Overexpression of dominant negative forms of IKKalpha and -beta demonstrates that only IKKbeta is the target for PKC and calcineurin .

Overexpression of dominant negative forms of <protein>IKKalpha and -beta</protein> demonstrates that only <protein>IKKbeta</protein> is the target for <protein>PKC</protein> and <protein>calcineurin</protein> .


These results indicate that within the TCR /CD3 signal transduction pathway both PKC and calcineurin are required for the effective activation of the IKK complex and NF-kappaB in T lymphocytes .

These results indicate that within the <protein>TCR</protein> <protein>/CD3</protein> signal transduction pathway both <protein>PKC</protein> and <protein>calcineurin</protein> are required for the effective activation of the <protein>IKK complex</protein> and <protein>NF-kappaB</protein> in <cell_type>T lymphocytes</cell_type> .


Diminished responses to IL-13 by human monocytes differentiated in vitro : role of the IL-13Ralpha1 chain and STAT6 .

Diminished responses to <protein>IL-13</protein> by human monocytes differentiated in vitro : role of the <protein>IL-13Ralpha1 chain</protein> and <protein>STAT6</protein> .


The primary IL-13 receptor complex on human monocytes is believed to be a heterodimer comprised of the IL-4R alpha chain and the IL-2R gamma chain ( gamma ( c ) ) -like molecule , IL-13R alpha1 .

The primary <protein>IL-13 receptor complex</protein> on <cell_type>human monocytes</cell_type> is believed to be a heterodimer comprised of the <protein>IL-4R alpha chain</protein> and the <protein>IL-2R gamma chain</protein> <protein>( gamma ( c ) ) -like molecule</protein> , <protein>IL-13R alpha1</protein> .


mRNA levels for IL-13R alpha1 , but not IL-4R alpha , were markedly decreased in in vitro monocyte-derived macrophages ( MDMac ) , and with increasing time of monocytes in culture correlated with the loss of IL-13 regulation of lipopolysaccharide-induced TNF-alpha production .

<rna>mRNA</rna> levels for <protein>IL-13R alpha1</protein> , but not <protein>IL-4R alpha</protein> , were markedly decreased in <cell_line>in vitro monocyte-derived macrophages</cell_line> ( <cell_line>MDMac</cell_line> ) , and with increasing time of <cell_type>monocytes</cell_type> in culture correlated with the loss of <protein>IL-13</protein> regulation of lipopolysaccharide-induced <protein>TNF-alpha</protein> production .


Analysis of cell lines Daudi and THP-1 that differentially express gamma ( c ) and IL-13R alpha1 showed that IL-13 can activate STAT6 in IL-13R alpha1-positive THP-1 cells but not in gamma ( c ) -positive , IL-13R alpha1-negative Daudi cells .

Analysis of <cell_line>cell lines</cell_line> <cell_line>Daudi</cell_line> and <cell_line>THP-1</cell_line> that differentially express <protein>gamma ( c )</protein> and <protein>IL-13R alpha1</protein> showed that <protein>IL-13</protein> can activate <protein>STAT6</protein> in <cell_line>IL-13R alpha1-positive THP-1 cells</cell_line> but not in <cell_line>gamma ( c ) -positive , IL-13R alpha1-negative Daudi cells</cell_line> .


IL-13 activation of STAT6 was reduced in MDMac which was associated with diminished IL-13 -induced expression of CD23 and MHC class II .

<protein>IL-13</protein> activation of <protein>STAT6</protein> was reduced in <cell_line>MDMac</cell_line> which was associated with diminished <protein>IL-13</protein> -induced expression of <protein>CD23</protein> and <protein>MHC class II</protein> .


However , with reduced IL-13R alpha1 expression and low nuclear STAT6 activity , some IL-13 -induced responses were unaltered in magnitude in MDMac .

However , with reduced <protein>IL-13R alpha1</protein> expression and low nuclear <protein>STAT6</protein> activity , some <protein>IL-13</protein> -induced responses were unaltered in magnitude in <cell_line>MDMac</cell_line> .


In the absence of functional IL-13R alpha1 and gamma ( c ) , IL-13 must signal through an alternative receptor complex on MDMac .

In the absence of functional <protein>IL-13R alpha1</protein> and <protein>gamma ( c )</protein> , <protein>IL-13</protein> must signal through an alternative <protein>receptor complex</protein> on <cell_line>MDMac</cell_line> .


Experiments with a blocking antibody to IL-4R alpha showed that this chain remains an essential component of the IL-13 receptor complex on MDMac .

Experiments with a blocking antibody to <protein>IL-4R alpha</protein> showed that this chain remains an essential component of the <protein>IL-13 receptor complex</protein> on <cell_line>MDMac</cell_line> .


The p53 paradox in the pathogenesis of tumor progression .

The <protein>p53</protein> paradox in the pathogenesis of tumor progression .


Recent evidence suggests that the p53 molecule appears in two different forms : the mutant p53 that stimulates tumor progression , and wild type p53 that inhibits tumor progression .

Recent evidence suggests that the <protein>p53 molecule</protein> appears in two different forms : the <protein>mutant p53</protein> that stimulates tumor progression , and <protein>wild type p53</protein> that inhibits tumor progression .


In addition , it has been established that tumor necrosis factor-alpha ( TNF-alpha ) can activate the expression of wild type p53 in concert with the nuclear transcription factor , NF-kappa B .

In addition , it has been established that <protein>tumor necrosis factor-alpha</protein> ( <protein>TNF-alpha</protein> ) can activate the expression of <protein>wild type p53</protein> in concert with the <protein>nuclear transcription factor</protein> , <protein>NF-kappa B</protein> .


Both TNF-alpha and NF-kappa B are also involved in the stimulation of the pathway that leads to the expression of major histocompatibility complex ( MHC ) class I molecules and , hence , antigen presentation to the T cells .

Both <protein>TNF-alpha</protein> and <protein>NF-kappa B</protein> are also involved in the stimulation of the pathway that leads to the expression of <protein>major histocompatibility complex ( MHC ) class I molecules</protein> and , hence , antigen presentation to the <cell_type>T cells</cell_type> .


In this paper we shall advance the hypothesis that : ( i ) TNF-alpha indirectly controls immune surveillance ; and ( ii ) TNF-alpha controls DNA repair and tumor suppression through the regulation of wild type p53 .

In this paper we shall advance the hypothesis that : ( i ) <protein>TNF-alpha</protein> indirectly controls immune surveillance ; and ( ii ) <protein>TNF-alpha</protein> controls DNA repair and tumor suppression through the regulation of <protein>wild type p53</protein> .


Thus , it is hypothesized that elevated TNF-alpha is primarily responsible for promoting tumor progression .

Thus , it is hypothesized that elevated <protein>TNF-alpha</protein> is primarily responsible for promoting tumor progression .


The promoter and 5 ' flanking sequences controlling human B29 gene expression .

The <dna>promoter</dna> and <dna>5 ' flanking sequences</dna> controlling human <dna>B29 gene</dna> expression .


The product of the B-cell-specific B29 gene ( B29 , Ig beta , CD79b ) is essential for Ig-mediated B-cell activation via the B-cell antigen receptor complex ( BCR ) on human and murine B lymphocytes .

The product of the <dna>B-cell-specific B29 gene</dna> ( <protein>B29</protein> , <protein>Ig beta</protein> , <protein>CD79b</protein> ) is essential for Ig-mediated B-cell activation via the <protein>B-cell antigen receptor complex</protein> ( <protein>BCR</protein> ) on <cell_type>human and murine B lymphocytes</cell_type> .


To better understand the regulation of this pivotal gene , we have analyzed the human genomic DNA sequence upstream of the B29 ATG start codon for transcriptional control activity .

To better understand the regulation of this pivotal gene , we have analyzed the <dna>human genomic DNA sequence</dna> upstream of the <protein>B29</protein> <dna>ATG start codon</dna> for transcriptional control activity .


The human B29 gene lacks either a TATA or a CAAT box and transcription is initiated at multiple sites .

The human <dna>B29 gene</dna> lacks either a <dna>TATA or a CAAT box</dna> and transcription is initiated at <dna>multiple sites</dna> .


The minimal promoter of the human B29 gene is contained within a 193-bp region 5 ' of these multiple start sites .

The minimal promoter of the <dna>human B29 gene</dna> is contained within a <dna>193-bp region</dna> 5 ' of these <dna>multiple start sites</dna> .


This minimal promoter exhibits B-cell-specific activity and contains SP1 , ETS , OCT , and IKAROS/LYF-1 transcription factor motifs .

This <dna>minimal promoter</dna> exhibits B-cell-specific activity and contains <dna>SP1 , ETS , OCT , and IKAROS/LYF-1 transcription factor motifs</dna> .


All these motifs are strikingly conserved in sequence and placement relative to the previously characterized murine B29 promoter .

All these motifs are strikingly conserved in sequence and placement relative to the previously characterized <dna>murine B29 promoter</dna> .


Additional upstream gene segments dramatically affected B29 minimal promoter activity .

Additional upstream gene segments dramatically affected <protein>B29</protein> <dna>minimal promoter</dna> activity .


A newly identified motif called the B29 conserved sequence ( BCS ) , found upstream of both human and murine B29 promoters , appears to stimulate B29 transcription through a novel mechanism .

A newly identified motif called the <dna>B29 conserved sequence</dna> ( <dna>BCS</dna> ) , found upstream of both <dna>human and murine B29 promoters</dna> , appears to stimulate <protein>B29</protein> transcription through a novel mechanism .


A single BCS had little effect either on the minimal B29 promoter or on a heterologous promoter .

A single <dna>BCS</dna> had little effect either on the minimal <dna>B29 promoter</dna> or on a heterologous promoter .


Instead , the BCS stimulated transcription by counteracting 5 ' negative regulatory DNA sequences that block the activity of the B29 minimal promoter in its absence .

Instead , the <dna>BCS</dna> stimulated transcription by counteracting <dna>5 ' negative regulatory DNA sequences</dna> that block the activity of the <protein>B29</protein> <dna>minimal promoter</dna> in its absence .


These findings indicate that B29 gene expression is controlled by the complex interplay of positive and negative regulatory elements .

These findings indicate that <dna>B29 gene</dna> expression is controlled by the complex interplay of positive and <dna>negative regulatory elements</dna> .


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

A novel <protein>immunosuppressive factor</protein> in bovine colostrum blocks activation of the <dna>interleukin 2 gene enhancer</dna> at the <dna>NFAT site</dna> .


A factor in bovine colostrum ( colostrum inhibitory factor , CIF ) inhibits interleukin 2 ( IL2 ) production in activated T helper cells by blocking the accumulation of IL2 mRNA .

A factor in bovine colostrum ( <protein>colostrum inhibitory factor</protein> , <protein>CIF</protein> ) inhibits <protein>interleukin 2</protein> ( <protein>IL2</protein> ) production in activated <cell_type>T helper cells</cell_type> by blocking the accumulation of <rna>IL2 mRNA</rna> .


To determine whether CIF blocks at the level of IL2 transcription , we introduced reporter plasmids into the human T leukemia cell line Jurkat by transient transfection .

To determine whether <protein>CIF</protein> blocks at the level of <protein>IL2</protein> transcription , we introduced reporter plasmids into the <cell_line>human T leukemia cell line Jurkat</cell_line> by transient transfection .


These contained the luciferase gene under the control of either the human IL2 upstream enhancer region ( segments -326 to +45 ) or three repeats of the NFAT element contained within it ( segments -255 to -285 ) .

These contained the <dna>luciferase gene</dna> under the control of either the <dna>human IL2 upstream enhancer</dna> region ( segments -326 to +45 ) or three repeats of the <dna>NFAT element</dna> contained within it ( segments -255 to -285 ) .


