Purified human peripheral T cells or Jurkat T cells were stimulated with Chinese hamster ovary ( CHO ) cells expressing either human CD80 ( CHO-CD80 ) or human CD86 ( CHO-CD86 ) or with anti-CD28 monoclonal antibody ( mAb ) .

<cell_line>Purified human peripheral T cells</cell_line> or <cell_line>Jurkat T cells</cell_line> were stimulated with Chinese hamster ovary ( CHO ) cells expressing either <protein>human CD80</protein> ( <protein>CHO-CD80</protein> ) or <protein>human CD86</protein> ( <protein>CHO-CD86</protein> ) or with <protein>anti-CD28 monoclonal antibody</protein> ( mAb ) .


In the presence of phorbol 12-myristate 13-acetate , both CHO-CD80 and CHO-CD86 , like anti-CD28 mAb , were capable of stimulating cytokine production from both human peripheral T cells and Jurkat T cells .

In the presence of phorbol 12-myristate 13-acetate , both <protein>CHO-CD80</protein> and <protein>CHO-CD86</protein> , like <protein>anti-CD28 mAb</protein> , were capable of stimulating <protein>cytokine</protein> production from both <cell_type>human peripheral T cells</cell_type> and <cell_line>Jurkat T cells</cell_line> .


Both CHO-CD80 and CHO-CD86 , in the presence of anti-CD3 mAb , costimulated NFAT -dependent transcriptional activation .

Both <protein>CHO-CD80</protein> and <protein>CHO-CD86</protein> , in the presence of <protein>anti-CD3 mAb</protein> , costimulated <protein>NFAT</protein> -dependent transcriptional activation .


Several intracellular signaling proteins , such as CBL and VAV , were phosphorylated on tyrosine in response to CD80 , CD86 , and anti-CD28 mAb .

Several intracellular <protein>signaling proteins</protein> , such as <protein>CBL</protein> and <protein>VAV</protein> , were phosphorylated on tyrosine in response to <protein>CD80</protein> , <protein>CD86</protein> , and <protein>anti-CD28 mAb</protein> .


Surprisingly , although stimulation of Jurkat T cells with either CHO-CD80 or anti-CD28 mAb resulted in robust tyrosine phosphorylation of CD28 itself , ligation with CHO-CD86 was unable to induce detectable CD28 tyrosyl phosphorylation over a range of stimulation conditions .

Surprisingly , although stimulation of <cell_line>Jurkat T cells</cell_line> with either <protein>CHO-CD80</protein> or <protein>anti-CD28 mAb</protein> resulted in robust tyrosine phosphorylation of <protein>CD28</protein> itself , ligation with <protein>CHO-CD86</protein> was unable to induce detectable <protein>CD28</protein> tyrosyl phosphorylation over a range of stimulation conditions .


In addition , the association of phosphoinositide 3-kinase with CD28 and enhanced tyrosine phosphorylation of phospholipase Cgamma were seen after anti-CD28 mAb and CHO-CD80 stimulation but to a much lesser extent after CHO-CD86 stimulation .

In addition , the association of <protein>phosphoinositide 3-kinase</protein> with <protein>CD28</protein> and enhanced tyrosine phosphorylation of phospholipase Cgamma were seen after anti-CD28 mAb and CHO-CD80 stimulation but to a much lesser extent after <protein>CHO-CD86</protein> stimulation .


Thus , ligation of CD28 with either CD80 or CD86 leads to shared early signal transduction events such as the tyrosine phosphorylation of CBL and VAV , to NFAT -mediated transcriptional activation , and to the costimulation of interleukin-2 and granulocyte-macrophage colony-stimulating factor production .

Thus , ligation of <protein>CD28</protein> with either <protein>CD80</protein> or <protein>CD86</protein> leads to shared early signal transduction events such as the tyrosine phosphorylation of <protein>CBL</protein> and <protein>VAV</protein> , to <protein>NFAT</protein> -mediated transcriptional activation , and to the costimulation of <protein>interleukin-2</protein> and <protein>granulocyte-macrophage colony-stimulating factor</protein> production .


However , CD80 and CD86 also induce distinct signal transduction pathways including the tyrosine phosphorylation of CD28 and phospholipase Cgamma1 and the SH2 -dependent association of phosphoinositide 3-kinase with CD28 .

However , <protein>CD80</protein> and <protein>CD86</protein> also induce distinct signal transduction pathways including the tyrosine phosphorylation of <protein>CD28</protein> and <protein>phospholipase Cgamma1</protein> and the <protein>SH2</protein> -dependent association of <protein>phosphoinositide 3-kinase</protein> with <protein>CD28</protein> .


These quantitative , if not qualitative , differences between signaling initiated by these two ligands for CD28 may contribute to functional differences ( e.g. Th1 or Th2 differentiation ) in T cell responses .

These quantitative , if not qualitative , differences between signaling initiated by these two ligands for <protein>CD28</protein> may contribute to functional differences ( e.g. Th1 or Th2 differentiation ) in T cell responses .


Binding of HMG-I ( Y ) elicits structural changes in a silencer of the human beta-globin gene .

Binding of <protein>HMG-I ( Y )</protein> elicits structural changes in a silencer of the <dna>human beta-globin gene</dna> .


Proteins involved in repression of the human beta-globin gene may be useful in the treatment of sickle cell anemia , in conjunction with therapy to reactivate fetal globin genes .

Proteins involved in repression of the <dna>human beta-globin gene</dna> may be useful in the treatment of sickle cell anemia , in conjunction with therapy to reactivate <dna>fetal globin genes</dna> .


If there is a reciprocal elevation of gamma-globin expression upon repression , this approach could be useful in additional hemoglobinopathies .

If there is a reciprocal elevation of <protein>gamma-globin</protein> expression upon repression , this approach could be useful in additional hemoglobinopathies .


We previously showed that repression of the beta-globin gene appears to be mediated through two DNA sequences , silencers I and II , and identified a protein termed BP1 which binds to both silencer sequences .

We previously showed that repression of the beta-globin gene appears to be mediated through two <dna>DNA sequences</dna> , <dna>silencers I and II</dna> , and identified a protein termed <protein>BP1</protein> which binds to both silencer sequences .


In this study , we cloned two cDNAs encoding proteins which bind to an oligonucleotide in silencer I containing a BP1 binding site .

In this study , we cloned two <dna>cDNAs</dna> encoding proteins which bind to an oligonucleotide in <dna>silencer I</dna> containing a <protein>BP1</protein> binding site .


These cDNAs correspond to HMG-I and HMG-Y , isoforms regarded as architectural proteins .

These <dna>cDNAs</dna> correspond to <protein>HMG-I</protein> and <protein>HMG-Y</protein> , isoforms regarded as <protein>architectural proteins</protein> .


We demonstrate that binding of HMG-I ( Y ) to this oligonucleotide causes bending/flexure of the DNA .

We demonstrate that binding of <protein>HMG-I ( Y )</protein> to this oligonucleotide causes bending/flexure of the DNA .


HMG-I ( Y ) also binds to a second oligonucleotide containing a BP1 binding site located in a negative control region upstream of the delta-globin gene , suggesting a role for HMG-I ( Y ) in repression of adult globin genes .

<protein>HMG-I ( Y )</protein> also binds to a second oligonucleotide containing a <dna>BP1 binding site</dna> located in a <dna>negative control region</dna> upstream of the <dna>delta-globin gene</dna> , suggesting a role for <protein>HMG-I ( Y )</protein> in repression of adult globin genes .


Expression studies revealed that HMG-I ( Y ) is ubiquitously expressed in human tissues that do not express beta-globin , being present in 48 of 50 tissues and six hematopoietic cell lines examined .

Expression studies revealed that <protein>HMG-I ( Y )</protein> is ubiquitously expressed in human tissues that do not express <protein>beta-globin</protein> , being present in 48 of 50 tissues and six <cell_line>hematopoietic cell lines</cell_line> examined .


Furthermore , HMG-I ( Y ) expression is down-regulated during differentiation of primary erythroid cells .

Furthermore , <protein>HMG-I ( Y )</protein> expression is down-regulated during differentiation of <cell_type>primary erythroid cells</cell_type> .


We present a model in which HMG-I ( Y ) alters DNA conformation to allow binding of repressor proteins , and in which the relative amount of HMG-I ( Y ) helps to determine the repressive state of the beta-globin gene .

We present a model in which <protein>HMG-I ( Y )</protein> alters DNA conformation to allow binding of <protein>repressor proteins</protein> , and in which the relative amount of HMG-I ( Y ) helps to determine the repressive state of the <dna>beta-globin gene</dna> .


Interactions between the class II transactivator and CREB binding protein increase transcription of major histocompatibility complex class II genes .

Interactions between the <protein>class II transactivator</protein> and <protein>CREB binding protein</protein> increase transcription of <dna>major histocompatibility complex class II genes</dna> .


Class II major histocompatibility ( class II ) genes are regulated in a B-cell-specific and gamma interferon-inducible fashion .

<dna>Class II major histocompatibility ( class II ) genes</dna> are regulated in a B-cell-specific and gamma interferon-inducible fashion .


The master switch for the expression of these genes is the class II transactivator ( CIITA ) .

The master switch for the expression of these genes is the <protein>class II transactivator</protein> ( <protein>CIITA</protein> ) .


In this report , we demonstrate that one of the functions of CIITA is to recruit the CREB binding protein ( CBP ) to class II promoters .

In this report , we demonstrate that one of the functions of <protein>CIITA</protein> is to recruit the <protein>CREB binding protein</protein> ( <protein>CBP</protein> ) to <dna>class II promoters</dna> .


Not only functional but also specific binding interactions between CIITA and CBP were demonstrated .

Not only functional but also specific binding interactions between <protein>CIITA</protein> and <protein>CBP</protein> were demonstrated .


Moreover , a dominant negative form of CBP decreased the activity of class II promoters and levels of class II determinants on the surface of cells .

Moreover , a dominant negative form of <protein>CBP</protein> decreased the activity of <dna>class II promoters</dna> and levels of <protein>class II determinants</protein> on the surface of cells .


Finally , the inhibition of class II gene expression by the glucocorticoid hormone could be attributed to the squelching of CBP by the glucocorticoid receptor .

Finally , the inhibition of <dna>class II gene</dna> expression by the glucocorticoid hormone could be attributed to the squelching of <protein>CBP</protein> by the <protein>glucocorticoid receptor</protein> .


We conclude that CBP , a histone acetyltransferase , plays an important role in the transcription of class II genes .

We conclude that <protein>CBP</protein> , a <protein>histone acetyltransferase</protein> , plays an important role in the transcription of <dna>class II genes</dna> .


Role of cyclic AMP response element-binding protein in cyclic AMP inhibition of NF-kappaB -mediated transcription .

Role of <protein>cyclic AMP response element-binding protein</protein> in cyclic AMP inhibition of <protein>NF-kappaB</protein> -mediated transcription .


The NF-kappaB family of transcription factors regulates the inducible expression of a variety of genes .

The <protein>NF-kappaB</protein> family of transcription factors regulates the inducible expression of a variety of genes .


Recently , we showed that elevation of intracellular cyclic AMP inhibits NF-kappaB -mediated transcription in human monocytes and endothelial cells without preventing nuclear translocation of NF-kappaB complexes .

Recently , we showed that elevation of intracellular cyclic AMP inhibits <protein>NF-kappaB</protein> -mediated transcription in <cell_type>human monocytes</cell_type> and <cell_type>endothelial cells</cell_type> without preventing nuclear translocation of <protein>NF-kappaB complexes</protein> .


The present study examined the molecular mechanism of this inhibition .


We hypothesized that activation of the protein kinase A signaling pathway may inhibit NF-kappaB -mediated transcription by phosphorylating proteins , such as cAMP response element-binding protein ( CREB ) , which compete for limiting amounts of the coactivator CBP .

