <dna>interleukin-2 enhancer</dna> in <cell_line>Jurkat T lymphocytes</cell_line> resistant to FK506 and cyclosporin A .


Thus a primary effect of these immunosuppressive reagents is to control the subcellular localization of the NF-AT family of transcription factors .

Thus a primary effect of these immunosuppressive reagents is to control the subcellular localization of the <protein>NF-AT</protein> family of <protein>transcription factors</protein> .


Autoregulation of the NF-kappa B transactivator RelA ( p65 ) by multiple cytoplasmic inhibitors containing ankyrin motifs .

Autoregulation of the <protein>NF-kappa B transactivator</protein> <protein>RelA</protein> ( <protein>p65</protein> ) by multiple <protein>cytoplasmic inhibitors</protein> containing <protein>ankyrin motifs</protein> .


RelA ( p65 ) functions as the critical transactivating component of the heterodimeric p50-p65 NF-kappa B complex and contains a high-affinity binding site for its cytoplasmic inhibitor , I kappa B alpha .

<protein>RelA</protein> ( <protein>p65</protein> ) functions as the critical transactivating component of the <protein>heterodimeric p50-p65 NF-kappa B complex</protein> and contains a <protein>high-affinity binding site</protein> for its <protein>cytoplasmic inhibitor</protein> , <protein>I kappa B alpha</protein> .


After cellular activation , I kappa B alpha is rapidly degraded in concert with the induced nuclear translocation of NF-kappa B .

After cellular activation , <protein>I kappa B alpha</protein> is rapidly degraded in concert with the induced nuclear translocation of <protein>NF-kappa B</protein> .


The present study demonstrates that tumor necrosis factor alpha -induced degradation of I kappa B alpha in human T cells is preceded by its rapid phosphorylation in vivo .

The present study demonstrates that <protein>tumor necrosis factor alpha</protein> -induced degradation of <protein>I kappa B alpha</protein> in <cell_type>human T cells</cell_type> is preceded by its rapid phosphorylation in vivo .


However , these effects on I kappa B alpha result in nuclear mobilization of only a fraction of the entire cytoplasmic pool of RelA .

However , these effects on <protein>I kappa B alpha</protein> result in nuclear mobilization of only a fraction of the entire cytoplasmic pool of <protein>RelA</protein> .


Subsequent studies have revealed that ( i ) cytoplasmic RelA is stably associated not only with I kappa B alpha but also with other ankyrin motif-rich proteins including the products of the NF-kappa B2 ( p100 ) and NF-kappa B1 ( p105 ) genes ; ( ii ) in contrast to RelA -I kappa B alpha , RelA-p100 cytoplasmic complexes are not dissociated following tumor necrosis factor alpha activation ; ( iii ) p100 functions as a potent inhibitor of RelA -mediated transcription in vivo ; ( iv ) the interaction of RelA and p100 involves the conserved Rel homology domain of both proteins but not the nuclear localization signal of RelA , which is required for I kappa B alpha binding ; ( v ) p100 inhibition of RelA function requires the C-terminal ankyrin motif domain , which mediates cytoplasmic retention of RelA ; and ( vi ) as observed with I kappa B alpha , nuclear RelA stimulates p100 mRNA and protein expression .

Subsequent studies have revealed that ( i ) <protein>cytoplasmic RelA</protein> is stably associated not only with <protein>I kappa B alpha</protein> but also with other <protein>ankyrin motif-rich proteins</protein> including the products of the <dna>NF-kappa B2 ( p100 ) and NF-kappa B1 ( p105 ) genes</dna> ; ( ii ) in contrast to <protein>RelA</protein> <protein>-I kappa B alpha</protein> , <protein>RelA-p100 cytoplasmic complexes</protein> are not dissociated following <protein>tumor necrosis factor alpha</protein> activation ; ( iii ) <protein>p100</protein> functions as a potent inhibitor of <protein>RelA</protein> -mediated transcription in vivo ; ( iv ) the interaction of <protein>RelA</protein> and <protein>p100</protein> involves the <protein>conserved Rel homology domain</protein> of both proteins but not the <protein>nuclear localization signal</protein> of <protein>RelA</protein> , which is required for <protein>I kappa B alpha</protein> binding ; ( v ) <protein>p100</protein> inhibition of <protein>RelA</protein> function requires the <protein>C-terminal ankyrin motif domain</protein> , which mediates cytoplasmic retention of <protein>RelA</protein> ; and ( vi ) as observed with <protein>I kappa B alpha</protein> , nuclear <protein>RelA</protein> stimulates <rna>p100 mRNA</rna> and protein expression .


