Regulation of low shear flow-induced HAEC VCAM-1 expression and monocyte adhesion .

Regulation of low shear flow-induced <cell_line>HAEC</cell_line> <protein>VCAM-1</protein> expression and monocyte adhesion .


We recently reported that prolonged exposure of human aortic endothelial cells ( HAEC ) to low shear stress flow patterns is associated with a sustained increase in the activated form of the transcriptional regulator nuclear factor-kappaB ( NF-kappaB ) .

We recently reported that prolonged exposure of <cell_type>human aortic endothelial cells</cell_type> ( <cell_line>HAEC</cell_line> ) to low shear stress flow patterns is associated with a sustained increase in the activated form of the transcriptional regulator <protein>nuclear factor-kappaB</protein> ( <protein>NF-kappaB</protein> ) .


Here we investigate the hypothesis that low shear-induced activation of NF-kappaB is responsible for enhanced expression of vascular cell adhesion molecule ( VCAM-1 ) resulting in augmented endothelial cell-monocyte ( EC-Mn ) adhesion and that this activation is dependent on intracellular oxidant activity .

Here we investigate the hypothesis that low shear-induced activation of <protein>NF-kappaB</protein> is responsible for enhanced expression of <protein>vascular cell adhesion molecule</protein> ( <protein>VCAM-1</protein> ) resulting in augmented endothelial cell-monocyte ( EC-Mn ) adhesion and that this activation is dependent on intracellular oxidant activity .


Before exposure to low shear ( 2 dyn/cm2 ) for 6 h , HAEC were preincubated with or without the antioxidants pyrrolidine dithiocarbamate ( PDTC ) or N-acetyl-L-cysteine ( NAC ) .

Before exposure to low shear ( 2 dyn/cm2 ) for 6 h , <cell_line>HAEC</cell_line> were preincubated with or without the antioxidants pyrrolidine dithiocarbamate ( PDTC ) or N-acetyl-L-cysteine ( NAC ) .


PDTC strongly inhibited low shear-induced activation of NF-kappaB , expression of VCAM-1 , and EC-Mn adhesion .

PDTC strongly inhibited low shear-induced activation of <protein>NF-kappaB</protein> , expression of <protein>VCAM-1</protein> , and EC-Mn adhesion .


Paradoxically , NAC exerted a positive effect on low shear-induced VCAM-1 expression and EC-Mn adhesion and only slightly downregulated NF-kappaB activation .

Paradoxically , NAC exerted a positive effect on low shear-induced <protein>VCAM-1</protein> expression and EC-Mn adhesion and only slightly downregulated <protein>NF-kappaB</protein> activation .


However , cytokine-induced NF-kappaB activation and VCAM-1 expression are blocked by both PDTC and NAC .

However , cytokine-induced <protein>NF-kappaB</protein> activation and <protein>VCAM-1</protein> expression are blocked by both PDTC and NAC .


These data suggest that NF-kappaB plays a key role in low shear-induced VCAM-1 expression and that pathways mediating low shear- and cytokine-induced EC-Mn adhesion may be differentially regulated .

These data suggest that <protein>NF-kappaB</protein> plays a key role in low shear-induced <protein>VCAM-1</protein> expression and that pathways mediating low shear- and cytokine-induced EC-Mn adhesion may be differentially regulated .


Nuclear factor-90 of activated T-cells : A double-stranded RNA -binding protein and substrate for the double-stranded RNA-dependent protein kinase , PKR .

<protein>Nuclear factor-90</protein> of <cell_type>activated T-cells</cell_type> : A <rna>double-stranded RNA</rna> -binding protein and substrate for the <protein>double-stranded RNA-dependent protein kinase</protein> , <protein>PKR</protein> .


NFAT transcription factors play a central role in initiating T-cell activation through the induction of immediate-early T-cell specific genes including interleukin-2 ( IL-2 ) .

NFAT transcription factors play a central role in initiating T-cell activation through the induction of immediate-early T-cell specific genes including interleukin-2 ( <protein>IL-2</protein> ) .


NFAT transcription factors bind to a sequence in the IL-2 enhancer known as the antigen receptor response element 2 ( ARRE-2 ) .

NFAT transcription factors bind to a sequence in the <dna>IL-2 enhancer</dna> known as the <dna>antigen receptor response element 2</dna> ( <dna>ARRE-2</dna> ) .


Multiple proteins exhibiting ARRE-2 binding activity have been isolated , including a heterodimer from stimulated T-cell nuclear extracts consisting of Mr = 90 000 ( NF90 ) and Mr = 45 000 ( NF45 ) subunits .

Multiple proteins exhibiting <dna>ARRE-2</dna> binding activity have been isolated , including a <protein>heterodimer</protein> from stimulated T-cell nuclear extracts consisting of Mr = 90 000 ( <protein>NF90</protein> ) and Mr = 45 000 ( <protein>NF45</protein> ) subunits .


The subunits of this heterodimer have been cloned , and NF90 was found to encode a protein containing two domains that are predicted to form motifs capable of binding to double-stranded RNA .

The subunits of this <protein>heterodimer</protein> have been cloned , and <protein>NF90</protein> was found to encode a protein containing two domains that are predicted to form motifs capable of binding to <rna>double-stranded RNA</rna> .


Using in vitro translated polypeptides , we have demonstrated that NF90 specifically binds to double-stranded RNA .

Using in vitro translated polypeptides , we have demonstrated that <protein>NF90</protein> specifically binds to <rna>double-stranded RNA</rna> .


Furthermore , NF90 was phosphorylated in a double-stranded RNA -dependent manner likely by the interferon-induced , double-stranded RNA-dependent protein kinase , PKR .

Furthermore , <protein>NF90</protein> was phosphorylated in a <rna>double-stranded RNA</rna> -dependent manner likely by the interferon-induced , <protein>double-stranded RNA-dependent protein kinase</protein> , <protein>PKR</protein> .


The NF90 protein was found to be expressed not only in T-cells , but also in nonimmune HeLa cells .

The <protein>NF90</protein> protein was found to be expressed not only in <cell_type>T-cells</cell_type> , but also in <cell_line>nonimmune HeLa cells</cell_line> .


In HeLa cells , the protein was almost exclusively localized to the ribosome salt wash fraction of cell lysates .

In <cell_line>HeLa cells</cell_line> , the protein was almost exclusively localized to the ribosome salt wash fraction of cell lysates .


NF-kappaB activation is required for C5a-induced interleukin-8 gene expression in mononuclear cells .

<protein>NF-kappaB</protein> activation is required for <dna>C5a-induced interleukin-8 gene</dna> expression in <cell_type>mononuclear cells</cell_type> .


C5a , a potent peptide chemoattractant , stimulates interleukin-8 ( IL-8 ) secretion from peripheral blood mononuclear cells ( PBMC ) .

C5a , a potent peptide chemoattractant , stimulates <protein>interleukin-8</protein> ( <protein>IL-8</protein> ) secretion from <cell_type>peripheral blood mononuclear cells</cell_type> ( <cell_type>PBMC</cell_type> ) .


Experiments were conducted to understand the mechanisms for C5a-induced IL-8 production , which was 14-fold greater than that in unstimulated cells by 2 hours .

Experiments were conducted to understand the mechanisms for C5a-induced <protein>IL-8</protein> production , which was 14-fold greater than that in <cell_type>unstimulated cells</cell_type> by 2 hours .


IL-8 secretion was accompanied by accumulation of IL-8 mRNA in the cytosol and by nuclear expression of a kappaB DNA binding activity within 30 minutes .

<protein>IL-8</protein> secretion was accompanied by accumulation of <rna>IL-8 mRNA</rna> in the cytosol and by nuclear expression of a <protein>kappaB</protein> DNA binding activity within 30 minutes .


AP-1 but not NF-IL-6 DNA binding activity was also detected in C5a-stimulated PBMC ; however , its delayed expression ( maximal at 4 hours ) suggested a less important role in the rapid production of IL-8 .

<protein>AP-1</protein> but not <protein>NF-IL-6</protein> DNA binding activity was also detected in <cell_line>C5a-stimulated PBMC</cell_line> ; however , its delayed expression ( maximal at 4 hours ) suggested a less important role in the rapid production of <protein>IL-8</protein> .


The correlation between C5a-induced kappaB binding activity and IL-8 gene expression was examined in the RAW264.7 macrophage cells using reporter genes directed by the kappaB sequence from IkappaBalpha and IL-8 promoter regions .

The correlation between C5a-induced <protein>kappaB</protein> binding activity and <protein>IL-8</protein> gene expression was examined in the <cell_line>RAW264.7 macrophage cells</cell_line> using reporter genes directed by the <dna>kappaB sequence</dna> from <protein>IkappaBalpha</protein> and <dna>IL-8 promoter regions</dna> .


C5a-induced reporter gene expression was abolished by introducing mutations into the kappaB sites and by coexpression of a dominant negative IkappaBalpha construct resistant to agonist-induced phosphorylation .

C5a-induced <dna>reporter gene</dna> expression was abolished by introducing mutations into the <dna>kappaB sites</dna> and by coexpression of a <dna>dominant negative IkappaBalpha construct</dna> resistant to agonist-induced phosphorylation .


Pertussis toxin , which ADP-ribosylates the Gi proteins known to couple to the C5a receptor , produced minimal inhibition of C5a-induced IL-8 expression and had little effect on C5a-induced calcium mobilization in RAW264.7 cells .

Pertussis toxin , which ADP-ribosylates the <protein>Gi proteins</protein> known to couple to the <protein>C5a receptor</protein> , produced minimal inhibition of C5a-induced <protein>IL-8</protein> expression and had little effect on C5a-induced calcium mobilization in <cell_line>RAW264.7 cells</cell_line> .


These results suggest that NF-kappaB activation is required for C5a-induced IL-8 gene expression and that this response is mediated primarily through a pertussis toxin-insensitive pathway .

These results suggest that <protein>NF-kappaB</protein> activation is required for C5a-induced <protein>IL-8</protein> gene expression and that this response is mediated primarily through a pertussis toxin-insensitive pathway .


Increased glucocorticoid receptor beta in airway cells of glucocorticoid-insensitive asthma .

Increased <protein>glucocorticoid receptor beta</protein> in <cell_type>airway cells</cell_type> of glucocorticoid-insensitive asthma .


Glucocorticoid ( GC ) -insensitive asthma is a challenging clinical problem that can be associated with life-threatening disease progression .


The molecular basis of GC insensitivity is unknown .


Alternative splicing of the GC receptor ( GCR ) pre-mRNA generates a second GCR , termed GCRbeta , which does not bind GC but antagonizes the transactivating activity of the classic GCR .

Alternative splicing of the <rna>GC receptor ( GCR ) pre-mRNA</rna> generates a second <protein>GCR</protein> , termed <protein>GCRbeta</protein> , which does not bind GC but antagonizes the transactivating activity of the classic <protein>GCR</protein> .


Thus increased expression of GCRbeta could account for glucocorticoid insensitivity .

Thus increased expression of <protein>GCRbeta</protein> could account for glucocorticoid insensitivity .


Bronchoalveolar lavage ( BAL ) cells and peripheral blood mononuclear cells ( PBMC ) were examined for GCRbeta immunoreactivity using a GCRbeta-specific antibody by immunohistochemical staining .

<cell_type>Bronchoalveolar lavage ( BAL ) cells</cell_type> and peripheral blood <cell_type>mononuclear cells</cell_type> ( <cell_type>PBMC</cell_type> ) were examined for <protein>GCRbeta</protein> immunoreactivity using a <protein>GCRbeta-specific antibody</protein> by immunohistochemical staining .


Cell localization of GCRbeta expression was performed using a double immunostaining technique .

Cell localization of <protein>GCRbeta</protein> expression was performed using a double immunostaining technique .