We hypothesized that activation of the <protein>protein kinase A</protein> signaling pathway may inhibit <protein>NF-kappaB</protein> -mediated transcription by phosphorylating proteins , such as <protein>cAMP response element-binding protein</protein> ( <protein>CREB</protein> ) , which compete for limiting amounts of the coactivator <protein>CBP</protein> .


Here , we show that the amino-terminal region ( amino acids 1-450 ) of CBP specifically interacts with the carboxyl-terminal region ( amino acids 286-551 ) of NF-kappaB p65 ( RelA ) both in vitro and in vivo .

Here , we show that the <protein>amino-terminal region</protein> ( <protein>amino acids 1-450</protein> ) of <protein>CBP</protein> specifically interacts with the <protein>carboxyl-terminal region</protein> ( <protein>amino acids 286-551</protein> ) of <protein>NF-kappaB</protein> <protein>p65</protein> ( <protein>RelA</protein> ) both in vitro and in vivo .


Functional studies using human endothelial cells demonstrated that overexpression of CBP rescued cAMP inhibition of NF-kappaB -mediated transcription and transcription mediated by a chimeric protein , GAL4-p65 ( 286-551 ) , which contained the GAL4 DNA binding domain fused to the carboxyl-terminal region of p65 ( amino acids 286-551 ) .

Functional studies using human <cell_type>endothelial cells</cell_type> demonstrated that overexpression of <protein>CBP</protein> rescued cAMP inhibition of <protein>NF-kappaB</protein> -mediated transcription and transcription mediated by a <protein>chimeric protein</protein> , <protein>GAL4-p65 ( 286-551 )</protein> , which contained the <protein>GAL4 DNA binding domain</protein> fused to the <protein>carboxyl-terminal region</protein> of <protein>p65</protein> ( <protein>amino acids 286-551</protein> ) .


In contrast , overexpression of CREB inhibited GAL4 -p65 ( 286-551 ) -mediated transcription .

In contrast , overexpression of <protein>CREB</protein> inhibited <protein>GAL4</protein> -p65 ( 286-551 ) -mediated transcription .


These results suggest that activation of the protein kinase A pathway inhibits NF-kappaB transcription by phosphorylating CREB , which competes with p65 for limiting amounts of CBP .

These results suggest that activation of the <protein>protein kinase A</protein> pathway inhibits <protein>NF-kappaB</protein> transcription by phosphorylating <protein>CREB</protein> , which competes with <protein>p65</protein> for limiting amounts of <protein>CBP</protein> .


Glucocorticoid receptors in cord blood lymphocytes of healthy neonates and of preterms suffering from respiratory distress syndrome .

<protein>Glucocorticoid receptors</protein> in <cell_type>cord blood lymphocytes</cell_type> of healthy neonates and of preterms suffering from respiratory distress syndrome .


We measured the number of glucocorticoid receptors ( GR ) in cord blood lymphocytes and the binding affinity ( Kd ) in 15 term and in 20 preterm babies .

We measured the number of <protein>glucocorticoid receptors</protein> ( <protein>GR</protein> ) in <cell_type>cord blood lymphocytes</cell_type> and the binding affinity ( Kd ) in 15 term and in 20 preterm babies .


Thirteen preterms of the latter group received prenatal steroid treatment .


Seven preterms developed neonatal respiratory distress syndrome ( NRDS ) .


The number of GR and the Kd were similar in the term and preterm ( with and without NRDS ) babies .

The number of <protein>GR</protein> and the Kd were similar in the term and preterm ( with and without NRDS ) babies .


The maximum ( 3H ) -thymidine incorporation into DNA of cord blood lymphocytes from all preterms , with or without NRDS was suppressed when compared to that from term babies or adults .

The maximum ( 3H ) -thymidine incorporation into DNA of <cell_type>cord blood lymphocytes</cell_type> from all preterms , with or without NRDS was suppressed when compared to that from term babies or adults .


This could partly be explained by the antenatal steroid treatment .


Sensitivity ( ID50 ) of the lymphocytes for the inhibitory effect of dexamethasone was the same in all groups .

Sensitivity ( ID50 ) of the <cell_type>lymphocytes</cell_type> for the inhibitory effect of dexamethasone was the same in all groups .


In this study on the number and function of GR in lymphocytes , we were unable to find a relation between the functionality of the GR and the development of NRDS .

In this study on the number and function of <protein>GR</protein> in <cell_type>lymphocytes</cell_type> , we were unable to find a relation between the functionality of the <protein>GR</protein> and the development of NRDS .


Inefficient termination of antigen responses in NF-ATp -deficient mice .

Inefficient termination of antigen responses in <protein>NF-ATp</protein> -deficient mice .


In order to elucidate the role of NF-ATp , one of the most prominent members of family of NF-AT transcription factors in peripheral T lymphocytes , in T cell activation and differentiation we created NF-ATp -deficient mice by gene targeting .

In order to elucidate the role of <protein>NF-ATp</protein> , one of the most prominent members of family of <protein>NF-AT transcription factors</protein> in <cell_type>peripheral T lymphocytes</cell_type> , in T cell activation and differentiation we created <protein>NF-ATp</protein> -deficient mice by gene targeting .


Such NF-ATp -/- mice are born and appear to develop a normal immune system .

Such <protein>NF-ATp</protein> -/- mice are born and appear to develop a normal immune system .


Apart from clear-cut defects in the synthesis of mRNAs for Th2-type lymphokines , such as IL-4 , IL-5 , IL-10 and IL-13 , in primary and secondary stimulations of spleen cells in vitro , of a distinct impaired deletion of V beta 11+/CD4+ T lymphocytes from these mice was detected after superantigen injection .

Apart from clear-cut defects in the synthesis of <rna>mRNAs</rna> for <protein>Th2-type lymphokines</protein> , such as <protein>IL-4</protein> , <protein>IL-5</protein> , <protein>IL-10</protein> and <protein>IL-13</protein> , in primary and secondary stimulations of <cell_type>spleen cells</cell_type> in vitro , of a distinct impaired deletion of <cell_line>V beta 11+/CD4+ T lymphocytes</cell_line> from these mice was detected after superantigen injection .


Moreover , NF-ATp -/- mice older than 6 weeks show an 2-5 fold increase in number of lymphocytes .

Moreover , <protein>NF-ATp</protein> -/- mice older than 6 weeks show an 2-5 fold increase in number of <cell_type>lymphocytes</cell_type> .


This is correlated with an increased expression of activation markers CD44 and CD69 and decreased expression of CD62 .

This is correlated with an increased expression of <protein>activation markers</protein> <protein>CD44</protein> and <protein>CD69</protein> and decreased expression of <protein>CD62</protein> .


AML and Ets proteins regulate the I alpha1 germ-line promoter .

<protein>AML and Ets proteins</protein> regulate the <dna>I alpha1 germ-line promoter</dna> .


The immunoglobulin heavy chain ( IgH ) class switch recombination of B lymphocytes preferentially targets unrearranged IgH genes that have already been rendered transcriptionally active .

The <protein>immunoglobulin heavy chain</protein> ( <protein>IgH</protein> ) class switch recombination of <cell_type>B lymphocytes</cell_type> preferentially targets unrearranged <dna>IgH genes</dna> that have already been rendered transcriptionally active .


Transcription of the germ-line IgH genes is controlled by intervening ( I ) regions upstream of their switch regions .

Transcription of the <dna>germ-line IgH genes</dna> is controlled by <dna>intervening ( I ) regions</dna> upstream of their switch regions .


The I alpha1 promoter activates transcription of the human germ-line C alpha1 gene for IgA1 and mediates the transforming growth factor ( TGF ) -beta1 responsiveness of this locus .

The I alpha1 promoter activates transcription of the <dna>human germ-line C alpha1 gene</dna> for <protein>IgA1</protein> and mediates the <protein>transforming growth factor ( TGF ) -beta1</protein> responsiveness of this locus .


Here we show that the I alpha1 promoter contains several binding sites for the AML/PEBP2/CBF family of transcription factors and that AML and Ets proteins are major regulators of the basal and TGF-beta-inducible promoter activity .

Here we show that the I alpha1 promoter contains several binding sites for the <protein>AML/PEBP2/CBF family of transcription factors</protein> and that <protein>AML and Ets proteins</protein> are major regulators of the basal and TGF-beta-inducible promoter activity .


Our data constitute a starting point for studies to elucidate the molecular mechanism by which TGF-beta regulates IgA production .

Our data constitute a starting point for studies to elucidate the molecular mechanism by which <protein>TGF-beta</protein> regulates <protein>IgA</protein> production .


Daidzein and genistein glucuronides in vitro are weakly estrogenic and activate human natural killer cells at nutritionally relevant concentrations .

Daidzein and genistein glucuronides in vitro are weakly estrogenic and activate <cell_type>human natural killer cells</cell_type> at nutritionally relevant concentrations .


Daidzein and genistein glucuronides ( DG and GG ) , major isoflavone metabolites , may be partly responsible for biological effects of isoflavones , such as estrogen receptor binding and natural killer cell ( NK ) activation or inhibition .

Daidzein and genistein glucuronides ( DG and GG ) , major isoflavone metabolites , may be partly responsible for biological effects of isoflavones , such as estrogen receptor binding and <cell_type>natural killer cell</cell_type> ( NK ) activation or inhibition .


DG and GG were synthesized using 3-methylcholanthrene-induced rat liver microsomes .


The Km and Vmax for daidzein and genistein were 9.0 and 7.7 micromol/L , and 0.7 and 1.6 micromol/ ( mg protein. min ) , respectively .


The absence of ultraviolet absorbance maxima shifts in the presence of sodium acetate confirmed that the synthesized products were 7-O-glucuronides .


DG and GG were further purified by a Sephadex LH-20 column .


DG and GG competed with the binding of 17beta- ( 3H ) estradiol to estrogen receptors of B6D2F1 mouse uterine cytosol .

DG and GG competed with the binding of 17beta- ( 3H ) estradiol to <protein>estrogen receptors</protein> of B6D2F1 mouse uterine cytosol .


The concentrations required for 50 % displacement of 17beta- ( 3H ) estradiol ( CB50 ) were : 17beta-estradiol , 1.34 nmol/L ; diethylstilbestrol , 1.46 nmol/L ; daidzein , 1.6 micromol/L ; DG , 14.7 micromol/L ; genistein , 0.154 micromol/L ; GG , 7.27 micromol/L .


In human peripheral blood NK cells , genistein at < 0.5 micromol/L and DG and GG at 0.1-10 micromol/L enhanced NK cell-mediated K562 cancer cell killing significantly ( P < 0.05 ) .

In <cell_type>human peripheral blood NK cells</cell_type> , genistein at < 0.5 micromol/L and DG and GG at 0.1-10 micromol/L enhanced NK cell-mediated <cell_line>K562 cancer cell</cell_line> killing significantly ( P < 0.05 ) .


At > 0.5 micromol/L , genistein inhibited NK cytotoxicity significantly ( P < 0.05 ) .


The glucuronides only inhibited NK cytotoxicity at 50 micromol/L .


Isoflavones , and especially the isoflavone glucuronides , enhanced activation of NK cells by interleukin-2 ( IL-2 ) , additively .

Isoflavones , and especially the isoflavone glucuronides , enhanced activation of <cell_type>NK cells</cell_type> by <protein>interleukin-2</protein> ( <protein>IL-2</protein> ) , additively .


At physiological concentrations , DG and GG were weakly estrogenic , and they activated human NK cells in nutritionally relevant concentrations in vitro , probably at a site different from IL-2 action .

At physiological concentrations , DG and GG were weakly estrogenic , and they activated <cell_type>human NK cells</cell_type> in nutritionally relevant concentrations in vitro , probably at a site different from <protein>IL-2</protein> action .


Cross-linking of CD44 on rheumatoid synovial cells up-regulates VCAM-1 .