Expression of luciferase in these cells was induced by phorbol myristate acetate plus a calcium ionophore .

Expression of <protein>luciferase</protein> in these cells was induced by phorbol myristate acetate plus a calcium ionophore .


CIF inhibited induction of either construct as did cyclosporine , which is known to block activation of the NFAT element .

<protein>CIF</protein> inhibited induction of either construct as did cyclosporine , which is known to block activation of the <protein>NFAT element</protein> .


CIF failed to inhibit several other enhancer elements .

<protein>CIF</protein> failed to inhibit several other <dna>enhancer elements</dna> .


The NFAT-controlled luciferase gene system distinguishes CIF from other T cell inhibitory activities present in colostrum , in particular , TGF beta 1 and TGF beta 2 and the glucocorticoids .

The <dna>NFAT-controlled luciferase gene system</dna> distinguishes <protein>CIF</protein> from other T cell inhibitory activities present in colostrum , in particular , <protein>TGF beta 1</protein> and <protein>TGF beta 2</protein> and the glucocorticoids .


Stably transfected Jurkat cells behaved similarly to the transiently transfected ones with respect to inhibition by CIF and cyclosporine .

Stably transfected Jurkat cells behaved similarly to the transiently transfected ones with respect to inhibition by <protein>CIF</protein> and cyclosporine .


The NFAT -luc assay is a useful technique for the rapid , sensitive measurement of CIF or other immunosuppressants with a similar mode of action .

The <protein>NFAT</protein> -luc assay is a useful technique for the rapid , sensitive measurement of <protein>CIF</protein> or other immunosuppressants with a similar mode of action .


Heat shock induces HIV-1 replication in chronically infected promyelocyte cell line OM10.1 .

Heat shock induces HIV-1 replication in chronically infected <cell_line>promyelocyte cell line OM10.1</cell_line> .


A long period of clinical latency before development of symptoms is characteristic of human immunodeficiency virus type 1 ( HIV-1 ) infection .


OM10.1 , a promyelocyte cell line latently infected with HIV-1 , has been developed as a model for studying the mechanism of viral latency and the activation of virus expression .

<cell_line>OM10.1</cell_line> , a <cell_line>promyelocyte cell line</cell_line> latently infected with HIV-1 , has been developed as a model for studying the mechanism of viral latency and the activation of virus expression .


We found that this latently infected cell line with heat shock at 42 degrees C for 2 h resulted in a high level of HIV-1 production without addition of any cytokines .

We found that this latently infected cell line with heat shock at 42 degrees C for 2 h resulted in a high level of HIV-1 production without addition of any <protein>cytokines</protein> .


The mechanism of activation was analyzed by using anti-TNF-alpha antibody and various inhibitors .

The mechanism of activation was analyzed by using <protein>anti-TNF-alpha antibody</protein> and various inhibitors .


Although the TNF-alpha level in culture supernatants was below the sensitivity of an ELISA assay system , addition of anti- TNF-alpha antibody in culture medium could partially suppress the heat shock induced HIV-1 production .

Although the <protein>TNF-alpha</protein> level in culture supernatants was below the sensitivity of an ELISA assay system , addition of anti- <protein>TNF-alpha</protein> antibody in culture medium could partially suppress the heat shock induced HIV-1 production .


Staurosporine ( PKC inhibitor ) , pentoxifylline ( NF-kappa B inhibitor ) , and Ro5-3335 ( HIV-1 Tat inhibitor ) also inhibited significantly the heat shock induced virus activation .

Staurosporine ( PKC inhibitor ) , pentoxifylline ( NF-kappa B inhibitor ) , and Ro5-3335 ( HIV-1 <protein>Tat</protein> inhibitor ) also inhibited significantly the heat shock induced virus activation .


In particular , staurosporine achieved approximately 90 % inhibition of the HIV-1 antigen expression in heat shock-treated OM10.1 at a non-toxic concentration .

In particular , staurosporine achieved approximately 90 % inhibition of the HIV-1 antigen expression in <cell_line>heat shock-treated OM10.1</cell_line> at a non-toxic concentration .


Although the mechanism of HIV-1 activation with heat shock has not been fully elucidated yet , it is presumed PKC plays an important role in HIV-1 activation .

Although the mechanism of HIV-1 activation with heat shock has not been fully elucidated yet , it is presumed <protein>PKC</protein> plays an important role in HIV-1 activation .


Thus , the present observations will provide a further insight into the pathogenesis of HIV-1 infections .


Activation of the CDC42 effector N-WASP by the Shigella flexneri IcsA protein promotes actin nucleation by Arp2/3 complex and bacterial actin -based motility .

Activation of the <protein>CDC42 effector</protein> <protein>N-WASP</protein> by the <protein>Shigella flexneri IcsA protein</protein> promotes <protein>actin</protein> nucleation by <protein>Arp2/3 complex</protein> and bacterial <protein>actin</protein> -based motility .


To propel itself in infected cells , the pathogen Shigella flexneri subverts the Cdc42 -controlled machinery responsible for actin assembly during filopodia formation .

To propel itself in <cell_type>infected cells</cell_type> , the pathogen Shigella flexneri subverts the <protein>Cdc42</protein> -controlled machinery responsible for <protein>actin</protein> assembly during filopodia formation .


Using a combination of bacterial motility assays in platelet extracts with Escherichia coli expressing the Shigella IcsA protein and in vitro analysis of reconstituted systems from purified proteins , we show here that the bacterial protein IcsA binds N-WASP and activates it in a Cdc42 -like fashion .

Using a combination of bacterial motility assays in platelet extracts with Escherichia coli expressing the <protein>Shigella IcsA protein</protein> and in vitro analysis of reconstituted systems from <protein>purified proteins</protein> , we show here that the <protein>bacterial protein IcsA</protein> binds <protein>N-WASP</protein> and activates it in a <protein>Cdc42</protein> -like fashion .


Dramatic stimulation of actin assembly is linked to the formation of a ternary IcsA-N-WASP-Arp2/3 complex , which nucleates actin polymerization .

Dramatic stimulation of <protein>actin</protein> assembly is linked to the formation of a ternary <protein>IcsA-N-WASP-Arp2/3 complex</protein> , which nucleates <protein>actin</protein> polymerization .


The Arp2/3 complex is essential in initiation of actin assembly and Shigella movement , as previously observed for Listeria monocytogenes .

The <protein>Arp2/3 complex</protein> is essential in initiation of <protein>actin</protein> assembly and Shigella movement , as previously observed for Listeria monocytogenes .


Activation of N-WASP by IcsA unmasks two domains acting together in insertional actin polymerization .

Activation of <protein>N-WASP</protein> by <protein>IcsA</protein> unmasks two domains acting together in insertional <protein>actin</protein> polymerization .


The isolated COOH-terminal domain of N-WASP containing a verprolin-homology region , a cofilin-homology sequence , and an acidic terminal segment ( VCA ) interacts with G-actin in a unique profilin-like functional fashion .

The isolated <protein>COOH-terminal domain</protein> of <protein>N-WASP</protein> containing a <protein>verprolin-homology region</protein> , a cofilin-homology sequence , and an <protein>acidic terminal segment</protein> ( <protein>VCA</protein> ) interacts with <protein>G-actin</protein> in a unique profilin-like functional fashion .


Hence , when N-WASP is activated , its COOH-terminal domain feeds barbed end growth of filaments and lowers the critical concentration at the bacterial surface .

Hence , when <protein>N-WASP</protein> is activated , its <protein>COOH-terminal domain</protein> feeds barbed end growth of filaments and lowers the critical concentration at the bacterial surface .


On the other hand , the NH ( 2 ) -terminal domain of N-WASP interacts with F-actin , mediating the attachment of the actin tail to the bacterium surface .

On the other hand , the <protein>NH ( 2 ) -terminal domain</protein> of <protein>N-WASP</protein> interacts with <protein>F-actin</protein> , mediating the attachment of the <protein>actin tail</protein> to the bacterium surface .


VASP is not involved in Shigella movement , and the function of profilin does not require its binding to proline-rich regions .

VASP is not involved in Shigella movement , and the function of <protein>profilin</protein> does not require its binding to <protein>proline-rich regions</protein> .


c-Maf induces monocytic differentiation and apoptosis in bipotent myeloid progenitors .

<protein>c-Maf</protein> induces monocytic differentiation and apoptosis in <cell_type>bipotent myeloid progenitors</cell_type> .


The transcriptional mechanisms that drive colony-forming unit granulocyte-macrophage ( CFU-GM ) myeloid progenitors to differentiate into cells of either the granulocytic or monocytic lineage are not fully understood .

The transcriptional mechanisms that drive <cell_line>colony-forming unit granulocyte-macrophage ( CFU-GM ) myeloid progenitors</cell_line> to differentiate into cells of either the <cell_type>granulocytic or monocytic lineage</cell_type> are not fully understood .


We have shown that the c-Maf and c-Myb transcription factors physically interact in myeloid cells to form inhibitory complexes that hinder transactivation of c-Myb target genes through direct binding to Myb consensus sites .

We have shown that the <protein>c-Maf</protein> and <protein>c-Myb transcription factors</protein> physically interact in myeloid cells to form inhibitory complexes that hinder transactivation of <dna>c-Myb target genes</dna> through direct binding to <dna>Myb consensus sites</dna> .


These complexes arise in a developmentally regulated pattern , peaking at the promyelocyte stage , or in cell model systems , appearing soon after the induction of monocytic differentiation .

These complexes arise in a developmentally regulated pattern , peaking at the <cell_type>promyelocyte stage</cell_type> , or in <cell_line>cell model systems</cell_line> , appearing soon after the induction of monocytic differentiation .


We wished to determine if this developmentally related interaction is a consequence of myeloid differentiation or an intrinsic differentiating stimulus .


Because the elevated Myb : Maf status seen in differentiating cells can be recapitulated by overexpression of c-Maf in myeloid cell lines , we inducibly expressed the c-Maf cDNA in 2 bipotent human myeloid progenitor cells .

Because the elevated <protein>Myb : Maf</protein> status seen in <cell_type>differentiating cells</cell_type> can be recapitulated by overexpression of <protein>c-Maf</protein> in myeloid cell lines , we inducibly expressed the <dna>c-Maf cDNA</dna> in 2 bipotent <cell_type>human myeloid progenitor cells</cell_type> .


Elevated levels of c-Maf protein led to marked increases in Myb : Maf complexes and the accumulation of monocyte/macrophage cells , followed by eventual programmed cell death .

Elevated levels of <protein>c-Maf protein</protein> led to marked increases in <protein>Myb : Maf complexes</protein> and the accumulation of <cell_type>monocyte/macrophage cells</cell_type> , followed by eventual programmed cell death .


Analysis of targets that could mediate these phenotypic changes indicated that c-Maf likely plays a key role in myeloid cell development through dual mechanisms ; inhibition of a select set of c-Myb regulated targets , such as Bcl-2 and CD13/APN , coupled with the activation of as yet undefined differentiation-promoting genes .

Analysis of targets that could mediate these phenotypic changes indicated that <protein>c-Maf</protein> likely plays a key role in myeloid cell development through dual mechanisms ; inhibition of a select set of <protein>c-Myb</protein> regulated targets , such as <protein>Bcl-2</protein> and <protein>CD13/APN</protein> , coupled with the activation of as yet undefined <dna>differentiation-promoting genes</dna> .


AML1 ( CBFalpha2 ) cooperates with B cell-specific activating protein ( BSAP/PAX5 ) in activation of the B cell-specific BLK gene promoter .

<protein>AML1 ( CBFalpha2 )</protein> cooperates with <protein>B cell-specific activating protein ( BSAP/PAX5 )</protein> in activation of the <dna>B cell-specific BLK gene promoter</dna> .