Patients with GC-insensitive asthma expressed a significantly higher number of GCRbeta-immunoreactive cells in their BAL and peripheral blood than GC-sensitive asthmatics or normal control subjects .

Patients with GC-insensitive asthma expressed a significantly higher number of <cell_type>GCRbeta-immunoreactive cells</cell_type> in their BAL and peripheral blood than GC-sensitive asthmatics or normal control subjects .


Furthermore , GCRbeta expression in GC-insensitive asthma was particularly high in airway T cells , which are thought to play a major role in the pathogenesis of asthma .

Furthermore , <protein>GCRbeta</protein> expression in GC-insensitive asthma was particularly high in airway <cell_type>T cells</cell_type> , which are thought to play a major role in the pathogenesis of asthma .


We also examined the expression of GCRbeta in specimens from the airways of patients with chronic bronchitis .

We also examined the expression of <protein>GCRbeta</protein> in specimens from the airways of patients with chronic bronchitis .


In chronic bronchitis , few cells were GCRbeta-positive and their numbers did not differ significantly from normal control subjects .

In chronic bronchitis , few cells were <cell_type>GCRbeta-positive</cell_type> and their numbers did not differ significantly from normal control subjects .


We conclude that GC-insensitive asthma is associated with increased expression of GCRbeta in airway T cells .

We conclude that GC-insensitive asthma is associated with increased expression of <protein>GCRbeta</protein> in airway <cell_type>T cells</cell_type> .


Activation of the Janus kinase 3 -STAT5a pathway after CD40 triggering of human monocytes but not of resting B cells .

Activation of the <protein>Janus kinase 3</protein> <protein>-STAT5a</protein> pathway after <protein>CD40</protein> triggering of <cell_type>human monocytes</cell_type> but not of <cell_type>resting B cells</cell_type> .


CD40 /CD40 ligand interactions play a key role in the immune responses of B lymphocytes , monocytes , and dendritic cells .

<protein>CD40</protein> <protein>/CD40</protein> ligand interactions play a key role in the immune responses of <cell_type>B lymphocytes</cell_type> , <cell_type>monocytes</cell_type> , and <cell_type>dendritic cells</cell_type> .


The signal transduction events triggered by cross-linking of the CD40 receptor have been widely studied in B cell lines , but little is known about signaling following CD40 stimulation of monocytes and resting tonsillar B cells .

The signal transduction events triggered by cross-linking of the <protein>CD40 receptor</protein> have been widely studied in <cell_line>B cell lines</cell_line> , but little is known about signaling following <protein>CD40</protein> stimulation of <cell_type>monocytes</cell_type> and <cell_line>resting tonsillar B cells</cell_line> .


Therefore , we studied the CD40 pathway in highly purified human monocytes and resting B cells .

Therefore , we studied the <protein>CD40</protein> pathway in highly purified <cell_type>human monocytes</cell_type> and <cell_type>resting B cells</cell_type> .


After CD40 triggering , a similar activation of the NF-kappaB ( but not of the AP-1 ) transcription factor complex occurred in both cell preparations .

After <protein>CD40</protein> triggering , a similar activation of the <protein>NF-kappaB</protein> ( but not of the <protein>AP-1</protein> ) <protein>transcription factor complex</protein> occurred in both cell preparations .


However , the components of the NF-kappaB complexes were different in monocytes and B cells , because p50 is part of the NF-kappaB complex induced by CD40 triggering in both monocytes and B cells , whereas p65 was only induced in B cells . In contrast , although the Janus kinase 3 tyrosine kinase was associated with CD40 molecules in both monocytes and resting B cells , Janus kinase 3 phosphorylation induction was observed only in CD40-activated monocytes , with subsequent induction of STAT5a DNA binding activity in the nucleus .

However , the components of the <protein>NF-kappaB complexes</protein> were different in <cell_type>monocytes</cell_type> and B cells , because <protein>p50</protein> is part of the <protein>NF-kappaB complex</protein> induced by <protein>CD40</protein> triggering in both <cell_type>monocytes</cell_type> and <cell_type>B cells</cell_type> , whereas <protein>p65</protein> was only induced in <cell_type>B cells</cell_type> . In contrast , although the <protein>Janus kinase 3 tyrosine kinase</protein> was associated with <protein>CD40</protein> molecules in both <cell_type>monocytes</cell_type> and <cell_type>resting B cells</cell_type> , <protein>Janus kinase 3</protein> phosphorylation induction was observed only in <cell_line>CD40-activated monocytes</cell_line> , with subsequent induction of <protein>STAT5a</protein> DNA binding activity in the nucleus .


These results suggest that the activation signals in human B cells and monocytes differ following CD40 stimulation .

These results suggest that the activation signals in <cell_type>human B cells</cell_type> and <cell_type>monocytes</cell_type> differ following <protein>CD40</protein> stimulation .


This observation is consistent with the detection of normal CD40 -induced monocyte activation in patients with CD40 ligand+ hyper IgM syndrome in whom a defect in CD40 -induced B cell activation has been reported .

This observation is consistent with the detection of normal <protein>CD40</protein> -induced monocyte activation in patients with <protein>CD40</protein> ligand+ hyper <protein>IgM</protein> syndrome in whom a defect in <protein>CD40</protein> -induced B cell activation has been reported .


Genetic evidence for an additional factor required for erythropoietin -induced signal transduction .

Genetic evidence for an additional factor required for <protein>erythropoietin</protein> -induced signal transduction .


Erythropoietin ( EPO ) and its receptor ( EPOR ) are required for the development of mature erythrocytes .

<protein>Erythropoietin</protein> ( <protein>EPO</protein> ) and its receptor ( <protein>EPOR</protein> ) are required for the development of mature erythrocytes .


After binding of ligand , the EPOR activates a variety of signaling pathways that ultimately control cellular proliferation , survival , and specific gene expression .

After binding of ligand , the <protein>EPOR</protein> activates a variety of signaling pathways that ultimately control cellular proliferation , survival , and specific gene expression .


Although erythroid progenitors appear to be the principal EPO-responsive cell type in vivo due to the restricted expression of the EPOR , many growth factor-dependent cell lines expressing the EPOR can respond to EPO by activating many or all of these pathways .

Although <cell_type>erythroid progenitors</cell_type> appear to be the principal <cell_type>EPO-responsive cell type</cell_type> in vivo due to the restricted expression of the <protein>EPOR</protein> , many <cell_line>growth factor-dependent cell lines</cell_line> expressing the <protein>EPOR</protein> can respond to <protein>EPO</protein> by activating many or all of these pathways .


In the present study , we have identified a cellular context ( the interleukin-2 [ IL-2 ] -dependent HT-2 line ) in which the EPO stimulation of the EPOR fails to support cellular proliferation , STAT-5 induction , or MAPK activation , despite efficient phosphorylation of the EPOR and JAK2 and inhibition of apoptosis after withdrawal of IL-2 .

In the present study , we have identified a cellular context ( the <cell_line>interleukin-2 [ IL-2 ] -dependent HT-2 line</cell_line> ) in which the <protein>EPO</protein> stimulation of the <protein>EPOR</protein> fails to support cellular proliferation , <protein>STAT-5</protein> induction , or <protein>MAPK</protein> activation , despite efficient phosphorylation of the <protein>EPOR</protein> and <protein>JAK2</protein> and inhibition of apoptosis after withdrawal of <protein>IL-2</protein> .


Interestingly , when we fused HT-2 cells expressing the EPOR with Ba/F3 cells in a complementation assay , the resulting hybridomas proliferated and potently activated STAT-5 and MAPK in response to EPO .

Interestingly , when we fused <cell_line>HT-2 cells</cell_line> expressing the <protein>EPOR</protein> with <cell_line>Ba/F3 cells</cell_line> in a complementation assay , the resulting <cell_line>hybridomas</cell_line> proliferated and potently activated <protein>STAT-5</protein> and <protein>MAPK</protein> in response to <protein>EPO</protein> .


These data indicate that an unidentified cellular factor is needed to mediate signaling by the EPOR .

These data indicate that an <protein>unidentified cellular factor</protein> is needed to mediate signaling by the <protein>EPOR</protein> .


Moreover , Ba/F3 cells apparently express this factor ( s ) and somatic fusions can , therefore , confer EPO -responsiveness to HT-2 cells that lack this factor .

Moreover , <cell_line>Ba/F3 cells</cell_line> apparently express this factor ( s ) and somatic fusions can , therefore , confer <protein>EPO</protein> -responsiveness to <cell_line>HT-2 cells</cell_line> that lack this factor .


Novel therapies for inflammatory bowel disease .


Looking back at successes and failures in newer approaches to treating IBD , it is tempting -- although still difficult -- to draw conclusions about pathogenesis .


When a therapy proves effective , do clinicians truly know how it works ? Even with a therapy as specific as anti-TNF antibody , it is not clear if the benefit is attributable to simple binding and clearance of TNF-alpha or to binding on the cell surface and subsequent deletion of the activated macrophage .

When a therapy proves effective , do clinicians truly know how it works ? Even with a therapy as specific as <protein>anti-TNF antibody</protein> , it is not clear if the benefit is attributable to simple binding and clearance of <protein>TNF-alpha</protein> or to binding on the cell surface and subsequent deletion of the <cell_type>activated macrophage</cell_type> .


When a drug appears to be less effective than preclinical models suggest , can failures in effectiveness from delivery or dosing be differentiated ? The disappointing results of clinical trials with IL-10 -- so at odds with the prediction of benefit from animal models -- bring into question the validity of those models as well as the soundness of design of the clinical trials on which efficacy of IL-10 is judged .

When a drug appears to be less effective than preclinical models suggest , can failures in effectiveness from delivery or dosing be differentiated ? The disappointing results of clinical trials with <protein>IL-10</protein> -- so at odds with the prediction of benefit from animal models -- bring into question the validity of those models as well as the soundness of design of the clinical trials on which efficacy of <protein>IL-10</protein> is judged .


The variability of response even to the most narrowly targeted agents suggests that these diseases are far more heterogeneous in humans than in their murine counterparts .


Clinicians are only just beginning to recognize subclinical markers of response , and it may soon be possible to predict response on the basis of genetic composition .


For the moment , however , the field of pharmacogenetics is embryonic .


Challenges in developing new therapeutic strategies include not only identifying novel agents , but also improving the definitions of clinical endpoints and defining efficacy at the biologic level .


Only through considered evaluation of clinical evidence may clinicians determine which therapies should remain novelties and which should become an accepted part of the armamentarium .


Potent and stable attenuation of live-HIV-1 by gain of a proteolysis-resistant inhibitor of NF-kappaB ( IkappaB-alphaS32/36A ) and the implications for vaccine development .

Potent and stable attenuation of live-HIV-1 by gain of a <protein>proteolysis-resistant inhibitor</protein> of <protein>NF-kappaB</protein> ( <protein>IkappaB-alphaS32/36A</protein> ) and the implications for vaccine development .


Live-attenuated human immunodeficiency viruses ( HIVs ) are candidates for Acquired Immunodeficiency Syndrome ( AIDS ) vaccine .


Based on the simian immunodeficiency virus ( SIV ) model for AIDS , loss-of-function ( e.g. deletion of accessory genes such as nef ) has been forwarded as a primary approach for creating enfeebled , but replication-competent , HIV-1/SIV .

Based on the simian immunodeficiency virus ( SIV ) model for AIDS , loss-of-function ( e.g. deletion of accessory genes such as <dna>nef</dna> ) has been forwarded as a primary approach for creating enfeebled , but replication-competent , HIV-1/SIV .


Regrettably , recent evidence suggests that loss-of-function alone is not always sufficient to prevent the emergence of virulent mutants .