Cross-linking of <protein>CD44</protein> on <cell_type>rheumatoid synovial cells</cell_type> up-regulates <protein>VCAM-1</protein> .


CD44 is a ubiquitous molecule also known as hyaluronic acid or homing receptor .


However , the cellular functions and its role in inflammation , for example , rheumatoid synovitis , are currently unknown .


In this study , we propose a novel function for CD44 .


Using synovial cells from rheumatoid arthritis ( RA ) patients , we demonstrated that CD44 cross-linking and binding to hyaluronan augmented VCAM-1 expression and subsequently VCAM-1 -mediated cell adhesion .

Using synovial cells from rheumatoid arthritis ( RA ) patients , we demonstrated that CD44 cross-linking and binding to hyaluronan augmented <protein>VCAM-1</protein> expression and subsequently <protein>VCAM-1</protein> -mediated cell adhesion .


Briefly , we found that 1 ) rheumatoid synovial cells highly expressed CD44 ; 2 ) cross-linking of CD44 markedly but transiently augmented VCAM-1 expression and its mRNA transcription much more than did IL-1beta and TNF-alpha ; 3 ) hyaluronan , especially when fragmented , also up-regulated VCAM-1 ; 4 ) CD44 activated the transcription factor AP-1 ; and 5 ) the integrin -dependent adhesive function of RA synovial cells to T cells was also amplified by CD44 cross-linking .

Briefly , we found that 1 ) <cell_type>rheumatoid synovial cells</cell_type> highly expressed <protein>CD44</protein> ; 2 ) cross-linking of <protein>CD44</protein> markedly but transiently augmented <protein>VCAM-1</protein> expression and its mRNA transcription much more than did <protein>IL-1beta</protein> and TNF-alpha ; 3 ) hyaluronan , especially when fragmented , also up-regulated <protein>VCAM-1</protein> ; 4 ) <protein>CD44</protein> activated the transcription factor AP-1 ; and 5 ) the <protein>integrin</protein> -dependent adhesive function of <cell_type>RA synovial cells</cell_type> to <cell_type>T cells</cell_type> was also amplified by <protein>CD44</protein> cross-linking .


These results indicate that the adhesion of RA synovial cells to matrices such as hyaluronic acid through CD44 could up-regulate VCAM-1 expression and VCAM-1 -mediated adhesion to T cells , which might in turn cause activation of T cells and synovial cells in RA synovitis .

These results indicate that the adhesion of <cell_type>RA synovial cells</cell_type> to matrices such as hyaluronic acid through <protein>CD44</protein> could up-regulate <protein>VCAM-1</protein> expression and <protein>VCAM-1</protein> -mediated adhesion to <cell_type>T cells</cell_type> , which might in turn cause activation of <cell_type>T cells</cell_type> and <cell_type>synovial cells</cell_type> in RA synovitis .


We therefore propose that such cross-talking among distinct adhesion molecules may be involved in the pathogenesis of inflammation , including RA synovitis .


NF-kappaB activation is a critical regulator of human granulocyte apoptosis in vitro .

<protein>NF-kappaB</protein> activation is a critical regulator of human granulocyte apoptosis in vitro .


During beneficial inflammation , potentially tissue-damaging granulocytes undergo apoptosis before being cleared by phagocytes in a non-phlogistic manner .

During beneficial inflammation , potentially <cell_type>tissue-damaging granulocytes</cell_type> undergo apoptosis before being cleared by phagocytes in a non-phlogistic manner .


Here we show that the rate of constitutive apoptosis in human neutrophils and eosinophils is greatly accelerated in both a rapid and concentration-dependent manner by the fungal metabolite gliotoxin , but not by its inactive analog methylthiogliotoxin .

Here we show that the rate of constitutive apoptosis in <cell_type>human neutrophils</cell_type> and <cell_type>eosinophils</cell_type> is greatly accelerated in both a rapid and concentration-dependent manner by the fungal metabolite gliotoxin , but not by its inactive analog methylthiogliotoxin .


This induction of apoptosis was abolished by the caspase inhibitor zVAD-fmk , correlated with the inhibition of nuclear factor-kappa B ( NF-kappaB ) , and was mimicked by a cell permeable inhibitory peptide of NF-kappaB , SN-50 ; other NF-kappaB inhibitors , curcumin and pyrrolidine dithiocarbamate ; and the proteasome inhibitor , MG-132 .

This induction of apoptosis was abolished by the caspase inhibitor zVAD-fmk , correlated with the inhibition of <protein>nuclear factor-kappa B</protein> ( <protein>NF-kappaB</protein> ) , and was mimicked by a cell permeable inhibitory peptide of <protein>NF-kappaB</protein> , SN-50 ; other <protein>NF-kappaB</protein> inhibitors , curcumin and pyrrolidine dithiocarbamate ; and the proteasome inhibitor , MG-132 .


Gliotoxin also augmented dramatically the early ( 2-6 h ) pro-apoptotic effects of tumor necrosis factor-alpha ( TNF-alpha ) in neutrophils and unmasked the ability of TNF-alpha to induce eosinophil apoptosis .

Gliotoxin also augmented dramatically the early ( 2-6 h ) pro-apoptotic effects of <protein>tumor necrosis factor-alpha</protein> ( <protein>TNF-alpha</protein> ) in <cell_type>neutrophils</cell_type> and unmasked the ability of <protein>TNF-alpha</protein> to induce <cell_type>eosinophil</cell_type> apoptosis .


In neutrophils , TNF-alpha caused a gliotoxin-inhibitable activation of an inducible form of NF-kappaB , a response that may underlie the ability of TNF-alpha to delay apoptosis at later times ( 12-24 h ) and limit its early killing effect .

In <cell_type>neutrophils</cell_type> , <protein>TNF-alpha</protein> caused a gliotoxin-inhibitable activation of an inducible form of <protein>NF-kappaB</protein> , a response that may underlie the ability of <protein>TNF-alpha</protein> to delay apoptosis at later times ( 12-24 h ) and limit its early killing effect .


Furthermore , cycloheximide displayed a similar capacity to enhance TNF-alpha induced neutrophil apoptosis even at time points when cycloheximide alone had no pro-apoptotic effect , suggesting that NF-kappaB may regulate the production of protein ( s ) which protect neutrophils from the cytotoxic effects of TNF-alpha .

Furthermore , cycloheximide displayed a similar capacity to enhance <protein>TNF-alpha</protein> induced neutrophil apoptosis even at time points when cycloheximide alone had no pro-apoptotic effect , suggesting that <protein>NF-kappaB</protein> may regulate the production of protein ( s ) which protect <cell_type>neutrophils</cell_type> from the cytotoxic effects of <protein>TNF-alpha</protein> .


These data shed light on the biochemical and molecular mechanisms regulating human granulocyte apoptosis and , in particular , indicate that the transcription factor NF-kappaB plays a crucial role in regulating the physiological cell death pathway in granulocytes .

These data shed light on the biochemical and molecular mechanisms regulating human granulocyte apoptosis and , in particular , indicate that the <protein>transcription factor</protein> <protein>NF-kappaB</protein> plays a crucial role in regulating the physiological cell death pathway in <cell_type>granulocytes</cell_type> .


Role of Egr-2 in up-regulation of Fas ligand in normal T cells and aberrant double-negative lpr and gld T cells .

Role of <protein>Egr-2</protein> in up-regulation of <protein>Fas ligand</protein> in normal <cell_type>T cells</cell_type> and <cell_type>aberrant double-negative lpr and gld T cells</cell_type> .


We previously identified a Fas ligand regulatory element ( FLRE ) in the Fas ligand ( fasL ) promoter that binds Egr family proteins and demonstrated that Egr-3 ( PILOT ) but not Egr-1 ( NGFI-A , Krox-24 , Tis-8 , and Zif-268 ) induces transcription of fasL .

We previously identified a <dna>Fas ligand regulatory element</dna> ( <dna>FLRE</dna> ) in the <dna>Fas ligand ( fasL ) promoter</dna> that binds <protein>Egr family proteins</protein> and demonstrated that <protein>Egr-3</protein> ( <protein>PILOT</protein> ) but not <protein>Egr-1</protein> ( <protein>NGFI-A</protein> , <protein>Krox-24</protein> , <protein>Tis-8</protein> , and <protein>Zif-268</protein> ) induces transcription of <protein>fasL</protein> .


The aberrant CD4 ( - ) CD8 ( - ) T cells from lpr/lpr and gld/gld mice , which have mutations in the genes encoding Fas and FasL , respectively , have an activated phenotype and constitutively express high levels of fasL mRNA , prompting us to ask what role if any the FLRE and Egr family proteins have in this aberrant expression of fasL .

The <cell_type>aberrant CD4 ( - ) CD8 ( - ) T cells</cell_type> from lpr/lpr and gld/gld mice , which have mutations in the genes encoding <protein>Fas</protein> and <protein>FasL</protein> , respectively , have an activated phenotype and constitutively express high levels of <rna>fasL mRNA</rna> , prompting us to ask what role if any the <dna>FLRE</dna> and <protein>Egr family proteins</protein> have in this aberrant expression of <protein>fasL</protein> .


Unstimulated MRL-lpr/lpr and C3H-gld/gld CD4 ( - ) CD8 ( - ) T cells constitutively contained high levels of two proteins that bound to the FLRE .

Unstimulated <cell_line>MRL-lpr/lpr and C3H-gld/gld CD4 ( - ) CD8 ( - ) T cells</cell_line> constitutively contained high levels of two proteins that bound to the <dna>FLRE</dna> .


Supershift analysis revealed these proteins to be Egr-1 and Egr-2 ( Krox-20 ) ; Egr-3 was not detected .

Supershift analysis revealed these proteins to be <protein>Egr-1</protein> and <protein>Egr-2</protein> ( Krox-20 ) ; <protein>Egr-3</protein> was not detected .


Activation of normal lymph node cells resulted in increased expression of Egr-1 , -2 , and -3 .

Activation of <cell_type>normal lymph node cells</cell_type> resulted in increased expression of <protein>Egr-1 , -2 , and -3</protein> .


As with egr-3 , expression of egr-2 was blocked by cyclosporin A .

As with <dna>egr-3</dna> , expression of <dna>egr-2</dna> was blocked by cyclosporin A .


Although overexpressed Egr-1 was ineffective , overexpressed Egr-2 was as potent as Egr-3 in inducing fasL promoter-dependent reporter constructs in T cell hybridomas and HeLa cells , and both up-regulated endogenous fasL mRNA in HeLa cells .

Although overexpressed <protein>Egr-1</protein> was ineffective , overexpressed <protein>Egr-2</protein> was as potent as <protein>Egr-3</protein> in inducing <dna>fasL promoter-dependent reporter constructs</dna> in T cell hybridomas and HeLa cells , and both up-regulated endogenous <rna>fasL mRNA</rna> in <cell_line>HeLa cells</cell_line> .


FasL-dependent reporter constructs in MRL-lpr/lpr and C3H-gld/gld CD4 ( - ) CD8 ( - ) T cells were constitutively active , and this activity was largely prevented by mutation of the critical Egr family binding element .

<dna>FasL-dependent reporter constructs</dna> in <cell_line>MRL-lpr/lpr and C3H-gld/gld CD4 ( - ) CD8 ( - ) T cells</cell_line> were constitutively active , and this activity was largely prevented by mutation of the critical <dna>Egr family binding element</dna> .


Thus , Egr-2 , in addition to Egr-3 , regulates FasL expression in activated normal T cells , and Egr-2 is likely to play a direct role in aberrant fasL up-regulation in lpr/lpr and gld/gld CD4 ( - ) CD8 ( - ) T cells .

Thus , <protein>Egr-2</protein> , in addition to <protein>Egr-3</protein> , regulates <protein>FasL</protein> expression in activated normal <cell_type>T cells</cell_type> , and <protein>Egr-2</protein> is likely to play a direct role in aberrant <protein>fasL</protein> up-regulation in <cell_line>lpr/lpr and gld/gld CD4 ( - ) CD8 ( - ) T cells</cell_line> .


