The levels of secreted IL-1beta , TNF-alpha , and IL-6 during 24 h of stimulation were comparable to those induced by Escherichia coli lipopolysaccharide at 1 microg/ml .

The levels of secreted <protein>IL-1beta</protein> , <protein>TNF-alpha</protein> , and <protein>IL-6</protein> during 24 h of stimulation were comparable to those induced by Escherichia coli lipopolysaccharide at 1 microg/ml .


Fab fragment of anti-E. chaffeensis immunoglobulin G complexed with E. chaffeensis did not induce any of these three cytokines , indicating that ehrlichial binding is required for IL-1beta mRNA expression and that binding of the immune complex to the Fc gamma receptor is required for TNF-alpha and IL-6 mRNA expression and enhanced IL-1beta mRNA expression .

Fab fragment of <protein>anti-E. chaffeensis immunoglobulin G</protein> complexed with E. chaffeensis did not induce any of these three cytokines , indicating that ehrlichial binding is required for <rna>IL-1beta mRNA</rna> expression and that binding of the <protein>immune complex</protein> to the <protein>Fc gamma receptor</protein> is required for <protein>TNF-alpha</protein> and <rna>IL-6 mRNA</rna> expression and enhanced <rna>IL-1beta mRNA</rna> expression .


Furthermore , prolonged degradation of IkappaB-alpha and activation of NF-kappaB were demonstrated in THP-1 cells exposed to anti-E. chaffeensis serum and E. chaffeensis .

Furthermore , prolonged degradation of <protein>IkappaB-alpha</protein> and activation of <protein>NF-kappaB</protein> were demonstrated in <cell_line>THP-1 cells</cell_line> exposed to anti-E. chaffeensis serum and E. chaffeensis .


This result implies that development of anti-E. chaffeensis antibody in patients can result in the production of major proinflammatory cytokines , which may play an important role in the pathophysiology of ehrlichiosis and immune responses to it .

This result implies that development of <protein>anti-E. chaffeensis antibody</protein> in patients can result in the production of <protein>major proinflammatory cytokines</protein> , which may play an important role in the pathophysiology of ehrlichiosis and immune responses to it .


The class II trans-activator CIITA interacts with the TBP-associated factor TAFII32 .

The <protein>class II trans-activator CIITA</protein> interacts with the <protein>TBP-associated factor TAFII32</protein> .


The class II trans- activator ( CIITA ) is the main transcriptional co-activator for the expression of MHC class II proteins .

The <protein>class II trans- activator</protein> ( <protein>CIITA</protein> ) is the main transcriptional co-activator for the expression of <protein>MHC class II proteins</protein> .


Its N-terminal 125 amino acids function as an independent transcriptional activation domain .

Its <protein>N-terminal 125 amino acids</protein> function as an <protein>independent transcriptional activation domain</protein> .


Analyses of the primary amino acid sequence of the activation domain predict the presence of three alpha-helices , each with a high proportion of acidic residues .

Analyses of the <protein>primary amino acid sequence</protein> of the <protein>activation domain</protein> predict the presence of three <protein>alpha-helices</protein> , each with a high proportion of acidic residues .


Using site-directed mutagenesis , we found that two of these predicted alpha-helices are required for full transcriptional activation by CIITA .

Using site-directed mutagenesis , we found that two of these predicted <protein>alpha-helices</protein> are required for full transcriptional activation by <protein>CIITA</protein> .


Moreover , a CIITA protein in which both functional alpha-helices have been deleted displays a dominant negative phenotype .

Moreover , a <protein>CIITA protein</protein> in which both functional <protein>alpha-helices</protein> have been deleted displays a dominant negative phenotype .


This activation domain of CIITA interacts with the 32 kDa subunit of the general transcription complex TFIID , TAFII32 .

This <protein>activation domain</protein> of <protein>CIITA</protein> interacts with the <protein>32 kDa subunit</protein> of the <protein>general transcription complex</protein> <protein>TFIID</protein> , <protein>TAFII32</protein> .


Decreased transcriptional activation by N-terminal deletions of CIITA is correlated directly with their reduced binding to TAFII32 .

Decreased transcriptional activation by <protein>N-terminal deletions</protein> of <protein>CIITA</protein> is correlated directly with their reduced binding to <protein>TAFII32</protein> .


We conclude that interactions between TAFII32 and CIITA are responsible for activation of class II genes .

We conclude that interactions between <protein>TAFII32</protein> and <protein>CIITA</protein> are responsible for activation of <dna>class II genes</dna> .


Characterization of interleukin-10 receptor expression on B-cell chronic lymphocytic leukemia cells .

Characterization of <protein>interleukin-10 receptor</protein> expression on <cell_line>B-cell chronic lymphocytic leukemia cells</cell_line> .


B-cell chronic lymphocytic leukemia ( B-CLL ) cells accumulate in vivo in the G0/G1 phase of the cell cycle , suggesting that their malignant expansion is due , at least in part , to a delay in cell death .

<cell_line>B-cell chronic lymphocytic leukemia ( B-CLL ) cells</cell_line> accumulate in vivo in the G0/G1 phase of the cell cycle , suggesting that their malignant expansion is due , at least in part , to a delay in cell death .


However , the cellular or molecular factors responsible for a delay in B-CLL cell death are unknown .

However , the cellular or <protein>molecular factors</protein> responsible for a delay in B-CLL cell death are unknown .


B-CLL cells do express receptors for interferon-alpha ( IFN-alpha ) and IFN-gamma , and activation of both has been shown to promote B-CLL survival in vitro by preventing apoptosis .

B-CLL cells do express receptors for <protein>interferon-alpha</protein> ( <protein>IFN-alpha</protein> ) and <protein>IFN-gamma</protein> , and activation of both has been shown to promote B-CLL survival in vitro by preventing apoptosis .


The interleukin-10 ( IL-10 ) receptor is another member of the IFN receptor family , but its ligand , IL-10 , has been reported to induce apoptosis in B-CLL cells .

The <protein>interleukin-10 ( IL-10 ) receptor</protein> is another member of the <protein>IFN receptor family</protein> , but its ligand , <protein>IL-10</protein> , has been reported to induce apoptosis in <cell_line>B-CLL cells</cell_line> .


In the current study , we undertook a biochemical analysis of IL-10 receptor expression on freshly isolated B-CLL cells and characterized the functional responsiveness of IL-10 binding to its constitutively expressed receptor .

In the current study , we undertook a biochemical analysis of <protein>IL-10 receptor</protein> expression on freshly isolated <cell_line>B-CLL cells</cell_line> and characterized the functional responsiveness of <protein>IL-10</protein> binding to its <protein>constitutively expressed receptor</protein> .


We show that B-CLL cells bind IL-10 with significant specificity and express between 47 and 127 IL-10 receptor sites per cell , with a dissociation constant in the range of 168 to 426 x 10 ( -12 ) mol/L .

We show that <cell_line>B-CLL cells</cell_line> bind <protein>IL-10</protein> with significant specificity and express between 47 and 127 <protein>IL-10 receptor</protein> sites per cell , with a dissociation constant in the range of 168 to 426 x 10 ( -12 ) mol/L .


Ligand binding and activation of the IL-10 receptor expressed on B-CLL cells results in the phosphorylation of signal transducer and activator of transcription 1 ( STAT1 ) and STAT3 proteins .

Ligand binding and activation of the <protein>IL-10 receptor</protein> expressed on <cell_line>B-CLL cells</cell_line> results in the phosphorylation of <protein>signal transducer and activator of transcription 1</protein> ( <protein>STAT1</protein> ) and <protein>STAT3</protein> proteins .


This pattern of STAT protein phosphorylation is identical to IL-10 receptor activation on normal cells and similar to IFN-alpha ( STAT1 and STAT3 ) and IFN-gamma ( STAT1 ) receptor activation in CLL .

This pattern of <protein>STAT protein</protein> phosphorylation is identical to <protein>IL-10 receptor</protein> activation on <cell_type>normal cells</cell_type> and similar to <protein>IFN-alpha</protein> ( <protein>STAT1</protein> and <protein>STAT3</protein> ) and <protein>IFN-gamma</protein> ( <protein>STAT1</protein> ) receptor activation in <cell_line>CLL</cell_line> .


Further , in consecutive samples of fresh blood obtained from patients with B-CLL cells , the addition of IL-10 inhibited B-CLL proliferation , enhanced B-CLL differentiation , but did not induce apoptosis .

Further , in consecutive samples of fresh blood obtained from patients with <cell_line>B-CLL cells</cell_line> , the addition of <protein>IL-10</protein> inhibited <cell_line>B-CLL</cell_line> proliferation , enhanced <cell_line>B-CLL</cell_line> differentiation , but did not induce apoptosis .


Indeed , IL-10 , like IFN-gamma , was able to significantly reduce the amount of B-CLL cell death caused by hydrocortisone-induced apoptosis .

Indeed , <protein>IL-10</protein> , like <protein>IFN-gamma</protein> , was able to significantly reduce the amount of B-CLL cell death caused by hydrocortisone-induced apoptosis .


We conclude that cytokines , which signal through the interferon family of receptors , have comparable functional effects on B-CLL cells .

We conclude that <protein>cytokines</protein> , which signal through the <protein>interferon family</protein> of receptors , have comparable functional effects on <cell_line>B-CLL cells</cell_line> .


Involvement of the N-terminal region of the human mineralocorticoid receptor hormone-binding domain in agonist and antagonist binding as revealed by a new monoclonal antibody .

Involvement of the <protein>N-terminal region</protein> of the <protein>human mineralocorticoid receptor hormone-binding domain</protein> in agonist and antagonist binding as revealed by a <protein>new monoclonal antibody</protein> .


To gain a better understanding of the mechanism of binding to the human mineralocorticoid receptor ( hMR ) , we developed a new monoclonal antibody ( mAb ) raised against the hormone-binding domain ( HBD ) .

To gain a better understanding of the mechanism of binding to the <protein>human mineralocorticoid receptor</protein> ( <protein>hMR</protein> ) , we developed a new <protein>monoclonal antibody</protein> ( <protein>mAb</protein> ) raised against the <protein>hormone-binding domain</protein> ( <protein>HBD</protein> ) .


For this purpose , mice were immunized with a fusion protein including the sequence Thr729-Lys984 of hMR .

For this purpose , mice were immunized with a <protein>fusion protein</protein> including the sequence <protein>Thr729-Lys984</protein> of <protein>hMR</protein> .


After ELISA screening , mAb 18C7 was selected for its specificity towards the HBD .

After ELISA screening , <protein>mAb 18C7</protein> was selected for its specificity towards the <protein>HBD</protein> .


This antibody recognized both the denatured and native MR forms , as well as the hetero-oligomeric MR form and the transformed MR state .

This antibody recognized both the denatured and <protein>native MR forms</protein> , as well as the <protein>hetero-oligomeric MR form</protein> and the <protein>transformed MR state</protein> .


By using several HBD subfragments , the mAb 18C7 epitope was located in the N-terminal region of the HBD from Thr729 to Leu765 .

By using several <protein>HBD subfragments</protein> , the <protein>mAb 18C7 epitope</protein> was located in the <protein>N-terminal region</protein> of the <protein>HBD</protein> from <protein>Thr729 to Leu765</protein> .


We then studied the effect of the antibody on aldosterone and progesterone binding to the hMR .

We then studied the effect of the antibody on aldosterone and progesterone binding to the <protein>hMR</protein> .