AML1 plays a critical role during hematopoiesis and chromosomal translocations involving AML1 are commonly associated with different forms of leukemia , including pre-B acute lymphoblastic leukemia .

<protein>AML1</protein> plays a critical role during hematopoiesis and chromosomal translocations involving <protein>AML1</protein> are commonly associated with different forms of leukemia , including pre-B acute lymphoblastic leukemia .


To understand the function of AML1 during B cell differentiation , we analyzed regulatory regions of B cell-specific genes for potential AML1-binding sites and have identified a putative AML1-binding site in the promoter of the B cell-specific tyrosine kinase gene , blk .

To understand the function of <protein>AML1</protein> during <cell_type>B cell</cell_type> differentiation , we analyzed <dna>regulatory regions</dna> of <dna>B cell-specific genes</dna> for potential <dna>AML1-binding sites</dna> and have identified a putative <dna>AML1-binding site</dna> in the promoter of the <dna>B cell-specific tyrosine kinase gene , blk</dna> .


Gel mobility shift assays and transient transfection assays demonstrate that AML1 binds specifically to this site in the blk promoter and this binding site is important for blk promoter activity .

Gel mobility shift assays and transient transfection assays demonstrate that <protein>AML1</protein> binds specifically to this site in the <dna>blk promoter</dna> and this binding site is important for <dna>blk</dna> promoter activity .


Furthermore , in vitro binding analysis revealed that the AML1 runt DNA-binding domain physically interacts with the paired DNA-binding domain of BSAP , a B cell-specific transcription factor .

Furthermore , in vitro binding analysis revealed that the <dna>AML1 runt DNA-binding domain</dna> physically interacts with the <dna>paired DNA-binding domain</dna> of <protein>BSAP</protein> , a <protein>B cell-specific transcription factor</protein> .


BSAP has been shown previously to be important for B cell-specific regulation of the blk gene .

<protein>BSAP</protein> has been shown previously to be important for B cell-specific regulation of the <dna>blk</dna> gene .


Physical interaction of AML1 with BSAP correlates with functional cooperativity in transfection studies where AML1 and BSAP synergistically activate blk promoter transcription by more than 50-fold .

Physical interaction of <protein>AML1</protein> with <protein>BSAP</protein> correlates with functional cooperativity in transfection studies where <protein>AML1</protein> and <protein>BSAP</protein> synergistically activate <dna>blk</dna> promoter transcription by more than 50-fold .


These results demonstrate physical and functional interactions between AML1 and BSAP and suggest that AML1 is an important factor for regulating a critical B cell-specific gene , blk .

These results demonstrate physical and functional interactions between <protein>AML1</protein> and <protein>BSAP</protein> and suggest that <protein>AML1</protein> is an important factor for regulating a critical <dna>B cell-specific gene</dna> , <dna>blk</dna> .


Transcriptional inhibition by interleukin-6 of the class A macrophage scavenger receptor in macrophages derived from human peripheral monocytes and the THP-1 monocytic cell line .

Transcriptional inhibition by <protein>interleukin-6</protein> of the <protein>class A macrophage scavenger receptor</protein> in <cell_type>macrophages</cell_type> derived from <cell_type>human peripheral monocytes</cell_type> and the <cell_line>THP-1 monocytic cell line</cell_line> .


Expression of the class A macrophage scavenger receptor ( MSR ) contributes to the uptake of modified low density lipoproteins ( LDL ) by macrophages and transformation of these cells into lipid-laden foam cells , which characterize atherosclerosis .

Expression of the <protein>class A macrophage scavenger receptor ( MSR )</protein> contributes to the uptake of <protein>modified low density lipoproteins ( LDL )</protein> by <cell_type>macrophages</cell_type> and transformation of these cells into <cell_type>lipid-laden foam cells</cell_type> , which characterize atherosclerosis .


Many environmental factors , in particular , proinflammatory cytokines and growth factors , can exert regulatory effects on MSR expression , whereas intracellular accumulation of cholesterol itself does not influence MSR levels to any considerable extent .

Many environmental factors , in particular , <protein>proinflammatory cytokines</protein> and <protein>growth factors</protein> , can exert regulatory effects on <protein>MSR</protein> expression , whereas intracellular accumulation of cholesterol itself does not influence <protein>MSR</protein> levels to any considerable extent .


In the present study , by using an in vitro model , we examined whether stimulation with interleukin-6 ( IL-6 ) , an immunoregulatory , multipotential cytokine , modulates the expression and activities of the MSR in macrophages .

In the present study , by using an in vitro model , we examined whether stimulation with <protein>interleukin-6 ( IL-6 )</protein> , an <protein>immunoregulatory</protein> , <protein>multipotential cytokine</protein> , modulates the expression and activities of the <protein>MSR</protein> in <cell_type>macrophages</cell_type> .


When treated with IL-6 , macrophages derived from peripheral monocytes and phorbol 12-myristate 13-acetate ( PMA ) -differentiated THP-1 monocytic cells showed significantly reduced uptake and/or binding of the MSR ligand , acetylated LDL .

When treated with <protein>IL-6</protein> , <cell_type>macrophages</cell_type> derived from <cell_type>peripheral monocytes</cell_type> and <cell_line>phorbol 12-myristate 13-acetate ( PMA ) -differentiated THP-1 monocytic cells</cell_line> showed significantly reduced uptake and/or binding of the <protein>MSR</protein> ligand , <protein>acetylated LDL</protein> .


This effect was paralleled by a reduction in the expression of MSR protein and mRNA .

This effect was paralleled by a reduction in the expression of <protein>MSR</protein> protein and <rna>mRNA</rna> .


Analysis of MSR promoter activity in THP-1 cells transfected with an MSR promoter-reporter gene construct demonstrated decreased activity of the MSR promoter in IL-6 -treated THP-1 macrophages .

Analysis of <dna>MSR promoter</dna> activity in <cell_line>THP-1 cells</cell_line> transfected with an <dna>MSR promoter-reporter gene construct</dna> demonstrated decreased activity of the <dna>MSR promoter</dna> in <protein>IL-6</protein> -treated <cell_line>THP-1</cell_line> <cell_type>macrophages</cell_type> .


Electrophoretic mobility gel shift assay also showed a reduction in the binding of a transcription factor to the MSR promoter AP-1/ets elements in IL-6 -treated cells .

Electrophoretic mobility gel shift assay also showed a reduction in the binding of a <protein>transcription factor</protein> to the <dna>MSR promoter AP-1/ets elements</dna> in <protein>IL-6</protein> -treated cells .


Thus , exposure to IL-6 may inhibit expression of the class A MSR in differentiated macrophages at transcriptional levels .

Thus , exposure to <protein>IL-6</protein> may inhibit expression of the <protein>class A MSR</protein> in differentiated <cell_type>macrophages</cell_type> at transcriptional levels .


This result suggests that this cytokine may modulate foam cell formation during atherogenesis .

This result suggests that this <protein>cytokine</protein> may modulate foam cell formation during atherogenesis .


Retinoblastoma protein expression leads to reduced Oct-1 DNA binding activity and enhances interleukin-8 expression .

Retinoblastoma protein expression leads to reduced <protein>Oct-1</protein> DNA binding activity and enhances <protein>interleukin-8</protein> expression .


Tumor cell lines with a defective retinoblastoma gene are unable to transcribe the HLA class II genes in response to IFN-gamma treatment , and reconstitution of functional Rb rescues IFN-gamma-induced class II gene expression .

Tumor cell lines with a defective <dna>retinoblastoma gene</dna> are unable to transcribe the <dna>HLA class II genes</dna> in response to <protein>IFN-gamma</protein> treatment , and reconstitution of functional <protein>Rb</protein> rescues IFN-gamma-induced <dna>class II gene</dna> expression .


However , the molecular mechanism of Rb rescue of the class II genes is unknown .

However , the molecular mechanism of <protein>Rb</protein> rescue of the <dna>class II genes</dna> is unknown .


We have examined the effect of Rb expression on the activation of the promoter for HLA-DRA , the prototype class II gene .

We have examined the effect of <protein>Rb</protein> expression on the activation of the promoter for <dna>HLA-DRA</dna> , the <dna>prototype class II gene</dna> .


Oct-1 , a POU domain transcription factor , was identified as a repressor of HLA-DRA promoter activity in the Rb-defective cells .

<protein>Oct-1</protein> , a <protein>POU domain transcription factor</protein> , was identified as a repressor of <dna>HLA-DRA</dna> promoter activity in the <cell_type>Rb-defective cells</cell_type> .


Rb expression led to phosphorylation of Oct-1 , thus relieving its repressive effect .

<protein>Rb</protein> expression led to phosphorylation of <protein>Oct-1</protein> , thus relieving its repressive effect .


Oct-1 has also been shown to repress interleukin 8 promoter activity .

<protein>Oct-1</protein> has also been shown to repress <protein>interleukin 8</protein> promoter activity .


Consistent with reduced levels of Oct-1 DNA binding activity in the Rb-transformed cell lines , interleukin 8 expression is higher in these cell lines .

Consistent with reduced levels of <protein>Oct-1</protein> DNA binding activity in the <cell_line>Rb-transformed cell lines</cell_line> , <protein>interleukin 8</protein> expression is higher in these cell lines .


Cell activation and apoptosis by bacterial lipoproteins through toll-like receptor-2 .

Cell activation and apoptosis by <protein>bacterial lipoproteins</protein> through <protein>toll-like receptor-2</protein> .


Apoptosis is implicated in the generation and resolution of inflammation in response to bacterial pathogens .


All bacterial pathogens produce lipoproteins ( BLPs ) , which trigger the innate immune response .

All bacterial pathogens produce <protein>lipoproteins ( BLPs )</protein> , which trigger the innate immune response .


BLPs were found to induce apoptosis in THP-1 monocytic cells through human Toll-like receptor-2 ( hTLR2 ) .

<protein>BLPs</protein> were found to induce apoptosis in <cell_line>THP-1 monocytic cells</cell_line> through <protein>human Toll-like receptor-2 ( hTLR2 )</protein> .


BLPs also initiated apoptosis in an epithelial cell line transfected with hTLR2 .

<protein>BLPs</protein> also initiated apoptosis in an <cell_line>epithelial cell line</cell_line> transfected with <protein>hTLR2</protein> .


In addition , BLPs stimulated nuclear factor-kappaB , a transcriptional activator of multiple host defense genes , and activated the respiratory burst through hTLR2 .

In addition , <protein>BLPs</protein> stimulated <protein>nuclear factor-kappaB</protein> , a <protein>transcriptional activator</protein> of <dna>multiple host defense genes</dna> , and activated the respiratory burst through <protein>hTLR2</protein> .


Thus , hTLR2 is a molecular link between microbial products , apoptosis , and host defense mechanisms .

Thus , <protein>hTLR2</protein> is a molecular link between microbial products , apoptosis , and host defense mechanisms .


Possible differences in the mechanism ( s ) of action of different glucocorticoid hormone compounds .


Different glucocorticoid hormones ( GCH ) show differences in the intensity and in the kinetics of their immunomodulating activity .


The mechanism ( s ) of action of GCH is under investigation , but is has been noted that they exert immune activity via the genomic pathway .


We have studied the effects of prednisone ( PDN ) , deflazacort ( DFC ) , and dexamethasone ( DXM ) on the production of cytokines ( IL-2 , IL-6 , TNF-alpha , IL-10 ) by peripheral T lymphocytes , and the effects on the inhibition of NF-kB DNA binding activity by activated Jurkat cell line .

We have studied the effects of prednisone ( PDN ) , deflazacort ( DFC ) , and dexamethasone ( DXM ) on the production of <protein>cytokines</protein> ( <protein>IL-2</protein> , <protein>IL-6</protein> , <protein>TNF-alpha</protein> , <protein>IL-10</protein> ) by <cell_type>peripheral T lymphocytes</cell_type> , and the effects on the inhibition of <protein>NF-kB</protein> DNA binding activity by <cell_line>activated Jurkat cell line</cell_line> .