New strategies that attenuate via mechanisms distinct from loss-of-function are needed for enhancing the safety phenotype of viral genome .

New strategies that attenuate via mechanisms distinct from loss-of-function are needed for enhancing the safety phenotype of <dna>viral genome</dna> .


Here , we propose gain-of-function to be used simultaneously with loss-of-function as a novel approach for attenuating HIV-1 .


We have constructed an HIV-1 genome carrying the cDNA of a proteolysis-resistant nuclear factor-kappaB inhibitor ( IkappaB-alphaS32/36A ) in the nef region .

We have constructed an <dna>HIV-1 genome</dna> carrying the <dna>cDNA</dna> of a <protein>proteolysis-resistant nuclear factor-kappaB inhibitor</protein> ( <protein>IkappaB-alphaS32/36A</protein> ) in the <dna>nef region</dna> .


HIV-1 expressing IkappaB-alphaS32/36A down-regulates viral expression and is highly attenuated in both Jurkat and peripheral blood mononuclear cells .

HIV-1 expressing <protein>IkappaB-alphaS32/36A</protein> down-regulates viral expression and is highly attenuated in both <cell_line>Jurkat</cell_line> and <cell_type>peripheral blood mononuclear cells</cell_type> .


We provide formal proof that the phenotypic and attenuating characteristics of IkappaB-alphaS32/36A permit its stable maintenance in a live , replicating HIV-1 despite 180 days of forced ex vivo passaging in tissue culture .

We provide formal proof that the phenotypic and attenuating characteristics of <protein>IkappaB-alphaS32/36A</protein> permit its stable maintenance in a live , replicating HIV-1 despite 180 days of forced ex vivo passaging in tissue culture .


As compared with other open-reading frames embedded into HIV/SIV genome , this degree of stability is unprecedented .

As compared with other <dna>open-reading frames</dna> embedded into <dna>HIV/SIV genome</dna> , this degree of stability is unprecedented .


Thus , IkappaB-alphaS32/36A offers proof-of-principle that artifactually gained functions , when used to attenuate the replication of live HIV-1 , can be stable .

Thus , <protein>IkappaB-alphaS32/36A</protein> offers proof-of-principle that artifactually gained functions , when used to attenuate the replication of live HIV-1 , can be stable .


These findings illustrate gain-of-function as a feasible strategy for developing safer live-attenuated HIVs to be tested as candidates for AIDS vaccine .


Multiple NF-ATc isoforms with individual transcriptional properties are synthesized in T lymphocytes .

Multiple <protein>NF-ATc isoforms</protein> with individual transcriptional properties are synthesized in <cell_type>T lymphocytes</cell_type> .


The transcription factor NF-ATc that controls gene expression in T lymphocytes and embryonic cardiac cells is expressed in three prominent isoforms .

The <protein>transcription factor</protein> <protein>NF-ATc</protein> that controls gene expression in <cell_type>T lymphocytes</cell_type> and <cell_type>embryonic cardiac cells</cell_type> is expressed in three prominent isoforms .


This is due to alternative splice/polyadenylation events that lead to the predominant synthesis of two long isoforms in naive T cells and a shorter NF-ATc isoform in effector T cells .

This is due to alternative splice/polyadenylation events that lead to the predominant synthesis of two long isoforms in <cell_type>naive T cells</cell_type> and a shorter <protein>NF-ATc isoform</protein> in <cell_type>effector T cells</cell_type> .


Whereas the previously described isoform NF-ATc/A contains a relatively short C terminus , the longer isoforms , B and C , span extra C-terminal peptides of 128 and 246 aa , respectively .

Whereas the previously described <protein>isoform NF-ATc/A</protein> contains a relatively short <protein>C terminus</protein> , the longer isoforms , <protein>B</protein> and <protein>C</protein> , span extra C-terminal peptides of 128 and 246 aa , respectively .


We show here that in addition to the strong N-terminal trans-activation domain , TAD-A , which is common to all three NF-ATc isoforms , NF-ATc/C contains a second trans-activation domain , TAD-B , in its C-terminal peptide .

We show here that in addition to the <protein>strong N-terminal trans-activation domain</protein> , <protein>TAD-A</protein> , which is common to all three <protein>NF-ATc isoforms</protein> , <protein>NF-ATc/C</protein> contains a second <protein>trans-activation domain</protein> , <protein>TAD-B</protein> , in its C-terminal peptide .


Various stimuli of T cells that induce the activity of TAD-A also enhance the activity of TAD-B , but , unlike TAD-A , TAD-B remains unphosphorylated by protein from 12-O-tetradecanoyl 12-phorbol 13-acetate-stimulated T cells .

Various stimuli of <cell_type>T cells</cell_type> that induce the activity of <protein>TAD-A</protein> also enhance the activity of <protein>TAD-B</protein> , but , unlike <protein>TAD-A</protein> , <protein>TAD-B</protein> remains unphosphorylated by protein from <cell_line>12-O-tetradecanoyl 12-phorbol 13-acetate-stimulated T cells</cell_line> .


The shorter C-terminal peptide of isoform NF-ATc/B exerts a suppressive transcriptional effect .

The shorter C-terminal peptide of isoform <protein>NF-ATc/B</protein> exerts a suppressive transcriptional effect .


These properties of NF-ATc/B and -C might be of importance for gene regulation in naive T lymphocytes in which NF-ATc/B and -C are predominantly synthesized .

These properties of <protein>NF-ATc/B and -C</protein> might be of importance for gene regulation in <cell_type>naive T lymphocytes</cell_type> in which <protein>NF-ATc/B and -C</protein> are predominantly synthesized .


Selection and long-term persistence of reactive CTL clones during an EBV chronic response are determined by avidity , CD8 variable contribution compensating for differences in TCR affinities .

Selection and long-term persistence of reactive <cell_line>CTL clones</cell_line> during an EBV chronic response are determined by avidity , <protein>CD8</protein> variable contribution compensating for differences in <protein>TCR</protein> affinities .


Recent studies have suggested that the diversity of TCR repertoire after primary immunization is conserved in memory T cells and that a progressive narrowing of this repertoire may take place during recall infections .

Recent studies have suggested that the diversity of <protein>TCR</protein> repertoire after primary immunization is conserved in <cell_type>memory T cells</cell_type> and that a progressive narrowing of this repertoire may take place during recall infections .


It now remains to be investigated which parameters determine the repertoire of the memory response and possibly restrict its diversity after subsequent antigenic challenges .


To address this question , we took advantage of a panel of CD8+ T cell clones from the joint of a rheumatoid arthritis patient and selected for their reactivity against a single MHC/peptide complex .

To address this question , we took advantage of a panel of <cell_line>CD8+ T cell clones</cell_line> from the joint of a rheumatoid arthritis patient and selected for their reactivity against a <protein>single MHC/peptide complex</protein> .


Characterization of both TCR chains documented a great diversity among those clones and the persistence of clonotypes over a 2-yr period .

Characterization of both <protein>TCR chains</protein> documented a great diversity among those clones and the persistence of clonotypes over a 2-yr period .


Strikingly , despite the observed repertoire heterogeneity , all clones displayed a narrow range of MHC /peptide density requirements in cytotoxicity assays ( ED50 between 9 and 36 nM ) .

Strikingly , despite the observed repertoire heterogeneity , all clones displayed a narrow range of <protein>MHC</protein> /peptide density requirements in cytotoxicity assays ( ED50 between 9 and 36 nM ) .


TCR affinities were then indirectly estimated by blocking CD8 interaction with an anti-CD8 mAb .

<protein>TCR</protein> affinities were then indirectly estimated by blocking <protein>CD8</protein> interaction with an <protein>anti-CD8 mAb</protein> .


We found a wide range of TCR affinities among the different clonotypes that segregated with Vbeta usage .

We found a wide range of <protein>TCR</protein> affinities among the different clonotypes that segregated with Vbeta usage .


We thus propose that during an in vivo chronic response , a narrow range of avidity of the TCR-CD8 complex conditions long-term clonotype persistence , and that the level of CD8 contribution is adjusted to keep clonotypes with variable TCR affinities within this avidity window .

We thus propose that during an in vivo chronic response , a narrow range of avidity of the <protein>TCR-CD8 complex</protein> conditions long-term clonotype persistence , and that the level of <protein>CD8</protein> contribution is adjusted to keep clonotypes with variable <protein>TCR</protein> affinities within this avidity window .


Nuclear localization and formation of beta-catenin-lymphoid enhancer factor 1 complexes are not sufficient for activation of gene expression .

Nuclear localization and formation of <protein>beta-catenin-lymphoid enhancer factor 1 complexes</protein> are not sufficient for activation of gene expression .


In response to activation of the Wnt signaling pathway , beta-catenin accumulates in the nucleus , where it cooperates with LEF/TCF ( for lymphoid enhancer factor and T-cell factor ) transcription factors to activate gene expression .

In response to activation of the Wnt signaling pathway , <protein>beta-catenin</protein> accumulates in the nucleus , where it cooperates with <protein>LEF/TCF</protein> ( for <protein>lymphoid enhancer factor and T-cell factor</protein> ) <protein>transcription factors</protein> to activate gene expression .


The mechanisms by which beta-catenin undergoes this shift in location and participates in activation of gene transcription are unknown .

The mechanisms by which <protein>beta-catenin</protein> undergoes this shift in location and participates in activation of gene transcription are unknown .


We demonstrate here that beta-catenin can be imported into the nucleus independently of LEF/TCF binding , and it may also be exported from nuclei .

We demonstrate here that <protein>beta-catenin</protein> can be imported into the nucleus independently of <protein>LEF/TCF</protein> binding , and it may also be exported from nuclei .


We have introduced a small deletion within beta-catenin ( Delta19 ) that disrupts binding to LEF-1 , E-cadherin , and APC but not axin .

We have introduced a small deletion within <protein>beta-catenin</protein> ( <dna>Delta19</dna> ) that disrupts binding to <protein>LEF-1</protein> , <protein>E-cadherin</protein> , and <protein>APC</protein> but not <protein>axin</protein> .


This Delta19 beta-catenin mutant localizes to the nucleus because it may not be efficiently sequestered in the cytoplasm .

This <protein>Delta19 beta-catenin mutant</protein> localizes to the nucleus because it may not be efficiently sequestered in the cytoplasm .


The nuclear localization of Delta19 definitively demonstrates that the mechanisms by which beta-catenin localizes in the nucleus are completely independent of LEF/TCF factors .

The nuclear localization of <dna>Delta19</dna> definitively demonstrates that the mechanisms by which <protein>beta-catenin</protein> localizes in the nucleus are completely independent of <protein>LEF/TCF factors</protein> .


beta-Catenin and LEF-1 complexes can activate reporter gene expression in a transformed T-lymphocyte cell line ( Jurkat ) but not in normal T lymphocytes , even though both factors are nuclear .

<protein>beta-Catenin</protein> and <protein>LEF-1 complexes</protein> can activate reporter gene expression in a <cell_line>transformed T-lymphocyte cell line</cell_line> ( <cell_line>Jurkat</cell_line> ) but not in <cell_type>normal T lymphocytes</cell_type> , even though both factors are nuclear .


Thus , localization of both factors to the nucleus is not sufficient for activation of gene expression .


Excess beta-catenin can squelch reporter gene activation by LEF-1-beta-catenin complexes but not activation by the transcription factor VP16 .

Excess <protein>beta-catenin</protein> can squelch reporter gene activation by <protein>LEF-1-beta-catenin complexes</protein> but not activation by the <protein>transcription factor</protein> <protein>VP16</protein> .


Taken together , these data suggest that a third component is necessary for gene activation and that this third component may vary with cell type .


Control of lymphocyte development by the Ikaros gene family .