When 18C7 was incubated with liganded MR , it was able to partly displace ( 20 % ) the hormone from its binding site .

When <protein>18C7</protein> was incubated with <protein>liganded MR</protein> , it was able to partly displace ( 20 % ) the hormone from its binding site .


When 18C7 was incubated with MR before aldosterone or progesterone , the antibody inhibited 75-80 % of the binding .

When <protein>18C7</protein> was incubated with <protein>MR</protein> before aldosterone or progesterone , the antibody inhibited 75-80 % of the binding .


The effect of 18C7 on the binding was similar with both hormones .

The effect of <protein>18C7</protein> on the binding was similar with both hormones .


A sucrose gradient analysis indicated the simultaneous presence of two kinds of receptor complexes : the steroid-MR complex and the antibody-MR complex .

A sucrose gradient analysis indicated the simultaneous presence of two kinds of receptor complexes : the <protein>steroid-MR complex</protein> and the <protein>antibody-MR complex</protein> .


After its associated proteins , especially the heat-shock protein hsp90 , had been cross-linked with the hMR by dimethylpimelimidate , 18C7 was still able to react with the receptor .

After its associated proteins , especially the <protein>heat-shock protein hsp90</protein> , had been cross-linked with the <protein>hMR</protein> by dimethylpimelimidate , <protein>18C7</protein> was still able to react with the receptor .


Our results indicated that the epitope recognized by 18C7 was directly implicated in hormone binding .

Our results indicated that the <protein>epitope</protein> recognized by <protein>18C7</protein> was directly implicated in hormone binding .


The lack of steroid binding of HBD mutants with the Thr729-Leu765 sequence deleted [ Jalaguier , Mesnier , Leger and Auzou ( 1996 ) J.Steroid Biochem.Mol.Biol.57 , 43-50 ] supports this hypothesis .

The lack of steroid binding of <protein>HBD</protein> mutants with the <protein>Thr729-Leu765 sequence</protein> deleted [ Jalaguier , Mesnier , Leger and Auzou ( 1996 ) J.Steroid Biochem.Mol.Biol.57 , 43-50 ] supports this hypothesis .


Because of the similar behaviours of aldosterone and progesterone , we conclude that the N-terminal Thr729-Leu765 region of the HBD is similarly involved in the binding of both hormones .

Because of the similar behaviours of aldosterone and progesterone , we conclude that the <protein>N-terminal Thr729-Leu765 region</protein> of the <protein>HBD</protein> is similarly involved in the binding of both hormones .


Molecular cloning of SLAP-130 , an SLP-76-associated substrate of the T cell antigen receptor-stimulated protein tyrosine kinases .

Molecular cloning of <protein>SLAP-130</protein> , an <protein>SLP-76-associated substrate</protein> of the <protein>T cell antigen receptor-stimulated protein tyrosine kinases</protein> .


Previous work has demonstrated that SLP-76 , a Grb2-associated tyrosine-phosphorylated protein , augments Interleukin-2 promoter activity when overexpressed in the Jurkat T cell line .

Previous work has demonstrated that <protein>SLP-76</protein> , a <protein>Grb2-associated tyrosine-phosphorylated protein</protein> , augments <dna>Interleukin-2 promoter</dna> activity when overexpressed in the <cell_line>Jurkat T cell line</cell_line> .


This activity requires regions of SLP-76 that mediate protein-protein interactions with other molecules in T cells , suggesting that SLP-76 -associated proteins also function to regulate signal transduction .

This activity requires regions of <protein>SLP-76</protein> that mediate protein-protein interactions with other molecules in <cell_type>T cells</cell_type> , suggesting that <protein>SLP-76</protein> -associated proteins also function to regulate signal transduction .


Here we describe the molecular cloning of SLAP-130 , a SLP-76-associated phosphoprotein of 130 kDa .

Here we describe the molecular cloning of <protein>SLAP-130</protein> , a <protein>SLP-76-associated phosphoprotein</protein> of 130 kDa .


We demonstrate that SLAP-130 is hematopoietic cell-specific and associates with the SH2 domain of SLP-76 .

We demonstrate that <protein>SLAP-130</protein> is hematopoietic cell-specific and associates with the <protein>SH2 domain</protein> of <protein>SLP-76</protein> .


Additionally , we show that SLAP-130 is a substrate of the T cell antigen receptor-induced protein tyrosine kinases .

Additionally , we show that <protein>SLAP-130</protein> is a substrate of the <protein>T cell antigen receptor-induced protein</protein> <protein>tyrosine kinases</protein> .


Interestingly , we find that in contrast to SLP-76 , overexpression of SLAP-130 diminishes T cell antigen receptor -induced activation of the interleukin-2 promoter in Jurkat T cells and interferes with the augmentation of interleukin-2 promoter activity seen when SLP-76 is overexpressed in these cells .

Interestingly , we find that in contrast to <protein>SLP-76</protein> , overexpression of <protein>SLAP-130</protein> diminishes <protein>T cell antigen receptor</protein> -induced activation of the <dna>interleukin-2 promoter</dna> in <cell_line>Jurkat T cells</cell_line> and interferes with the augmentation of <dna>interleukin-2 promoter</dna> activity seen when <protein>SLP-76</protein> is overexpressed in these cells .


These data suggest that SLP-76 recruits a negative regulator , SLAP-130 , as well as positive regulators of signal transduction in T cells .

These data suggest that <protein>SLP-76</protein> recruits a negative regulator , <protein>SLAP-130</protein> , as well as positive regulators of signal transduction in <cell_type>T cells</cell_type> .


Molecular actions of prolactin in the immune system .


The immunoregulatory properties of prolactin , a pituitary peptide hormone , have received renewed attention .

The <protein>immunoregulatory</protein> properties of prolactin , a pituitary peptide hormone , have received renewed attention .


The prolactin receptor , a member of the hematopoietin/cytokine receptor superfamily , is ubiquitously expressed by cells in the immune system .

The <protein>prolactin receptor</protein> , a member of the <protein>hematopoietin/cytokine receptor superfamily</protein> , is ubiquitously expressed by cells in the immune system .


Certain subpopulations of lymphocytes synthesize and secrete biologically active prolactin , which suggests that prolactin can act as an autocrine and/or paracrine factor to modulate the activities of cells of the immune system .

Certain subpopulations of <cell_type>lymphocytes</cell_type> synthesize and secrete biologically active prolactin , which suggests that prolactin can act as an autocrine and/or paracrine factor to modulate the activities of cells of the immune system .


This review focuses on the molecular actions of prolactin in the immune system .


Emphasis is given to recent information about the molecular mechanisms of prolactin receptor signal transduction , and the signaling molecules and prolactin-inducible target genes that participate in these responses .

Emphasis is given to recent information about the molecular mechanisms of prolactin receptor signal transduction , and the <protein>signaling molecules</protein> and <dna>prolactin-inducible target genes</dna> that participate in these responses .


In particular , the prolactin-inducible interferon regulatory factor-1 gene and its roles in mediating diverse immune responses .

In particular , the <dna>prolactin-inducible interferon regulatory factor-1 gene</dna> and its roles in mediating diverse immune responses .


Transcriptional regulation during myelopoiesis .


The coordinated production of all blood cells from a common stem cell is a highly regulated process involving successive stages of commitment and differentiation .

The coordinated production of all <cell_type>blood cells</cell_type> from a <cell_type>common stem cell</cell_type> is a highly regulated process involving successive stages of commitment and differentiation .


From analyses of mice deficient in transcription factor genes and from the characterizations of chromosome breakpoints in human leukemias , it has become evident that transcription factors are important regulators of hematopoiesis .

From analyses of mice deficient in <dna>transcription factor genes</dna> and from the characterizations of <dna>chromosome breakpoints</dna> in human leukemias , it has become evident that <protein>transcription factors</protein> are important regulators of hematopoiesis .


During myelopoiesis , which includes the development of granulocytic and monocytic lineages , transcription factors from several families are active , including AML1/CBF beta , C/EBP , Ets , c-Myb , HOX , and MZF-1 .

During myelopoiesis , which includes the development of <cell_type>granulocytic and monocytic lineages</cell_type> , <protein>transcription factors</protein> from several families are active , including <dna>AML1/CBF beta</dna> , <dna>C/EBP</dna> , <dna>Ets</dna> , <dna>c-Myb</dna> , <dna>HOX</dna> , and <dna>MZF-1</dna> .


Few of these factors are expressed exclusively in myeloid cells ; instead it appears that they cooperatively regulate transcription of myeloid-specific genes .

Few of these factors are expressed exclusively in <cell_type>myeloid cells</cell_type> ; instead it appears that they cooperatively regulate transcription of <dna>myeloid-specific genes</dna> .


Here we discuss recent advances in transcriptional regulation during myelopoiesis .


Activation of transcription factor NF-kappa B by phagocytic stimuli in human neutrophils .

Activation of transcription factor <protein>NF-kappa B</protein> by phagocytic stimuli in human neutrophils .


Phagocytosis represents an important physiological trigger for the inducible expression of several genes in human neutrophils .

Phagocytosis represents an important physiological trigger for the inducible expression of several genes in <cell_type>human neutrophils</cell_type> .


Here , we report that a DNA-binding activity primarily consisting of the classical NF-kappa B heterodimer , p50/RelA , is induced in phagocytosing neutrophils .

Here , we report that a DNA-binding activity primarily consisting of the classical <protein>NF-kappa B heterodimer</protein> , <protein>p50/RelA</protein> , is induced in <cell_type>phagocytosing neutrophils</cell_type> .


Under these conditions , NF-kappa B activation was found to be a rapid and transient response , reaching a maximum by 10-15 min , and returning to near-basal levels by 30 min .

Under these conditions , <protein>NF-kappa B</protein> activation was found to be a rapid and transient response , reaching a maximum by 10-15 min , and returning to near-basal levels by 30 min .


In neutrophils undergoing the phagocytosis of opsonized yeasts , the onset of NF-kappa B activation was paralleled by a decline in immunoreactive I kappa B-alpha protein levels , and the cellular I kappa B -alpha pool was replenished by 30 min , in agreement with our gel shift data .

In neutrophils undergoing the phagocytosis of opsonized yeasts , the onset of <protein>NF-kappa B</protein> activation was paralleled by a decline in immunoreactive <protein>I kappa B-alpha</protein> protein levels , and the cellular <protein>I kappa B</protein> -alpha pool was replenished by 30 min , in agreement with our gel shift data .


We conclude that NF-kappa B activation could constitute one of the mechanisms whereby the expression of kappa B-responsive genes is enhanced in phagocytosing neutrophils .

We conclude that <protein>NF-kappa B</protein> activation could constitute one of the mechanisms whereby the expression of <dna>kappa B-responsive genes</dna> is enhanced in <cell_type>phagocytosing neutrophils</cell_type> .


To our knowledge , this represents the first demonstration that phagocytic stimuli can induce NF-kappa B activation in human neutrophils .

To our knowledge , this represents the first demonstration that phagocytic stimuli can induce <protein>NF-kappa B</protein> activation in <cell_type>human neutrophils</cell_type> .


Induction of human immunodeficiency virus type 1 expression in monocytic cells by Cryptococcus neoformans and Candida albicans .

Induction of human immunodeficiency virus type 1 expression in <cell_type>monocytic cells</cell_type> by Cryptococcus neoformans and Candida albicans .