The activity of the CCAAT-box binding factor NF-Y is modulated through the regulated expression of its A subunit during monocyte to macrophage differentiation : regulation of tissue-specific genes through a ubiquitous transcription factor .

The activity of the <protein>CCAAT-box binding factor NF-Y</protein> is modulated through the regulated expression of its <protein>A subunit</protein> during monocyte to macrophage differentiation : regulation of <dna>tissue-specific genes</dna> through a ubiquitous <protein>transcription factor</protein> .


In this study , we analyzed the regulation of NF-Y expression during human monocyte to macrophage maturation .

In this study , we analyzed the regulation of <protein>NF-Y</protein> expression during human monocyte to macrophage maturation .


NF-Y is a ubiquitous and evolutionarily conserved transcription factor that binds specifically to the CCAAT motif present in the 5 ' promoter region of a wide variety of genes .

<protein>NF-Y</protein> is a ubiquitous and evolutionarily conserved <protein>transcription factor</protein> that binds specifically to the <dna>CCAAT motif</dna> present in the <dna>5 ' promoter region</dna> of a wide variety of genes .


We show here that in circulating monocytes , NF-Y binding activity is not detected on the CCAAT motif present in the promoters of genes such as major histocompatibility complex ( MHC ) class II , gp91-phox , mig , and fibronectin , whereas during macrophage differentiation , a progressive increase in NF-Y binding activity is observed on these promoters .

We show here that in <cell_type>circulating monocytes</cell_type> , <protein>NF-Y</protein> binding activity is not detected on the <dna>CCAAT motif</dna> present in the <dna>promoters of genes</dna> such as <dna>major histocompatibility complex ( MHC ) class II</dna> , <dna>gp91-phox</dna> , <dna>mig</dna> , and fibronectin , whereas during macrophage differentiation , a progressive increase in <protein>NF-Y</protein> binding activity is observed on these promoters .


Analysis of NF-Y subunit expression indicates that the absence of NF-Y activity in circulating monocytes is caused by a lack of the A subunit .

Analysis of <protein>NF-Y</protein> subunit expression indicates that the absence of <protein>NF-Y</protein> activity in <cell_type>circulating monocytes</cell_type> is caused by a lack of the <protein>A subunit</protein> .


Furthermore , addition of the recombinant NF-Y A subunit restores NF-Y binding .

Furthermore , addition of the recombinant <protein>NF-Y</protein> <protein>A subunit</protein> restores <protein>NF-Y</protein> binding .


We show that the lack of NF-YA protein is due to posttranscriptional regulation and not to a specific proteolytic activity .

We show that the lack of <protein>NF-YA protein</protein> is due to posttranscriptional regulation and not to a specific proteolytic activity .


In fact , NF-YA mRNA is present at the same level at all days of monocyte cultivation , whereas the protein is absent in freshly isolated monocytes but is progressively synthesized during the maturation process .

In fact , <rna>NF-YA mRNA</rna> is present at the same level at all days of monocyte cultivation , whereas the protein is absent in <cell_type>freshly isolated monocytes</cell_type> but is progressively synthesized during the maturation process .


We thus conclude that the NF-Y A subunit plays a relevant role in activating transcription of genes highly expressed in mature monocytes .

We thus conclude that the <protein>NF-Y</protein> <protein>A subunit</protein> plays a relevant role in activating transcription of genes highly expressed in <cell_type>mature monocytes</cell_type> .


In line with this conclusion , we show that the cut/CDP protein , a transcriptional repressor that inhibits gpc91-phox gene expression by preventing NF-Y binding to the CAAT box , is absent in monocytes .

In line with this conclusion , we show that the <protein>cut/CDP protein</protein> , a transcriptional repressor that inhibits <dna>gpc91-phox gene</dna> expression by preventing <protein>NF-Y</protein> binding to the <dna>CAAT box</dna> , is absent in <cell_type>monocytes</cell_type> .


Interdomain B in ZAP-70 regulates but is not required for ZAP-70 signaling function in lymphocytes .

<protein>Interdomain B</protein> in <protein>ZAP-70</protein> regulates but is not required for <protein>ZAP-70</protein> signaling function in <cell_type>lymphocytes</cell_type> .


The protein tyrosine kinase ZAP-70 plays an important role in T-cell activation and development .

The protein tyrosine kinase <protein>ZAP-70</protein> plays an important role in T-cell activation and development .


After T-cell receptor stimulation , ZAP-70 associates with the receptor and is phosphorylated on many tyrosines , including Y292 , Y315 , and Y319 within interdomain B .

After <protein>T-cell receptor</protein> stimulation , <protein>ZAP-70</protein> associates with the receptor and is phosphorylated on many tyrosines , including Y292 , Y315 , and Y319 within interdomain B .


Previously , we demonstrated that Y292 negatively regulates ZAP-70 function and that Y315 positively regulates ZAP-70 function by interacting with Vav .

Previously , we demonstrated that Y292 negatively regulates <protein>ZAP-70</protein> function and that Y315 positively regulates <protein>ZAP-70</protein> function by interacting with <protein>Vav</protein> .


Recent studies have suggested that Y319 also positively regulate ZAP-70 function .

Recent studies have suggested that Y319 also positively regulate <protein>ZAP-70</protein> function .


Paradoxically , removal of interdomain B ( to create the construct designated Delta ) , containing the Y292 , Y315 , and Y319 sites , did not eliminate the ability of ZAP-70 to induce multiple gene reporters in Syk-deficient DT-40 B cells and ZAP-70/Syk-deficient Jurkat cells .

Paradoxically , removal of <protein>interdomain B</protein> ( to create the construct designated <protein>Delta</protein> ) , containing the Y292 , Y315 , and Y319 sites , did not eliminate the ability of <protein>ZAP-70</protein> to induce <dna>multiple gene reporters</dna> in <cell_line>Syk-deficient DT-40 B cells</cell_line> and <cell_line>ZAP-70/Syk-deficient Jurkat cells</cell_line> .


Here we show that Delta still utilizes the same pathways as wild-type ZAP-70 to mediate NF-AT induction .

Here we show that <protein>Delta</protein> still utilizes the same pathways as wild-type <protein>ZAP-70</protein> to mediate <protein>NF-AT</protein> induction .


This is manifested by the ability of Delta to restore induction of calcium fluxes and mitogen-activated protein kinase activation and by the ability of dominant negative Ras and FK506 to block the induction of NF-AT activity mediated by Delta .

This is manifested by the ability of <protein>Delta</protein> to restore induction of calcium fluxes and <protein>mitogen-activated protein kinase</protein> activation and by the ability of dominant negative <protein>Ras</protein> and <protein>FK506</protein> to block the induction of <protein>NF-AT</protein> activity mediated by <protein>Delta</protein> .


Biochemically we show that the stimulated tyrosine phosphorylation of Vav , Shc , and ZAP-70 itself is diminished , whereas that of Slp-76 is increased in cells reconstituted with Delta .

Biochemically we show that the stimulated tyrosine phosphorylation of <protein>Vav</protein> , <protein>Shc</protein> , and <protein>ZAP-70</protein> itself is diminished , whereas that of <protein>Slp-76</protein> is increased in cells reconstituted with <protein>Delta</protein> .


Deletion of interdomain B did not affect the ability of ZAP-70 to bind to the receptor .

Deletion of <protein>interdomain B</protein> did not affect the ability of <protein>ZAP-70</protein> to bind to the receptor .


The in vitro kinase activity of ZAP-70 lacking interdomain B was markedly reduced , but the kinase activity was still required for the protein 's in vivo activity .

The in vitro kinase activity of <protein>ZAP-70</protein> lacking <protein>interdomain B</protein> was markedly reduced , but the kinase activity was still required for the protein 's in vivo activity .


Based on these data , we concluded that interdomain B regulates but is not required for ZAP-70 signaling function leading to cellular responses .

Based on these data , we concluded that <protein>interdomain B</protein> regulates but is not required for <protein>ZAP-70</protein> signaling function leading to cellular responses .


Differential induction of DNA-binding activities following CD19 cross-linking in human B lineage cells .

Differential induction of <protein>DNA-binding activities</protein> following <protein>CD19</protein> cross-linking in <cell_type>human B lineage cells</cell_type> .


The B cell-specific cell surface molecule CD19 is expressed at all stages of B cell development , including normal plasma cells , and mediates signal transduction via interaction with cytoplasmic effector proteins .

The <protein>B cell-specific cell surface molecule</protein> <protein>CD19</protein> is expressed at all stages of B cell development , including normal plasma cells , and mediates signal transduction via interaction with <protein>cytoplasmic effector proteins</protein> .


Cross-linking CD19 on early human B lineage cells induces the formation of a CD19/Vav/phosphatidylinositol-3 kinase complex , tyrosine phosphorylation of CD19 and Vav , and activation of the Ras pathway .

Cross-linking <protein>CD19</protein> on early <cell_type>human B lineage cells</cell_type> induces the formation of a <protein>CD19/Vav/phosphatidylinositol-3 kinase complex</protein> , tyrosine phosphorylation of <protein>CD19</protein> and <protein>Vav</protein> , and activation of the <protein>Ras</protein> pathway .


To further explore the ramifications of CD19 signaling , the current study examined whether phosphorylation of Elk-1 , activation of activator protein-1 ( AP-1 ) , or activation of nuclear factor-kappaB ( NF-kappaB ) transcription factors occurred following CD19 cross-linking .

To further explore the ramifications of <protein>CD19</protein> signaling , the current study examined whether phosphorylation of <protein>Elk-1</protein> , activation of <protein>activator protein-1</protein> ( <protein>AP-1</protein> ) , or activation of nuclear factor-kappaB ( <protein>NF-kappaB</protein> ) <protein>transcription factors</protein> occurred following <protein>CD19</protein> cross-linking .


The cells used were the BLIN-1 pre-B cell line expressing low levels of cell surface mu heavy chain associated with surrogate light chain and the 1E8 immature B cell line expressing cell surface mu/kappa .

The cells used were the <cell_line>BLIN-1 pre-B cell line</cell_line> expressing low levels of <protein>cell surface mu heavy chain</protein> associated with <protein>surrogate light chain</protein> and the 1E8 immature B cell line expressing <protein>cell surface mu/kappa</protein> .


Lysates from CD19 cross-linked 1E8 cells induced robust phosphorylation of an Elk-1 fusion protein in vitro , whereas no phosphorylation of Elk-1 fusion protein occurred using lysates from CD19 cross-linked BLIN-1 cells .

Lysates from <protein>CD19</protein> cross-linked <cell_line>1E8 cells</cell_line> induced robust phosphorylation of an <protein>Elk-1 fusion protein</protein> in vitro , whereas no phosphorylation of <protein>Elk-1 fusion protein</protein> occurred using lysates from <cell_line>CD19 cross-linked BLIN-1 cells</cell_line> .


An electrophoretic mobility shift assay employing AP-1 and NF-kappaB consensus oligonucleotides was used to demonstrate that AP-1 -binding activity increased , while constitutive NF-kappaB -binding activity was not enhanced , following 2 h of CD19 cross-linking in 1E8 cells . Supershift experiments revealed that JunD and c-Fos proteins mediated anti- CD19 induced AP-1 -binding activity in 1E8 cells .

An electrophoretic mobility shift assay employing <protein>AP-1</protein> and <protein>NF-kappaB</protein> consensus oligonucleotides was used to demonstrate that <protein>AP-1</protein> -binding activity increased , while constitutive <protein>NF-kappaB</protein> -binding activity was not enhanced , following 2 h of <protein>CD19</protein> cross-linking in <cell_line>1E8 cells</cell_line> . Supershift experiments revealed that <protein>JunD</protein> and <protein>c-Fos proteins</protein> mediated anti- <protein>CD19</protein> induced <protein>AP-1</protein> -binding activity in <cell_line>1E8 cells</cell_line> .