Control of lymphocyte development by the <dna>Ikaros gene family</dna> .


Lymphoid cell differentiation relies on precisely orchestrated gene activation and repression events .


Gene targeting studies have demonstrated crucial roles for the transcription factors Ikaros and Aiolos in regulating multiple stages of B and T cell development .

Gene targeting studies have demonstrated crucial roles for the <protein>transcription factors</protein> <protein>Ikaros</protein> and <protein>Aiolos</protein> in regulating multiple stages of B and T cell development .


Recent experiments suggest that Ikaros and Aiolos set B cell antigen-receptor ( BCR ) - and TCR-mediated signaling thresholds and that the molecules exist within T cells in nuclear complexes that contain nucleosome remodeling and histone deacetylase activities .

Recent experiments suggest that <protein>Ikaros</protein> and <protein>Aiolos</protein> set B cell antigen-receptor ( BCR ) - and TCR-mediated signaling thresholds and that the molecules exist within <cell_type>T cells</cell_type> in <protein>nuclear complexes</protein> that contain nucleosome remodeling and <protein>histone deacetylase</protein> activities .


Transcriptional targeting of retroviral vectors to the erythroblastic progeny of transduced hematopoietic stem cells .

Transcriptional targeting of retroviral vectors to the <cell_type>erythroblastic progeny</cell_type> of <cell_line>transduced hematopoietic stem cells</cell_line> .


Targeted expression to specific tissues or cell lineages is a necessary feature of a gene therapy vector for many clinical applications , such as correction of hemoglobinopathies or thalassemias by transplantation of genetically modified hematopoietic stem cells .

Targeted expression to specific tissues or cell lineages is a necessary feature of a gene therapy vector for many clinical applications , such as correction of hemoglobinopathies or thalassemias by transplantation of <cell_type>genetically modified hematopoietic stem cells</cell_type> .


We developed retroviral vectors in which the constitutive viral enhancer in the U3 region of the 3 ' LTR is replaced by an autoregulatory enhancer of the erythroid-specific GATA-1 transcription factor gene .

We developed retroviral vectors in which the <dna>constitutive viral enhancer</dna> in the <dna>U3 region</dna> of the <dna>3 ' LTR</dna> is replaced by an <dna>autoregulatory enhancer</dna> of the <dna>erythroid-specific GATA-1 transcription factor gene</dna> .


The replaced enhancer is propagated to the 5 ' LTR upon integration into the target cell genome .

The <dna>replaced enhancer</dna> is propagated to the <dna>5 ' LTR</dna> upon integration into the <dna>target cell genome</dna> .


The modified vectors were used to transduce human hematopoietic cell lines , cord blood-derived CD34 ( + ) stem/progenitor cells , and murine bone marrow repopulating stem cells .

The modified vectors were used to transduce <cell_line>human hematopoietic cell lines</cell_line> , <cell_type>cord blood-derived CD34 ( + ) stem/progenitor cells</cell_type> , and <cell_line>murine bone marrow repopulating stem cells</cell_line> .


The expression of appropriate reporter genes ( triangle upLNGFR , EGFP ) was analyzed in the differentiated progeny of transduced stem cells in vitro , in liquid culture as well as in clonogenic assay , and in vivo , after bone marrow transplantation in lethally irradiated mice .

The expression of appropriate <dna>reporter genes</dna> ( <dna>triangle upLNGFR</dna> , <dna>EGFP</dna> ) was analyzed in the differentiated progeny of <cell_line>transduced stem cells</cell_line> in vitro , in liquid culture as well as in clonogenic assay , and in vivo , after bone marrow transplantation in lethally irradiated mice .


The GATA-1 autoregulatory enhancer effectively restricts the expression of the LTR-driven proviral transcription unit to the erythroblastic progeny of both human progenitors and mouse-repopulating stem cells .

The <dna>GATA-1 autoregulatory enhancer</dna> effectively restricts the expression of the <dna>LTR-driven proviral transcription unit</dna> to the <cell_type>erythroblastic progeny</cell_type> of both <cell_type>human progenitors</cell_type> and <cell_line>mouse-repopulating stem cells</cell_line> .


Packaging of viral particles , integration into the target genome , and stability of the integrated provirus are not affected by the LTR modification .

Packaging of viral particles , integration into the target genome , and stability of the integrated provirus are not affected by the <dna>LTR</dna> modification .


Enhancer replacement is therefore an effective strategy to target expression of a retroviral transgene to a specific progeny of transduced hematopoietic stem cells .

Enhancer replacement is therefore an effective strategy to target expression of a <dna>retroviral transgene</dna> to a specific progeny of <cell_line>transduced hematopoietic stem cells</cell_line> .


Regulation of Fas ligand expression and cell death by apoptosis-linked gene 4 .

Regulation of <protein>Fas ligand</protein> expression and cell death by <dna>apoptosis-linked gene 4</dna> .


Programmed cell death is a process required for the normal development of an organism .


One of the best understood apoptotic pathways occurs in T lymphocytes and is mediated by Fas /Fas ligand ( FasL ) interaction .

One of the best understood apoptotic pathways occurs in <cell_type>T lymphocytes</cell_type> and is mediated by <protein>Fas</protein> <protein>/Fas ligand</protein> ( <protein>FasL</protein> ) interaction .


During studies of apoptosis induced by T cell-receptor engagement , we identified ALG-4F , a truncated transcript that prevents T cell-receptor-induced FasL upregulation and cell death .

During studies of apoptosis induced by T cell-receptor engagement , we identified <protein>ALG-4F</protein> , a <protein>truncated transcript</protein> that prevents T cell-receptor-induced <protein>FasL</protein> upregulation and cell death .


Overexpression of full-length ALG-4 induced transcription of FasL and , consequently , apoptosis .

Overexpression of <protein>full-length ALG-4</protein> induced transcription of <protein>FasL</protein> and , consequently , apoptosis .


These results indicate that ALG-4 is necessary and sufficient for FasL expression .

These results indicate that <protein>ALG-4</protein> is necessary and sufficient for <protein>FasL</protein> expression .


Fas/ FasL interaction initiates cell death in many other systems , and its dysregulation is a mechanism by which several pathologic conditions arise .

Fas/ <protein>FasL</protein> interaction initiates cell death in many other systems , and its dysregulation is a mechanism by which several pathologic conditions arise .


Understanding the molecular mechanisms of FasL regulation could be very useful in elucidating how these diseases develop and in identifying potential therapeutic targets .

Understanding the molecular mechanisms of <protein>FasL</protein> regulation could be very useful in elucidating how these diseases develop and in identifying potential therapeutic targets .


The Epstein-Barr virus oncoprotein latent membrane protein 1 engages the tumor necrosis factor receptor-associated proteins TRADD and receptor-interacting protein ( RIP ) but does not induce apoptosis or require RIP for NF-kappaB activation .

The <protein>Epstein-Barr virus oncoprotein latent membrane protein 1</protein> engages the <protein>tumor necrosis factor receptor-associated proteins</protein> <protein>TRADD</protein> and <protein>receptor-interacting protein</protein> ( <protein>RIP</protein> ) but does not induce apoptosis or require <protein>RIP</protein> for <protein>NF-kappaB</protein> activation .


A site in the Epstein-Barr virus ( EBV ) transforming protein LMP1 that constitutively associates with the tumor necrosis factor receptor 1 ( TNFR1 ) -associated death domain protein TRADD to mediate NF-kappaB and c-Jun N-terminal kinase activation is critical for long-term lymphoblastoid cell proliferation .

A site in the <protein>Epstein-Barr virus ( EBV ) transforming protein</protein> <protein>LMP1</protein> that constitutively associates with the <protein>tumor necrosis factor receptor 1 ( TNFR1 ) -associated death domain protein</protein> <protein>TRADD</protein> to mediate <protein>NF-kappaB</protein> and <protein>c-Jun N-terminal kinase</protein> activation is critical for long-term lymphoblastoid cell proliferation .


We now find that LMP1 signaling through TRADD differs from TNFR1 signaling through TRADD .

We now find that <protein>LMP1</protein> signaling through <protein>TRADD</protein> differs from <protein>TNFR1</protein> signaling through <protein>TRADD</protein> .


LMP1 needs only 11 amino acids to activate NF-kappaB or synergize with TRADD in NF-kappaB activation , while TNFR1 requires approximately 70 residues .

<protein>LMP1</protein> needs only 11 amino acids to activate <protein>NF-kappaB</protein> or synergize with <protein>TRADD</protein> in <protein>NF-kappaB</protein> activation , while <protein>TNFR1</protein> requires approximately 70 residues .


Further , LMP1 does not require TRADD residues 294 to 312 for NF-kappaB activation , while TNFR1 requires TRADD residues 296 to 302 .

Further , <protein>LMP1</protein> does not require <protein>TRADD</protein> residues 294 to 312 for <protein>NF-kappaB</protein> activation , while <protein>TNFR1</protein> requires <protein>TRADD</protein> residues 296 to 302 .


LMP1 is partially blocked for NF-kappaB activation by a TRADD mutant consisting of residues 122 to 293 .

<protein>LMP1</protein> is partially blocked for <protein>NF-kappaB</protein> activation by a <protein>TRADD</protein> mutant consisting of <protein>residues 122 to 293</protein> .


Unlike TNFR1 , LMP1 can interact directly with receptor-interacting protein ( RIP ) and stably associates with RIP in EBV-transformed lymphoblastoid cell lines .

Unlike <protein>TNFR1</protein> , <protein>LMP1</protein> can interact directly with <protein>receptor-interacting protein</protein> ( <protein>RIP</protein> ) and stably associates with <protein>RIP</protein> in <cell_line>EBV-transformed lymphoblastoid cell lines</cell_line> .


Surprisingly , LMP1 does not require RIP for NF-kappaB activation .

Surprisingly , <protein>LMP1</protein> does not require <protein>RIP</protein> for <protein>NF-kappaB</protein> activation .


Despite constitutive association with TRADD or RIP , LMP1 does not induce apoptosis in EBV-negative Burkitt lymphoma or human embryonic kidney 293 cells .

Despite constitutive association with <protein>TRADD</protein> or <protein>RIP</protein> , <protein>LMP1</protein> does not induce apoptosis in <cell_line>EBV-negative Burkitt lymphoma or human embryonic kidney 293 cells</cell_line> .


These results add a different perspective to the molecular interactions through which LMP1 , TRADD , and RIP participate in B-lymphocyte activation and growth .

These results add a different perspective to the molecular interactions through which <protein>LMP1</protein> , <protein>TRADD</protein> , and <protein>RIP</protein> participate in B-lymphocyte activation and growth .


Inhibition of NF-kappa B activity in human T lymphocytes induces caspase-dependent apoptosis without detectable activation of caspase-1 and -3 .

Inhibition of <protein>NF-kappa B</protein> activity in human T lymphocytes induces caspase-dependent apoptosis without detectable activation of <protein>caspase-1 and -3</protein> .


NF-kappa B is involved in the transcriptional control of various genes that act as extrinsic and intrinsic survival factors for T cells .

<protein>NF-kappa B</protein> is involved in the transcriptional control of <dna>various genes</dna> that act as extrinsic and intrinsic survival factors for <cell_type>T cells</cell_type> .


Our findings show that suppression of NF-kappa B activity with cell-permeable SN50 peptide , which masks the nuclear localization sequence of NF-kappa B1 dimers and prevents their nuclear localization , induces apoptosis in resting normal human PBL .

Our findings show that suppression of <protein>NF-kappa B</protein> activity with cell-permeable SN50 peptide , which masks the nuclear localization sequence of <protein>NF-kappa B1 dimers</protein> and prevents their nuclear localization , induces apoptosis in <cell_type>resting normal human PBL</cell_type> .