Because candidiasis and cryptococcosis are common in human immunodeficiency virus ( HIV ) -infected persons , the effect of Cryptococcus neoformans and Candida albicans on HIV expression in monocytic cells was examined .

Because candidiasis and cryptococcosis are common in human immunodeficiency virus ( HIV ) -infected persons , the effect of Cryptococcus neoformans and Candida albicans on HIV expression in <cell_type>monocytic cells</cell_type> was examined .


Stimulation of the latently HIV-infected myelomonocytic cell line OM-10.1 with C. neoformans and C. albicans in the presence of pooled human serum caused a ratio-dependent increase in HIV production .

Stimulation of the latently <cell_line>HIV-infected myelomonocytic cell line</cell_line> OM-10.1 with C. neoformans and C. albicans in the presence of pooled human serum caused a ratio-dependent increase in HIV production .


Induction of HIV by C. neoformans was enhanced by anti-capsular antibody , while induction by both organisms was inhibited by anti-TNF-alpha antibody .

Induction of HIV by C. neoformans was enhanced by <protein>anti-capsular antibody</protein> , while induction by both organisms was inhibited by <protein>anti-TNF-alpha antibody</protein> .


In THP-1 cells transfected with HIV plasmid constructs , both organisms induced transcription from the HIV long terminal repeat that was dependent on intact NF-kappaB binding sequences .

In THP-1 cells transfected with <dna>HIV plasmid constructs</dna> , both organisms induced transcription from the <dna>HIV long terminal repeat</dna> that was dependent on intact <dna>NF-kappaB binding sequences</dna> .


Thus , C. neoformans and C. albicans enhance HIV expression in monocytic cells through a TNF-alpha- and NF-kappaB-dependent mechanism .

Thus , C. neoformans and C. albicans enhance HIV expression in <cell_type>monocytic cells</cell_type> through a TNF-alpha- and NF-kappaB-dependent mechanism .


In HIV-infected patients , such enhancement may further impair host immunity and could accelerate the course of HIV disease .


Nuclear levels of NF-kappaB correlate with syncytium-forming capacity of 8e51 cells , expressing a defective HIV virus .

Nuclear levels of <protein>NF-kappaB</protein> correlate with syncytium-forming capacity of <cell_line>8e51 cells</cell_line> , expressing a defective HIV virus .


The double NF-kappaB site identified in the LTR of the human immunodeficiency virus-1 ( HIV-1 ) has been demonstrated to be necessary for efficient viral transcription .

The <dna>double NF-kappaB site</dna> identified in the <dna>LTR</dna> of the human immunodeficiency virus-1 ( HIV-1 ) has been demonstrated to be necessary for efficient viral transcription .


In this report we present the characterisation of NF-kappaB subunits engaged in complexes binding to the HIV-1 NF-kappaB site in human 8e51 T-cells , that harbour a defective HIV-1 .

In this report we present the characterisation of <protein>NF-kappaB subunits</protein> engaged in complexes binding to the <dna>HIV-1 NF-kappaB site</dna> in <cell_line>human 8e51 T-cells</cell_line> , that harbour a defective HIV-1 .


At least four different specific NF-kappaB complexes are present in the nucleus of these cells .

At least four different specific <protein>NF-kappaB complexes</protein> are present in the nucleus of these cells .


With the use of specific antibodies we have determined the composition of each complex using electrophoretic mobility shift assays .

With the use of <protein>specific antibodies</protein> we have determined the composition of each complex using electrophoretic mobility shift assays .


The results show the presence of several NF-kappaB family members , with the transactivating RelA being engaged in multiple complexes .

The results show the presence of several <protein>NF-kappaB family members ,</protein> with the transactivating <protein>RelA</protein> being engaged in multiple complexes .


The importance of NF-kappaB complexes in viral functions has been established comparing the level of NF-kappaB DNA-binding complexes with syncytia-forming activity of 8e51 cells .

The importance of <protein>NF-kappaB complexes</protein> in viral functions has been established comparing the level of NF-kappaB DNA-binding complexes with syncytia-forming activity of <cell_line>8e51 cells</cell_line> .


In fact , 8e51 cells that had almost lost their syncytia-forming capacity were found to contain at least 10 times less active NF-kappaB DNA-binding complex than the actively fusing cells .

In fact , <cell_line>8e51 cells</cell_line> that had almost lost their syncytia-forming capacity were found to contain at least 10 times less active <protein>NF-kappaB DNA-binding complex</protein> than the <cell_type>actively fusing cells</cell_type> .


The correlation is specific as the level of at least three other transcription factors did not change .

The correlation is specific as the level of at least three other <protein>transcription factors</protein> did not change .


c-Myb and Ets proteins synergize to overcome transcriptional repression by ZEB .

<dna>c-Myb</dna> and <protein>Ets proteins</protein> synergize to overcome transcriptional repression by <protein>ZEB</protein> .


The Zfh family of zinc finger/homeodomain proteins was first identified in Drosophila where it is required for differentiation of tissues such as the central nervous system and muscle .

The <protein>Zfh family</protein> of <protein>zinc finger/homeodomain proteins</protein> was first identified in Drosophila where it is required for differentiation of tissues such as the central nervous system and muscle .


ZEB , a vertebrate homolog of Zfh-1 , binds a subset of E boxes and blocks myogenesis through transcriptional repression of muscle genes .

<protein>ZEB</protein> , a vertebrate homolog of <protein>Zfh-1</protein> , binds a subset of <dna>E boxes</dna> and blocks myogenesis through transcriptional repression of <dna>muscle genes</dna> .


We present evidence here that ZEB also has an important role in controlling hematopoietic gene transcription .

We present evidence here that <protein>ZEB</protein> also has an important role in controlling <dna>hematopoietic gene</dna> transcription .


Two families of transcription factors that are required for normal hematopoiesis are c-Myb and Ets .

Two families of <protein>transcription factors</protein> that are required for normal hematopoiesis are <protein>c-Myb</protein> and <protein>Ets</protein> .


These factors act synergistically to activate transcription , and this synergy is required for transcription of at least several important hematopoietic genes .

These factors act synergistically to activate transcription , and this synergy is required for transcription of at least several important <dna>hematopoietic genes</dna> .


ZEB blocks the activity of c-Myb and Ets individually , but together the factors synergize to resist this repression .

<protein>ZEB</protein> blocks the activity of <protein>c-Myb</protein> and <protein>Ets</protein> individually , but together the factors synergize to resist this repression .


Such repression imposes a requirement for both c-Myb and Ets for transcriptional activity , providing one explanation for why synergy between these factors is important .

Such repression imposes a requirement for both <protein>c-Myb</protein> and <protein>Ets</protein> for transcriptional activity , providing one explanation for why synergy between these factors is important .


The balance between repression by ZEB and transcriptional activation by c-Myb /Ets provides a flexible regulatory mechanism for controlling gene expression in hematopoietic cells .

The balance between repression by <protein>ZEB</protein> and transcriptional activation by <protein>c-Myb</protein> <protein>/Ets</protein> provides a flexible regulatory mechanism for controlling gene expression in <cell_type>hematopoietic cells</cell_type> .


We demonstrate that one target of this positive/negative regulation in vivo is the alpha4 integrin , which play a key role in normal hematopoiesis and function of mature leukocytes .

We demonstrate that one target of this positive/negative regulation in vivo is the <protein>alpha4 integrin</protein> , which play a key role in normal hematopoiesis and function of <cell_type>mature leukocytes</cell_type> .


Comparison of the transactivation domains of Stat5 and Stat6 in lymphoid cells and mammary epithelial cells .

Comparison of the <protein>transactivation domains</protein> of <protein>Stat5</protein> and <protein>Stat6</protein> in <cell_type>lymphoid cells</cell_type> and <cell_type>mammary epithelial cells</cell_type> .


Stat ( signal transducers and activators of transcription ) and Jak ( Janus kinases ) proteins are central components in the signal transduction events in hematopoietic and epithelial cells .

<protein>Stat</protein> ( <protein>signal transducers and activators of transcription</protein> ) and <protein>Jak</protein> ( <protein>Janus kinases</protein> ) proteins are central components in the signal transduction events in <cell_type>hematopoietic and epithelial cells</cell_type> .


They are rapidly activated by various cytokines , hormones , and growth factors .

They are rapidly activated by various <protein>cytokines</protein> , hormones , and growth factors .


Upon ligand binding and cytokine receptor dimerization , Stat proteins are phosphorylated on tyrosine residues by Jak kinases .

Upon ligand binding and <protein>cytokine receptor</protein> dimerization , <protein>Stat proteins</protein> are phosphorylated on tyrosine residues by <protein>Jak kinases</protein> .


Activated Stat proteins form homo- or heterodimers , translocate to the nucleus , and induce transcription from responsive genes .

Activated <protein>Stat proteins</protein> form homo- or heterodimers , translocate to the nucleus , and induce transcription from responsive genes .


Stat5 and Stat6 are transcription factors active in mammary epithelial cells and immune cells .

<protein>Stat5</protein> and <protein>Stat6</protein> are <protein>transcription factors</protein> active in <cell_type>mammary epithelial cells</cell_type> and immune cells .


Prolactin activates Stat5 , and interleukin-4 ( IL-4 ) activates Stat6 .

Prolactin activates <protein>Stat5</protein> , and <protein>interleukin-4</protein> ( <protein>IL-4</protein> ) activates <protein>Stat6</protein> .


Both cytokines are able to stimulate cell proliferation , differentiation , and survival .

Both <protein>cytokines</protein> are able to stimulate cell proliferation , differentiation , and survival .


We investigated the transactivation potential of Stat6 and found that it is not restricted to lymphocytes .

We investigated the transactivation potential of <protein>Stat6</protein> and found that it is not restricted to lymphocytes .


IL-4 -dependent activation of Stat6 was also observed in HC11 mammary epithelial cells .

<protein>IL-4</protein> -dependent activation of <protein>Stat6</protein> was also observed in <cell_line>HC11 mammary epithelial cells</cell_line> .


In these cells , Stat6 activation led to the induction of the beta-casein gene promoter .

In these cells , <protein>Stat6</protein> activation led to the induction of the <dna>beta-casein gene promoter</dna> .


The induction of this promoter was confirmed in COS7 cells .

The induction of this promoter was confirmed in <cell_line>COS7 cells</cell_line> .


The glucocorticoid receptor was able to further enhance IL-4 -induced gene transcription through the action of Stat6 .

The <protein>glucocorticoid receptor</protein> was able to further enhance <protein>IL-4</protein> -induced gene transcription through the action of <protein>Stat6</protein> .


Deletion analysis of the carboxyl-terminal region of Stat6 and recombination of this region with a heterologous DNA binding domain allowed the delimitation and characterization of the transactivation domain of Stat6 .

Deletion analysis of the <protein>carboxyl-terminal region</protein> of <protein>Stat6</protein> and recombination of this region with a <protein>heterologous DNA binding domain</protein> allowed the delimitation and characterization of the transactivation domain of <protein>Stat6</protein> .


The potencies of the transactivation domains of Stat5 , Stat6 , and viral protein VP16 were compared .

The potencies of the <protein>transactivation domains</protein> of <protein>Stat5</protein> , <protein>Stat6</protein> , and <protein>viral protein VP16</protein> were compared .


Stat6 had a transactivation domain which was about 10-fold stronger than that of Stat5 .