In contrast , CD19 cross-linking in BLIN-1 cells resulted in the induction of NF-kappaB , but had no apparent effect on AP-1 -binding activity .

In contrast , <protein>CD19</protein> cross-linking in <cell_line>BLIN-1 cells</cell_line> resulted in the induction of <protein>NF-kappaB</protein> , but had no apparent effect on <protein>AP-1</protein> -binding activity .


These data suggest that CD19 -mediated signal transduction activates different transcription factors at juxtaposed stages of B cell development that may culminate in the activation or suppression of distinct sets of genes .

These data suggest that <protein>CD19</protein> -mediated signal transduction activates different <protein>transcription factors</protein> at juxtaposed stages of B cell development that may culminate in the activation or suppression of distinct sets of <dna>genes</dna> .


Oxidants , transcription factors , and intestinal inflammation .

Oxidants , <protein>transcription factors</protein> , and intestinal inflammation .


It is now well appreciated that chronic gut inflammation is characterized by enhanced production of reactive metabolites of oxygen and nitrogen .


Some of these oxidants are known to modulate the expression of a variety of genes that are involved in the immune and inflammatory responses .


For example , certain oxidants are known to activate the nuclear transcription factor kappa B , which regulates the expression of a variety of different adhesion molecules , cytokines , and enzymes .

For example , certain oxidants are known to activate the <protein>nuclear transcription factor kappa B</protein> , which regulates the expression of a variety of different <protein>adhesion molecules</protein> , <protein>cytokines</protein> , and <protein>enzymes</protein> .


Oxidants are also known to activate another transcription factor , activator protein-1 .

Oxidants are also known to activate another <protein>transcription factor</protein> , <protein>activator protein-1</protein> .


This transcription factor is composed of products from the fos and jun proto-oncogene family and is believed to be important in regulating cell growth and proliferation .

This <protein>transcription factor</protein> is composed of products from the <protein>fos and jun proto-oncogene family</protein> and is believed to be important in regulating cell growth and proliferation .


Finally , oxidants are believed to promote intestinal epithelial cell apoptosis , and the B-cell lymphoma/leukemia-2 gene product is believed to inhibit this phenomenon in an antioxidant-dependent manner .

Finally , oxidants are believed to promote <cell_type>intestinal epithelial cell</cell_type> apoptosis , and the <protein>B-cell lymphoma/leukemia-2 gene product</protein> is believed to inhibit this phenomenon in an antioxidant-dependent manner .


Taken together , these observations suggest that nontoxic concentrations of reactive metabolites of oxygen and nitrogen play an important role in regulating the expression of genes involved in the inflammatory response and in modulating apoptosis .


Identification of target genes of the lymphoid-specific transcription factor Oct2 .

Identification of <dna>target genes</dna> of the <protein>lymphoid-specific transcription factor Oct2</protein> .


The Oct2 transcription factor is expressed predominantly in B lymphocytes and plays an essential role during the terminal phase of B cell differentiation .

The <protein>Oct2</protein> <protein>transcription factor</protein> is expressed predominantly in <cell_type>B lymphocytes</cell_type> and plays an essential role during the terminal phase of B cell differentiation .


The regulatory regions of several genes specifically expressed in B cells contain functional binding sites for Oct2 .

The <dna>regulatory regions</dna> of several genes specifically expressed in <cell_type>B cells</cell_type> contain <dna>functional binding sites</dna> for <protein>Oct2</protein> .


Nevertheless , none of the genes originally thought to be regulated by Oct2 were affected in their expression in Oct2-deficient B cells .

Nevertheless , none of the genes originally thought to be regulated by <protein>Oct2</protein> were affected in their expression in <cell_line>Oct2-deficient B cells</cell_line> .


In an attempt to find such elusive Oct2 target genes and to understand the molecular function of Oct2 in B cell development , we isolated cDNAs for Oct2 target genes .

In an attempt to find such elusive <protein>Oct2</protein> <dna>target genes</dna> and to understand the molecular function of <protein>Oct2</protein> in B cell development , we isolated <dna>cDNAs</dna> for <protein>Oct2</protein> <dna>target genes</dna> .


So far , we have identified five potential targets for Oct2 : the membrane glycoprotein CD36 , the cysteine-rich secreted protein 3 ( CRISP-3 ) , a mouse homolog of the human monocyte/neutrophil elastase inhibitor ( mEI ) and two unknown cDNA sequences Nov1 and Nov2 .

So far , we have identified five potential targets for <protein>Oct2</protein> : the <protein>membrane glycoprotein</protein> <protein>CD36</protein> , the <protein>cysteine-rich secreted protein 3</protein> ( <protein>CRISP-3</protein> ) , a mouse homolog of the <protein>human monocyte/neutrophil elastase inhibitor</protein> ( <protein>mEI</protein> ) and two unknown <dna>cDNA sequences</dna> <dna>Nov1</dna> and <dna>Nov2</dna> .


These target genes show quite distinct expression patterns demonstrating that transcription factors in addition to Oct2 are involved in their regulation .

These <dna>target genes</dna> show quite distinct expression patterns demonstrating that <protein>transcription factors</protein> in addition to <protein>Oct2</protein> are involved in their regulation .


Whereas CD36 and mEI were expressed in all hematopoetic cell lines containing Oct2 , .

Whereas <protein>CD36</protein> and <protein>mEI</protein> were expressed in all <cell_line>hematopoetic cell lines</cell_line> containing <protein>Oct2</protein> , .


CRISP-3 is pre-B cell-specific , Nov1 is plasma B cell-specific and Nov2 is B cell-specifically expressed .

<protein>CRISP-3</protein> is pre-B cell-specific , <dna>Nov1</dna> is plasma B cell-specific and <dna>Nov2</dna> is B cell-specifically expressed .


RFX-B is the gene responsible for the most common cause of the bare lymphocyte syndrome , an MHC class II immunodeficiency [ published erratum appears in Immunity 1999 Mar ; 10 ( 3 ) : 399 ]

<dna>RFX-B</dna> is the gene responsible for the most common cause of the bare lymphocyte syndrome , an <protein>MHC class II</protein> immunodeficiency [ published erratum appears in Immunity 1999 Mar ; 10 ( 3 ) : 399 ]


The bare lymphocyte syndrome ( BLS ) is characterized by the absence of MHC class II transcription and humoral- and cellular-mediated immune responses to foreign antigens .

The bare lymphocyte syndrome ( BLS ) is characterized by the absence of <protein>MHC class II</protein> transcription and humoral- and cellular-mediated immune responses to foreign antigens .


Three of the four BLS genetic complementation groups have defects in the activity of the MHC class II transcription factor RFX .

Three of the four BLS genetic complementation groups have defects in the activity of the <protein>MHC class II transcription factor</protein> <protein>RFX</protein> .


We have purified the RFX complex and sequenced its three subunits .

We have purified the <protein>RFX complex</protein> and sequenced its three subunits .


The sequence of the smallest subunit describes a novel gene , termed RFX-B .

The sequence of the smallest subunit describes a novel gene , termed <dna>RFX-B</dna> .


RFX-B complements the predominant BLS complementation group ( group B ) and was found to be mutant in cell lines from this BLS group .

<dna>RFX-B</dna> complements the predominant <protein>BLS complementation group</protein> ( <protein>group B</protein> ) and was found to be mutant in <cell_line>cell lines</cell_line> from this BLS group .


The protein has no known DNA-binding domain but does contain three ankyrin repeats that are likely to be important in protein-protein interactions .

The protein has no known <protein>DNA-binding domain</protein> but does contain three ankyrin repeats that are likely to be important in protein-protein interactions .


Binding of c-Rel to STAT5 target sequences in HTLV-I-transformed T cells .

Binding of <protein>c-Rel</protein> to <dna>STAT5 target sequences</dna> in <cell_type>HTLV-I-transformed T cells</cell_type> .


The type I human T-cell leukemia virus ( HTLV-I ) induces abnormal growth and subsequent transformation of T cells , which is associated with the development of an acute T-cell malignancy termed adult T-cell leukemia .

The type I human T-cell leukemia virus ( HTLV-I ) induces abnormal growth and subsequent transformation of <cell_type>T cells</cell_type> , which is associated with the development of an acute T-cell malignancy termed adult T-cell leukemia .


A characteristic of HTLV-I-transformed T cells is the constitutive nuclear expression of NF-kappaB/Rel family of transcription factors , which appears to be essential for the growth of these transformed cells .

A characteristic of <cell_type>HTLV-I-transformed T cells</cell_type> is the constitutive nuclear expression of <protein>NF-kappaB/Rel family</protein> of transcription factors , which appears to be essential for the growth of these <cell_type>transformed cells</cell_type> .


Although NF-kappaB/Rel factors are known to induce the expression of T-cell growth factor interleukin ( IL ) -2 , it is unclear how they participate in the IL-2 -independent growth of HTLV-I-transformed cells .

Although <protein>NF-kappaB/Rel factors</protein> are known to induce the expression of <protein>T-cell growth factor interleukin ( IL ) -2</protein> , it is unclear how they participate in the <protein>IL-2</protein> -independent growth of <cell_line>HTLV-I-transformed cells</cell_line> .


In this study , we show that certain NF-kappaB/Rel members , predominantly c-Rel , interact with enhancer sequences for STAT5 , a key transcription factor mediating IL-2 -induced T-cell proliferation .

In this study , we show that certain <protein>NF-kappaB/Rel members</protein> , predominantly <protein>c-Rel</protein> , interact with enhancer sequences for <protein>STAT5</protein> , a key transcription factor mediating <protein>IL-2</protein> -induced T-cell proliferation .


Reporter gene assays reveal that the binding of c-Rel to the STAT5 site present in the Fc gammaR1 gene leads to potent transactivation of this enhancer .

Reporter gene assays reveal that the binding of <protein>c-Rel</protein> to the <protein>STAT5</protein> site present in the <dna>Fc gammaR1 gene</dna> leads to potent transactivation of this enhancer .


Binding of c-Rel to the Fc gammaR1 STAT site also occurs in human peripheral blood T cells immortalized with HTLV-I in vitro and is correlated with enhanced levels of proliferation of these cells .

Binding of <protein>c-Rel</protein> to the <dna>Fc gammaR1 STAT site</dna> also occurs in human <cell_type>peripheral blood T cells</cell_type> immortalized with HTLV-I in vitro and is correlated with enhanced levels of proliferation of these cells .


These results raise the possibility that NF-kappaB/Rel may participate in the growth control of HTLV-I-transformed T cells by regulating genes driven by both kappaB and certain STAT enhancers .

These results raise the possibility that <protein>NF-kappaB/Rel</protein> may participate in the growth control of <cell_line>HTLV-I-transformed T cells</cell_line> by regulating genes driven by both <dna>kappaB</dna> and certain <dna>STAT enhancers</dna> .


Activation-dependent transcriptional regulation of the human Fas promoter requires NF-kappaB p50-p65 recruitment .

Activation-dependent transcriptional regulation of the <dna>human Fas promoter</dna> requires <protein>NF-kappaB p50-p65</protein> recruitment .


Fas ( CD95 ) and Fas ligand ( CD95L ) are an interacting receptor-ligand pair required for immune homeostasis .

<protein>Fas</protein> ( <protein>CD95</protein> ) and <protein>Fas ligand</protein> ( <protein>CD95L</protein> ) are an interacting receptor-ligand pair required for immune homeostasis .


Lymphocyte activation results in the upregulation of Fas expression and the acquisition of sensitivity to FasL -mediated apoptosis .

Lymphocyte activation results in the upregulation of <protein>Fas</protein> expression and the acquisition of sensitivity to <protein>FasL</protein> -mediated apoptosis .


Although Fas upregulation is central to the preservation of immunologic tolerance , little is known about the molecular machinery underlying this process .