These findings thus reveal the presence of a second inducible autoregulated inhibitory pathway that helps ensure the rapid but transient action of nuclear NF-kappa B .

These findings thus reveal the presence of a second inducible autoregulated inhibitory pathway that helps ensure the rapid but transient action of <protein>nuclear NF-kappa B</protein> .


Effects of CD45 on NF-kappa B .

Effects of <protein>CD45</protein> on <protein>NF-kappa B</protein> .


Implications for replication of HIV-1 .


Increased levels of replication of the HIV type 1 are observed after the activation of infected T cells through the TCR .

Increased levels of replication of the HIV type 1 are observed after the activation of <cell_type>infected T cells</cell_type> through the <protein>TCR</protein> .


However , anti- CD45 antibodies inhibit these effects in cells from infected individuals .

However , anti- <protein>CD45</protein> antibodies inhibit these effects in cells from infected individuals .


In this study , we examined interrelationships between CD45 and HIV-1 further .

In this study , we examined interrelationships between <protein>CD45</protein> and HIV-1 further .


We measured effects on the HIV-1 LTR in T cell lines that were stimulated with antibodies against CD45 and in those that lacked the expression of CD45 on their surfaces .

We measured effects on the <dna>HIV-1 LTR</dna> in <cell_line>T cell lines</cell_line> that were stimulated with antibodies against <protein>CD45</protein> and in those that lacked the expression of <protein>CD45</protein> on their surfaces .


First , anti-CD45 antibodies did not affect basal but decreased activated levels of expression from the HIV-1 LTR .

First , <protein>anti-CD45 antibodies</protein> did not affect basal but decreased activated levels of expression from the <dna>HIV-1 LTR</dna> .


Second , T cells , which lack CD45 and can not signal via the TCR , supported higher levels of viral replication and gene expression .

Second , <cell_type>T cells</cell_type> , which lack <protein>CD45</protein> and can not signal via the <protein>TCR</protein> , supported higher levels of viral replication and gene expression .


This was due to the presence of active NF-kappa B complexes in the nucleus of CD45 -T cells .

This was due to the presence of active <protein>NF-kappa B complexes</protein> in the nucleus of <protein>CD45</protein> -T cells .


Additionally , infected T cells displayed lower levels of CD45 on their surfaces .

Additionally , <cell_type>infected T cells</cell_type> displayed lower levels of <protein>CD45</protein> on their surfaces .


Thus , CD45 plays an active role in the physiology of T cells and in the replication of HIV-1 .

Thus , <protein>CD45</protein> plays an active role in the physiology of <cell_type>T cells</cell_type> and in the replication of HIV-1 .


Calcineurin potentiates activation of the granulocyte-macrophage colony-stimulating factor gene in T cells : involvement of the conserved lymphokine element 0 .

<protein>Calcineurin</protein> potentiates activation of the <dna>granulocyte-macrophage colony-stimulating factor gene</dna> in <cell_type>T cells</cell_type> : involvement of the <dna>conserved lymphokine element 0</dna> .


Granulocyte-macrophage colony-stimulating factor ( GM-CSF ) and interleukin-2 ( IL-2 ) are produced by stimulation with phorbol-12-myristate acetate ( PMA ) and calcium ionophore ( A23187 ) in human T cell leukemia Jurkat cells .

<protein>Granulocyte-macrophage colony-stimulating factor</protein> ( <protein>GM-CSF</protein> ) and <protein>interleukin-2</protein> ( <protein>IL-2</protein> ) are produced by stimulation with phorbol-12-myristate acetate ( PMA ) and calcium ionophore ( A23187 ) in <cell_line>human T cell leukemia Jurkat cells</cell_line> .


The expression of GM-CSF and IL-2 is inhibited by immunosuppressive drugs such as cyclosporin A ( CsA ) and FK506 .

The expression of <protein>GM-CSF</protein> and <protein>IL-2</protein> is inhibited by immunosuppressive drugs such as cyclosporin A ( CsA ) and FK506 .