<protein>Stat6</protein> had a transactivation domain which was about 10-fold stronger than that of <protein>Stat5</protein> .


In pre-B cells ( Ba/F3 ) , the transactivation domain of Stat6 was IL-4 regulated , independently from its DNA binding function .

In <cell_type>pre-B cells</cell_type> ( Ba/F3 ) , the transactivation domain of <protein>Stat6</protein> was <protein>IL-4</protein> regulated , independently from its DNA binding function .


ETS1 , NFkappaB and AP1 synergistically transactivate the human GM-CSF promoter .

<protein>ETS1</protein> , <protein>NFkappaB</protein> and <protein>AP1</protein> synergistically transactivate the <dna>human GM-CSF promoter</dna> .


Activation of helper T cells results in coordinate expression of a number of cytokines involved in differentiation , proliferation and activation of the haematopoietic system .

Activation of <cell_type>helper T cells</cell_type> results in coordinate expression of a number of <protein>cytokines</protein> involved in differentiation , proliferation and activation of the haematopoietic system .


Granulocyte-macrophage colony stimulating factor ( GM-CSF ) is one such cytokine , whose increased expression results mostly from increases in transcription .

<protein>Granulocyte-macrophage colony stimulating factor</protein> ( <protein>GM-CSF</protein> ) is one such <protein>cytokine</protein> , whose increased expression results mostly from increases in transcription .


Cis-acting elements with NFkappaB , AP1 and ETS-like binding motifs have been identified in the promoter region of the GM-CSF gene , and are important or essential for transcriptional activity following T cell activation .

<dna>Cis-acting elements</dna> with <dna>NFkappaB , AP1 and ETS-like binding motifs</dna> have been identified in the promoter region of the <dna>GM-CSF gene</dna> , and are important or essential for transcriptional activity following T cell activation .


ETS1 is a transcription factor of the ETS family that is expressed in T cells .

<protein>ETS1</protein> is a <protein>transcription factor</protein> of the <protein>ETS family</protein> that is expressed in <cell_type>T cells</cell_type> .


We have previously shown that ETS1 can transactivate GM-CSF in Jurkat T cells , but only after the cells have been stimulated by treatment with PMA and ionomycin , agents that mimic T cell activation .

We have previously shown that <protein>ETS1</protein> can transactivate <protein>GM-CSF</protein> in <cell_line>Jurkat T cells</cell_line> , but only after the cells have been stimulated by treatment with PMA and ionomycin , agents that mimic T cell activation .


Thus we proposed that ETS1 , which is expressed constitutively in Jurkat cells , may act in concert with PMA/ionomycin inducible factors .

Thus we proposed that <protein>ETS1</protein> , which is expressed constitutively in <cell_line>Jurkat cells</cell_line> , may act in concert with <protein>PMA/ionomycin inducible factors</protein> .


Here we show that ETS1 can transactivate a GM-CSF reporter construct in unstimulated Jurkat cells , providing that either NFkappaB or AP1 transcription factors are supplied by co-transfection .

Here we show that <protein>ETS1</protein> can transactivate a <dna>GM-CSF reporter construct</dna> in unstimulated <cell_line>Jurkat cells</cell_line> , providing that either <protein>NFkappaB</protein> or <protein>AP1</protein> <protein>transcription factors</protein> are supplied by co-transfection .


We confirm that binding of endogenous NFkappaB and AP1 is induced following PMA/ionomycin treatment of T cells .

We confirm that binding of endogenous <protein>NFkappaB</protein> and <protein>AP1</protein> is induced following PMA/ionomycin treatment of <cell_type>T cells</cell_type> .


Transactivation by ETS1 , NFkappaB and AP1 is synergistic , and mutation of the individual binding sites reveals that the transcriptional activities of these factors are interdependent .

Transactivation by <protein>ETS1</protein> , <protein>NFkappaB</protein> and <protein>AP1</protein> is synergistic , and mutation of the individual binding sites reveals that the transcriptional activities of these factors are interdependent .


Our results suggest that constitutive ETS1 , and inducible NFkappaB and AP1 , cooperate as part of a higher order transcriptional complex in activated T cells .

Our results suggest that constitutive <protein>ETS1</protein> , and inducible <protein>NFkappaB</protein> and <protein>AP1</protein> , cooperate as part of a higher order <protein>transcriptional complex</protein> in activated <cell_type>T cells</cell_type> .


Abnormal T lymphocyte development induced by targeted overexpression of IkappaB alpha .

Abnormal <cell_type>T lymphocyte</cell_type> development induced by targeted overexpression of <protein>IkappaB alpha</protein> .


A role in thymic maturation for factors of the NF-kappaB family has long been suspected , but not yet proven .

A role in thymic maturation for factors of the <protein>NF-kappaB family</protein> has long been suspected , but not yet proven .


Transgenic mice with a lymphocyte-specific defect in NF-kappaB activation were produced by targeted expression of human IkappaB alpha .

Transgenic mice with a lymphocyte-specific defect in <protein>NF-kappaB</protein> activation were produced by targeted expression of human <protein>IkappaB alpha</protein> .


The thymic cellularity of these mice was significantly decreased .


The proportion of mature , TCRhigh thymocytes of the alphabeta lineage was reduced , and the remaining TCRhigh population contained an unusually high proportion of double-positive cells .

The proportion of mature , <cell_type>TCRhigh thymocytes</cell_type> of the <cell_type>alphabeta lineage</cell_type> was reduced , and the remaining <cell_type>TCRhigh population</cell_type> contained an unusually high proportion of <cell_line>double-positive cells</cell_line> .


This defect in maturation resulted in a transgene dose-dependent reduction in peripheral T lymphocytes , with the CD8 lineage being more severely affected .

This defect in maturation resulted in a transgene dose-dependent reduction in <cell_type>peripheral T lymphocytes</cell_type> , with the <cell_type>CD8 lineage</cell_type> being more severely affected .


These data provide direct evidence for the involvement of NF-kappaB/Rel family proteins in late stages of T lymphocyte development , coincident with positive and negative selection .

These data provide direct evidence for the involvement of <protein>NF-kappaB/Rel family proteins</protein> in late stages of <cell_type>T lymphocyte</cell_type> development , coincident with positive and negative selection .


CD40 is a functional activation antigen and B7-independent T cell costimulatory molecule on normal human lung fibroblasts .

<protein>CD40</protein> is a functional <protein>activation antigen</protein> and <protein>B7-independent T cell costimulatory molecule</protein> on normal <cell_type>human lung fibroblasts</cell_type> .


CD40 is an important signaling and activation Ag found on certain bone marrow-derived cells .

<protein>CD40</protein> is an important signaling and activation Ag found on certain <cell_type>bone marrow-derived cells</cell_type> .


Recently , CD40 also has been shown to be expressed by mesenchymal cells , including human fibroblasts .

Recently , <protein>CD40</protein> also has been shown to be expressed by <cell_type>mesenchymal cells</cell_type> , including <cell_type>human fibroblasts</cell_type> .


Little is known about the role of CD40 in fibroblasts .

Little is known about the role of <protein>CD40</protein> in <cell_type>fibroblasts</cell_type> .


The current study investigates the hypothesis that CD40 expressed on lung fibroblasts is an activation structure and mechanism for interaction with hemopoietic cells .

The current study investigates the hypothesis that <protein>CD40</protein> expressed on <cell_type>lung fibroblasts</cell_type> is an activation structure and mechanism for interaction with <cell_type>hemopoietic cells</cell_type> .


Communication between resident tissue fibroblasts and T cells is necessary for normal wound healing , and can be pathologic , resulting in tissue fibrosis .

Communication between resident tissue <cell_type>fibroblasts</cell_type> and <cell_type>T cells</cell_type> is necessary for normal wound healing , and can be pathologic , resulting in tissue fibrosis .


Signaling through CD40 with soluble CD40 ligand stimulated fibroblast activation , as evidenced by mobilization of nuclear factor-kappaB and by induction of the proinflammatory and chemoattractant cytokines IL-6 and IL-8 .

Signaling through <protein>CD40</protein> with soluble <protein>CD40</protein> ligand stimulated fibroblast activation , as evidenced by mobilization of nuclear factor-kappaB and by induction of the proinflammatory and chemoattractant <protein>cytokines</protein> <protein>IL-6</protein> and <protein>IL-8</protein> .


IFN-gamma-primed lung fibroblasts costimulate T lymphocyte proliferation utilizing CD40 , but not the well-studied costimulatory molecules B7-1 and B7-2 .

<cell_line>IFN-gamma-primed lung fibroblasts</cell_line> costimulate T lymphocyte proliferation utilizing <protein>CD40</protein> , but not the well-studied <protein>costimulatory molecules</protein> <protein>B7-1</protein> and <protein>B7-2</protein> .


Data reported herein support the hypothesis that cognate interactions between tissue fibroblasts and infiltrating T lymphocytes , via the CD40 /CD40L pathway , augment inflammation and may promote fibrogenesis by activating both cell types .

Data reported herein support the hypothesis that cognate interactions between <cell_type>tissue fibroblasts</cell_type> and infiltrating T lymphocytes , via the <protein>CD40</protein> <protein>/CD40L</protein> pathway , augment inflammation and may promote fibrogenesis by activating both cell types .


Biphasic control of NF-kappa B activation induced by the triggering of HLA-DR antigens expressed on B cells .

Biphasic control of <protein>NF-kappa B</protein> activation induced by the triggering of <protein>HLA-DR antigens</protein> expressed on <cell_type>B cells</cell_type> .


The regulation of NF-kappa B activation following the triggering of HLA-DR antigens by mAb L243 has been studied at various times in Raji cells .

The regulation of <protein>NF-kappa B</protein> activation following the triggering of <protein>HLA-DR antigens</protein> by <protein>mAb L243</protein> has been studied at various times in <cell_line>Raji cells</cell_line> .


Electrophoretic mobility shift assays demonstrated a strong increase of NF-kappa B DNA binding after triggering of HLA-DR antigens .

Electrophoretic mobility shift assays demonstrated a strong increase of <protein>NF-kappa B</protein> DNA binding after triggering of <protein>HLA-DR antigens</protein> .


Using TNF-alpha-activity neutralizing antibodies , the authors demonstrated that the upregulation of NF-kappa B was found to depend , at later time point , on an autocrine effect of TNF-alpha secreted following triggering of HLA-DR antigens .

Using <protein>TNF-alpha-activity neutralizing antibodies</protein> , the authors demonstrated that the upregulation of <protein>NF-kappa B</protein> was found to depend , at later time point , on an autocrine effect of <protein>TNF-alpha</protein> secreted following triggering of <protein>HLA-DR antigens</protein> .


In contrast , it was found to be TNF-alpha independent in the early time point .

In contrast , it was found to be <protein>TNF-alpha</protein> independent in the early time point .


Moreover , the upregulation of NF-kappa B binding activity is regulated by the triggering of selected epitopes of HLA-DR antigens .

Moreover , the upregulation of <protein>NF-kappa B</protein> binding activity is regulated by the triggering of selected <protein>epitopes</protein> of <protein>HLA-DR antigens</protein> .


In fact , mAb L243 but not the staphylococcal superantigens , staphylococcal exotoxin toxic shock syndrome toxin-I or staphylococcal enterotoxin B , regulate the NF-kappa B binding activity .