The data obtained show that the three GCH molecules exert an immunosuppression on cytokine production by T lymphocytes and a strong decrease in the nuclear translocation of NF-kB in Jurkat cells ; moreover , ( a ) not all the cytokines investigated were affected , and not with the same intensity , by the three GCH and ( b ) DXM inhibited the binding activity of NF-kB less than that of DFC and PDN .

The data obtained show that the three GCH molecules exert an immunosuppression on <protein>cytokine</protein> production by <cell_type>T lymphocytes</cell_type> and a strong decrease in the nuclear translocation of <protein>NF-kB</protein> in <cell_line>Jurkat cells</cell_line> ; moreover , ( a ) not all the <protein>cytokines</protein> investigated were affected , and not with the same intensity , by the three GCH and ( b ) DXM inhibited the binding activity of <protein>NF-kB</protein> less than that of DFC and PDN .


These data are in agreement with the concept that different GCH compounds might differ in their binding and affinity properties , tissue-specific metabolism , and interaction with transcription factor .

These data are in agreement with the concept that different GCH compounds might differ in their binding and affinity properties , tissue-specific metabolism , and interaction with <protein>transcription factor</protein> .


Distinctive expression pattern of the BCL-6 protein in nodular lymphocyte predominance Hodgkin 's disease .

Distinctive expression pattern of the <protein>BCL-6 protein</protein> in nodular lymphocyte predominance Hodgkin 's disease .


The BCL-6 gene encoding a nuclear-located Kruppel-type zinc finger protein is rearranged in about 30 % diffuse large B-cell lymphomas and is expressed predominantly in normal germinal center B cells and related lymphomas .

The <rna>BCL-6 gene</rna> encoding a <protein>nuclear-located Kruppel-type zinc finger protein</protein> is rearranged in about 30 % diffuse large <cell_type>B-cell lymphomas</cell_type> and is expressed predominantly in normal <cell_type>germinal center B cells</cell_type> and related lymphomas .


These findings suggest that BCL-6 may play a role in regulating differentiation of normal germinal center B cells and that its deregulated expression caused by rearrangements may contribute to lymphomagenesis .

These findings suggest that <protein>BCL-6</protein> may play a role in regulating differentiation of normal <cell_type>germinal center B cells</cell_type> and that its deregulated expression caused by rearrangements may contribute to lymphomagenesis .


This prompted us to investigate the expression of the BCL-6 protein in Hodgkin 's disease ( HD ) , focusing on the nodular lymphocyte predominance subtype ( NLPHD ) , which differs from classical HD by virtue of the B-cell nature of the malignant cell population ( so-called L & H cells ) and its relationship with germinal centers .

This prompted us to investigate the expression of the <protein>BCL-6 protein</protein> in Hodgkin 's disease ( HD ) , focusing on the nodular lymphocyte predominance subtype ( NLPHD ) , which differs from classical HD by virtue of the B-cell nature of the <cell_type>malignant cell population</cell_type> ( so-called <cell_type>L & H cells</cell_type> ) and its relationship with germinal centers .


Forty-one HD samples ( 19 NLPHD , 12 nodular sclerosis , and 10 mixed cellularity ) were immunostained with the monoclonal antibodies PG-B6 and PG-B6p that react with a fixative-sensitive and a formalin-resistant epitope on the aminoterminal region of the BCL-6 gene product , respectively .

Forty-one HD samples ( 19 NLPHD , 12 nodular sclerosis , and 10 mixed cellularity ) were immunostained with the <protein>monoclonal antibodies</protein> <protein>PG-B6</protein> and <protein>PG-B6p</protein> that react with a fixative-sensitive and a <protein>formalin-resistant epitope</protein> on the <protein>aminoterminal region</protein> of the <protein>BCL-6 gene product</protein> , respectively .


Strong nuclear positivity for the BCL-6 protein was detected in tumor ( L & H ) cells in all cases of NLPHD .

Strong nuclear positivity for the <protein>BCL-6 protein</protein> was detected in <cell_type>tumor ( L & H ) cells</cell_type> in all cases of NLPHD .


In contrast , BCL-6 was expressed only in a small percentage of Hodgkin and Reed-Sternberg cells in about 30 % of classical HD cases .

In contrast , <protein>BCL-6</protein> was expressed only in a small percentage of <cell_line>Hodgkin and Reed-Sternberg cells</cell_line> in about 30 % of classical HD cases .


Notably , the nuclei of reactive CD3+/CD4+ T cells nearby to and rosetting around L & H cells in NLPHD were also strongly BCL-6 + , but lacked CD40 ligand ( CD40L ) expression .

Notably , the nuclei of <cell_type>reactive CD3+/CD4+ T cells</cell_type> nearby to and rosetting around <cell_type>L & H cells</cell_type> in NLPHD were also strongly <protein>BCL-6</protein> + , but lacked <protein>CD40 ligand</protein> ( <protein>CD40L</protein> ) expression .


This staining pattern clearly differed from that of classical HD , whose cellular background was made up of CD3+/CD4+ T cells showing the BCL-6 -/ CD40L + phenotype .

This staining pattern clearly differed from that of classical HD , whose cellular background was made up of <cell_type>CD3+/CD4+ T cells</cell_type> showing the <protein>BCL-6</protein> -/ <protein>CD40L</protein> + phenotype .


These results further support the concept that NLPHD is an histogenetically distinct , B-cell-derived subtype of HD and suggest a role for BCL-6 in its development .

These results further support the concept that NLPHD is an histogenetically distinct , B-cell-derived subtype of HD and suggest a role for <protein>BCL-6</protein> in its development .


Selenium-mediated inhibition of transcription factor NF-kappa B and HIV-1 LTR promoter activity .

Selenium-mediated inhibition of <protein>transcription factor NF-kappa B</protein> and <dna>HIV-1 LTR promoter</dna> activity .


The eukaryotic transcription factor NF-kappa B is involved in the inducible expression of various inflammatory genes as well as in HIV-1 replication .

The <protein>eukaryotic transcription factor NF-kappa B</protein> is involved in the inducible expression of various <dna>inflammatory genes</dna> as well as in HIV-1 replication .


Activation of NF-kappa B is induced by prooxidants and several stimuli eliciting oxidative stress , such as cytokines , lipopolysaccharide , UV irradiation and other mediators .

Activation of <protein>NF-kappa B</protein> is induced by prooxidants and several stimuli eliciting oxidative stress , such as <protein>cytokines</protein> , lipopolysaccharide , UV irradiation and other mediators .


Various antioxidants inhibit NF-kappa B activation in response to these stimuli .

Various antioxidants inhibit <protein>NF-kappa B</protein> activation in response to these stimuli .


In this study , we have investigated the effects of selenium , an integral component of glutathione peroxidase ( GPX ) , on NF-kappa B activation .

In this study , we have investigated the effects of selenium , an integral component of <protein>glutathione peroxidase</protein> ( <protein>GPX</protein> ) , on <protein>NF-kappa B</protein> activation .


In selenium-deprived Jurkat and ESb-L T lymphocytes , supplementation of selenium led to a substantial increase of GPX activity .

In <cell_line>selenium-deprived Jurkat</cell_line> and <cell_line>ESb-L T lymphocytes</cell_line> , supplementation of selenium led to a substantial increase of <protein>GPX</protein> activity .


Analysis of DNA binding revealed that NF-kappa B activation in response to TNF was significantly inhibited under these conditions .

Analysis of DNA binding revealed that <protein>NF-kappa B</protein> activation in response to <protein>TNF</protein> was significantly inhibited under these conditions .


Likewise , reporter gene assays using luciferase constructs driven by the HIV-1 long terminal repeat showed a dose-dependent inhibition of NF-kappa B controlled gene expression by selenium .

Likewise , reporter gene assays using luciferase constructs driven by the <dna>HIV-1 long terminal repeat</dna> showed a dose-dependent inhibition of <protein>NF-kappa B</protein> controlled gene expression by selenium .


The effects of selenium were specific for NF-kappa B , since the activity of the transcription factor AP-1 was not suppressed .

The effects of selenium were specific for <protein>NF-kappa B</protein> , since the activity of the transcription factor <protein>AP-1</protein> was not suppressed .


These data suggest that selenium supplementation may be used to modulate the expression of NF-kappa B target genes and HIV-1 .

These data suggest that selenium supplementation may be used to modulate the expression of <dna>NF-kappa B target genes</dna> and HIV-1 .


The involvement of p23 , hsp90 , and immunophilins in the assembly of progesterone receptor complexes .

The involvement of <protein>p23</protein> , <protein>hsp90</protein> , and <protein>immunophilins</protein> in the assembly of <protein>progesterone receptor complexes</protein> .


To better understand the assembly mechanism for the progesterone receptor ( PR ) , we have developed cell-free systems for studying interactions of PR , hsp90 , and other associated proteins .

To better understand the assembly mechanism for the <protein>progesterone receptor</protein> ( <protein>PR</protein> ) , we have developed cell-free systems for studying interactions of <protein>PR</protein> , <protein>hsp90</protein> , and other <protein>associated proteins</protein> .


When PR is incubated in rabbit reticulocyte lysate , its association with hsp90 , hsp70 , the three immunophilins FKBP54 , FKBP52 and CyP-40 , and with p23 is observed .

When <protein>PR</protein> is incubated in rabbit reticulocyte lysate , its association with <protein>hsp90</protein> , <protein>hsp70</protein> , the three <protein>immunophilins</protein> <protein>FKBP54</protein> , <protein>FKBP52</protein> and <protein>CyP-40</protein> , and with <protein>p23</protein> is observed .


These interactions require ATP/Mg2+ and when ATP is limiting the PR complex is altered to one containing the proteins p60 and p48 , but lacking immunophilins and p23 .

These interactions require ATP/Mg2+ and when ATP is limiting the <protein>PR</protein> complex is altered to one containing the proteins <protein>p60</protein> and <protein>p48</protein> , but lacking <protein>immunophilins</protein> and <protein>p23</protein> .


We have studied two pre-formed hsp90 complexes that may participate in the assembly of PR complexes .

We have studied two <protein>pre-formed hsp90 complexes</protein> that may participate in the assembly of <protein>PR complexes</protein> .


One contains hsp90 bound to hsp70 and p60 and this complex forms spontaneously in the absence of ATP .

One contains <protein>hsp90</protein> bound to <protein>hsp70</protein> and <protein>p60</protein> and this complex forms spontaneously in the absence of ATP .


A second complex contains hsp90 bound to p23 plus the three immunophilins and some hsp70 .

A second complex contains <protein>hsp90</protein> bound to <protein>p23</protein> plus the three <protein>immunophilins</protein> and some <protein>hsp70</protein> .


The formation of this complex requires ATP .


In further studies we have shown that purified hsp90 can bind to purified p23 and this interaction requires both ATP and molybdate .

In further studies we have shown that purified <protein>hsp90</protein> can bind to purified <protein>p23</protein> and this interaction requires both ATP and molybdate .


This explains , in part , the known effects of ATP and molybdate on assembly of PR complexes .

This explains , in part , the known effects of ATP and molybdate on assembly of <protein>PR complexes</protein> .


Increased IkappaB expression and diminished nuclear NF-kappaB in human mononuclear cells following hydrocortisone injection .

Increased <protein>IkappaB</protein> expression and diminished <protein>nuclear NF-kappaB</protein> in <cell_type>human mononuclear cells</cell_type> following hydrocortisone injection .


We have recently demonstrated that hydrocortisone and other glucocorticoids inhibit reactive oxygen species ( ROS ) generation by mononuclear ( MNC ) and polymorphonuclear leucocytes ( PMNL ) .