Inhibition of NF-kappa B resulted in the externalization of phosphatidylserine , induction of DNA breaks , and morphological changes consistent with apoptosis .

Inhibition of <protein>NF-kappa B</protein> resulted in the externalization of phosphatidylserine , induction of DNA breaks , and morphological changes consistent with apoptosis .


DNA fragmentation was efficiently blocked by the caspase inhibitor Z-VAD-fmk and partially blocked by Ac-DEVD-fmk , suggesting that SN50-mediated apoptosis is caspase -dependent .

DNA fragmentation was efficiently blocked by the <protein>caspase inhibitor</protein> <protein>Z-VAD-fmk</protein> and partially blocked by <protein>Ac-DEVD-fmk</protein> , suggesting that SN50-mediated apoptosis is <protein>caspase</protein> -dependent .


Interestingly , apoptosis induced by NF-kappa B suppression , in contrast to that induced by TPEN ( N , N , N ' , N'-tetrakis [ 2-pyridylmethyl ] ethylenediamine ) or soluble Fas ligand ( CD95 ) , was observed in the absence of active death effector proteases caspase-1 -like ( IL-1 converting enzyme ) , caspase-3 -like ( CPP32/Yama/apopain ) , and caspase-6 -like and without cleavage of caspase-3 substrates poly ( ADP-ribose ) polymerase and DNA fragmentation factor-45 .

Interestingly , apoptosis induced by <protein>NF-kappa B</protein> suppression , in contrast to that induced by TPEN ( N , N , N ' , N'-tetrakis [ 2-pyridylmethyl ] ethylenediamine ) or <protein>soluble Fas ligand</protein> ( <protein>CD95</protein> ) , was observed in the absence of <protein>active death effector proteases</protein> <protein>caspase-1</protein> -like ( <protein>IL-1 converting enzyme</protein> ) , <protein>caspase-3</protein> -like ( <protein>CPP32/Yama/apopain</protein> ) , and <protein>caspase-6</protein> -like and without cleavage of <protein>caspase-3 substrates</protein> <protein>poly ( ADP-ribose ) polymerase</protein> and <protein>DNA fragmentation factor-45</protein> .


These findings suggest either low level of activation is required or that different caspases are involved .

These findings suggest either low level of activation is required or that different <protein>caspases</protein> are involved .


Preactivation of T cells resulting in NF-kappa B nuclear translocation protected cells from SN50-induced apoptosis .

Preactivation of <cell_type>T cells</cell_type> resulting in <protein>NF-kappa B</protein> nuclear translocation protected cells from SN50-induced apoptosis .


Our findings demonstrate an essential role of NF-kappa B in survival of naive PBL .

Our findings demonstrate an essential role of <protein>NF-kappa B</protein> in survival of <cell_type>naive PBL</cell_type> .


Signal transduction pathways in normal human monocytes stimulated by cytokines and mediators : comparative study with normal human neutrophils or transformed cells and the putative roles in functionality and cell biology .

Signal transduction pathways in <cell_type>normal human monocytes</cell_type> stimulated by <protein>cytokines</protein> and mediators : comparative study with <cell_type>normal human neutrophils</cell_type> or <cell_line>transformed cells</cell_line> and the putative roles in functionality and cell biology .


Granulocyte-macrophage colony-stimulating factor ( GM-CSF ) and interleukin ( IL ) -3 induced tyrosine phosphorylation of 92-kDa protein in normal human monocytes .

<protein>Granulocyte-macrophage colony-stimulating factor</protein> ( <protein>GM-CSF</protein> ) and <protein>interleukin ( IL ) -3</protein> induced tyrosine phosphorylation of <protein>92-kDa protein</protein> in <cell_type>normal human monocytes</cell_type> .


We identified this 92-kDa protein as STAT5 , but not as STATs1 , 3 , and 6 nor c-fes and vav protooncogene products , and demonstrated its translocation to the nucleus , enhancement of specific DNA binding capacity , and potentiation of trancriptional activity by GM-CSF .

We identified this <protein>92-kDa protein</protein> as <protein>STAT5</protein> , but not as <protein>STATs1 , 3 , and 6</protein> nor <protein>c-fes</protein> and <protein>vav protooncogene products</protein> , and demonstrated its translocation to the nucleus , enhancement of specific DNA binding capacity , and potentiation of trancriptional activity by <protein>GM-CSF</protein> .


N-formyl-methionyl-leucyl-phenylalanine ( FMLP ) and phorbol myristate acetate ( PMA ) induced tyrosine phosphorylation of 42- and 44-kDa proteins , which were identified as extracellular signal-regulated kinase ( ERK ) , in human monocytes .

N-formyl-methionyl-leucyl-phenylalanine ( FMLP ) and phorbol myristate acetate ( PMA ) induced tyrosine phosphorylation of <protein>42- and 44-kDa proteins</protein> , which were identified as <protein>extracellular signal-regulated kinase</protein> ( <protein>ERK</protein> ) , in <cell_type>human monocytes</cell_type> .


In marked contrast to neutrophils and MO7e cells , GM-CSF did not induce tyrosine phosphorylation and activation of ERK in monocytes .

In marked contrast to <cell_type>neutrophils</cell_type> and <cell_type>MO7e cells</cell_type> , <protein>GM-CSF</protein> did not induce tyrosine phosphorylation and activation of <protein>ERK</protein> in <cell_type>monocytes</cell_type> .


Among upstream signaling molecules of ERK , Shc was constitutively associated with Grb2 and was not tyrosine-phosphorylated by GM-CSF and FMLP , and Sos1 and c-Raf-1 were not phosphorylated by GM-CSF , IL-3 , TNF , and FMLP in monocytes , whereas all these signaling molecules were affected and/or utilized by GM-CSF in MO7e cells .

Among <protein>upstream signaling molecules</protein> of <protein>ERK</protein> , <protein>Shc</protein> was constitutively associated with Grb2 and was not tyrosine-phosphorylated by <protein>GM-CSF</protein> and FMLP , and <protein>Sos1</protein> and <protein>c-Raf-1</protein> were not phosphorylated by <protein>GM-CSF</protein> , IL-3 , TNF , and FMLP in <cell_type>monocytes</cell_type> , whereas all these signaling molecules were affected and/or utilized by <protein>GM-CSF</protein> in <cell_type>MO7e cells</cell_type> .


In contrast to neutrophils , p38 was constitutively phosphorylated and agonist-dependent phosphorylation and activation was not detected in human monocytes .

In contrast to <cell_type>neutrophils</cell_type> , <protein>p38</protein> was constitutively phosphorylated and agonist-dependent phosphorylation and activation was not detected in <cell_type>human monocytes</cell_type> .


Superoxide release stimulated by FMLP was inhibited partially by PD98059 or SB203580 , a specific inhibitor of ERK or p38 pathway , and was almost completely inhibited by the combination of both inhibitors , whereas PMA-induced superoxide release was resistant to these two inhibitors in monocytes .

Superoxide release stimulated by FMLP was inhibited partially by PD98059 or SB203580 , a specific inhibitor of ERK or p38 pathway , and was almost completely inhibited by the combination of both inhibitors , whereas PMA-induced superoxide release was resistant to these two inhibitors in <cell_type>monocytes</cell_type> .


PD98059 inhibited GM-CSF -dependent proliferation of MO7e cells .

PD98059 inhibited <protein>GM-CSF</protein> -dependent proliferation of <cell_type>MO7e cells</cell_type> .


Present results indicate trancriptional roles of STAT5 and functional roles of ERK and/or p38 in normal human monocytes stimulated by physiological receptor-mediated agonists GM-CSF and FMLP .

Present results indicate trancriptional roles of <protein>STAT5</protein> and functional roles of <protein>ERK</protein> and/or <protein>p38</protein> in <cell_type>normal human monocytes</cell_type> stimulated by physiological receptor-mediated agonists <protein>GM-CSF</protein> and FMLP .


Possible roles of ERK in proliferation of transformed cells were also suggested .

Possible roles of <protein>ERK</protein> in proliferation of <cell_line>transformed cells</cell_line> were also suggested .


Aiolos transcription factor controls cell death in T cells by regulating Bcl-2 expression and its cellular localization .

<protein>Aiolos</protein> <protein>transcription factor</protein> controls cell death in <cell_type>T cells</cell_type> by regulating <protein>Bcl-2</protein> expression and its cellular localization .


We searched for proteins that interact with Ras in interleukin ( IL ) -2-stimulated or IL-2-deprived cells , and found that the transcription factor Aiolos interacts with Ras .

We searched for proteins that interact with <protein>Ras</protein> in <cell_line>interleukin ( IL ) -2-stimulated or IL-2-deprived cells</cell_line> , and found that the <protein>transcription factor</protein> <protein>Aiolos</protein> interacts with <protein>Ras</protein> .


The Ras -Aiolos interaction was confirmed in vitro and in vivo by co-immunoprecipitation .

The <protein>Ras</protein> <protein>-Aiolos</protein> interaction was confirmed in vitro and in vivo by co-immunoprecipitation .


Indirect immunofluorescence shows that IL-2 controls the cellular distribution of Aiolos and induces its tyrosine phosphorylation , required for dissociation from Ras .

Indirect immunofluorescence shows that <protein>IL-2</protein> controls the cellular distribution of <protein>Aiolos</protein> and induces its tyrosine phosphorylation , required for dissociation from <protein>Ras</protein> .


We also identified functional Aiolos-binding sites in the Bcl-2 promoter , which are able to activate the luciferase reporter gene .

We also identified functional <dna>Aiolos-binding sites</dna> in the <dna>Bcl-2 promoter</dna> , which are able to activate the <dna>luciferase reporter gene</dna> .


Mutation of Aiolos-binding sites within the Bcl-2 promoter inhibits transactivation of the reporter gene luciferase , suggesting direct control of Bcl-2 expression by Aiolos .

Mutation of <dna>Aiolos-binding sites</dna> within the <dna>Bcl-2 promoter</dna> inhibits transactivation of the <dna>reporter gene luciferase</dna> , suggesting direct control of <protein>Bcl-2</protein> expression by <protein>Aiolos</protein> .


Co-transfection experiments confirm that Aiolos induces Bcl-2 expression and prevents apoptosis in IL-2-deprived cells .

Co-transfection experiments confirm that <protein>Aiolos</protein> induces <protein>Bcl-2</protein> expression and prevents apoptosis in <cell_line>IL-2-deprived cells</cell_line> .


We propose a model for the regulation of Bcl-2 expression via Aiolos .

We propose a model for the regulation of <protein>Bcl-2</protein> expression via <protein>Aiolos</protein> .


Binding of YY1 and Oct1 to a novel element that downregulates expression of IL-5 in human T cells .

Binding of <protein>YY1</protein> and <protein>Oct1</protein> to a novel element that downregulates expression of <protein>IL-5</protein> in human <cell_type>T cells</cell_type> .


BACKGROUND : IL-5 controls development of eosinophilia and has been shown to be involved in the pathogenesis of allergic diseases .

BACKGROUND : <protein>IL-5</protein> controls development of eosinophilia and has been shown to be involved in the pathogenesis of allergic diseases .


In both atopic and nonatopic asthma , elevated IL-5 has been detected in peripheral blood and the airways .

In both atopic and nonatopic asthma , elevated <protein>IL-5</protein> has been detected in peripheral blood and the airways .


IL-5 is produced mainly by activated T cells , and its expression is regulated at the transcriptional level .

<protein>IL-5</protein> is produced mainly by activated <cell_type>T cells</cell_type> , and its expression is regulated at the transcriptional level .


OBJECTIVE : This study focuses on the functional analysis of the human IL-5 ( hIL-5 ) promoter and characterization of cis -regulatory elements and transcription factors involved in the suppression of IL-5 transcription in T cells .