In fact , <protein>mAb L243</protein> but not the <protein>staphylococcal superantigens</protein> , <protein>staphylococcal exotoxin toxic shock syndrome toxin-I</protein> or <protein>staphylococcal enterotoxin B</protein> , regulate the <protein>NF-kappa B</protein> binding activity .


Regulation of inosine-5'-monophosphate dehydrogenase type II gene expression in human T cells .

Regulation of <protein>inosine-5'-monophosphate dehydrogenase type II</protein> gene expression in <cell_type>human T cells</cell_type> .


Role for a novel 5 ' palindromic octamer sequence .

Role for a novel <dna>5 ' palindromic octamer sequence</dna> .


Expression of the gene encoding human inosine- 5'-monophosphate dehydrogenase ( IMPDH ) type II , an enzyme catalyzing the rate-limiting step in the generation of guanine nucleotides , is increased more than 10-fold in activated peripheral blood T lymphocytes and is required for T cell activation .

Expression of the gene encoding <protein>human inosine- 5'-monophosphate dehydrogenase ( IMPDH ) type II</protein> , an <protein>enzyme</protein> catalyzing the rate-limiting step in the generation of guanine nucleotides , is increased more than 10-fold in <cell_type>activated peripheral blood T lymphocytes</cell_type> and is required for T cell activation .


We have examined the 5'-regulatory sequences that are important for the transcriptional regulation of this gene in T cells .

We have examined the <dna>5'-regulatory sequences</dna> that are important for the transcriptional regulation of this gene in <cell_type>T cells</cell_type> .


DNase I mapping of genomic DNA identified a hypersensitive element near the transcription initiation site .

<protein>DNase I</protein> mapping of <dna>genomic DNA</dna> identified a <dna>hypersensitive element</dna> near the <dna>transcription initiation site</dna> .


Fine mapping by in vivo footprinting demonstrated five transcription factor binding sites that are occupied in both resting and activated peripheral blood T lymphocytes ; these are tandem CRE motifs , a Sp1 site , an overlapping Egr-1/Sp1 site , and a novel palindromic octamer sequence ( POS ) .

Fine mapping by in vivo footprinting demonstrated five transcription factor binding sites that are occupied in both resting and <cell_type>activated peripheral blood T lymphocytes</cell_type> ; these are <dna>tandem CRE motifs</dna> , a <dna>Sp1 site</dna> , an overlapping <dna>Egr-1/Sp1 site</dna> , and a novel <dna>palindromic octamer sequence</dna> ( <dna>POS</dna> ) .


The tandem CRE and POS sites are of major functional importance as judged by mutational and electrophoretic mobility shift analyses .

The tandem <dna>CRE and POS sites</dna> are of major functional importance as judged by mutational and electrophoretic mobility shift analyses .


These data provide evidence that expression of the human IMPDH type II gene is predominantly regulated by the nuclear factors ATF-2 and an as yet unidentified POS-binding protein .

These data provide evidence that expression of the <dna>human IMPDH type II gene</dna> is predominantly regulated by the <protein>nuclear factors</protein> <protein>ATF-2</protein> and an as yet unidentified <protein>POS-binding protein</protein> .


Additional major protein-DNA interactions do not occur within the promoter region after T lymphocyte activation , indicating a requirement for additional protein-protein interactions and/or post-translational modifications of pre-bound transcription factors to account for the observed increase in IMPDH type II gene expression .

Additional major protein-DNA interactions do not occur within the promoter region after T lymphocyte activation , indicating a requirement for additional protein-protein interactions and/or post-translational modifications of <protein>pre-bound transcription factors</protein> to account for the observed increase in IMPDH type II gene expression .


Distinct mechanisms for N-acetylcysteine inhibition of cytokine-induced E-selectin and VCAM-1 expression .

Distinct mechanisms for N-acetylcysteine inhibition of <protein>cytokine-induced E-selectin</protein> and <protein>VCAM-1</protein> expression .


We have examined the effects of N-acetyl-L-cysteine ( NAC ) , a well-characterized , thiol-containing antioxidant , on agonist-induced monocytic cell adhesion to endothelial cells ( EC ) .

We have examined the effects of N-acetyl-L-cysteine ( NAC ) , a well-characterized , thiol-containing antioxidant , on agonist-induced monocytic cell adhesion to <cell_type>endothelial cells</cell_type> ( <cell_type>EC</cell_type> ) .


NAC inhibited interleukin-1 ( IL-1 beta ) -induced , but not basal , adhesion with 50 % inhibition at approximately 20 mM .

NAC inhibited <protein>interleukin-1</protein> ( <protein>IL-1 beta</protein> ) -induced , but not basal , adhesion with 50 % inhibition at approximately 20 mM .


Monocytic cell adhesion to EC in response to tumor necrosis factor-alpha ( TNF-alpha ) , lipopolysaccharide ( LPS ) , alpha-thrombin , or phorbol 12-myristate 13-acetate ( PMA ) was similarly inhibited by NAC .

Monocytic cell adhesion to <cell_type>EC</cell_type> in response to <protein>tumor necrosis factor-alpha</protein> ( <protein>TNF-alpha</protein> ) , lipopolysaccharide ( LPS ) , <protein>alpha-thrombin</protein> , or phorbol 12-myristate 13-acetate ( PMA ) was similarly inhibited by NAC .


Unlike published studies with pyrrolidinedithiocarbamate , which specifically inhibited vascular cell adhesion molecule 1 ( VCAM-1 ) , NAC inhibited IL-1 beta-induced mRNA and cell surface expression of both E-selectin and VCAM-1 .

Unlike published studies with pyrrolidinedithiocarbamate , which specifically inhibited <protein>vascular cell adhesion molecule 1</protein> ( <protein>VCAM-1</protein> ) , NAC inhibited <rna>IL-1 beta-induced mRNA</rna> and cell surface expression of both <protein>E-selectin</protein> and <protein>VCAM-1</protein> .


NAC had no effect on the half-life of E-selectin or VCAM-1 mRNA .

NAC had no effect on the half-life of <rna>E-selectin or VCAM-1 mRNA</rna> .


Although NAC reduced nuclear factor-kappa B ( NF-kappa B ) activation in EC as measured by gel-shift assays using an oligonucleotide probe corresponding to the consensus NF-kappa B binding sites of the VCAM-1 gene ( VCAM-NF-kappa B ) , the antioxidant had no appreciable effect when an oligomer corresponding to the consensus NF-kappa B binding site of the E-selectin gene ( E-selectin -NF-kappa B ) was used .

Although NAC reduced <protein>nuclear factor-kappa B</protein> ( <protein>NF-kappa B</protein> ) activation in <cell_type>EC</cell_type> as measured by gel-shift assays using an <dna>oligonucleotide probe</dna> corresponding to the consensus <dna>NF-kappa B binding sites</dna> of the <dna>VCAM-1 gene</dna> ( <protein>VCAM-NF-kappa B</protein> ) , the antioxidant had no appreciable effect when an oligomer corresponding to the <dna>consensus NF-kappa B binding site</dna> of the <dna>E-selectin gene</dna> ( <protein>E-selectin</protein> <protein>-NF-kappa B</protein> ) was used .


Because NF-kappa B has been reported to be redox sensitive , we studied the effects of NAC on the EC redox environment .

Because <protein>NF-kappa B</protein> has been reported to be redox sensitive , we studied the effects of NAC on the <cell_type>EC</cell_type> redox environment .


NAC caused an expected dramatic increase in the reduced glutathione ( GSH ) levels in EC .

NAC caused an expected dramatic increase in the reduced glutathione ( GSH ) levels in <cell_type>EC</cell_type> .


In vitro studies demonstrated that whereas the binding affinity of NF-kappa B to the VCAM-NF-kappa B oligomer peaked at a GSH-to-oxidized glutathione ( GSSG ) ratio of approximately 200 and decreased at higher ratios , the binding to the E-selectin-NF-kappa B oligomer appeared relatively unaffected even at ratios > 400 , i.e. , those achieved in EC treated with 40 mM NAC .

In vitro studies demonstrated that whereas the binding affinity of <protein>NF-kappa B</protein> to the <dna>VCAM-NF-kappa B oligomer</dna> peaked at a GSH-to-oxidized glutathione ( GSSG ) ratio of approximately 200 and decreased at higher ratios , the binding to the <dna>E-selectin-NF-kappa B oligomer</dna> appeared relatively unaffected even at ratios > 400 , i.e. , those achieved in <cell_type>EC</cell_type> treated with 40 mM NAC .


These results suggest that NF-kappa B binding to its consensus sequences in the VCAM-1 and E-selectin gene exhibits marked differences in redox sensitivity , allowing for differential gene expression regulated by the same transcription factor .

These results suggest that <protein>NF-kappa B</protein> binding to its consensus sequences in the <protein>VCAM-1</protein> and <dna>E-selectin gene</dna> exhibits marked differences in redox sensitivity , allowing for differential gene expression regulated by the same transcription factor .


Our data also demonstrate that NAC increases the GSH-to-GSSG ratio within the EC suggesting one possible mechanism through which this antioxidant inhibits agonist-induced monocyte adhesion to EC .

Our data also demonstrate that NAC increases the GSH-to-GSSG ratio within the <cell_type>EC</cell_type> suggesting one possible mechanism through which this antioxidant inhibits agonist-induced monocyte adhesion to <cell_type>EC</cell_type> .


Opposite effects of the acute promyelocytic leukemia PML-retinoic acid receptor alpha ( RAR alpha ) and PLZF-RAR alpha fusion proteins on retinoic acid signalling .

Opposite effects of the <protein>acute promyelocytic leukemia PML-retinoic acid receptor alpha</protein> ( <protein>RAR alpha</protein> ) and <protein>PLZF-RAR alpha fusion proteins</protein> on retinoic acid signalling .


Fusion proteins involving the retinoic acid receptor alpha ( RAR alpha ) and the PML or PLZF nuclear protein are the genetic markers of acute promyelocytic leukemias ( APLs ) .

Fusion proteins involving the retinoic acid <protein>receptor alpha</protein> ( <protein>RAR alpha</protein> ) and the <protein>PML</protein> or <protein>PLZF nuclear protein</protein> are the genetic markers of acute promyelocytic leukemias ( APLs ) .


APLs with the PML-RAR alpha or the PLZF-RAR alpha fusion protein are phenotypically indistinguishable except that they differ in their sensitivity to retinoic acid ( RA ) -induced differentiation : PML-RAR alpha blasts are sensitive to RA and patients enter disease remission after RA treatment , while patients with PLZF-RAR alpha do not .

APLs with the <protein>PML-RAR alpha</protein> or the <protein>PLZF-RAR alpha fusion protein</protein> are phenotypically indistinguishable except that they differ in their sensitivity to retinoic acid ( RA ) -induced differentiation : <protein>PML-RAR alpha</protein> blasts are sensitive to RA and patients enter disease remission after RA treatment , while patients with <protein>PLZF-RAR alpha</protein> do not .


We here report that ( i ) like PML-RAR alpha expression , PLZF-RAR alpha expression blocks terminal differentiation of hematopoietic precursor cell lines ( U937 and HL-60 ) in response to different stimuli ( vitamin D3 , transforming growth factor beta1 , and dimethyl sulfoxide ) ; ( ii ) PML-RAR alpha , but not PLZF-RAR alpha , increases RA sensitivity of hematopoietic precursor cells and restores RA sensitivity of RA-resistant hematopoietic cells ; ( iii ) PML-RAR alpha and PLZF-RAR alpha have similar RA binding affinities ; and ( iv ) PML-RAR alpha enhances the RA response of RA target genes ( those for RAR beta , RAR gamma , and transglutaminase type II [ TGase ] ) in vivo , while PLZF-RAR alpha expression has either no effect ( RAR beta ) or an inhibitory activity ( RAR gamma and type II TGase ) .