We have recently demonstrated that hydrocortisone and other glucocorticoids inhibit reactive oxygen species ( ROS ) generation by <cell_type>mononuclear ( MNC ) and polymorphonuclear leucocytes</cell_type> ( <cell_type>PMNL</cell_type> ) .


Since NF-kappaB/ IkappaB system regulates the transcription of proinflammatory genes , including those responsible for ROS generation , we tested the hypothesis that hydrocortisone may stimulate IkappaB production thus inhibiting NF-kappaB translocation from the cytosol into the nucleus in MNC , in vivo .

Since NF-kappaB/ <protein>IkappaB</protein> system regulates the transcription of <protein>proinflammatory genes</protein> , including those responsible for ROS generation , we tested the hypothesis that hydrocortisone may stimulate <protein>IkappaB</protein> production thus inhibiting <protein>NF-kappaB</protein> translocation from the cytosol into the nucleus in <cell_type>MNC</cell_type> , in vivo .


One hundred milligram of hydrocortisone was injected intravenously into 4 normal subjects .


Blood samples were obtained prior to the injection and at 1 , 2 , 4 , 8 and 24 hr after the injection .


Nuclear extracts and total cell lysates were prepared from MNC by standard techniques .

Nuclear extracts and total cell lysates were prepared from <cell_type>MNC</cell_type> by standard techniques .


IkappaB levels in MNC homogenates increased at 1 hr , peaked at 2-4 hr , started to decrease at 8 hr , and returned to baseline levels at 24 hr .

<protein>IkappaB</protein> levels in <cell_type>MNC</cell_type> homogenates increased at 1 hr , peaked at 2-4 hr , started to decrease at 8 hr , and returned to baseline levels at 24 hr .


NF-kappaB in MNC nuclear extracts decreased at 1 hr , reached a nadir at 4 hr , gradually increased at 8 hr and returned back to baseline levels at 24 hr .

<protein>NF-kappaB</protein> in <cell_type>MNC</cell_type> nuclear extracts decreased at 1 hr , reached a nadir at 4 hr , gradually increased at 8 hr and returned back to baseline levels at 24 hr .


The total protein content of NF-kappaB subunit ( P65 ) in MNC lysates also showed a decrease following hydrocortisone injection .

The total protein content of <protein>NF-kappaB subunit</protein> ( <protein>P65</protein> ) in <cell_type>MNC</cell_type> lysates also showed a decrease following hydrocortisone injection .


This decrease was observed at 2 hr , reached a nadir at 4 hr , and returned to baseline levels at 24 hr .


ROS generation inhibition paralleled NF-kappaB levels in the nucleus .

ROS generation inhibition paralleled <protein>NF-kappaB</protein> levels in the nucleus .


It was inhibited at 1 hr , reached a nadir at 2-4 hr , started to increase at 8 hr , and returned to basal levels at 24 hr .


Our data demonstrate that hydrocortisone induces IkappaB and suppresses NF-kappaB expression in MNC in parallel .

Our data demonstrate that hydrocortisone induces <protein>IkappaB</protein> and suppresses <protein>NF-kappaB</protein> expression in <cell_type>MNC</cell_type> in parallel .


IkappaB further reduces the translocation of NF-kappaB into the nucleus thus preventing the expression of proinflammatory genes .

<protein>IkappaB</protein> further reduces the translocation of <protein>NF-kappaB</protein> into the nucleus thus preventing the expression of <dna>proinflammatory genes</dna> .


Glucocorticoids induce apoptosis in human monocytes : potential role of IL-1 beta .

Glucocorticoids induce apoptosis in <cell_type>human monocytes</cell_type> : potential role of <protein>IL-1 beta</protein> .


Glucocorticoids ( GC ) are potent anti-inflammatory and immunosuppressive agents that act on a variety of immune cells , including monocytes and macrophages .

Glucocorticoids ( GC ) are potent anti-inflammatory and immunosuppressive agents that act on a variety of <cell_type>immune cells</cell_type> , including <cell_type>monocytes</cell_type> and <cell_type>macrophages</cell_type> .


However , the exact cellular mechanisms underlying this anti-inflammatory capacity are still unknown .


In our study , we determined the induction of apoptosis by GC in human monocytes .

In our study , we determined the induction of apoptosis by GC in <cell_type>human monocytes</cell_type> .


Peripheral blood monocytes were isolated by density centrifugation methods with a purity of > 90 % and were cultured in RPMI 1640 medium .

<cell_type>Peripheral blood monocytes</cell_type> were isolated by density centrifugation methods with a purity of > 90 % and were cultured in RPMI 1640 medium .


Monocyte apoptosis was determined by four independent methods , including annexin-V staining , TUNEL , DNA-laddering , and typical morphology by means of transmission electron microscopy .


TNF-alpha and IL-1beta were measured by ELISA .

<protein>TNF-alpha</protein> and <protein>IL-1beta</protein> were measured by ELISA .


GC receptor was blocked with mifepristone .

<protein>GC receptor</protein> was blocked with mifepristone .


Caspase 3 was inhibited with caspase-3 inhibitor ( DEVD-CHO ) .


Stimulation with different GC at therapeutic concentrations resulted in monocyte apoptosis in a time- and dose-dependent manner .


Necrosis was excluded by propidium iodide staining .


Proinflammatory cytokines such as IL-1beta and TNF-alpha were down-regulated by GC treatment .

<protein>Proinflammatory cytokines</protein> such as <protein>IL-1beta</protein> and <protein>TNF-alpha</protein> were down-regulated by GC treatment .


Continuous treatment of monocytes with IL-1beta , but not with TNF-alpha , could almost completely prevent GC-induced cell death .

Continuous treatment of <cell_type>monocytes</cell_type> with <protein>IL-1beta</protein> , but not with <protein>TNF-alpha</protein> , could almost completely prevent GC-induced cell death .


The addition of mifepristone or caspase-3 inhibitor could partially abrogate GC-induced apoptosis as well as GC-induced inhibition of IL-1beta .

The addition of mifepristone or caspase-3 inhibitor could partially abrogate GC-induced apoptosis as well as GC-induced inhibition of <protein>IL-1beta</protein> .


This is the first study to demonstrate induction of apoptosis by GC in human monocytes .

This is the first study to demonstrate induction of apoptosis by GC in <cell_type>human monocytes</cell_type> .


GC-induced monocyte apoptosis may be partially mediated through effects on IL-1beta production .

GC-induced monocyte apoptosis may be partially mediated through effects on <protein>IL-1beta</protein> production .


It is conceivable that GC exert their anti-inflammatory capacity in various diseases , at least in part , by the induction of apoptosis in monocytes .

It is conceivable that GC exert their anti-inflammatory capacity in various diseases , at least in part , by the induction of apoptosis in <cell_type>monocytes</cell_type> .


Differential effects of lipopolysaccharide and tumor necrosis factor on monocytic IkappaB kinase signalsome activation and IkappaB proteolysis .

Differential effects of lipopolysaccharide and <protein>tumor necrosis factor</protein> on <protein>monocytic IkappaB kinase</protein> signalsome activation and <protein>IkappaB</protein> proteolysis .


The inflammatory mediators lipopolysaccharide ( LPS ) and tumor necrosis factor ( TNF ) are potent activators of NF-kappaB .

The inflammatory mediators lipopolysaccharide ( LPS ) and <protein>tumor necrosis factor</protein> ( <protein>TNF</protein> ) are potent activators of <protein>NF-kappaB</protein> .


This study compared the effect of these stimuli on endogenous IkappaB kinase ( IKK ) signalsome activation and IkappaB phosphorylation/proteolysis in human monocytic cells and investigated the role of the signalsome proteins IKK-alpha , IKK-beta , NF-kappaB-inducing kinase ( NIK ) , IKK-gamma ( NF-kappaB essential modulator ) , and IKK complex-associated protein .

This study compared the effect of these stimuli on endogenous <protein>IkappaB kinase</protein> ( <protein>IKK</protein> ) signalsome activation and <protein>IkappaB</protein> phosphorylation/proteolysis in <cell_type>human monocytic cells</cell_type> and investigated the role of the <protein>signalsome proteins</protein> <protein>IKK-alpha</protein> , <protein>IKK-beta</protein> , <protein>NF-kappaB-inducing kinase</protein> ( <protein>NIK</protein> ) , <protein>IKK-gamma</protein> ( <protein>NF-kappaB</protein> essential modulator ) , and <protein>IKK complex-associated protein</protein> .


Kinase assays showed that TNF elicited a rapid but short-lived induction of IKK activity with a 3-fold greater effect on IKK-alpha than on IKK-beta , peaking at 5 min .

Kinase assays showed that <protein>TNF</protein> elicited a rapid but short-lived induction of <protein>IKK</protein> activity with a 3-fold greater effect on <protein>IKK-alpha</protein> than on <protein>IKK-beta</protein> , peaking at 5 min .


In contrast , LPS predominantly stimulated IKK-beta activity , which slowly increased , peaking at 30 min .

In contrast , LPS predominantly stimulated <protein>IKK-beta</protein> activity , which slowly increased , peaking at 30 min .


A second peak was observed at a later time point following LPS stimulation , which consisted of both IKK-alpha and -beta activity .


The endogenous levels of the signalsome components were unaffected by stimulation .


Furthermore , our studies showed association of the IKK-alpha/beta heterodimer with NIK , IkappaB-alpha and -epsilon in unstimulated cells .

Furthermore , our studies showed association of the <protein>IKK-alpha/beta heterodimer</protein> with <protein>NIK</protein> , <protein>IkappaB-alpha</protein> and <protein>-epsilon</protein> in <cell_type>unstimulated cells</cell_type> .


Exposure to LPS or TNF led to differential patterns of IkappaB-alpha and IkappaB-epsilon disappearance from and reassembly with the signalsome , whereas IKK-alpha , IKK-beta , and NIK remained complex-associated .

Exposure to LPS or <protein>TNF</protein> led to differential patterns of <protein>IkappaB-alpha</protein> and <protein>IkappaB-epsilon</protein> disappearance from and reassembly with the signalsome , whereas <protein>IKK-alpha</protein> , <protein>IKK-beta</protein> , and <protein>NIK</protein> remained complex-associated .


NIK can not phosphorylate IkappaB-alpha directly , but it appears to be a functionally important subunit , because mutated NIK inhibited stimulus-induced kappaB -dependent transcription more effectively than mutated IKK-alpha or -beta .

<protein>NIK</protein> can not phosphorylate <protein>IkappaB-alpha</protein> directly , but it appears to be a functionally important subunit , because mutated <protein>NIK</protein> inhibited stimulus-induced <protein>kappaB</protein> -dependent transcription more effectively than mutated <protein>IKK-alpha</protein> or <protein>-beta</protein> .


Overexpression of IKK complex-associated protein inhibited stimulus-mediated transcription , whereas NF-kappaB essential modulator enhanced it .

Overexpression of <protein>IKK complex-associated protein</protein> inhibited stimulus-mediated transcription , whereas <protein>NF-kappaB</protein> essential modulator enhanced it .


The understanding of LPS- and TNF-induced signaling may allow the development of specific strategies to treat sepsis-associated disease .


Tumor necrosis factor alpha decreases , and interleukin-10 increases , the sensitivity of human monocytes to dexamethasone : potential regulation of the glucocorticoid receptor .

<protein>Tumor necrosis factor alpha</protein> decreases , and <protein>interleukin-10</protein> increases , the sensitivity of <cell_type>human monocytes</cell_type> to dexamethasone : potential regulation of the <protein>glucocorticoid receptor</protein> .


Resistance to glucocorticoid therapy has been observed in patients with autoimmune/inflammatory diseases and may be related to the inflammatory process itself .