OBJECTIVE : This study focuses on the functional analysis of the <dna>human IL-5 ( hIL-5 ) promoter</dna> and characterization of cis -regulatory elements and transcription factors involved in the suppression of <protein>IL-5</protein> transcription in <cell_type>T cells</cell_type> .


METHODS : Methods used in this study include DNase I footprint assays , electrophoretic mobility shift assays , and functional analysis by mammalian cell transfection involving deletion analysis and site-directed mutagenesis .

METHODS : Methods used in this study include <protein>DNase I</protein> footprint assays , electrophoretic mobility shift assays , and functional analysis by mammalian cell transfection involving deletion analysis and site-directed mutagenesis .


RESULTS : We identified 5 protein binding regions ( BRs ) located within the proximal hIL-5 promoter .

RESULTS : We identified 5 <dna>protein binding regions</dna> ( <dna>BRs</dna> ) located within the <dna>proximal hIL-5 promoter</dna> .


Functional analysis indicates that the BRs are involved in control of hIL-5 promoter activity .

Functional analysis indicates that the <dna>BRs</dna> are involved in control of <dna>hIL-5 promoter</dna> activity .


Two of these regions , BR3 and BR4 located at positions -102 to -73 , have not previously been described as regulators of IL-5 expression in T cells .

Two of these regions , <dna>BR3</dna> and <dna>BR4</dna> located at positions -102 to -73 , have not previously been described as regulators of <protein>IL-5</protein> expression in <cell_type>T cells</cell_type> .


We show that the BR3 sequence contains a novel negative regulatory element located at positions -90 to -79 of the hIL-5 promoter , which binds Oct1 , octamer-like , and YY1 nuclear factors .

We show that the <dna>BR3</dna> sequence contains a novel <dna>negative regulatory element</dna> located at positions -90 to -79 of the <dna>hIL-5 promoter</dna> , which binds <protein>Oct1</protein> , <protein>octamer-like</protein> , and <protein>YY1 nuclear factors</protein> .


Substitution mutations , which abolished binding of these proteins to the BR3 sequence , significantly increased hIL-5 promoter activity in activated T cells .

Substitution mutations , which abolished binding of these proteins to the <dna>BR3 sequence</dna> , significantly increased <dna>hIL-5 promoter</dna> activity in <cell_type>activated T cells</cell_type> .


CONCLUSION : We suggest that Oct1 , YY1 , and octamer-like factors binding to the -90/-79 sequence within the proximal IL-5 promoter are involved in suppression of IL-5 transcription in T cells .

CONCLUSION : We suggest that <protein>Oct1</protein> , <protein>YY1</protein> , and <protein>octamer-like factors</protein> binding to the <dna>-90/-79 sequence</dna> within the <dna>proximal IL-5 promoter</dna> are involved in suppression of <protein>IL-5</protein> transcription in <cell_type>T cells</cell_type> .


New immunosuppressive drug PNU156804 blocks IL-2 -dependent proliferation and NF-kappa B and AP-1 activation .

New immunosuppressive drug PNU156804 blocks <protein>IL-2</protein> -dependent proliferation and NF-kappa B and AP-1 activation .


We had previously shown that the drug undecylprodigiosin ( UP ) blocks human lymphocyte proliferation in vitro .

We had previously shown that the drug undecylprodigiosin ( UP ) blocks <cell_type>human lymphocyte</cell_type> proliferation in vitro .


We have now investigated the mechanism of action of a new analogue of UP , PNU156804 , which shows a more favorable activity profile than UP in mice .


We demonstrate here that the biological effect of PNU156804 in vitro is indistinguishable from UP : PNU156804 blocks human T cell proliferation in mid-late G1 , as determined by cell cycle analysis , expression of cyclins , and cyclin-dependent kinases and retinoblastoma phosphorylation .

We demonstrate here that the biological effect of PNU156804 in vitro is indistinguishable from UP : PNU156804 blocks <cell_type>human T cell</cell_type> proliferation in mid-late G1 , as determined by cell cycle analysis , expression of <protein>cyclins</protein> , and <protein>cyclin-dependent kinases</protein> and retinoblastoma phosphorylation .


In addition , we show that PNU156804 does not block significantly the induction of either IL-2 or IL-2R alpha- and gamma-chains but inhibits IL-2 -dependent T cell proliferation .

In addition , we show that PNU156804 does not block significantly the induction of either <protein>IL-2</protein> or <protein>IL-2R alpha- and gamma-chains</protein> but inhibits <protein>IL-2</protein> -dependent T cell proliferation .


We have investigated several molecular pathways that are known to be activated by IL-2 in T cells .

We have investigated several molecular pathways that are known to be activated by <protein>IL-2</protein> in <cell_type>T cells</cell_type> .


We show that PNU156804 does not inhibit c-myc and bcl-2 mRNA induction .


On the other hand , PNU156804 efficiently inhibits the activation of the NF-kappa B and AP-1 transcription factors .

On the other hand , PNU156804 efficiently inhibits the activation of the <protein>NF-kappa B</protein> and <protein>AP-1</protein> transcription factors .


PNU156804 inhibition of NF-kappa B activation is due to the inhibition of the degradation of I kappa B-alpha and I kappa B-beta .

PNU156804 inhibition of <protein>NF-kappa B</protein> activation is due to the inhibition of the degradation of <protein>I kappa B-alpha</protein> and <protein>I kappa B-beta</protein> .


PNU156804 action is restricted to some signaling pathways ; it does not affect NF-kappa B activation by PMA in T cells but blocks that induced by CD40 cross-linking in B lymphocytes .

PNU156804 action is restricted to some signaling pathways ; it does not affect <protein>NF-kappa B</protein> activation by PMA in <cell_type>T cells</cell_type> but blocks that induced by <protein>CD40</protein> cross-linking in <cell_type>B lymphocytes</cell_type> .


We conclude that the prodigiosin family of immunosuppressants is a new family of molecules that show a novel target specificity clearly distinct from that of other immunosuppressive drugs such as cyclosporin A , FK506 , and rapamycin .


Human T-cell leukemia virus type 1 Tax protein induces the expression of STAT1 and STAT5 genes in T-cells .

<protein>Human T-cell leukemia virus type 1 Tax protein</protein> induces the expression of <dna>STAT1 and STAT5 genes</dna> in <cell_type>T-cells</cell_type> .


Human T-cell leukemia virus type 1 ( HTLV-1 ) Tax transforms normal T-cells in the presence of interleukin ( IL ) -2 in vitro .

<protein>Human T-cell leukemia virus type 1 ( HTLV-1 ) Tax</protein> transforms normal <cell_type>T-cells</cell_type> in the presence of <protein>interleukin ( IL ) -2</protein> in vitro .


STAT is a family of transcription factors that play a pivotal role in cytokine-induced functions of a various type of cells .

<protein>STAT</protein> is a family of <protein>transcription factors</protein> that play a pivotal role in cytokine-induced functions of a various type of cells .


We investigated the involvement of STATs in the transformation of T-cells by HTLV-1 .

We investigated the involvement of <protein>STATs</protein> in the transformation of <cell_type>T-cells</cell_type> by HTLV-1 .


HTLV-1-transformed T-cell lines expressed higher amounts of STAT1 , STAT3 and STAT5 RNA and proteins than virus-negative T cells .

<cell_line>HTLV-1-transformed T-cell lines</cell_line> expressed higher amounts of <rna>STAT1 , STAT3 and STAT5 RNA</rna> and proteins than <cell_type>virus-negative T cells</cell_type> .


The expression of STAT1 and STAT5 in a human T-cell line was induced by Tax .

The expression of <protein>STAT1</protein> and <protein>STAT5</protein> in a <cell_line>human T-cell line</cell_line> was induced by <protein>Tax</protein> .


IL-2 induced the DNA binding activity of STAT3 and STAT5 of a HTLV-1-transformed cell line and then stimulated its proliferation .

<protein>IL-2</protein> induced the DNA binding activity of <protein>STAT3</protein> and <protein>STAT5</protein> of a HTLV-1-transformed cell line and then stimulated its proliferation .


In contrast , IL-2 did neither in a cell line lacking STAT3 and STAT5 .

In contrast , <protein>IL-2</protein> did neither in a cell line lacking <protein>STAT3</protein> and <protein>STAT5</protein> .


The expression of STAT1 , STAT3 and STAT5 mRNAs were also induced by a T-cell mitogen in normal human peripheral blood mononuclear cells .

The expression of <rna>STAT1 , STAT3 and STAT5 mRNAs</rna> were also induced by a <protein>T-cell mitogen</protein> in normal <cell_type>human peripheral blood mononuclear cells</cell_type> .


Our results suggest that the induction of STAT1 and STAT5 by Tax enhances cytokine -induced functions of virus-infected T-cells , hence the induction may play a role in IL-2 -dependent transformation steps of T-cells by HTLV-1 .

Our results suggest that the induction of <protein>STAT1</protein> and <protein>STAT5</protein> by <protein>Tax</protein> enhances <protein>cytokine</protein> -induced functions of virus-infected <cell_type>T-cells</cell_type> , hence the induction may play a role in <protein>IL-2</protein> -dependent transformation steps of <cell_type>T-cells</cell_type> by HTLV-1 .


Transcriptional control of the IL-5 gene by human helper T cells : IL-5 synthesis is regulated independently from IL-2 or IL-4 synthesis .

Transcriptional control of the <protein>IL-5</protein> gene by <cell_type>human helper T cells</cell_type> : <protein>IL-5</protein> synthesis is regulated independently from <protein>IL-2</protein> or <protein>IL-4</protein> synthesis .


BACKGROUND : IL-5 is fundamentally involved in eosinophilic inflammation .

BACKGROUND : <protein>IL-5</protein> is fundamentally involved in eosinophilic inflammation .


Control of IL-5 production may be effective for the management of allergic diseases .

Control of <protein>IL-5</protein> production may be effective for the management of allergic diseases .


OBJECTIVE : We aimed to find the transcriptional mechanisms that regulate the IL-5 gene to selectively control IL-5 synthesis .

OBJECTIVE : We aimed to find the transcriptional mechanisms that regulate the <protein>IL-5</protein> gene to selectively control <protein>IL-5</protein> synthesis .


METHODS : Allergen-specific T-cell clones and T-cell hybridomas were established from the peripheral blood lymphocytes of patients with asthma , and the transcriptional regulation of the IL-5 gene was investigated with transient transfection and electrophoretic mobility shift analysis .

METHODS : <cell_line>Allergen-specific T-cell clones</cell_line> and <cell_line>T-cell hybridomas</cell_line> were established from the <cell_type>peripheral blood lymphocytes</cell_type> of patients with asthma , and the transcriptional regulation of the <dna>IL-5 gene</dna> was investigated with transient transfection and electrophoretic mobility shift analysis .


RESULTS : A human IL-5 promoter/enhancer-luciferase gene construct , pIL-5 ( -511 ) Luc , was transcribed on activation of IL-5-producing T-cell clones , but not IL-5-nonproducing clones .

RESULTS : A human <dna>IL-5 promoter/enhancer-luciferase gene construct</dna> , <dna>pIL-5 ( -511 ) Luc</dna> , was transcribed on activation of <cell_line>IL-5-producing T-cell clones</cell_line> , but not <cell_line>IL-5-nonproducing clones</cell_line> .


pIL-5 ( -511 ) Luc was transcribed by T-cell hybridomas derived from fusion between IL-5-producing T-cell clones and an IL-5 gene-nonexpressing T-cell line , but not by hybridomas derived from IL-5-nonproducing T-cell clones .