We here report that ( i ) like <protein>PML-RAR alpha</protein> expression , <protein>PLZF-RAR alpha</protein> expression blocks terminal differentiation of hematopoietic precursor cell lines ( <cell_line>U937</cell_line> and <cell_line>HL-60</cell_line> ) in response to different stimuli ( vitamin D3 , transforming growth factor beta1 , and dimethyl sulfoxide ) ; ( ii ) <protein>PML-RAR alpha</protein> , but not <protein>PLZF-RAR alpha</protein> , increases RA sensitivity of <cell_type>hematopoietic precursor cells</cell_type> and restores RA sensitivity of <cell_type>RA-resistant hematopoietic cells</cell_type> ; ( iii ) <protein>PML-RAR alpha</protein> and <protein>PLZF-RAR alpha</protein> have similar RA binding affinities ; and ( iv ) <protein>PML-RAR alpha</protein> enhances the RA response of <dna>RA target genes</dna> ( those for <protein>RAR beta</protein> , <protein>RAR gamma</protein> , and <protein>transglutaminase type II</protein> [ <protein>TGase</protein> ] ) in vivo , while <protein>PLZF-RAR alpha</protein> expression has either no effect ( <protein>RAR beta</protein> ) or an inhibitory activity ( <protein>RAR gamma</protein> and <protein>type II TGase</protein> ) .


These data demonstrate that PML-RAR alpha and PLZF-RAR alpha have similar ( inhibitory ) effects on RA-independent differentiation and opposite ( stimulatory or inhibitory ) effects on RA-dependent differentiation and that they behave in vivo as RA-dependent enhancers or inhibitors of RA-responsive genes , respectively .

These data demonstrate that <protein>PML-RAR alpha</protein> and <protein>PLZF-RAR alpha</protein> have similar ( inhibitory ) effects on RA-independent differentiation and opposite ( stimulatory or inhibitory ) effects on RA-dependent differentiation and that they behave in vivo as <dna>RA-dependent enhancers</dna> or inhibitors of <dna>RA-responsive genes</dna> , respectively .


Their different activities on the RA signalling pathway might underlie the different responses of PML-RAR alpha and PLZF-RAR alpha APLs to RA treatment .

Their different activities on the RA signalling pathway might underlie the different responses of <protein>PML-RAR alpha</protein> and <protein>PLZF-RAR alpha</protein> APLs to RA treatment .


The PLZF-RAR alpha fusion protein contains an approximately 120-amino-acid N-terminal motif ( called the POZ domain ) , which is also found in a variety of zinc finger proteins and a group of poxvirus proteins and which mediates protein-protein interactions .

The <protein>PLZF-RAR alpha fusion protein</protein> contains an approximately <protein>120-amino-acid N-terminal motif</protein> ( called the <protein>POZ domain</protein> ) , which is also found in a variety of zinc finger proteins and a group of <protein>poxvirus proteins</protein> and which mediates protein-protein interactions .


Deletion of the PLZF POZ domain partially abrogated the inhibitory effect of PLZF-RAR alpha on RA-induced differentiation and on RA-mediated type II TGase up-regulation , suggesting that POZ-mediated protein interactions might be responsible for the inhibitory transcriptional activities of PLZF-RAR alpha .

Deletion of the <protein>PLZF POZ domain</protein> partially abrogated the inhibitory effect of <protein>PLZF-RAR alpha</protein> on RA-induced differentiation and on RA-mediated <protein>type II TGase</protein> up-regulation , suggesting that POZ-mediated protein interactions might be responsible for the inhibitory transcriptional activities of <protein>PLZF-RAR alpha</protein> .


A human homologue of the Drosophila Toll protein signals activation of adaptive immunity [ see comments ]

A human homologue of the <protein>Drosophila Toll protein</protein> signals activation of adaptive immunity [ see comments ]


Induction of the adaptive immune response depends on the expression of co-stimulatory molecules and cytokines by antigen-presenting cells .

Induction of the adaptive immune response depends on the expression of <protein>co-stimulatory molecules</protein> and <protein>cytokines</protein> by <cell_type>antigen-presenting cells</cell_type> .


The mechanisms that control the initial induction of these signals upon infection are poorly understood .


It has been proposed that their expression is controlled by the non-clonal , or innate , component of immunity that preceded in evolution the development of an adaptive immune system in vertebrates .


We report here the cloning and characterization of a human homologue of the Drosophila toll protein ( Toll ) which has been shown to induce the innate immune response in adult Drosophila .

We report here the cloning and characterization of a <protein>human homologue</protein> of the <protein>Drosophila toll protein</protein> ( <protein>Toll</protein> ) which has been shown to induce the innate immune response in adult Drosophila .


Like Drosophila Toll , human Toll is a type I transmembrane protein with an extracellular domain consisting of a leucine-rich repeat ( LRR ) domain , and a cytoplasmic domain homologous to the cytoplasmic domain of the human interleukin ( IL ) -1 receptor .

Like Drosophila <protein>Toll</protein> , <protein>human Toll</protein> is a type I transmembrane protein with an extracellular domain consisting of a <protein>leucine-rich repeat ( LRR ) domain</protein> , and a cytoplasmic domain homologous to the <protein>cytoplasmic domain</protein> of the <protein>human interleukin ( IL ) -1 receptor</protein> .


Both Drosophila Toll and the IL-1 receptor are known to signal through the NF-kappaB pathway .

Both Drosophila <protein>Toll</protein> and the IL-1 receptor are known to signal through the <protein>NF-kappaB</protein> pathway .


We show that a constitutively active mutant of human Toll transfected into human cell lines can induce the activation of NF-kappaB and the expression of NF-kappaB-controlled genes for the inflammatory cytokines IL-1 , IL-6 and IL-8 , as well as the expression of the co-stimulatory molecule B7.1 , which is required for the activation of naive T cells .

We show that a constitutively active mutant of <protein>human Toll</protein> transfected into human cell lines can induce the activation of <protein>NF-kappaB</protein> and the expression of <dna>NF-kappaB-controlled genes</dna> for the <protein>inflammatory cytokines</protein> <protein>IL-1</protein> , <protein>IL-6</protein> and <protein>IL-8</protein> , as well as the expression of the <protein>co-stimulatory molecule</protein> <protein>B7.1</protein> , which is required for the activation of <cell_type>naive T cells</cell_type> .


Role of ascorbate in the activation of NF-kappaB by tumour necrosis factor-alpha in T-cells .

Role of ascorbate in the activation of <protein>NF-kappaB</protein> by <protein>tumour necrosis factor-alpha</protein> in <cell_type>T-cells</cell_type> .


The first product of ascorbate oxidation , the ascorbate free radical ( AFR ) , acts in biological systems mainly as an oxidant , and through its role in the plasma membrane redox system exerts different effects on the cell .


We have investigated the role of ascorbate , AFR and dehydroascorbate ( DHA ) in the activation of the NF-kappaB transcription factor in Jurkat T-cells stimulated by tumour necrosis factor-alpha ( TNF-alpha ) .

We have investigated the role of ascorbate , AFR and dehydroascorbate ( DHA ) in the activation of the <protein>NF-kappaB transcription factor</protein> in <cell_line>Jurkat T-cells</cell_line> stimulated by <protein>tumour necrosis factor-alpha</protein> ( <protein>TNF-alpha</protein> ) .


Here we show , by electrophoretic mobility shift assays , that ascorbate increases the binding of NF-kappaB to DNA in TNF-alpha-stimulated Jurkat cells .

Here we show , by electrophoretic mobility shift assays , that ascorbate increases the binding of <protein>NF-kappaB</protein> to DNA in <cell_line>TNF-alpha-stimulated Jurkat cells</cell_line> .


The ability of ascorbate to enhance cytoplasmic inhibitory IkBalpha protein degradation correlates completely with its capacity to induce NF-kappaB binding to DNA and to potentiate NF-kappaB -mediated transactivation of the HIV-1 long terminal repeat promoter in TNF-alpha-stimulated Jurkat cells but not in cells stimulated with PMA plus ionomycin .

The ability of ascorbate to enhance cytoplasmic inhibitory <protein>IkBalpha protein</protein> degradation correlates completely with its capacity to induce <protein>NF-kappaB</protein> binding to DNA and to potentiate <protein>NF-kappaB</protein> -mediated transactivation of the <dna>HIV-1 long terminal repeat promoter</dna> in <cell_line>TNF-alpha-stimulated Jurkat cells</cell_line> but not in cells stimulated with PMA plus ionomycin .


AFR behaves like ascorbate , while DHA and ascorbate phosphate do not affect TNF-alpha -mediated NF-kappaB activation .

AFR behaves like ascorbate , while DHA and ascorbate phosphate do not affect <protein>TNF-alpha</protein> -mediated <protein>NF-kappaB</protein> activation .


These results provide new evidence for a possible relationship between the activation of the electron-transport system at the plasma membrane by ascorbate or its free radical and redox-dependent gene transcription in T-cells .

These results provide new evidence for a possible relationship between the activation of the electron-transport system at the plasma membrane by ascorbate or its free radical and redox-dependent gene transcription in <cell_type>T-cells</cell_type> .


Human cytomegalovirus induces interleukin-8 production by a human monocytic cell line , THP-1 , through acting concurrently on AP-1- and NF-kappaB-binding sites of the interleukin-8 gene .

Human cytomegalovirus induces <protein>interleukin-8</protein> production by a <cell_line>human monocytic cell line</cell_line> , <cell_line>THP-1</cell_line> , through acting concurrently on <dna>AP-1- and NF-kappaB-binding sites</dna> of the <dna>interleukin-8 gene</dna> .


Cytomegalovirus ( CMV ) infection induced interleukin-8 ( IL-8 ) gene transcription in a human monocytic cell line , THP-1 cells , leading to IL-8 secretion .

Cytomegalovirus ( CMV ) infection induced <protein>interleukin-8</protein> ( <protein>IL-8</protein> ) gene transcription in a <cell_line>human monocytic cell line</cell_line> , <cell_line>THP-1 cells</cell_line> , leading to <protein>IL-8</protein> secretion .


The functional analysis of the IL-8 gene revealed that both AP-1- and NF-kappaB factor-binding elements were involved in conferring the responsiveness to CMV .

The functional analysis of the <dna>IL-8 gene</dna> revealed that both <dna>AP-1- and NF-kappaB factor-binding elements</dna> were involved in conferring the responsiveness to CMV .


Moreover , electrophoretic mobility shift assays demonstrated that CMV induced the formation of NF-kappaB and AP-1 complexes .

Moreover , electrophoretic mobility shift assays demonstrated that CMV induced the formation of <protein>NF-kappaB</protein> and <protein>AP-1 complexes</protein> .


These results suggest that CMV activates these transcriptional factors , resulting in IL-8 gene expression .

These results suggest that CMV activates these <protein>transcriptional factors</protein> , resulting in <protein>IL-8</protein> gene expression .


Construction and biological characterization of an interleukin-12 fusion protein ( Flexi-12 ) : delivery to acute myeloid leukemic blasts using adeno-associated virus .