To investigate the events involved in activation-induced Fas upregulation , we have examined mRNA accumulation , fas promoter activity , and protein expression in the Jurkat T-cell line treated with phorbol myristate acetate and ionomycin ( P/I ) , pharmacological mimics of T-cell receptor activation .

To investigate the events involved in activation-induced <protein>Fas</protein> upregulation , we have examined mRNA accumulation , fas promoter activity , and protein expression in the <cell_line>Jurkat T-cell line</cell_line> treated with phorbol myristate acetate and ionomycin ( P/I ) , pharmacological mimics of <protein>T-cell receptor</protein> activation .


Although resting Jurkat cells express Fas , Fas mRNA was induced approximately 10-fold in 2 h upon P/I stimulation .

Although <cell_line>resting Jurkat cells</cell_line> express <protein>Fas</protein> , <rna>Fas mRNA</rna> was induced approximately 10-fold in 2 h upon P/I stimulation .


Using sequential deletion mutants of the human fas promoter in transient transfection assays , we identified a 47-bp sequence ( positions -306 to -260 relative to the ATG ) required for activation-driven fas upregulation .

Using <dna>sequential deletion mutants</dna> of the <dna>human fas promoter</dna> in transient transfection assays , we identified a <dna>47-bp sequence</dna> ( <dna>positions -306 to -260</dna> relative to the ATG ) required for activation-driven fas upregulation .


Sequence analysis revealed the presence of a previously unrecognized composite binding site for both the Sp1 and NF-kappaB transcription factors at positions -295 to -286 .

Sequence analysis revealed the presence of a previously <dna>unrecognized composite binding site</dna> for both the <protein>Sp1</protein> and <protein>NF-kappaB</protein> <protein>transcription factors</protein> at <dna>positions -295 to -286</dna> .


Electrophoretic mobility shift assay ( EMSA ) and supershift analyses of this region documented constitutive binding of Sp1 in unactivated nuclear extracts and inducible binding of p50-p65 NF-kappaB heterodimers after P/I activation .

Electrophoretic mobility shift assay ( EMSA ) and supershift analyses of this region documented constitutive binding of <protein>Sp1</protein> in unactivated nuclear extracts and inducible binding of <protein>p50-p65 NF-kappaB heterodimers</protein> after P/I activation .


Sp1 and NF-kappaB transcription factor binding was shown to be mutually exclusive by EMSA displacement studies with purified recombinant Sp1 and recombinant p50 .

<protein>Sp1</protein> and <protein>NF-kappaB</protein> transcription factor binding was shown to be mutually exclusive by EMSA displacement studies with purified recombinant <protein>Sp1</protein> and recombinant <protein>p50</protein> .


The functional contribution of the kappaB-Sp1 composite site in P/I-inducible fas promoter activation was verified by using kappaB- Sp1 concatamers ( -295 to -286 ) in a thymidine kinase promoter-driven reporter construct and native promoter constructs in Jurkat cells overexpressing IkappaB-alpha .

The functional contribution of the <dna>kappaB-Sp1 composite site</dna> in <dna>P/I-inducible fas promoter</dna> activation was verified by using kappaB- <protein>Sp1</protein> concatamers ( <dna>-295 to -286</dna> ) in a <dna>thymidine kinase promoter-driven reporter construct</dna> and <dna>native promoter constructs</dna> in <cell_line>Jurkat cells</cell_line> overexpressing <protein>IkappaB-alpha</protein> .


Site-directed mutagenesis of the critical guanine nucleotides in the kappaB-Sp1 element documented the essential role of this site in activation-dependent fas promoter induction .

Site-directed mutagenesis of the critical guanine nucleotides in the <dna>kappaB-Sp1 element</dna> documented the essential role of this site in activation-dependent fas promoter induction .


Evidence for repression of IL-2 gene activation in anergic T cells .

Evidence for repression of <dna>IL-2 gene</dna> activation in <cell_type>anergic T cells</cell_type> .


The induction of clonal anergy in a T cell inhibits IL-2 secretion because of the development of a proximal signal transduction defect .

The induction of clonal anergy in a T cell inhibits <protein>IL-2</protein> secretion because of the development of a proximal signal transduction defect .


Fusion of anergic murine T cells to human Jurkat T leukemia cells and formation of heterokaryons failed to result in a complementation of this signaling defect and restoration of murine IL-2 mRNA inducibility .

Fusion of <cell_type>anergic murine T cells</cell_type> to <cell_type>human Jurkat T leukemia cells</cell_type> and formation of <cell_line>heterokaryons</cell_line> failed to result in a complementation of this signaling defect and restoration of <rna>murine IL-2 mRNA</rna> inducibility .


Instead , signal transduction to the human IL-2 gene became disrupted .

Instead , signal transduction to the human <dna>IL-2 gene</dna> became disrupted .


Heterokaryons formed by the fusion of anergic murine T cells to normal murine T cells also failed to accumulate intracellular IL-2 protein in response to stimulation either with the combination of CD3 and CD28 mAbs or with ionomycin plus a protein kinase C -activating phorbol ester .

<cell_line>Heterokaryons</cell_line> formed by the fusion of <cell_type>anergic murine T cells</cell_type> to <cell_type>normal murine T cells</cell_type> also failed to accumulate <protein>intracellular IL-2 protein</protein> in response to stimulation either with the combination of <protein>CD3</protein> and <protein>CD28 mAbs</protein> or with ionomycin plus a <protein>protein kinase C</protein> -activating phorbol ester .


The results argue against a loss-of-function signaling defect as the sole basis for clonal anergy induction and document the presence of a dominant-acting repressor molecule that inhibits signal transduction to the IL-2 gene within viable anergic T cells .

The results argue against a loss-of-function signaling defect as the sole basis for clonal anergy induction and document the presence of a <protein>dominant-acting repressor molecule</protein> that inhibits signal transduction to the <dna>IL-2 gene</dna> within viable <cell_type>anergic T cells</cell_type> .


Comparison of HTLV-I basal transcription and expression of CREB/ATF-1/CREM family members in peripheral blood mononuclear cells and Jurkat T cells .

Comparison of HTLV-I basal transcription and expression of <protein>CREB/ATF-1/CREM family members</protein> in <cell_type>peripheral blood mononuclear cells</cell_type> and <cell_line>Jurkat T cells</cell_line> .


HTLV-I is the etiologic agent of adult T-cell leukemia/lymphoma and is associated with tropical spastic paraparesis/HTLV-I-associated myelopathy .


Following integration into the host cell genome , HTLV-I replication is regulated by both host and viral mechanisms that control transcription .


Low levels of viral transcription ( basal transcription ) occur before expression of the virally encoded Tax protein ( Tax -mediated transcription ) .

Low levels of viral transcription ( basal transcription ) occur before expression of the <protein>virally encoded Tax protein</protein> ( <protein>Tax</protein> -mediated transcription ) .


Members of the cyclic adenosine monophosphate ( cAMP ) response element binding ( CREB ) /activating transcription factor 1 ( ATF-1 ) family of transcription factors bind three 21-bp repeats ( Tax-responsive element-1 , or TRE-1 ) within the viral promoter and are important for basal and Tax-mediated transcription .

Members of the <protein>cyclic adenosine monophosphate ( cAMP ) response element binding ( CREB ) /activating transcription factor 1 ( ATF-1 ) family</protein> of <protein>transcription factors</protein> bind three <dna>21-bp repeats</dna> ( <dna>Tax-responsive element-1</dna> , or <dna>TRE-1</dna> ) within the <dna>viral promoter</dna> and are important for basal and Tax-mediated transcription .


Using mitogen stimulated and quiescent peripheral blood mononuclear cells ( PBMC ) and Jurkat cells , we compared differences in basal transcription and amounts and binding of transcription factors with TRE-1 .

Using mitogen stimulated and quiescent <cell_type>peripheral blood mononuclear cells</cell_type> ( <cell_type>PBMC</cell_type> ) and <cell_line>Jurkat cells</cell_line> , we compared differences in basal transcription and amounts and binding of <protein>transcription factors</protein> with <dna>TRE-1</dna> .


We demonstrate that amounts of transcriptionally active phosphorylated CREB protein ( P-CREB ) differ between activated PBMC and Jurkat cells .

We demonstrate that amounts of transcriptionally active <protein>phosphorylated CREB protein</protein> ( <protein>P-CREB</protein> ) differ between activated <cell_type>PBMC</cell_type> and <cell_line>Jurkat cells</cell_line> .


Following stimulation , P-CREB levels remain elevated in PBMC for up to 24 hours whereas CREB is dephosphorylated in Jurkat cells within 4 hours following stimulation .

Following stimulation , <protein>P-CREB</protein> levels remain elevated in <cell_type>PBMC</cell_type> for up to 24 hours whereas CREB is dephosphorylated in <cell_line>Jurkat cells</cell_line> within 4 hours following stimulation .


The differences in P-CREB levels between PBMC and Jurkat cells were directly correlated with basal transcription of HTLV-I in the two cell types .

The differences in <protein>P-CREB</protein> levels between <cell_type>PBMC</cell_type> and <cell_line>Jurkat cells</cell_line> were directly correlated with basal transcription of HTLV-I in the two cell types .


Using electrophoretic mobility shift assays , we determined that the pattern of band migration differed between the two cell types .


These data demonstrate that PBMC differentially regulate basal HTLV-I transcription compared with Jurkat T cells , and this differential regulation is due , in part to differential phosphorylation and binding of CREB/ATF-1 to TRE-1 in the HTLV-I promoter .

These data demonstrate that <cell_type>PBMC</cell_type> differentially regulate basal HTLV-I transcription compared with <cell_line>Jurkat T cells</cell_line> , and this differential regulation is due , in part to differential phosphorylation and binding of <protein>CREB/ATF-1</protein> to <dna>TRE-1</dna> in the HTLV-I promoter .


We demonstrate the utility of using primary lymphocyte models to study HTLV-I transcription in the context of cell signaling and suggest that activated PBMC maintain elevated levels of P-CREB , which promote basal HTLV-I transcription and enhance viral persistence in vivo .

We demonstrate the utility of using primary lymphocyte models to study HTLV-I transcription in the context of cell signaling and suggest that activated <cell_type>PBMC</cell_type> maintain elevated levels of <protein>P-CREB</protein> , which promote basal HTLV-I transcription and enhance viral persistence in vivo .


Anoxia/reoxygenation-induced tolerance with respect to polymorphonuclear leukocyte adhesion to cultured endothelial cells .

Anoxia/reoxygenation-induced tolerance with respect to polymorphonuclear leukocyte adhesion to <cell_line>cultured endothelial cells</cell_line> .


A nuclear factor-kappaB -mediated phenomenon .

A <protein>nuclear factor-kappaB</protein> -mediated phenomenon .


Exposing human umbilical vein endothelial cells ( HUVECs ) to anoxia/reoxygenation ( A/R ) results in an increase in polymorphonuclear leukocyte ( PMN ) adhesion to HUVECs .

Exposing <cell_line>human umbilical vein endothelial cells</cell_line> ( <cell_line>HUVECs</cell_line> ) to anoxia/reoxygenation ( A/R ) results in an increase in polymorphonuclear leukocyte ( <cell_type>PMN</cell_type> ) adhesion to <cell_line>HUVECs</cell_line> .


This A/R-induced hyperadhesion is completely prevented by a previous ( 24 hours earlier ) exposure of HUVECs to A/R .

This A/R-induced hyperadhesion is completely prevented by a previous ( 24 hours earlier ) exposure of <cell_line>HUVECs</cell_line> to A/R .


This phenomenon has been termed `` A/R tolerance. ''


Exposing HUVECs to A/R induces an increase in nuclear factor kappaB ( NF-kappaB ) in HUVEC nuclei within 4 hours .

Exposing <cell_line>HUVECs</cell_line> to A/R induces an increase in <protein>nuclear factor kappaB</protein> ( <protein>NF-kappaB</protein> ) in HUVEC nuclei within 4 hours .