<dna>pIL-5 ( -511 ) Luc</dna> was transcribed by <cell_line>T-cell hybridomas</cell_line> derived from fusion between <cell_line>IL-5-producing T-cell clones</cell_line> and an <cell_line>IL-5 gene-nonexpressing T-cell line</cell_line> , but not by <cell_line>hybridomas</cell_line> derived from <cell_line>IL-5-nonproducing T-cell clones</cell_line> .


IL-5 synthesis was not only induced by T-cell receptor stimulation but also by IL-2 receptor stimulation .

<protein>IL-5</protein> synthesis was not only induced by T-cell receptor stimulation but also by <protein>IL-2</protein> receptor stimulation .


Binding of NF-AT , NF-kappaB , and AP-1 was induced by T-cell receptor ( TcR ) stimulation , although there was no significant upregulation of binding by IL-2 stimulation .

Binding of <protein>NF-AT</protein> , <protein>NF-kappaB</protein> , and <protein>AP-1</protein> was induced by <protein>T-cell receptor</protein> ( <protein>TcR</protein> ) stimulation , although there was no significant upregulation of binding by <protein>IL-2</protein> stimulation .


CONCLUSION : IL-5 synthesis by human helper T cells is regulated at the transcriptional level .

CONCLUSION : <protein>IL-5</protein> synthesis by <cell_type>human helper T cells</cell_type> is regulated at the transcriptional level .


A unique transcriptional mechanism distinct from those regulating the IL-2 or IL-4 genes seems to control the IL-5 gene .

A unique transcriptional mechanism distinct from those regulating the <dna>IL-2 or IL-4 genes</dna> seems to control the <dna>IL-5 gene</dna> .


Selective regulation of IL-5 gene transcription may be useful for treating eosinophlic inflammation .

Selective regulation of <protein>IL-5</protein> gene transcription may be useful for treating eosinophlic inflammation .


Defining therapeutic targets by using adenovirus : blocking NF-kappaB inhibits both inflammatory and destructive mechanisms in rheumatoid synovium but spares anti-inflammatory mediators .

Defining therapeutic targets by using adenovirus : blocking <protein>NF-kappaB</protein> inhibits both inflammatory and destructive mechanisms in rheumatoid synovium but spares <protein>anti-inflammatory mediators</protein> .


The role of the transcription factor NF-kappaB in the pathogenesis of rheumatoid arthritis has long been a subject of controversy .

The role of the <protein>transcription factor</protein> <protein>NF-kappaB</protein> in the pathogenesis of rheumatoid arthritis has long been a subject of controversy .


We used an adenoviral technique of blocking NF-kappaB through overexpression of the inhibitory subunit IkappaBalpha , which has the advantage that it can be used in the diseased tissue itself , with > 90 % of the synovial macrophages , fibroblasts , and T cells infected .

We used an adenoviral technique of blocking <protein>NF-kappaB</protein> through overexpression of the <protein>inhibitory subunit</protein> <protein>IkappaBalpha</protein> , which has the advantage that it can be used in the diseased tissue itself , with > 90 % of the <cell_type>synovial macrophages</cell_type> , <cell_type>fibroblasts</cell_type> , and <cell_type>T cells</cell_type> infected .


We found that the spontaneous production of tumor necrosis factor alpha and other pro-inflammatory cytokines is NF-kappaB -dependent in rheumatoid synovial tissue , in contrast to the main anti-inflammatory mediators , like IL-10 and -11 , and the IL-1 receptor antagonist .

We found that the spontaneous production of <protein>tumor necrosis factor alpha</protein> and other <protein>pro-inflammatory cytokines</protein> is <protein>NF-kappaB</protein> -dependent in rheumatoid synovial tissue , in contrast to the main <protein>anti-inflammatory mediators</protein> , like <protein>IL-10 and -11</protein> , and the <protein>IL-1 receptor antagonist</protein> .


Of even more interest , IkappaBalpha overexpression inhibited the production of matrix metalloproteinases 1 and 3 while not affecting their tissue inhibitor .

Of even more interest , <protein>IkappaBalpha</protein> overexpression inhibited the production of <protein>matrix metalloproteinases 1 and 3</protein> while not affecting their tissue inhibitor .


Blocking NF-kappaB in the rheumatoid joint thus has a very beneficial profile , reducing both the inflammatory response and the tissue destruction .

Blocking <protein>NF-kappaB</protein> in the rheumatoid joint thus has a very beneficial profile , reducing both the inflammatory response and the tissue destruction .


The adenoviral technique described here has widespread applicability , allowing rapid testing of the effects of blocking a potential therapeutic target in either cultures of normal cells or in the diseased tissue itself .

The adenoviral technique described here has widespread applicability , allowing rapid testing of the effects of blocking a potential therapeutic target in either cultures of <cell_type>normal cells</cell_type> or in the diseased tissue itself .


Resistance to tumor necrosis factor induced apoptosis in vitro correlates with high metastatic capacity of cells in vivo .

Resistance to <protein>tumor necrosis factor</protein> induced apoptosis in vitro correlates with high metastatic capacity of cells in vivo .


TNF is one of the cytokines secreted by the cells of the immune system .

<protein>TNF</protein> is one of the <protein>cytokines</protein> secreted by the cells of the immune system .


Our data demonstrate that those cell lines lacking capability to form metastatic tumors in vivo are susceptible to TNF induced apoptosis in vitro .

Our data demonstrate that those <cell_line>cell lines</cell_line> lacking capability to form metastatic tumors in vivo are susceptible to <protein>TNF</protein> induced apoptosis in vitro .


However , cell lines with high metastatic potential are resistant to TNF in vitro .

However , <cell_line>cell lines</cell_line> with high metastatic potential are resistant to <protein>TNF</protein> in vitro .


Furthermore , the same cell lines were resistant to cytolytic action of other cytotoxic proteins secreted by LAK cells .

Furthermore , the same <cell_line>cell lines</cell_line> were resistant to cytolytic action of other <protein>cytotoxic proteins</protein> secreted by <cell_line>LAK cells</cell_line> .


Our data showed that TNF resistance in vitro correlates with the increased level of transcription factor NF-kappaB .

Our data showed that <protein>TNF</protein> resistance in vitro correlates with the increased level of <protein>transcription factor</protein> <protein>NF-kappaB</protein> .


This finding may provide a tool to improve current protocols of immunotherapy and insights to how tumor cells are or are not killed by LAK cells .

This finding may provide a tool to improve current protocols of immunotherapy and insights to how <cell_type>tumor cells</cell_type> are or are not killed by <cell_line>LAK cells</cell_line> .


Clonality analysis of granulocytes and T lymphocytes in healthy females by the PCR-based HUMARA method .

Clonality analysis of <cell_type>granulocytes</cell_type> and <cell_type>T lymphocytes</cell_type> in healthy females by the PCR-based <dna>HUMARA</dna> method .


Clonality analysis utilizing X-chromosome inactivation has been used in the study of various diseases , including hematological malignancies .

Clonality analysis utilizing <dna>X-chromosome</dna> inactivation has been used in the study of various diseases , including hematological malignancies .


The human androgen receptor gene ( HUMARA ) assay is the newest of such methods , and the majority of the female population can be assessed by this relatively simple procedure .

The <dna>human androgen receptor gene</dna> ( <dna>HUMARA</dna> ) assay is the newest of such methods , and the majority of the female population can be assessed by this relatively simple procedure .


One problem in using these clonality analysis methods , however , is that there may be significant variation in Lyonization in blood cells in normal individuals .

One problem in using these clonality analysis methods , however , is that there may be significant variation in Lyonization in <cell_type>blood cells</cell_type> in normal individuals .


To determine the diversity in X-chromosome methylation patterns , which reflect Lyonization , assessed by the HUMARA assay in the supposedly normal population , we analyzed granulocytes and T cells from 97 relatively young ( 18- to 35-year-old ) healthy female volunteers .

To determine the diversity in <dna>X-chromosome</dna> methylation patterns , which reflect Lyonization , assessed by the <dna>HUMARA</dna> assay in the supposedly normal population , we analyzed <cell_type>granulocytes</cell_type> and <cell_line>T cells</cell_line> from 97 relatively young ( 18- to 35-year-old ) healthy female volunteers .


We found that the methylation patterns in the two HUMARA alleles were distributed even more widely , both in granuloctyes and in T cells , than previously reported with other methods .

We found that the methylation patterns in the two <dna>HUMARA alleles</dna> were distributed even more widely , both in <cell_type>granuloctyes</cell_type> and in <cell_line>T cells</cell_line> , than previously reported with other methods .


We also found that the deviation of methylation in granulocytes and T cells was well correlated .

We also found that the deviation of methylation in <cell_type>granulocytes</cell_type> and <cell_line>T cells</cell_line> was well correlated .


Thus , we conclude that appropriate controls from the same individuals , such as T cells in the case of stem cell disorders , should always be employed to conclusively determine whether certain cells of hematopoietic origin are clonal .

Thus , we conclude that appropriate controls from the same individuals , such as <cell_line>T cells</cell_line> in the case of stem cell disorders , should always be employed to conclusively determine whether certain cells of hematopoietic origin are clonal .


Functional B-cell response in intrahepatic lymphoid follicles in chronic hepatitis C .


Intrahepatic lymphoid follicle ( ILF ) formation is one of the most characteristic and commonly observed histological features in patients with chronic hepatitis C .


However , little is known regarding whether follicles in the liver belong to functional lymphoid tissues , where B cells are activated , differentiated , and proliferated , or if the lymphocytes are merely infiltrated after recruitment from the secondary lymphoid organs .


To ascertain this possibility , we examined the expression of markers for B-cell activation , differentiation , and proliferation in ILFs in patients with chronic hepatitis C using surgically resected specimens , and compared them with specimens of perihepatic lymph nodes by an immunohistochemical technique .


Germinal center ( GC ) formation in the ILFs was frequently found in HCV-positive cases .


The distribution of immunoglobulin M ( IgM ) - , IgD- , and IgG-positive cells and the expression patterns of Ki-67 , CD23 , or bcl-2 and bcl-6 gene products in the follicles with GC formation in the liver of patients with chronic hepatitis C were similar to those of lymph nodes , indicating that B cells are activated , proliferated , and differentiated in the ILFs with GC formation in patients with chronic hepatitis C .

The distribution of immunoglobulin M ( IgM ) - , IgD- , and IgG-positive cells and the expression patterns of Ki-67 , CD23 , or bcl-2 and bcl-6 gene products in the follicles with GC formation in the liver of patients with chronic hepatitis C were similar to those of lymph nodes , indicating that <cell_type>B cells</cell_type> are activated , proliferated , and differentiated in the ILFs with GC formation in patients with chronic hepatitis C .


Oligoclonal expansion of B cells in the livers with ILFs was confirmed by an analysis of immunoglobulin heavy chain ( IgH ) gene rearrangement determined by polymerase chain reaction ( PCR ) .

Oligoclonal expansion of <cell_type>B cells</cell_type> in the livers with ILFs was confirmed by an analysis of <dna>immunoglobulin heavy chain ( IgH ) gene</dna> rearrangement determined by polymerase chain reaction ( PCR ) .


These data strongly suggest that ILFs with GC formation , which are frequently found in patients with chronic hepatitis C , may functionally be the same as those found in lymph nodes with respect to B-cell expansion and maturation .


Downregulation of Wilms ' tumor gene ( WT1 ) is not a prerequisite for erythroid or megakaryocytic differentiation of the leukemic cell line K562 .

Downregulation of <dna>Wilms ' tumor gene</dna> ( <dna>WT1</dna> ) is not a prerequisite for erythroid or megakaryocytic differentiation of the <cell_line>leukemic cell line K562</cell_line> .


The Wilms ' tumor gene ( WT1 ) encodes a transcription factor of the zinc finger type .

The <dna>Wilms ' tumor gene</dna> ( <dna>WT1</dna> ) encodes a <protein>transcription factor</protein> of the <protein>zinc finger type</protein> .