Construction and biological characterization of an <protein>interleukin-12 fusion protein</protein> ( <protein>Flexi-12</protein> ) : delivery to <cell_type>acute myeloid leukemic blasts</cell_type> using adeno-associated virus .


Interleukin-12 ( IL-12 ) is a cytokine that exhibits pleiotropic effects on lymphocytes and natural killer cells and has been shown to have promise for the immunotherapy of cancer .

<protein>Interleukin-12</protein> ( <protein>IL-12</protein> ) is a <protein>cytokine</protein> that exhibits pleiotropic effects on <cell_type>lymphocytes</cell_type> and <cell_type>natural killer cells</cell_type> and has been shown to have promise for the immunotherapy of cancer .


The combination of the immune costimulatory molecule B7.1 and IL-12 has been shown to be synergistic for T cell activation .

The combination of the <protein>immune costimulatory molecule</protein> <protein>B7.1</protein> and <protein>IL-12</protein> has been shown to be synergistic for T cell activation .


By transfecting tumor cells with both IL-12 and B7.1 cDNAs , it may be possible to use these modified targets as vaccines .

By transfecting tumor cells with both <dna>IL-12 and B7.1 cDNAs</dna> , it may be possible to use these modified targets as vaccines .


A major obstacle in designing a vector to deliver these genes results from the structure of IL-12 .

A major obstacle in designing a vector to deliver these genes results from the structure of <protein>IL-12</protein> .


Functional IL-12 is a heterodimer composed of two distinct subunits that are encoded by separate genes on different chromosomes .

Functional <protein>IL-12</protein> is a <protein>heterodimer</protein> composed of two distinct subunits that are encoded by separate genes on different chromosomes .


Production of functional IL-12 requires the coordinated expression of both genes .

Production of <protein>functional IL-12</protein> requires the coordinated expression of both genes .


This presents several problems in vectors , particularly those in which additional genes , either a co-stimulatory gene or a selectable marker , are inserted .

This presents several problems in vectors , particularly those in which additional genes , either a <dna>co-stimulatory gene</dna> or a selectable marker , are inserted .


Therefore , we have constructed a single cDNA that encodes a single-chain protein , called Flexi-12 , which retains all of the biological characteristics of recombinant IL-12 ( rIL-12 ) .

Therefore , we have constructed a single <dna>cDNA</dna> that encodes a <protein>single-chain protein</protein> , called <protein>Flexi-12</protein> , which retains all of the biological characteristics of recombinant <protein>IL-12</protein> ( <protein>rIL-12</protein> ) .


The monomeric polypeptide Flexi-12 is able to induce the proliferation of phytohemagglutinin ( PHA ) blasts , induce PHA blasts to secrete interferon-gamma ( IFN-gamma ) and additionally , by preincubation , enhance the killing of K562 targets by PBLs .

The monomeric polypeptide <protein>Flexi-12</protein> is able to induce the proliferation of <cell_line>phytohemagglutinin ( PHA ) blasts</cell_line> , induce <cell_line>PHA blasts</cell_line> to secrete <protein>interferon-gamma</protein> ( <protein>IFN-gamma</protein> ) and additionally , by preincubation , enhance the killing of K562 targets by <cell_type>PBLs</cell_type> .


These phenomena are in a dose-dependent manner comparable to that seen with rIL-12 .

These phenomena are in a dose-dependent manner comparable to that seen with <protein>rIL-12</protein> .


We have also shown that tyrosine phosphorylation of the STAT 4 transcription factor , which has been shown to be unique to the IL-12 signaling pathway , occurs with Flexi-12 at levels similar to those seen with rIL-12 .

We have also shown that tyrosine phosphorylation of the <protein>STAT 4 transcription factor</protein> , which has been shown to be unique to the <protein>IL-12</protein> signaling pathway , occurs with <protein>Flexi-12</protein> at levels similar to those seen with <protein>rIL-12</protein> .


We have packaged Flexi-12 into a recombinant adeno-associated virus ( AAV ) and used this vector to infect acute myeloid leukemic ( AML ) blasts .

We have packaged <protein>Flexi-12</protein> into a recombinant adeno-associated virus ( AAV ) and used this vector to infect <cell_type>acute myeloid leukemic ( AML ) blasts</cell_type> .


Infected AML blasts produced between 2 and 6 ng of IL-12/10 ( 6 ) cells per ml per 48 hr .

Infected <cell_type>AML blasts</cell_type> produced between 2 and 6 ng of IL-12/10 ( 6 ) cells per ml per 48 hr .


These studies also confirm that AAV is an efficient delivery vehicle for cytokines to leukemic cells .

These studies also confirm that AAV is an efficient delivery vehicle for <protein>cytokines</protein> to leukemic cells .


Direct analysis of these modified cells acting as tumor vaccines is underway .


Plasma sialyltransferase levels in psychiatric disorders as a possible indicator of HPA axis function .

Plasma <protein>sialyltransferase</protein> levels in psychiatric disorders as a possible indicator of HPA axis function .


A dysfunction in the regulation of the hypothalamic-pituitary-adrenal ( HPA ) axis , possibly attributed to a change in glucocorticoid receptor ( GR ) functionality , has been implicated in depression .

A dysfunction in the regulation of the hypothalamic-pituitary-adrenal ( HPA ) axis , possibly attributed to a change in <protein>glucocorticoid receptor</protein> ( <protein>GR</protein> ) functionality , has been implicated in depression .


We have measured both lymphocyte GR receptor binding parameters and plasma sialyltransferase activity , as a biochemical marker of GR function , in two groups of patients suffering from depression or schizophrenia and in a group of age- and sex-matched controls .

We have measured both <protein>lymphocyte GR</protein> receptor binding parameters and plasma <protein>sialyltransferase</protein> activity , as a biochemical marker of <protein>GR</protein> function , in two groups of patients suffering from depression or schizophrenia and in a group of age- and sex-matched controls .


While there was a significant increase in plasma cortisol levels in the depressed group , there were no changes in the lymphocyte GR binding parameters ( K ( m ) and Bmax ) .

While there was a significant increase in plasma cortisol levels in the depressed group , there were no changes in the lymphocyte <protein>GR</protein> binding parameters ( K ( m ) and Bmax ) .


There was , however , a significant decrease in the plasma sialyltransferase : cortisol ratio in the depressed group suggesting an inability of the raised cortisol levels to induce enzyme expression and this ratio may provide a useful biochemical marker of cortisol receptor function .

There was , however , a significant decrease in the <protein>plasma sialyltransferase :</protein> cortisol ratio in the depressed group suggesting an inability of the raised cortisol levels to induce enzyme expression and this ratio may provide a useful biochemical marker of cortisol receptor function .


Although there was an increase in the plasma activity of the alpha 2 , 6 sialyltransferase isozyme in the schizophrenic group , no other changes were determined .

Although there was an increase in the plasma activity of the <protein>alpha 2 , 6 sialyltransferase isozyme</protein> in the schizophrenic group , no other changes were determined .


Therefore , while the total plasma sialyltransferase : cortisol ratio reflects HPA axis function , alterations in specific isozyme activity may also be associated with other CNS disease states .

Therefore , while the total <protein>plasma sialyltransferase</protein> : cortisol ratio reflects HPA axis function , alterations in specific isozyme activity may also be associated with other CNS disease states .


Regulation of CD95 ( Fas ) ligand expression by TCR -mediated signaling events .

Regulation of <protein>CD95 ( Fas ) ligand</protein> expression by <protein>TCR</protein> -mediated signaling events .


Stimulation of mature peripheral T cells by TCR engagement results in activation of signals that drive induction of cytokine gene expression and clonal expansion .

Stimulation of <cell_type>mature peripheral T cells</cell_type> by <protein>TCR</protein> engagement results in activation of signals that drive induction of <protein>cytokine</protein> gene expression and clonal expansion .


However , under some conditions , engagement of the TCR leads instead to apoptosis .

However , under some conditions , engagement of the <protein>TCR</protein> leads instead to apoptosis .


Recent studies demonstrate that TCR -stimulated apoptosis requires expression of CD95 ligand on activated T cells followed by an interaction between CD95 ligand and the CD95 receptor also expressed on this population .

Recent studies demonstrate that <protein>TCR</protein> -stimulated apoptosis requires expression of <protein>CD95 ligand</protein> on <cell_type>activated T cells</cell_type> followed by an interaction between <protein>CD95 ligand</protein> and the <protein>CD95 receptor</protein> also expressed on this population .


The experiments reported in this study were designed to address the signaling events triggered by TCR engagement that are important for regulating CD95 ligand gene expression .

The experiments reported in this study were designed to address the signaling events triggered by <protein>TCR</protein> engagement that are important for regulating <protein>CD95 ligand</protein> gene expression .


To approach this , we generated a luciferase reporter construct containing elements of the CD95 ligand promoter .

To approach this , we generated a <dna>luciferase reporter construct</dna> containing elements of the <protein>CD95 ligand</protein> promoter .


Using a previously described mutant of the Jurkat T cell line , we show that proximal signaling events dependent on the presence of the CD45 tyrosine phosphatase are required for TCR -stimulated CD95 ligand expression .

Using a previously described mutant of the <cell_line>Jurkat T cell line</cell_line> , we show that proximal signaling events dependent on the presence of the <protein>CD45 tyrosine phosphatase</protein> are required for <protein>TCR</protein> -stimulated <protein>CD95 ligand</protein> expression .


Transient transfection studies demonstrate further that TCR -stimulated activation of the Ras signaling pathway is required for optimal activation of CD95 ligand .

Transient transfection studies demonstrate further that <protein>TCR</protein> -stimulated activation of the <protein>Ras</protein> signaling pathway is required for optimal activation of <protein>CD95 ligand</protein> .


Next , in an effort to determine critical transcription factors that regulate CD95 ligand expression , we demonstrate a cyclosporin A-sensitive nuclear factor-AT response element in the promoter region of this gene that is critical for optimal CD95 ligand reporter activity in stimulated T cells .

Next , in an effort to determine critical <protein>transcription factors</protein> that regulate <protein>CD95 ligand</protein> expression , we demonstrate a <dna>cyclosporin A-sensitive nuclear factor-AT response element</dna> in the <dna>promoter region</dna> of this gene that is critical for optimal <protein>CD95 ligand</protein> reporter activity in <cell_type>stimulated T cells</cell_type> .


Together , these studies begin a dissection of the biochemical events that lead to expression of CD95 ligand , a required step for TCR -induced apoptosis .

Together , these studies begin a dissection of the biochemical events that lead to expression of <protein>CD95 ligand</protein> , a required step for <protein>TCR</protein> -induced apoptosis .


Reactive oxygen species and antioxidants in inflammatory diseases .


This paper aims to review the role of free radical-induced tissue damage and antioxidant defence mechanisms in inflammatory diseases that involve pathogenic processes similar to the periodontal diseases .


There is a clearly defined and substantial role for free radicals or reactive oxygen species ( ROS ) in periodontitis , but little research has been performed in this area .


This paper reviews the considerable data available relating ROS activity and antioxidant defence to inflammatory diseases and attempts to draw parallels with periodontitis , in an effort to stimulate more periodontal research in this important area .


The recent discovery of the transcription factor nuclear factor kappa B ( NF-kappa B ) is reviewed and several potential pathways for cytokine -induced periodontal tissue damage , mediated by NF-kappa B1 are discussed .