Interfering with either NF-kappaB activation ( proteasome inhibitor ) or translocation ( double-stranded oligonucleotides containing NF-kappaB binding sequence ) prevents the development of A/R tolerance ( ie , the increase in A/R-induced PMN adhesion to HUVECs is the same after the first and second A/R challenges ) .

Interfering with either <protein>NF-kappaB</protein> activation ( <protein>proteasome</protein> inhibitor ) or translocation ( double-stranded oligonucleotides containing <dna>NF-kappaB binding sequence</dna> ) prevents the development of A/R tolerance ( ie , the increase in A/R-induced PMN adhesion to <cell_line>HUVECs</cell_line> is the same after the first and second A/R challenges ) .


NO production by HUVECs is increased after the second A/R challenge , but not after the first A/R challenge .

NO production by <cell_line>HUVECs</cell_line> is increased after the second A/R challenge , but not after the first A/R challenge .


Inhibition of NO synthase ( NOS ) during the second A/R challenge prevents the development of A/R tolerance with respect to PMN adhesion .

Inhibition of <protein>NO synthase</protein> ( <protein>NOS</protein> ) during the second A/R challenge prevents the development of A/R tolerance with respect to PMN adhesion .


However , while HUVECs contained endothelial NOS protein , no inducible NOS was detected in either tolerant or nontolerant cells .

However , while <cell_line>HUVECs</cell_line> contained endothelial <protein>NOS protein</protein> , no inducible <protein>NOS</protein> was detected in either tolerant or <cell_type>nontolerant cells</cell_type> .


Further studies indicated that inhibition of GTP-cyclohydrolase I ( an enzyme involved in de novo synthesis of an important cofactor for NOS activity , tetrahydrobiopterin ) prevented the generation of NO in A/R-tolerant cells .

Further studies indicated that inhibition of <protein>GTP-cyclohydrolase I</protein> ( an enzyme involved in de novo synthesis of an important cofactor for <protein>NOS</protein> activity , tetrahydrobiopterin ) prevented the generation of NO in <cell_line>A/R-tolerant cells</cell_line> .


Extracellular generation of NO ( NO donor ) did not effect the hyperadhesion response induced by the initial A/R challenge .


A/R also induced an oxidant stress in naive HUVECs , but not in A/R-tolerant HUVECs .

A/R also induced an oxidant stress in naive <cell_line>HUVECs</cell_line> , but not in <cell_line>A/R-tolerant HUVECs</cell_line> .


Inhibition of NOS during the second A/R insult results in the generation of an oxidant stress similar to that observed after the first A/R challenge .

Inhibition of <protein>NOS</protein> during the second A/R insult results in the generation of an oxidant stress similar to that observed after the first A/R challenge .


Taken together , the findings of the present study are consistent with a role for NF-kappaB in the development of A/R tolerance ( with respect to PMN adhesion ) , perhaps by transcriptional regulation of GTP-cyclohydrolase .

Taken together , the findings of the present study are consistent with a role for <protein>NF-kappaB</protein> in the development of A/R tolerance ( with respect to <cell_type>PMN</cell_type> adhesion ) , perhaps by transcriptional regulation of <protein>GTP-cyclohydrolase</protein> .


The increased NO production during the second A/R insult reduces PMN adhesion most likely by reducing the intracellular oxidant stress induced by A/R .


Mildly oxidized low-density lipoproteins decrease early production of interleukin 2 and nuclear factor kappaB binding to DNA in activated T-lymphocytes .

Mildly oxidized <protein>low-density lipoproteins</protein> decrease early production of <protein>interleukin 2</protein> and <protein>nuclear factor kappaB</protein> binding to DNA in <cell_type>activated T-lymphocytes</cell_type> .


Activated T-lymphocytes are found early in atherosclerosis lesions , but little is known about their role .


Oxidized low-density lipoproteins ( oxLDLs ) are considered to be involved in the pathogenesis of the lesions , and we have previously demonstrated that oxLDLs inhibit not only interleukin ( IL ) -2-receptor expression on the surface of in vitro-activated T-lymphocytes but also their proliferation .

Oxidized <protein>low-density lipoproteins</protein> ( <protein>oxLDLs</protein> ) are considered to be involved in the pathogenesis of the lesions , and we have previously demonstrated that <protein>oxLDLs</protein> inhibit not only <protein>interleukin ( IL ) -2-receptor</protein> expression on the surface of in vitro-activated T-lymphocytes but also their proliferation .


We have now investigated the effect of oxLDLs on blast differentiation , on IL-2 synthesis and on the activation of the nuclear factor kappaB ( NF-kappaB ) system in activated lymphocytes .

We have now investigated the effect of <protein>oxLDLs</protein> on blast differentiation , on <protein>IL-2</protein> synthesis and on the activation of the <protein>nuclear factor kappaB</protein> ( <protein>NF-kappaB</protein> ) system in activated lymphocytes .


Mildly oxLDLs ( 50 and 100 microgram/ml ) decreased the number of lymphoblasts and the level of IL-2 concentration in the culture supernatants after activation of lymphocytes by phytohaemagglutinin and PMA+ionomycin .

Mildly <protein>oxLDLs</protein> ( 50 and 100 microgram/ml ) decreased the number of <cell_type>lymphoblasts</cell_type> and the level of IL-2 concentration in the culture supernatants after activation of <cell_type>lymphocytes</cell_type> by <protein>phytohaemagglutinin</protein> and PMA+ionomycin .


The inhibition of IL-2 production was observed in the CD3 ( + ) T-lymphocyte cytoplasm as early as 4 h after activation by PMA+ionomycin .

The inhibition of <protein>IL-2</protein> production was observed in the <protein>CD3</protein> ( + ) T-lymphocyte cytoplasm as early as 4 h after activation by PMA+ionomycin .


The study of NF-kappaB showed that oxLDLs led to a decrease of activation-induced p65/p50 NF-kappaB heterodimer binding to DNA , whereas the presence of the constitutive nuclear form of p50 dimer was unchanged .

The study of <protein>NF-kappaB</protein> showed that <protein>oxLDLs</protein> led to a decrease of <protein>activation-induced p65/p50</protein> <protein>NF-kappaB</protein> heterodimer binding to DNA , whereas the presence of the constitutive nuclear form of <protein>p50 dimer</protein> was unchanged .


This was correlated with an unchanged level of the active form of the cytosolic inhibitor protein IkappaB-alpha .

This was correlated with an unchanged level of the active form of the <protein>cytosolic inhibitor protein</protein> <protein>IkappaB-alpha</protein> .


Taken together , these observations suggest that the immunosuppressive effect of oxLDLs might operate via a dysregulation of the T-lymphocyte activation mechanisms .

Taken together , these observations suggest that the immunosuppressive effect of <protein>oxLDLs</protein> might operate via a dysregulation of the <cell_type>T-lymphocyte</cell_type> activation mechanisms .


The B29 ( immunoglobulin beta-chain ) gene is a genetic target for early B-cell factor .

The <dna>B29 ( immunoglobulin beta-chain ) gene</dna> is a genetic target for <protein>early B-cell factor</protein> .


Early B-cell factor ( EBF ) is a transcription factor suggested as essential for early B-lymphocyte development by findings in mice where the coding gene has been inactivated by homologous disruption .

<protein>Early B-cell factor</protein> ( <protein>EBF</protein> ) is a <protein>transcription factor</protein> suggested as essential for early <cell_type>B-lymphocyte</cell_type> development by findings in mice where the coding gene has been inactivated by homologous disruption .


This makes the identification of genetic targets for this transcription factor pertinent for the understanding of early B-cell development .

This makes the identification of genetic targets for this <protein>transcription factor</protein> pertinent for the understanding of early B-cell development .


The lack of B29 transcripts , coding for the beta subunit of the B-cell receptor complex , in pro-B cells from EBF -deficient mice suggested that B29 might be a genetic target for EBF .

The lack of <rna>B29 transcripts</rna> , coding for the <protein>beta subunit</protein> of the B-cell receptor complex , in <cell_type>pro-B cells</cell_type> from <protein>EBF</protein> -deficient mice suggested that B29 might be a genetic target for <protein>EBF</protein> .


We here present data suggesting that EBF interacts with three independent sites within the mouse B29 promoter .

We here present data suggesting that <protein>EBF</protein> interacts with three independent sites within the <dna>mouse B29 promoter</dna> .


Furthermore , ectopic expression of EBF in HeLa cells activated a B29 promoter-controlled reporter construct 13-fold and induced a low level of expression from the endogenous B29 gene .

Furthermore , ectopic expression of <protein>EBF</protein> in <cell_line>HeLa cells</cell_line> activated a <dna>B29 promoter-controlled reporter construct</dna> 13-fold and induced a low level of expression from the <dna>endogenous B29 gene</dna> .


Finally , mutations in the EBF binding sites diminished B29 promoter activity in pre-B cells while the same mutations did not have as striking an effect on the promoter function in B-cell lines of later differentiation stages .

Finally , mutations in the <protein>EBF</protein> binding sites diminished <dna>B29 promoter</dna> activity in <cell_type>pre-B cells</cell_type> while the same mutations did not have as striking an effect on the promoter function in <cell_line>B-cell lines</cell_line> of later differentiation stages .


These data suggest that the B29 gene is a genetic target for EBF in early B-cell development .

These data suggest that the <dna>B29 gene</dna> is a genetic target for <protein>EBF</protein> in early B-cell development .


Delta-opioid receptors expressed by Jurkat T cells enhance IL-2 secretion by increasing AP-1 complexes and activity of the NF-AT/AP-1-binding promoter element .

Delta-opioid receptors expressed by <cell_line>Jurkat T cells</cell_line> enhance <protein>IL-2</protein> secretion by increasing <protein>AP-1 complexes</protein> and activity of the <dna>NF-AT/AP-1-binding promoter element</dna> .


Recent molecular evidence points to transient and/or stage-specific expression of delta- and kappa-opioid receptors by thymic and peripheral T lymphocytes .

Recent molecular evidence points to transient and/or stage-specific expression of <protein>delta- and kappa-opioid receptors</protein> by <cell_type>thymic and peripheral T lymphocytes</cell_type> .


Since medical treatments or stress commonly increase opioid levels , it is important to understand the mechanisms by which opioids affect T lymphocyte functions .


We therefore created and studied a T cell line expressing the cloned delta-opioid receptor ( DOR1 ) .

We therefore created and studied a <cell_line>T cell line</cell_line> expressing the <protein>cloned delta-opioid receptor</protein> ( <protein>DOR1</protein> ) .


DOR1 ligation by a specific DOR1 agonist , deltorphin , augmented IL-2 secretion by synergizing with signals from TCR-CD3 and CD28 .

<protein>DOR1</protein> ligation by a specific <protein>DOR1</protein> agonist , deltorphin , augmented <protein>IL-2</protein> secretion by synergizing with signals from <protein>TCR-CD3</protein> and <protein>CD28</protein> .


Reporter gene constructs were used to map this effect of deltorphin to the AP-1- and NF-AT/AP-1-binding sites of the IL-2 promoter .

Reporter gene constructs were used to map this effect of deltorphin to the <dna>AP-1- and NF-AT/AP-1-binding sites</dna> of the <dna>IL-2 promoter</dna> .


Although DOR1 signaling increased [ Ca2+ ] i , deltorphin enhanced transcriptional activity of the NF-AT/AP-1-binding site via a mechanism independent of calcineurin and distinct from the effects of elevated [ Ca2+ ] i .

Although <protein>DOR1</protein> signaling increased [ Ca2+ ] i , deltorphin enhanced transcriptional activity of the <dna>NF-AT/AP-1-binding site</dna> via a mechanism independent of <protein>calcineurin</protein> and distinct from the effects of elevated [ Ca2+ ] i .


Deltorphin also increased accumulation of AP-1 transcription factor complexes , suggesting that DOR1 augments IL-2 secretion by increasing the AP-1 component of the NF-AT/AP-1 transcription factor .