The aim of this study was to examine the ability of tumor necrosis factor alpha ( TNFalpha , a proinflammatory cytokine ) and interleukin ( IL ) -10 ( an anti-inflammatory cytokine ) to differentially regulate the sensitivity of human monocytes /macrophages to glucocorticoids .

The aim of this study was to examine the ability of <protein>tumor necrosis factor alpha</protein> ( <protein>TNFalpha</protein> , a <protein>proinflammatory cytokine</protein> ) and <protein>interleukin ( IL ) -10</protein> ( an <protein>anti-inflammatory cytokine</protein> ) to differentially regulate the sensitivity of <cell_type>human monocytes</cell_type> <cell_type>/macrophages</cell_type> to glucocorticoids .


To accomplish this , we first analyzed the pattern of TNFalpha and IL-10 inhibition by dexamethasone in LPS-stimulated whole-blood cell cultures .

To accomplish this , we first analyzed the pattern of <protein>TNFalpha</protein> and IL-10 inhibition by dexamethasone in <cell_line>LPS-stimulated whole-blood cell cultures</cell_line> .


Second , we studied the modulation of the sensitivity of these cells to dexamethasone by preincubation with TNFalpha or IL-10 and measurement of LPS-stimulated IL-6 secretion .

Second , we studied the modulation of the sensitivity of these cells to dexamethasone by preincubation with <protein>TNFalpha</protein> or <protein>IL-10</protein> and measurement of LPS-stimulated <protein>IL-6</protein> secretion .


In addition , we evaluated the effect of dexamethasone on phorbolmyristate-acetate-stimulated IL-1 receptor antagonist secretion by the human monocytic cell line U937 .

In addition , we evaluated the effect of dexamethasone on phorbolmyristate-acetate-stimulated <protein>IL-1 receptor</protein> antagonist secretion by the <cell_line>human monocytic cell line U937</cell_line> .


Finally , we investigated whether the modulation of corticosensitivity in TNFalpha- and IL-10-pretreated U937 cells was related to a change of the glucocorticoid receptor concentration and affinity .

Finally , we investigated whether the modulation of corticosensitivity in <cell_line>TNFalpha- and IL-10-pretreated U937 cells</cell_line> was related to a change of the <protein>glucocorticoid receptor</protein> concentration and affinity .


Dexamethasone had different effects on LPS-induced TNFalpha and IL-10 secretion ; whereas it suppressed TNFalpha in a dose-dependent fashion , its effect on IL-10 secretion was biphasic , producing stimulation at lower , and inhibition at higher doses .

Dexamethasone had different effects on LPS-induced <protein>TNFalpha</protein> and <protein>IL-10</protein> secretion ; whereas it suppressed <protein>TNFalpha</protein> in a dose-dependent fashion , its effect on <protein>IL-10</protein> secretion was biphasic , producing stimulation at lower , and inhibition at higher doses .


The concentration of LPS employed influenced the effect of dexamethasone on IL-10 secretion ( P < 0.001 ) .

The concentration of LPS employed influenced the effect of dexamethasone on <protein>IL-10</protein> secretion ( P < 0.001 ) .


Pretreatment with TNFalpha diminished , and with IL-10 improved , the ability of dexamethasone to suppress IL-6 secretion in whole-blood cell cultures ( P < 0.01 for both ) and to enhance IL-1 receptor antagonist secretion by U937 cells ( P < 0.05 for both ) .

Pretreatment with <protein>TNFalpha</protein> diminished , and with <protein>IL-10</protein> improved , the ability of dexamethasone to suppress <protein>IL-6</protein> secretion in <cell_type>whole-blood cell cultures</cell_type> ( P < 0.01 for both ) and to enhance <protein>IL-1</protein> receptor antagonist secretion by U937 cells ( P < 0.05 for both ) .


TNFalpha decreased ( P < 0.001 ) , while IL-10 increased ( P < 0.001 ) , the concentration of dexamethasone binding sites in these cells , with no discernible effect on their binding affinity .

<protein>TNFalpha</protein> decreased ( P < 0.001 ) , while <protein>IL-10</protein> increased ( P < 0.001 ) , the concentration of dexamethasone binding sites in these cells , with no discernible effect on their binding affinity .


We conclude that glucocorticoids differentially modulate TNFalpha and IL-10 secretion by human monocytes in a LPS dose-dependent fashion and that the sensitivity of these cells to glucocorticoids is altered by TNFalpha or IL-10 pretreatment ; TNFalpha blocks their effects , whereas IL-10 acts synergistically with glucocorticoids .

We conclude that glucocorticoids differentially modulate <protein>TNFalpha</protein> and <protein>IL-10</protein> secretion by <cell_type>human monocytes</cell_type> in a LPS dose-dependent fashion and that the sensitivity of these cells to glucocorticoids is altered by <protein>TNFalpha</protein> or <protein>IL-10</protein> pretreatment ; <protein>TNFalpha</protein> blocks their effects , whereas <protein>IL-10</protein> acts synergistically with glucocorticoids .


This is accompanied by opposite glucocorticoid receptor changes , respectively opposing and favoring glucocorticoid actions .

This is accompanied by opposite <protein>glucocorticoid receptor</protein> changes , respectively opposing and favoring glucocorticoid actions .


This study suggests that the pattern of pro-/antiinflammatory cytokine secretion may alter the response of patients to glucocorticoid therapy .

This study suggests that the pattern of pro-/antiinflammatory <protein>cytokine</protein> secretion may alter the response of patients to glucocorticoid therapy .


Block of granulocytic differentiation of 32Dcl3 cells by AML1/ETO ( MTG8 ) but not by highly expressed Bcl-2 .

Block of granulocytic differentiation of <cell_line>32Dcl3 cells</cell_line> by <dna>AML1/ETO ( MTG8 )</dna> but not by highly expressed <protein>Bcl-2</protein> .


The chimeric gene , AML1/ETO ( MTG8 ) , generated in t ( 8 ; 21 ) acute myeloid leukemia enhances the expression of Bcl-2 .

The <dna>chimeric gene ,</dna> <dna>AML1/ETO ( MTG8 )</dna> , generated in <dna>t ( 8 ; 21 )</dna> acute myeloid leukemia enhances the expression of <protein>Bcl-2</protein> .


To evaluate whether this enhancement is the primary role of AML1/ETO in leukemogenesis , effects of over-expression of Bcl-2 in the murine myeloid precursor cell line , 32Dcl3 , were examined .

To evaluate whether this enhancement is the primary role of <dna>AML1/ETO</dna> in leukemogenesis , effects of over-expression of <protein>Bcl-2</protein> in the <cell_line>murine myeloid precursor cell line</cell_line> , <cell_line>32Dcl3</cell_line> , were examined .


When 32Dcl3 cells expressing exogenous Bcl-2 were induced to differentiate , the onset of morphological differentiation was delayed .

When <cell_line>32Dcl3 cells</cell_line> expressing exogenous <protein>Bcl-2</protein> were induced to differentiate , the onset of morphological differentiation was delayed .


However , even the cells expressing very high levels of exogenous Bcl-2 eventually underwent differentiation without a significant decrease in the synthesis of Bcl-2 .

However , even the cells expressing very high levels of exogenous <protein>Bcl-2</protein> eventually underwent differentiation without a significant decrease in the synthesis of <protein>Bcl-2</protein> .


On the contrary , 32Dcl3 cells stably expressing AML1/ETO were completely resistant to differentiation and continued to grow in the presence of G-CSF .

On the contrary , <cell_line>32Dcl3 cells</cell_line> stably expressing <dna>AML1/ETO</dna> were completely resistant to differentiation and continued to grow in the presence of <protein>G-CSF</protein> .


These results are consistent with the interpretation that stimulation of Bcl-2 expression is not the primary target of AML1/ETO .

These results are consistent with the interpretation that stimulation of <protein>Bcl-2</protein> expression is not the primary target of <dna>AML1/ETO</dna> .


Host defense mechanisms triggered by microbial lipoproteins through toll-like receptors .

Host defense mechanisms triggered by <protein>microbial lipoproteins</protein> through <protein>toll-like receptors</protein> .


The generation of cell-mediated immunity against many infectious pathogens involves the production of interleukin-12 ( IL-12 ) , a key signal of the innate immune system .

The generation of cell-mediated immunity against many infectious pathogens involves the production of <protein>interleukin-12</protein> ( <protein>IL-12</protein> ) , a key signal of the innate immune system .


Yet , for many pathogens , the molecules that induce IL-12 production by macrophages and the mechanisms by which they do so remain undefined .

Yet , for many pathogens , the molecules that induce <protein>IL-12</protein> production by <cell_type>macrophages</cell_type> and the mechanisms by which they do so remain undefined .


Here it is shown that microbial lipoproteins are potent stimulators of IL-12 production by human macrophages , and that induction is mediated by Toll-like receptors ( TLRs ) .

Here it is shown that <protein>microbial lipoproteins</protein> are potent stimulators of <protein>IL-12</protein> production by human <cell_type>macrophages</cell_type> , and that induction is mediated by <protein>Toll-like receptors</protein> ( <protein>TLRs</protein> ) .


Several lipoproteins stimulated TLR-dependent transcription of inducible nitric oxide synthase and the production of nitric oxide , a powerful microbicidal pathway .

Several lipoproteins stimulated TLR-dependent transcription of inducible <protein>nitric oxide synthase</protein> and the production of nitric oxide , a powerful microbicidal pathway .


Activation of TLRs by microbial lipoproteins may initiate innate defense mechanisms against infectious pathogens .

Activation of <protein>TLRs</protein> by <protein>microbial lipoproteins</protein> may initiate innate defense mechanisms against infectious pathogens .


Abnormal NF-kappa B activity in T lymphocytes from patients with systemic lupus erythematosus is associated with decreased p65-RelA protein expression .

Abnormal <protein>NF-kappa B</protein> activity in <cell_type>T lymphocytes</cell_type> from patients with systemic lupus erythematosus is associated with decreased <protein>p65-RelA</protein> protein expression .


Numerous cellular and biochemical abnormalities in immune regulation have been described in patients with systemic lupus erythematosus ( SLE ) , including surface Ag receptor -initiated signaling events and lymphokine production .

Numerous cellular and biochemical abnormalities in immune regulation have been described in patients with systemic lupus erythematosus ( SLE ) , including surface <protein>Ag receptor</protein> -initiated signaling events and <protein>lymphokine</protein> production .


Because NF-kappa B contributes to the transcription of numerous inflammatory genes and has been shown to be a molecular target of antiinflammatory drugs , we sought to characterize the functional role of the NF-kappa B protein complex in lupus T cells .

Because <protein>NF-kappa B</protein> contributes to the transcription of numerous <dna>inflammatory genes</dna> and has been shown to be a molecular target of antiinflammatory drugs , we sought to characterize the functional role of the <protein>NF-kappa B protein complex</protein> in <cell_type>lupus T cells</cell_type> .


Freshly isolated T cells from lupus patients , rheumatoid arthritis ( RA ) patients , and normal individuals were activated physiologically via the TCR with anti-CD3 and anti-CD28 Abs to assess proximal membrane signaling , and with PMA and a calcium ionophore ( A23187 ) to bypass membrane-mediated signaling events .

Freshly isolated <cell_type>T cells</cell_type> from lupus patients , rheumatoid arthritis ( RA ) patients , and normal individuals were activated physiologically via the <protein>TCR</protein> with <protein>anti-CD3</protein> and <protein>anti-CD28 Abs</protein> to assess proximal membrane signaling , and with PMA and a calcium ionophore ( A23187 ) to bypass membrane-mediated signaling events .


We measured the NF-kappa B binding activity in nuclear extracts by gel shift analysis .

We measured the <protein>NF-kappa B</protein> binding activity in nuclear extracts by gel shift analysis .