Earlier studies on the IL-2 gene expression showed that overexpression of calcineurin ( CN ) , a Ca2+/calmodulin-dependent protein phosphatase , can stimulate transcription from the IL-2 promoter through the NF-AT-binding site .

Earlier studies on the <protein>IL-2</protein> gene expression showed that overexpression of <protein>calcineurin</protein> ( <protein>CN</protein> ) , a <protein>Ca2+/calmodulin-dependent protein phosphatase</protein> , can stimulate transcription from the <dna>IL-2 promoter</dna> through the <dna>NF-AT-binding site</dna> .


In this study , we obtained evidence that transfection of the cDNAs for CN A ( catalytic ) and CN B ( regulatory ) subunits also augments transcription from the GM-CSF promoter and recovers the transcription inhibited by CsA .

In this study , we obtained evidence that transfection of the <dna>cDNAs</dna> for <protein>CN A ( catalytic ) and CN B ( regulatory ) subunits</protein> also augments transcription from the <dna>GM-CSF promoter</dna> and recovers the transcription inhibited by CsA .


The constitutively active type of the CN A subunit , which lacks the auto-inhibitory and calmodulin-binding domains , acts in synergy with PMA to activate transcription from the GM-CSF promoter .

The constitutively active type of the <protein>CN A subunit</protein> , which lacks the <protein>auto-inhibitory</protein> and <protein>calmodulin-binding domains</protein> , acts in synergy with PMA to activate transcription from the <dna>GM-CSF promoter</dna> .


We also found that the active CN partially replaces calcium ionophore in synergy with PMA to induce expression of endogenous GM-CSF and IL-2 .

We also found that the active <protein>CN</protein> partially replaces calcium ionophore in synergy with PMA to induce expression of endogenous <protein>GM-CSF</protein> and <protein>IL-2</protein> .


By multimerizing the regulatory elements of the GM-CSF promoter , we found that one of the target sites for the CN action is the conserved lymphokine element 0 ( CLE0 ) , located at positions between -54 and -40 .

By multimerizing the <dna>regulatory elements</dna> of the <dna>GM-CSF promoter</dna> , we found that one of the <dna>target sites</dna> for the <protein>CN</protein> action is the <dna>conserved lymphokine element 0</dna> ( <dna>CLE0</dna> ) , located at positions between -54 and -40 .


Mobility shift assays showed that the CLE0 sequence has an AP1-binding site and is associated with an NF-AT-like factor , termed NF-CLE0 gamma .

Mobility shift assays showed that the <dna>CLE0</dna> sequence has an <dna>AP1-binding site</dna> and is associated with an <protein>NF-AT-like factor</protein> , termed <protein>NF-CLE0 gamma</protein> .


NF- CLE0 gamma binding is induced by PMA/A23187 and is inhibited by treatment with CsA .

NF- <dna>CLE0</dna> gamma binding is induced by PMA/A23187 and is inhibited by treatment with CsA .


These results suggest that CN is involved in the coordinated induction of the GM-CSF and IL-2 genes and that the CLE0 sequence of the GM-CSF gene is a functional analogue of the NF-AT-binding site in the IL-2 promoter , which mediates signals downstream of T cell activation .

These results suggest that <protein>CN</protein> is involved in the coordinated induction of the <dna>GM-CSF and IL-2 genes</dna> and that the <dna>CLE0 sequence</dna> of the <dna>GM-CSF gene</dna> is a functional analogue of the <dna>NF-AT-binding site</dna> in the <dna>IL-2 promoter</dna> , which mediates signals downstream of T cell activation .


Alternate immune system targets for TCDD : lymphocyte stem cells and extrathymic T-cell development .

Alternate immune system targets for TCDD : <cell_type>lymphocyte stem cells</cell_type> and <cell_type>extrathymic T-cell</cell_type> development .


We here summarize evidence that thymic atrophy induced by 2 , 3 , 7 , 8-tetrachlorodibenzo-p-dioxin ( TCDD ) can be mediated , at least in part , by damage to extrathymic T-cell precursors in bone marrow and fetal liver .

We here summarize evidence that thymic atrophy induced by 2 , 3 , 7 , 8-tetrachlorodibenzo-p-dioxin ( TCDD ) can be mediated , at least in part , by damage to <cell_type>extrathymic T-cell precursors</cell_type> in bone marrow and fetal liver .