A high expression of WT1 has been detected in a range of acute leukemias , and WT1 is downregulated during induced differentiation of some leukemic cell lines .

A high expression of <dna>WT1</dna> has been detected in a range of acute leukemias , and <dna>WT1</dna> is downregulated during induced differentiation of some <cell_line>leukemic cell lines</cell_line> .


Overexpression of WT1 in some myeloid cell lines confers resistance to differentiation induction .

Overexpression of <dna>WT1</dna> in some <cell_line>myeloid cell lines</cell_line> confers resistance to differentiation induction .


These observations suggest that a high WT1 expression in hematopoietic cells is incompatible with differentiation .

These observations suggest that a high <dna>WT1</dna> expression in <cell_type>hematopoietic cells</cell_type> is incompatible with differentiation .


In this study , each of the four different isoforms of WT1 was constitutively overexpressed in the leukemic cell line K562 .

In this study , each of the four different isoforms of <dna>WT1</dna> was constitutively overexpressed in the <cell_line>leukemic cell line K562</cell_line> .


K562 cells express endogenous WT1 , which is downregulated as a response to induced differentiation along the erythroid and megakaryocytic pathways .

<cell_line>K562 cells</cell_line> express endogenous <dna>WT1</dna> , which is downregulated as a response to induced differentiation along the erythroid and megakaryocytic pathways .


We now demonstrate that a forced exogenous expression of the four different isoforms of WT1 in K562 does not affect the differentiation response , as judged by accumulation of hemoglobin in response to hemin or the expression of megakaryocytic cell surface markers in response to 12-O-tetradecanoylphorbol-13-acetate ( TPA ) .

We now demonstrate that a forced exogenous expression of the four different isoforms of <dna>WT1</dna> in <cell_line>K562</cell_line> does not affect the differentiation response , as judged by accumulation of <protein>hemoglobin</protein> in response to <protein>hemin</protein> or the expression of <protein>megakaryocytic cell surface markers</protein> in response to 12-O-tetradecanoylphorbol-13-acetate ( TPA ) .


We conclude that downregulation of WT1 during induced differentiation of K562 cells is not a prerequisite for erythroid or megakaryocytic differentiation of these cells .

We conclude that downregulation of <dna>WT1</dna> during induced differentiation of <cell_line>K562 cells</cell_line> is not a prerequisite for erythroid or megakaryocytic differentiation of these cells .


Association of the interleukin-4 receptor alpha chain with p47phox , an activator of the phagocyte NADPH oxidase in B cells .

Association of the <protein>interleukin-4 receptor alpha chain</protein> with p47phox , an activator of the <protein>phagocyte NADPH oxidase</protein> in <cell_type>B cells</cell_type> .


Interleukin ( IL ) -4 plays an important role in IgE synthesis in B cells and in Th2 differentiation in T cells .

<protein>Interleukin ( IL ) -4</protein> plays an important role in <protein>IgE</protein> synthesis in <cell_type>B cells</cell_type> and in Th2 differentiation in <cell_line>T cells</cell_line> .


IL-4 conducts its biological activities through binding to the IL-4 receptor ( IL-4R ) on the surface of target cells .

<protein>IL-4</protein> conducts its biological activities through binding to the <protein>IL-4 receptor</protein> ( <protein>IL-4R</protein> ) on the surface of <cell_type>target cells</cell_type> .


IL-4R are thought to be composed of the IL-4R alpha chain ( IL-4R alpha ) and either the IL-2R gamma chain or the IL-13R alpha chain .

<protein>IL-4R</protein> are thought to be composed of the <protein>IL-4R alpha chain</protein> ( <protein>IL-4R alpha</protein> ) and either the <protein>IL-2R gamma chain</protein> or the <protein>IL-13R alpha chain</protein> .


We have previously shown that the membrane-proximal portion in the cytoplasmic domain of the human IL-4R alpha ( hIL-4R alpha ) is critical for proliferation , generation of germline epsilon transcript , and activation of STAT6 , based on analyses of truncated hIL-4R alphas .

We have previously shown that the <protein>membrane-proximal portion</protein> in the <protein>cytoplasmic domain</protein> of the <protein>human IL-4R alpha</protein> ( <protein>hIL-4R alpha</protein> ) is critical for proliferation , generation of germline epsilon transcript , and activation of <protein>STAT6</protein> , based on analyses of <protein>truncated hIL-4R alphas</protein> .


In this study , we found that p47phox , an activator of the phagocyte NADPH oxidase , binds to this portion by the two-hybrid system .

In this study , we found that p47phox , an activator of the <protein>phagocyte NADPH oxidase</protein> , binds to this portion by the two-hybrid system .


Furthermore , we observed the association of p47phox with the hIL-4R alpha in B cells derived from a normal donor .

Furthermore , we observed the association of p47phox with the <protein>hIL-4R alpha</protein> in <cell_type>B cells</cell_type> derived from a normal donor .


These results suggest that p47phox is involved in the signal transduction of IL-4 in B cells .

These results suggest that p47phox is involved in the signal transduction of <protein>IL-4</protein> in <cell_type>B cells</cell_type> .


However , activation of STAT6 , CD23 expression , and IgE synthesis induced by IL-4 were not affected in p47phox-deficient patients , which raises the possibility that p47phox may be important in other signaling activities as well in B cells .

However , activation of <protein>STAT6</protein> , <protein>CD23</protein> expression , and <protein>IgE</protein> synthesis induced by <protein>IL-4</protein> were not affected in p47phox-deficient patients , which raises the possibility that p47phox may be important in other signaling activities as well in <cell_type>B cells</cell_type> .


Effects of diesel organic extracts on chemokine production by peripheral blood mononuclear cells .

Effects of diesel organic extracts on <protein>chemokine</protein> production by <cell_type>peripheral blood mononuclear cells</cell_type> .


BACKGROUND : Polyaromatic hydrocarbons ( PAHs ) associated with diesel exhaust particles ( DEPs ) are found in the atmospheric urban pollution .


Such compounds have been shown to favor IgE production , bronchial hyperresponsiveness , and airway inflammation .

Such compounds have been shown to favor <protein>IgE</protein> production , bronchial hyperresponsiveness , and airway inflammation .


Chemokines are a group of chemotactic cytokines involved in the recruitment of inflammatory cells .

Chemokines are a group of <protein>chemotactic cytokines</protein> involved in the recruitment of <cell_type>inflammatory cells</cell_type> .


OBJECTIVE : We investigated the effect of DEP-PAHs on the release and mRNA expression of IL-8 , MCP-1 , and RANTES by PBMCs obtained from healthy subjects .

OBJECTIVE : We investigated the effect of DEP-PAHs on the release and <rna>mRNA</rna> expression of <protein>IL-8</protein> , <protein>MCP-1</protein> , and <protein>RANTES</protein> by <cell_type>PBMCs</cell_type> obtained from healthy subjects .


METHODS : Protein production in supernatants was assessed by ELISA , and mRNA expression was evaluated by semiquantitative RT-PCR .

METHODS : Protein production in supernatants was assessed by ELISA , and <rna>mRNA</rna> expression was evaluated by semiquantitative RT-PCR .


RESULTS : Secretion of IL-8 and RANTES increased in a dose-dependent manner with increasing concentrations of DEP-PAHs ( range , 0.5 ng to 50 ng/mL ) .

RESULTS : Secretion of <protein>IL-8</protein> and <protein>RANTES</protein> increased in a dose-dependent manner with increasing concentrations of DEP-PAHs ( range , 0.5 ng to 50 ng/mL ) .


On the contrary , the release of MCP-1 was significantly inhibited , also in a dose-dependent manner .

On the contrary , the release of <protein>MCP-1</protein> was significantly inhibited , also in a dose-dependent manner .


Messenger RNA production coding for IL-8 , RANTES , and MCP-1 showed parallel variations to the production of the correspondent proteins .

<rna>Messenger RNA</rna> production coding for <protein>IL-8</protein> , <protein>RANTES</protein> , and <protein>MCP-1</protein> showed parallel variations to the production of the <protein>correspondent proteins</protein> .


Effects of DEP-PAHs became significant at 7 hours and up to 48 hours time culture for MCP-1 , and up to 24 hours time culture for IL-8 and RANTES .

Effects of DEP-PAHs became significant at 7 hours and up to 48 hours time culture for <protein>MCP-1</protein> , and up to 24 hours time culture for <protein>IL-8</protein> and <protein>RANTES</protein> .


Moreover , supernatants from DEP-PAH-activated cells , compared with those of controls , exhibited a significantly enhanced chemotactic activity for neutrophils and eosinophils , which was significantly inhibited by pretreatment with anti-IL-8 and anti-RANTES neutralizing antibodies , respectively .

Moreover , supernatants from <cell_type>DEP-PAH-activated cells</cell_type> , compared with those of controls , exhibited a significantly enhanced chemotactic activity for <cell_type>neutrophils</cell_type> and <cell_type>eosinophils</cell_type> , which was significantly inhibited by pretreatment with <protein>anti-IL-8</protein> and <protein>anti-RANTES</protein> neutralizing antibodies , respectively .


CONCLUSION : These findings suggest that the chemokine pathways are modulated by DEP-PAHs at the transcriptional level , reinforcing the idea that the development of inflammatory reactions might be affected by diesel exhaust emission .

CONCLUSION : These findings suggest that the <protein>chemokine</protein> pathways are modulated by DEP-PAHs at the transcriptional level , reinforcing the idea that the development of inflammatory reactions might be affected by diesel exhaust emission .


20-Epi analogues of 1 , 25-dihydroxyvitamin D3 are highly potent inducers of DRIP coactivator complex binding to the vitamin D3 receptor .

20-Epi analogues of 1 , 25-dihydroxyvitamin D3 are highly potent inducers of <protein>DRIP coactivator complex</protein> binding to the <protein>vitamin D3 receptor</protein> .


1 , 25-Dihydroxyvitamin D3 ( 1 , 25 ( OH ) 2D3 ) plays a major role in the stimulation of bone growth , mineralization , and intestinal calcium and phosphate absorption ; it also acts as a general inhibitor of cellular proliferation .


Several new , clinically relevant compounds dissociate antiproliferative and calcemic activities of 1 , 25 ( OH ) 2D3 , but the molecular basis for this has not been clearly elucidated .


Here , we tested whether the potency of one class of compounds , 20-epi analogues , to induce myeloid cell differentiation , is because of direct molecular effects on vitamin D receptor ( VDR ) .

Here , we tested whether the potency of one class of compounds , 20-epi analogues , to induce myeloid cell differentiation , is because of direct molecular effects on <protein>vitamin D receptor</protein> ( <protein>VDR</protein> ) .


We report that two 20-epi analogues , MC1627 and MC1288 , induced differentiation and transcription of p21 ( Waf1 , Cip1 ) , a key VDR target gene involved in growth inhibition , at a concentration 100-fold lower than that of 1 , 25 ( OH ) 2D3 .

We report that two 20-epi analogues , MC1627 and MC1288 , induced differentiation and transcription of <dna>p21</dna> ( <dna>Waf1</dna> , <dna>Cip1</dna> ) , a key <dna>VDR target gene</dna> involved in growth inhibition , at a concentration 100-fold lower than that of 1 , 25 ( OH ) 2D3 .


We compared this sensitivity to analogue effects on VDR interacting proteins : RXR , GRIP-1 , and DRIP205 , a subunit of the DRIP coactivator complex .

We compared this sensitivity to analogue effects on <protein>VDR</protein> interacting proteins : <protein>RXR</protein> , <protein>GRIP-1</protein> , and <protein>DRIP205</protein> , a subunit of the <protein>DRIP coactivator complex</protein> .