When compared with normal cells , the activation of NF-kappa B activity in SLE patients was significantly decreased in SLE , but not in RA , patients .

When compared with <cell_type>normal cells</cell_type> , the activation of <protein>NF-kappa B</protein> activity in SLE patients was significantly decreased in SLE , but not in RA , patients .


NF-kappa B binding activity was absent in several SLE patients who were not receiving any medication , including corticosteroids .

<protein>NF-kappa B</protein> binding activity was absent in several SLE patients who were not receiving any medication , including corticosteroids .


Also , NF-kappa B activity remained absent in follow-up studies .

Also , <protein>NF-kappa B</protein> activity remained absent in follow-up studies .


In supershift experiments using specific Abs , we showed that , in the group of SLE patients who displayed undetectable NF-kappa B activity , p65 complexes were not formed .

In supershift experiments using specific <protein>Abs</protein> , we showed that , in the group of SLE patients who displayed undetectable <protein>NF-kappa B</protein> activity , <protein>p65 complexes</protein> were not formed .


Finally , immunoblot analysis of nuclear extracts showed decreased or absent p65 protein levels .

Finally , immunoblot analysis of nuclear extracts showed decreased or absent <protein>p65 protein</protein> levels .


As p65 complexes are transcriptionally active in comparison to the p50 homodimer , this novel finding may provide insight on the origin of abnormal cytokine or other gene transcription in SLE patients .

As <protein>p65 complexes</protein> are transcriptionally active in comparison to the p50 homodimer , this novel finding may provide insight on the origin of <protein>abnormal cytokine</protein> or other gene transcription in SLE patients .


Signaling events induced by lipopolysaccharide-activated toll-like receptor 2 .

Signaling events induced by <protein>lipopolysaccharide-activated toll-like receptor 2</protein> .


Human Toll-like receptor 2 ( TLR2 ) is a signaling receptor that responds to LPS and activates NF-kappaB .

<protein>Human Toll-like receptor 2 ( TLR2 )</protein> is a <protein>signaling receptor</protein> that responds to LPS and activates <protein>NF-kappaB</protein> .


Here , we investigate further the events triggered by TLR2 in response to LPS .

Here , we investigate further the events triggered by <protein>TLR2</protein> in response to LPS .


We show that TLR2 associates with the high-affinity LPS binding protein membrane CD14 to serve as an LPS receptor complex , and that LPS treatment enhances the oligomerization of TLR2 .

We show that <protein>TLR2</protein> associates with the <protein>high-affinity LPS binding protein</protein> membrane <protein>CD14</protein> to serve as an <protein>LPS receptor complex</protein> , and that LPS treatment enhances the oligomerization of <protein>TLR2</protein> .


Concomitant with receptor oligomerization , the IL-1R-associated kinase ( IRAK ) is recruited to the TLR2 complex .

Concomitant with receptor oligomerization , the <protein>IL-1R-associated kinase ( IRAK )</protein> is recruited to the <protein>TLR2</protein> complex .


Intracellular deletion variants of TLR2 lacking C-terminal 13 or 141 aa fail to recruit IRAK , which is consistent with the inability of these mutants to transmit LPS cellular signaling .

Intracellular deletion variants of <protein>TLR2</protein> lacking C-terminal 13 or 141 aa fail to recruit <protein>IRAK</protein> , which is consistent with the inability of these mutants to transmit LPS cellular signaling .


Moreover , both deletion mutants could still form complexes with wild-type TLR2 and act in a dominant-negative ( DN ) fashion to block TLR2 -mediated signal transduction .

Moreover , both deletion mutants could still form complexes with <protein>wild-type TLR2</protein> and act in a dominant-negative ( DN ) fashion to block <protein>TLR2</protein> -mediated signal transduction .


DN constructs of myeloid differentiation protein , IRAK , TNF receptor-associated factor 6 , and NF-kappaB-inducing kinase , when coexpressed with TLR2 , abrogate TLR2 -mediated NF-kappaB activation .

DN constructs of <protein>myeloid differentiation protein</protein> , <protein>IRAK</protein> , <protein>TNF receptor-associated factor 6</protein> , and <protein>NF-kappaB-inducing kinase</protein> , when coexpressed with <protein>TLR2</protein> , abrogate <protein>TLR2</protein> -mediated <protein>NF-kappaB</protein> activation .


These results reveal a conserved signaling pathway for TLR2 and IL-1Rs and suggest a molecular mechanism for the inhibition of TLR2 by DN variants .

These results reveal a conserved signaling pathway for <protein>TLR2</protein> and <protein>IL-1Rs</protein> and suggest a molecular mechanism for the inhibition of <protein>TLR2</protein> by DN variants .


Tyrphostin AG-490 inhibits cytokine-mediated JAK3 /STAT5a/b signal transduction and cellular proliferation of antigen-activated human T cells .

Tyrphostin AG-490 inhibits cytokine-mediated <protein>JAK3</protein> <protein>/STAT5a/b</protein> signal transduction and cellular proliferation of antigen-activated human <cell_type>T cells</cell_type> .


Janus kinase 3 ( JAK3 ) is a cytoplasmic tyrosine kinase required for T cell development and activated by cytokines that utilize the interleukin-2 ( IL-2 ) receptor common gamma chain ( gamma ( c ) ) .

<protein>Janus kinase 3 ( JAK3 )</protein> is a <protein>cytoplasmic tyrosine kinase</protein> required for <cell_type>T cell</cell_type> development and activated by <protein>cytokines</protein> that utilize the <protein>interleukin-2 ( IL-2 ) receptor common gamma chain ( gamma ( c ) )</protein> .


Genetic inactivation of JAK3 is manifested as severe combined immunodeficiency disease ( SCID ) in humans and mice .

Genetic inactivation of <protein>JAK3</protein> is manifested as severe combined immunodeficiency disease ( SCID ) in humans and mice .


These findings have suggested that JAK3 represents a pharmacological target to control certain lymphoid-derived diseases .

These findings have suggested that <protein>JAK3</protein> represents a pharmacological target to control certain lymphoid-derived diseases .


Here we provide novel evidence that AG-490 potently inhibits the autokinase activity of JAK3 and tyrosine phosphorylation and DNA binding of signal transducer and activator of transcription 5a and 5b ( STAT5a/b ) .

Here we provide novel evidence that AG-490 potently inhibits the autokinase activity of <protein>JAK3</protein> and tyrosine phosphorylation and DNA binding of <protein>signal transducer and activator of transcription 5a and 5b ( STAT5a/b )</protein> .


Similar inhibitory effects were observed with other cytokines that use gamma ( c ) .

Similar inhibitory effects were observed with other <protein>cytokines</protein> that use <protein>gamma ( c )</protein> .


AG-490 also inhibited IL-2 -mediated proliferative growth in human T cells with an IC50 ) = 25 microM that was partially recoverable .

AG-490 also inhibited <protein>IL-2</protein> -mediated proliferative growth in human <cell_type>T cells</cell_type> with an IC50 ) = 25 microM that was partially recoverable .


Moreover , we demonstrate that this inhibitor prevented tetanus toxoid antigen-specific T cell proliferation and expansion but failed to block activation of Zap70 or p56Lck after anti-CD3 stimulation of human T cells .

Moreover , we demonstrate that this inhibitor prevented tetanus toxoid antigen-specific T cell proliferation and expansion but failed to block activation of <protein>Zap70</protein> or <protein>p56Lck</protein> after <protein>anti-CD3</protein> stimulation of human <cell_type>T cells</cell_type> .


Taken together , these findings suggest that AG-490 inhibits the JAK3 -mediated Type II signaling pathway but not the T cell receptor-derived Type I pathway and possesses therapeutic potential for T cell -derived pathologies such as graft-versus-host disease , allergy , and autoimmune disorders .

Taken together , these findings suggest that AG-490 inhibits the <protein>JAK3</protein> -mediated Type II signaling pathway but not the <cell_type>T cell</cell_type> receptor-derived Type I pathway and possesses therapeutic potential for <cell_type>T cell</cell_type> -derived pathologies such as graft-versus-host disease , allergy , and autoimmune disorders .


C/EBP beta in rheumatoid arthritis : correlation with inflammation , not disease specificity .

<protein>C/EBP beta</protein> in rheumatoid arthritis : correlation with inflammation , not disease specificity .


Rheumatoid arthritis synovial tissue was examined and compared with osteoarthritis tissue for the presence of the nuclear transcription factor C/EBP beta ( NF-IL-6 ) .

Rheumatoid arthritis synovial tissue was examined and compared with osteoarthritis tissue for the presence of the <protein>nuclear transcription factor C/EBP beta ( NF-IL-6 )</protein> .


The region ( lining or sublining ) , cell type , and subcellular distribution ( cytoplasmic or nuclear ) of the expression of C/EBP beta was characterized .

The region ( lining or sublining ) , cell type , and subcellular distribution ( cytoplasmic or nuclear ) of the expression of <protein>C/EBP beta</protein> was characterized .


Rheumatoid arthritis synovial fluid and blood and normal peripheral blood were also examined .


C/EBP beta was detected in the synovial lining and in sublining cells of synovial tissue from patients with both rheumatoid and osteoarthritis .

<protein>C/EBP beta</protein> was detected in the synovial lining and in <cell_type>sublining cells of synovial tissue</cell_type> from patients with both rheumatoid and osteoarthritis .


A significant ( P < 0.001 and < 0.05 , respectively ) increase in the percentage of cells with nuclear staining was seen in the lining layer , compared to cells in the sublining region , in rheumatoid and osteoarthritis .


In both diseases a strong correlation ( r = 0.79 , P < 0.001 ) was observed between the percentage of cells in the synovial lining that were positive for nuclear C/EBP beta and lining cell depth .

In both diseases a strong correlation ( r = 0.79 , P < 0.001 ) was observed between the percentage of cells in the synovial lining that were positive for <protein>nuclear C/EBP beta</protein> and lining cell depth .


Two-color immunohistochemistry demonstrated that both macrophages and fibroblast-like synoviocytes were positive for nuclear C/EBP beta .

Two-color immunohistochemistry demonstrated that both <cell_type>macrophages</cell_type> and <cell_type>fibroblast-like synoviocytes</cell_type> were positive for nuclear <protein>C/EBP beta</protein> .


The presence of C/EBP beta was confirmed by immunohistochemistry and Western blot analysis with isolated synovial fibroblasts .

The presence of <protein>C/EBP beta</protein> was confirmed by immunohistochemistry and Western blot analysis with isolated <cell_type>synovial fibroblasts</cell_type> .


Nuclear C/EBP beta was also detected in rheumatoid synovial fluid monocytes/macrophages , but not in lymphocytes or neutrophils .

Nuclear <protein>C/EBP beta</protein> was also detected in <cell_type>rheumatoid synovial fluid monocytes/macrophages , but not in lymphocytes or neutrophils</cell_type> .


Western blot analysis confirmed the presence of C/EBP beta in these cells .

Western blot analysis confirmed the presence of <protein>C/EBP beta</protein> in these cells .


The intensity of C/EBP beta staining was greater ( P < 0.001 ) in synovial fluid monocytes than in those from normal or rheumatoid peripheral blood .

The intensity of <protein>C/EBP beta</protein> staining was greater ( P < 0.001 ) in <cell_type>synovial fluid monocytes</cell_type> than in those from <cell_type>normal or rheumatoid peripheral blood</cell_type> .


In conclusion , the enhanced nuclear staining for C/EBP beta in the synovial lining , compared to the sublining , suggesting activation in the lining , and the positive correlation of lining layer depth with the percentage of cells in the lining positive for nuclear C/EBP beta , suggest a potential role for C/EBP beta in chronic inflammation .