Deltorphin also increased accumulation of <protein>AP-1 transcription factor complexes</protein> , suggesting that <protein>DOR1</protein> augments <protein>IL-2</protein> secretion by increasing the <protein>AP-1 component</protein> of the <protein>NF-AT/AP-1 transcription factor</protein> .


These results advance the molecular understanding of opioid effects on lymphocytes , and in addition , demonstrate regulation of IL-2 synthesis and secretion by the novel mechanism of receptor-mediated AP-1 induction .

These results advance the molecular understanding of opioid effects on <cell_type>lymphocytes</cell_type> , and in addition , demonstrate regulation of <protein>IL-2</protein> synthesis and secretion by the novel mechanism of receptor-mediated <protein>AP-1</protein> induction .


Induction of interleukin-12 p40 transcript by CD40 ligation via activation of nuclear factor-kappaB .

Induction of <protein>interleukin-12</protein> <rna>p40 transcript</rna> by <protein>CD40</protein> ligation via activation of <protein>nuclear factor-kappaB</protein> .


Interleukin-12 is produced in response to infection with bacteria or parasites or to bacterial constituents such as LPS in monocytes/macrophages and dendritic cells , and also generated by the interaction between activated T cells and antigen-presenting cells via CD40-CD40 ligand ( CD40L ) .

<protein>Interleukin-12</protein> is produced in response to infection with bacteria or parasites or to bacterial constituents such as LPS in <cell_type>monocytes/macrophages</cell_type> and <cell_type>dendritic cells</cell_type> , and also generated by the interaction between <cell_type>activated T cells</cell_type> and <cell_type>antigen-presenting cells</cell_type> via <protein>CD40-CD40 ligand</protein> ( <protein>CD40L</protein> ) .


So far , transcriptional analyses of p40 have been carried out only using bacterial constituents such as LPS as stimuli .

So far , transcriptional analyses of <protein>p40</protein> have been carried out only using bacterial constituents such as LPS as stimuli .


In the present study , we have characterized the transcriptional induction of p40 by CD40 ligation in a human B lymphoblastoid cell line , Daudi , and a human acute monocytic leukemia cell line , THP-1 .

In the present study , we have characterized the transcriptional induction of <protein>p40</protein> by <protein>CD40</protein> ligation in a <cell_line>human B lymphoblastoid cell line</cell_line> , <cell_line>Daudi</cell_line> , and a <cell_line>human acute monocytic leukemia cell line</cell_line> , <cell_line>THP-1</cell_line> .


These cells , stimulated by an agonistic monoclonal antibody against CD40 or by transfection with a CD40L expression vector , secreted p40 and showed enhanced p40 mRNA expression .

These cells , stimulated by an agonistic monoclonal antibody against <protein>CD40</protein> or by transfection with a <dna>CD40L expression vector</dna> , secreted <protein>p40</protein> and showed enhanced <rna>p40 mRNA</rna> expression .


Sequence analysis of the p40 promoter region identified two potential nuclear factor ( NF ) -kappaB binding sites conserved between mouse and human .

Sequence analysis of the <dna>p40 promoter region</dna> identified two <dna>potential nuclear factor ( NF ) -kappaB binding sites</dna> conserved between mouse and human .


Electrophoretic mobility shift assay revealed that the potential NF-kappaB binding sequence which is located around 120 bp upstream of the transcription initiation site in murine and human p40 genes formed an NF-kappaB complex with nuclear extract from Daudi cells stimulated by CD40 ligation .

Electrophoretic mobility shift assay revealed that the potential <dna>NF-kappaB binding sequence</dna> which is located around <dna>120 bp upstream</dna> of the <dna>transcription initiation site</dna> in <dna>murine and human p40 genes</dna> formed an <protein>NF-kappaB complex</protein> with nuclear extract from <cell_line>Daudi cells</cell_line> stimulated by <protein>CD40</protein> ligation .


Moreover , transfection of Daudi cells with the polymerized NF-kappaB binding sequence ligated to a thymidine kinase/chloramphenicol acetyltransferase ( CAT ) reporter plasmid greatly induced CAT activity , but transfection with the polymerized mutated NF-kappaB binding sequence did not .

Moreover , transfection of <cell_line>Daudi cells</cell_line> with the polymerized <dna>NF-kappaB binding sequence</dna> ligated to a <dna>thymidine kinase/chloramphenicol acetyltransferase ( CAT ) reporter plasmid</dna> greatly induced <protein>CAT</protein> activity , but transfection with the <dna>polymerized mutated NF-kappaB binding sequence</dna> did not .


These results suggest that the NF-kappaB binding site located around 120 bp upstream of the transcription initiation site in murine and human p40 promoter regions could be important for the p40 induction by CD40 ligation via activation of NF-kappaB .

These results suggest that the <dna>NF-kappaB binding site</dna> located around 120 bp upstream of the <dna>transcription initiation site</dna> in <dna>murine and human p40 promoter regions</dna> could be important for the <protein>p40</protein> induction by <protein>CD40</protein> ligation via activation of <protein>NF-kappaB</protein> .


High frequency of germ-line BRCA2 mutations among Hungarian male breast cancer patients without family history .

High frequency of germ-line <dna>BRCA2</dna> mutations among Hungarian male breast cancer patients without family history .


To determine the contribution of BRCA1 and BRCA2 mutations to the pathogenesis of male breast cancer in Hungary , the country with the highest male breast cancer mortality rates in continental Europe , a series of 18 male breast cancer patients and three patients with gynecomastia was analyzed for germ-line mutations in both BRCA1 and BRCA2 .

To determine the contribution of <dna>BRCA1</dna> and <dna>BRCA2</dna> mutations to the pathogenesis of male breast cancer in Hungary , the country with the highest male breast cancer mortality rates in continental Europe , a series of 18 male breast cancer patients and three patients with gynecomastia was analyzed for germ-line mutations in both <dna>BRCA1</dna> and <dna>BRCA2</dna> .


Although no germ-line BRCA1 mutation was observed , 6 of the 18 male breast cancer cases ( 33 % ) carried truncating mutations in the BRCA2 gene .

Although no germ-line <dna>BRCA1</dna> mutation was observed , 6 of the 18 male breast cancer cases ( 33 % ) carried truncating mutations in the <dna>BRCA2</dna> gene .


Unexpectedly , none of them reported a family history for breast/ovarian cancer .


Four of six truncating mutations were novel , and two mutations were recurrent .


Four patients ( 22 % ) had a family history of breast/ovarian cancer in at least one first- or second-degree relative ; however , no BRCA2 mutation was identified among them .

Four patients ( 22 % ) had a family history of breast/ovarian cancer in at least one first- or second-degree relative ; however , no <dna>BRCA2</dna> mutation was identified among them .


No mutation was identified in either of the genes in the gynecomastias .


These results provide evidence for a strong genetic component of male breast cancer in Hungary .


T-cell expression of the human GATA-3 gene is regulated by a non-lineage-specific silencer .

T-cell expression of the <dna>human GATA-3 gene</dna> is regulated by a <dna>non-lineage-specific silencer</dna> .


The GATA-3 transcription factor is required for development of the T-cell lineage and Th2 cytokine gene expression in CD4 T-cells .

The <protein>GATA-3 transcription factor</protein> is required for development of the <cell_line>T-cell lineage</cell_line> and <dna>Th2 cytokine gene</dna> expression in <cell_type>CD4 T-cells</cell_type> .


We have mapped the DNase-I-hypersensitive ( HS ) regions of the human GATA-3 gene in T-cells and non-T-cells and studied their transcriptional activities .

We have mapped the <dna>DNase-I-hypersensitive ( HS ) regions</dna> of the <dna>human GATA-3 gene</dna> in <cell_type>T-cells</cell_type> and <cell_type>non-T-cells</cell_type> and studied their transcriptional activities .


HS I-III , located 5 ' from the transcriptional initiation site , were found in hematopoietic and non-hematopoietic cells , whereas HS IV-VII , located 3 ' from the transcriptional start site , were exclusively observed in T-cells .

<dna>HS I-III</dna> , located 5 ' from the <dna>transcriptional initiation site</dna> , were found in <cell_type>hematopoietic and non-hematopoietic cells</cell_type> , whereas <dna>HS IV-VII</dna> , located 3 ' from the <dna>transcriptional start site</dna> , were exclusively observed in <cell_type>T-cells</cell_type> .


Among these hypersensitive sites , two transcriptional control elements were found , one in the first intron of the GATA-3 gene and the other between 8.3 and 5.9 kilobases 5 ' from the GATA-3 transcriptional initiation site .

Among these <dna>hypersensitive sites</dna> , two <dna>transcriptional control elements</dna> were found , one in the <dna>first intron</dna> of the <dna>GATA-3 gene</dna> and the other <dna>between 8.3 and 5.9 kilobases</dna> 5 ' from the <dna>GATA-3 transcriptional initiation site</dna> .


The first intron acted as a strong transcriptional activator in a position-dependent manner and with no cell-type specificity .

The <dna>first intron</dna> acted as a strong transcriptional activator in a position-dependent manner and with no cell-type specificity .


The upstream regulatory element could confer T-cell specificity to the GATA-3 promoter activity , and analysis of this region revealed a 707-base pair silencer that drastically inhibited GATA-3 promoter activity in non-T-cells .

The <dna>upstream regulatory element</dna> could confer T-cell specificity to the <dna>GATA-3 promoter</dna> activity , and analysis of this region revealed a <dna>707-base pair silencer</dna> that drastically inhibited <dna>GATA-3 promoter</dna> activity in <cell_type>non-T-cells</cell_type> .


Two CAGGTG E-boxes , located at the 5'- and 3'-ends of the silencer , were necessary for this silencer activity .

Two CAGGTG E-boxes , located at the <dna>5'- and 3'-ends</dna> of the silencer , were necessary for this silencer activity .


The 3'-CAGGTG E-box could bind USF proteins , the ubiquitous repressor ZEB , or the basic helix-loop-helix proteins E2A and HEB , and we showed that a competition between ZEB and E2A/HEB proteins is involved in the silencer activity .

The <dna>3'-CAGGTG E-box</dna> could bind <protein>USF proteins</protein> , the <protein>ubiquitous repressor</protein> <protein>ZEB</protein> , or the <protein>basic helix-loop-helix proteins</protein> <protein>E2A</protein> and <protein>HEB</protein> , and we showed that a competition between <protein>ZEB</protein> and <protein>E2A/HEB proteins</protein> is involved in the silencer activity .


Reactive oxygen intermediate-dependent NF-kappaB activation by interleukin-1beta requires 5-lipoxygenase or NADPH oxidase activity .

Reactive oxygen intermediate-dependent <protein>NF-kappaB</protein> activation by <protein>interleukin-1beta</protein> requires <protein>5-lipoxygenase</protein> or NADPH oxidase activity .


We previously reported that the role of reactive oxygen intermediates ( ROIs ) in NF-kappaB activation by proinflammatory cytokines was cell specific .

We previously reported that the role of reactive oxygen intermediates ( ROIs ) in <protein>NF-kappaB</protein> activation by <protein>proinflammatory cytokines</protein> was cell specific .


However , the sources for ROIs in various cell types are yet to be determined and might include 5-lipoxygenase ( 5-LOX ) and NADPH oxidase .

However , the sources for ROIs in various cell types are yet to be determined and might include <protein>5-lipoxygenase</protein> ( <protein>5-LOX</protein> ) and NADPH oxidase .


5-LOX and 5-LOX activating protein ( FLAP ) are coexpressed in lymphoid cells but not in monocytic or epithelial cells .

<protein>5-LOX</protein> and <protein>5-LOX activating protein</protein> ( <protein>FLAP</protein> ) are coexpressed in <cell_type>lymphoid cells</cell_type> but not in <cell_type>monocytic or epithelial cells</cell_type> .