In conclusion , the enhanced nuclear staining for <protein>C/EBP beta</protein> in the synovial lining , compared to the sublining , suggesting activation in the lining , and the positive correlation of lining layer depth with the percentage of cells in the lining positive for nuclear <protein>C/EBP beta</protein> , suggest a potential role for <protein>C/EBP beta</protein> in chronic inflammation .


The regulation of the production or activity of C/EBP beta , to inhibit inflammatory mediator expression by synovial macrophages and fibroblasts , offers a novel approach to therapeutic intervention .

The regulation of the production or activity of <protein>C/EBP beta</protein> , to inhibit inflammatory mediator expression by <cell_type>synovial macrophages</cell_type> and <cell_type>fibroblasts</cell_type> , offers a novel approach to therapeutic intervention .


Direct interaction of hematopoietic transcription factors PU.1 and GATA-1 : functional antagonism in erythroid cells .

Direct interaction of <protein>hematopoietic transcription factors PU.1 and GATA-1</protein> : functional antagonism in <cell_type>erythroid cells</cell_type> .


Malignant transformation usually inhibits terminal cell differentiation but the precise mechanisms involved are not understood .


PU.1 is a hematopoietic-specific Ets family transcription factor that is required for development of some lymphoid and myeloid lineages .

<protein>PU.1</protein> is a <protein>hematopoietic-specific Ets family transcription factor</protein> that is required for development of some <cell_type>lymphoid and myeloid lineages</cell_type> .


PU.1 can also act as an oncoprotein as activation of its expression in erythroid precursors by proviral insertion or transgenesis causes erythroleukemias in mice .

<protein>PU.1</protein> can also act as an <protein>oncoprotein</protein> as activation of its expression in erythroid precursors by proviral insertion or transgenesis causes erythroleukemias in mice .


Restoration of terminal differentiation in the mouse erythroleukemia ( MEL ) cells requires a decline in the level of PU.1 , indicating that PU.1 can block erythroid differentiation .

Restoration of terminal differentiation in the <cell_line>mouse erythroleukemia ( MEL ) cells</cell_line> requires a decline in the level of <protein>PU.1</protein> , indicating that <protein>PU.1</protein> can block erythroid differentiation .


Here we investigate the mechanism by which PU.1 interferes with erythroid differentiation .

Here we investigate the mechanism by which <protein>PU.1</protein> interferes with erythroid differentiation .


We find that PU.1 interacts directly with GATA-1 , a zinc finger transcription factor required for erythroid differentiation .

We find that <protein>PU.1</protein> interacts directly with <protein>GATA-1</protein> , a <protein>zinc finger transcription factor</protein> required for erythroid differentiation .


Interaction between PU.1 and GATA-1 requires intact DNA-binding domains in both proteins .

Interaction between <protein>PU.1</protein> and <protein>GATA-1</protein> requires intact <protein>DNA-binding domains</protein> in both proteins .


PU.1 represses GATA-1 -mediated transcriptional activation .

<protein>PU.1</protein> represses <protein>GATA-1</protein> -mediated transcriptional activation .


Both the DNA binding and transactivation domains of PU.1 are required for repression and both domains are also needed to block terminal differentiation in MEL cells .

Both the <protein>DNA binding</protein> and <protein>transactivation domains</protein> of <protein>PU.1</protein> are required for repression and both domains are also needed to block terminal differentiation in <cell_line>MEL cells</cell_line> .


We also show that ectopic expression of PU.1 in Xenopus embryos is sufficient to block erythropoiesis during normal development .

We also show that ectopic expression of <protein>PU.1</protein> in Xenopus embryos is sufficient to block erythropoiesis during normal development .


Furthermore , introduction of exogenous GATA-1 in both MEL cells and Xenopus embryos and explants relieves the block to erythroid differentiation imposed by PU.1 .

Furthermore , introduction of exogenous <protein>GATA-1</protein> in both <cell_line>MEL cells</cell_line> and Xenopus embryos and explants relieves the block to erythroid differentiation imposed by <protein>PU.1</protein> .


Our results indicate that the stoichiometry of directly interacting but opposing transcription factors may be a crucial determinant governing processes of normal differentiation and malignant transformation .

Our results indicate that the stoichiometry of directly interacting but opposing <protein>transcription factors</protein> may be a crucial determinant governing processes of normal differentiation and malignant transformation .


IL-12 induces IFN regulating factor-1 ( IRF-1 ) gene expression in human NK and T cells .

<protein>IL-12</protein> induces <protein>IFN regulating factor-1 ( IRF-1 )</protein> gene expression in human <cell_type>NK</cell_type> and <cell_type>T cells</cell_type> .


IL-12 is a critical immunoregulatory cytokine that promotes cell-mediated immune responses and the differentiation of naive CD4+ cells to Th1 cells ; however , relatively few IL-12 target genes have been identified .

<protein>IL-12</protein> is a <protein>critical immunoregulatory cytokine</protein> that promotes cell-mediated immune responses and the differentiation of naive <cell_type>CD4+ cells</cell_type> to <cell_type>Th1 cells</cell_type> ; however , relatively few <protein>IL-12</protein> target genes have been identified .


To better clarify the molecular basis of IL-12 action , we set out to characterize genes up-regulated by IL-12 , first by contrasting IL-12 -and IFN-alpha-inducible genes .

To better clarify the molecular basis of <protein>IL-12</protein> action , we set out to characterize genes up-regulated by <protein>IL-12</protein> , first by contrasting <dna>IL-12</dna> -and <dna>IFN-alpha-inducible genes</dna> .


We identified several genes up-regulated by IL-12 , namely , MIP-1alpha , MIP-1beta , IL-1RA , and IFN regulatory factor-1 ( IRF-1 ) .

We identified several genes up-regulated by <protein>IL-12</protein> , namely , <protein>MIP-1alpha</protein> , <protein>MIP-1beta</protein> , <protein>IL-1RA</protein> , and <protein>IFN regulatory factor-1 ( IRF-1 )</protein> .


IRF-1 is a transcription factor regulated by IFNs that is also essential for Th1 responses .

<protein>IRF-1</protein> is a <protein>transcription factor</protein> regulated by <protein>IFNs</protein> that is also essential for <cell_type>Th1</cell_type> responses .


We demonstrated that IL-12 directly up-regulates IRF-1 to the same extent as IFN-alpha in normal human T cells and in NK cells .

We demonstrated that <protein>IL-12</protein> directly up-regulates <protein>IRF-1</protein> to the same extent as <protein>IFN-alpha</protein> in normal human <cell_type>T cells</cell_type> and in <cell_type>NK cells</cell_type> .


We showed that IL-12 had a direct effect on IRF-1 , an effect not mediated indirectly by the induction of IFN-gamma production .

We showed that <protein>IL-12</protein> had a direct effect on <protein>IRF-1</protein> , an effect not mediated indirectly by the induction of <protein>IFN-gamma</protein> production .


Furthermore , IL-2 and IL-12 synergistically induced IRF-1 , whereas IFN-alpha and IL-12 did not .

Furthermore , <protein>IL-2</protein> and <protein>IL-12</protein> synergistically induced <protein>IRF-1</protein> , whereas <protein>IFN-alpha</protein> and <protein>IL-12</protein> did not .


The participation of STAT4 in the regulation of IRF-1 was demonstrated in two ways .

The participation of <protein>STAT4</protein> in the regulation of <protein>IRF-1</protein> was demonstrated in two ways .


First , STAT4 was required for the IL-12 -dependent transactivation of an IRF-1 reporter construct , and second , STAT4 binding to the IRF-1 promoter was shown using EMSA .

First , <protein>STAT4</protein> was required for the <protein>IL-12</protein> -dependent transactivation of an <dna>IRF-1 reporter construct</dna> , and second , <protein>STAT4</protein> binding to the <dna>IRF-1 promoter</dna> was shown using EMSA .


In contrast to IL-12 , no up-regulation of IRF-1 was found in IL-4-stimulated cells , and IL-4 did not block IL-12 -dependent up-regulation of IRF-1 .

In contrast to <protein>IL-12</protein> , no up-regulation of <protein>IRF-1</protein> was found in <cell_line>IL-4-stimulated cells</cell_line> , and <protein>IL-4</protein> did not block <protein>IL-12</protein> -dependent up-regulation of <protein>IRF-1</protein> .


Therefore , IRF-1 may be an important contributor to IL-12 signaling , and we speculate that the defective IL-12 responses seen in IRF -1-/- mice might be attributable , in part , to the absence of this transcription factor .

Therefore , <protein>IRF-1</protein> may be an important contributor to <protein>IL-12</protein> signaling , and we speculate that the defective <protein>IL-12</protein> responses seen in <protein>IRF</protein> -1-/- mice might be attributable , in part , to the absence of this <protein>transcription factor</protein> .


Targeted remodeling of human beta-globin promoter chromatin structure produces increased expression and decreased silencing .


The chromatin structure of the human beta-globin gene locus assumes a transcriptionally-active conformation in erythroid cells .

The chromatin structure of the <dna>human beta-globin gene locus</dna> assumes a transcriptionally-active conformation in <cell_type>erythroid cells</cell_type> .


One feature of this chromatin reorganization is the formation of DNase 1 hypersensitive sites in the regions of active globin gene promoters .

One feature of this chromatin reorganization is the formation of <dna>DNase 1 hypersensitive sites</dna> in the regions of <dna>active globin gene promoters</dna> .


This reorganization requires the globin locus control region and is associated with normal expression of the beta-like globin genes .

This reorganization requires the <dna>globin locus control region</dna> and is associated with normal expression of the <dna>beta-like globin genes</dna> .


To determine whether it is possible to artificially enhance the opening of the chromatin structure of a minimal beta-globin promoter , we placed a 101bp , erythroid-specific DNase 1 hypersensitive site-forming element ( HSFE ) immediately upstream of the beta-globin promoter and gene .

To determine whether it is possible to artificially enhance the opening of the chromatin structure of a <dna>minimal beta-globin promoter</dna> , we placed a 101bp , <dna>erythroid-specific DNase 1 hypersensitive site-forming element</dna> ( <dna>HSFE</dna> ) immediately upstream of the <dna>beta-globin promoter and gene</dna> .


This element includes binding sites for NF-E2 , AP-1 , GATA-1 and Sp-1 .

This element includes binding sites for <protein>NF-E2</protein> , <protein>AP-1</protein> , <protein>GATA-1</protein> and <protein>Sp-1</protein> .


Constructs were stably transfected into murine erythroleukemia cells and promoter chromatin structure and gene expression were analyzed .

Constructs were stably transfected into <cell_type>murine erythroleukemia cells</cell_type> and promoter chromatin structure and gene expression were analyzed .


The HSFE induced an area of enhanced DNase 1 hypersensitivity extending from the transcriptional start site to -300bp of the artificial promoter and significantly increased the proportion of beta-globin promoters in an open chromatin configuration .

The <dna>HSFE</dna> induced an area of enhanced <protein>DNase 1</protein> hypersensitivity extending from the transcriptional start site to -300bp of the <dna>artificial promoter</dna> and significantly increased the proportion of <dna>beta-globin promoters</dna> in an open chromatin configuration .


This remodeling of promoter chromatin structure resulted in 3-fold increases in beta-globin gene transcription and induction , and inhibited long-term beta-globin gene silencing .

This remodeling of promoter chromatin structure resulted in 3-fold increases in <dna>beta-globin gene</dna> transcription and induction , and inhibited long-term <dna>beta-globin gene</dna> silencing .


These results indicate that a relatively small cis-acting element is able to enhance remodeling of promoter chromatin structure resulting in increased beta-globin gene expression .

These results indicate that a relatively small <dna>cis-acting element</dna> is able to enhance remodeling of promoter chromatin structure resulting in increased <dna>beta-globin gene</dna> expression .