The recent discovery of the <protein>transcription factor nuclear factor kappa B</protein> ( <protein>NF-kappa B</protein> ) is reviewed and several potential pathways for <protein>cytokine</protein> -induced periodontal tissue damage , mediated by <protein>NF-kappa B1</protein> are discussed .


Emphasis is placed on cytokines that have been studied in periodontitis , principally TNF-alpha , IL-1 , IL-6 , IL-8 and beta-interferon .

Emphasis is placed on <protein>cytokines</protein> that have been studied in periodontitis , principally <protein>TNF-alpha</protein> , <protein>IL-1</protein> , <protein>IL-6</protein> , <protein>IL-8</protein> and <protein>beta-interferon</protein> .


The link between cellular production of such important mediators of inflammation and the antioxidant ( AO ) thiols , cysteine and reduced glutathione ( GSH ) , is discussed and it is hypothesised that NF-kappa B antagonists may offer important therapeutic benefits .

The link between cellular production of such important mediators of inflammation and the antioxidant ( AO ) thiols , cysteine and reduced glutathione ( GSH ) , is discussed and it is hypothesised that <protein>NF-kappa B</protein> antagonists may offer important therapeutic benefits .


Mutations in the TSC2 gene : analysis of the complete coding sequence using the protein truncation test ( PTT ) .

Mutations in the <dna>TSC2 gene</dna> : analysis of the complete coding sequence using the protein truncation test ( PTT ) .


Mutations in the TSC2 gene on chromosome 16p13.3 are responsible for approximately 50 % of familial tuberous sclerosis ( TSC ) .

Mutations in the <dna>TSC2 gene</dna> on <dna>chromosome 16p13.3</dna> are responsible for approximately 50 % of familial tuberous sclerosis ( TSC ) .


The gene has 41 small exons spanning 45 kb of genomic DNA and encoding a 5.5 kb mRNA .

The gene has <dna>41 small exons</dna> spanning <dna>45 kb</dna> of <dna>genomic DNA</dna> and encoding a <rna>5.5 kb mRNA</rna> .


Large germline deletions of TSC2 occur in < 5 % of cases , and a number of small intragenic mutations have been described .

Large germline deletions of <dna>TSC2</dna> occur in < 5 % of cases , and a number of small intragenic mutations have been described .


We analysed mRNA from 18 unrelated cases of TSC for TSC2 mutations using the protein truncation test ( PTT ) .

We analysed mRNA from 18 unrelated cases of TSC for <dna>TSC2 mutations</dna> using the protein truncation test ( PTT ) .


Three cases were predicted to be TSC2 mutations on the basis of linkage analysis or because a hamartoma from the patient showed loss of heterozygosity for 16p13.3 markers .

Three cases were predicted to be <dna>TSC2 mutations</dna> on the basis of linkage analysis or because a hamartoma from the patient showed loss of heterozygosity for <dna>16p13.3 markers</dna> .


Three overlapping PCR products , covering the complete coding sequence of mRNA , were generated from lymphoblastoid cell lines , translated into 35S-methionine labelled protein , and analysed by SDS-PAGE .

Three overlapping <dna>PCR products</dna> , covering the complete coding sequence of <rna>mRNA</rna> , were generated from <cell_line>lymphoblastoid cell lines</cell_line> , translated into <protein>35S-methionine labelled protein</protein> , and analysed by SDS-PAGE .


PCR products showing PTT shifts were directly sequenced , and mutations confirmed by restriction enzyme digestion where possible .

<dna>PCR products</dna> showing PTT shifts were directly sequenced , and mutations confirmed by restriction enzyme digestion where possible .


Six PTT shifts were identified .


Five of these were caused by mutations predicted to produce a truncated protein : ( i ) a sporadic case showed a 32 bp deletion in exon 11 , and a mutant mRNA without exon 11 was produced ; the normal exon 10 was also spliced out ; ( ii ) a sporadic case had a 1 bp deletion in exon 12 ( 1634delT ) ; ( iii ) a TSC2-linked mother and daughter pair had a G -- > T transversion in exon 23 ( G2715T ) introducing a cryptic splice site causing a 29 bp truncation of mRNA from exon 23 ; ( iv ) a sporadic case showed a 2 bp deletion in exon 36 ; ( v ) a sporadic case showed a 1 bp insertion disrupting the donor splice site of exon 37 ( 5007+2insA ) , resulting in the use of an upstream exonic cryptic splice site to cause a 29 bp truncation of mRNA from exon 37 .

Five of these were caused by mutations predicted to produce a <protein>truncated protein</protein> : ( i ) a sporadic case showed a <dna>32 bp deletion</dna> in <rna>exon 11</rna> , and a <rna>mutant mRNA</rna> without <rna>exon 11</rna> was produced ; the normal exon 10 was also spliced out ; ( ii ) a sporadic case had a <dna>1 bp deletion</dna> in <rna>exon 12</rna> ( <rna>1634delT</rna> ) ; ( iii ) a <rna>TSC2-linked mother and daughter pair</rna> had a G -- > T transversion in <rna>exon 23</rna> ( <rna>G2715T</rna> ) introducing a cryptic splice site causing a 29 bp truncation of mRNA from <rna>exon 23</rna> ; ( iv ) a sporadic case showed a <dna>2 bp deletion</dna> in <rna>exon 36</rna> ; ( v ) a sporadic case showed a <dna>1 bp insertion</dna> disrupting the donor splice site of <rna>exon 37</rna> ( <rna>5007+2insA</rna> ) , resulting in the use of an <rna>upstream exonic cryptic splice site</rna> to cause a <rna>29 bp truncation</rna> of <rna>mRNA</rna> from <rna>exon 37</rna> .


In one case , the PTT shift was explained by in-frame splicing out of exon 10 , in the presence of a normal exon 10 genomic sequence .

In one case , the PTT shift was explained by in-frame splicing out of <rna>exon 10</rna> , in the presence of a <dna>normal exon 10 genomic sequence</dna> .


Alternative splicing of exon 10 of the TSC2 gene may be a normal variant .

Alternative splicing of <rna>exon 10</rna> of the <dna>TSC2 gene</dna> may be a normal variant .


Three 3rd base substitution polymorphisms were also detected during direct sequencing of PCR products .

Three 3rd base substitution polymorphisms were also detected during direct sequencing of <dna>PCR products</dna> .


Confirmed mutations were identified in 28 % of the families studied and on the assumption that half of the sporadic cases should have TSC2 mutations , a crude estimate of the detection rate would be 60 % .

Confirmed mutations were identified in 28 % of the families studied and on the assumption that half of the sporadic cases should have <dna>TSC2 mutations</dna> , a crude estimate of the detection rate would be 60 % .


This compares favourably with other screening methods used for TSC2 , notably SSCP , and since PTT involves much less work it may be the method of choice .

This compares favourably with other screening methods used for <dna>TSC2</dna> , notably SSCP , and since PTT involves much less work it may be the method of choice .


alpha-Tocopheryl succinate inhibits monocytic cell adhesion to endothelial cells by suppressing NF-kappa B mobilization .

alpha-Tocopheryl succinate inhibits monocytic cell adhesion to <cell_type>endothelial cells</cell_type> by suppressing <protein>NF-kappa B</protein> mobilization .


The adherence of monocytes to activated endothelium is an early event in atherogenesis .

The adherence of <cell_type>monocytes</cell_type> to activated endothelium is an early event in atherogenesis .


Because antioxidants have been considered to be of antiatherosclerotic potential , we investigated the effects of alpha-tocopherol ( TCP ) and its acetate and succinate esters on monocyte adhesion to cytokine-stimulated human umbilical vein endothelial cells ( HUVEC ) .

Because antioxidants have been considered to be of antiatherosclerotic potential , we investigated the effects of alpha-tocopherol ( TCP ) and its acetate and succinate esters on monocyte adhesion to <cell_line>cytokine-stimulated human umbilical vein endothelial cells</cell_line> ( <cell_line>HUVEC</cell_line> ) .


Endothelial cells were treated with TCP , alpha-tocopherol acetate ( TCP acetate ) , or alpha-tocopheryl succinate ( TCP succinate ) before stimulation with tumor necrosis factor-alpha ( TNF-alpha ; 10 U/ml , 6 h ) or interleukin-1 beta ( IL-1 beta ; 10 U/ml , 6 h ) .

<cell_type>Endothelial cells</cell_type> were treated with TCP , alpha-tocopherol acetate ( TCP acetate ) , or alpha-tocopheryl succinate ( TCP succinate ) before stimulation with <protein>tumor necrosis factor-alpha</protein> ( <protein>TNF-alpha</protein> ; 10 U/ml , 6 h ) or <protein>interleukin-1 beta</protein> ( <protein>IL-1 beta</protein> ; 10 U/ml , 6 h ) .


Cytokine-stimulated cell surface expression of vascular cell adhesion molecule-1 ( VCAM-1 , CD106 ) and E-selectin ( ELAM-1 , CD62E ) , but not of intercellular adhesion molecule-1 ( ICAM-1 , CD54 ) , was time- and dose-dependently inhibited by TCP succinate but not by TCP or TCP acetate .

Cytokine-stimulated cell surface expression of <protein>vascular cell adhesion molecule-1</protein> ( <protein>VCAM-1</protein> , <protein>CD106</protein> ) and <protein>E-selectin</protein> ( <protein>ELAM-1</protein> , <protein>CD62E</protein> ) , but not of <protein>intercellular adhesion molecule-1</protein> ( <protein>ICAM-1</protein> , <protein>CD54</protein> ) , was time- and dose-dependently inhibited by TCP succinate but not by TCP or TCP acetate .


TCP succinate ( 200 microM , 24 h ) reduced TNF-induced VCAM-1 and E-selectin expression from a specific mean fluorescence intensity of 151 +/- 28 to 12 +/- 4 channels and from 225 +/- 38 to 79 +/- 21 channels , respectively .

TCP succinate ( 200 microM , 24 h ) reduced <protein>TNF-induced VCAM-1</protein> and <protein>E-selectin</protein> expression from a specific mean fluorescence intensity of 151 +/- 28 to 12 +/- 4 channels and from 225 +/- 38 to 79 +/- 21 channels , respectively .


Succinate alone had no effect .


Decreased adhesion molecule expression was associated with a reduction of monocytic cell adhesion .

Decreased <protein>adhesion molecule</protein> expression was associated with a reduction of monocytic cell adhesion .


TCP succinate ( 20 microM , 72 h ) , but not TCP ( 200 microM , 72 h ) , reduced U-937 cell adhesion to TNF-alpha-stimulated ( 10 U/ml , 6 h ) HUVEC by 30 % ( P < 0.025 ) and to IL-1 beta-stimulated HUVEC by 56 % ( P < 0.010 ) .

TCP succinate ( 20 microM , 72 h ) , but not TCP ( 200 microM , 72 h ) , reduced <cell_line>U-937 cell</cell_line> adhesion to <cell_line>TNF-alpha-stimulated ( 10 U/ml , 6 h ) HUVEC</cell_line> by 30 % ( P < 0.025 ) and to <cell_line>IL-1 beta-stimulated HUVEC</cell_line> by 56 % ( P < 0.010 ) .


Electrophoretic mobility-shift assays of HUVEC nuclear proteins revealed a decrease in TNF-alpha-stimulated nuclear factor-kappa B ( NF-kappa B ) activation after pretreatment of HUVEC with TCP succinate but not with TCP , TCP acetate , or succinate alone .

Electrophoretic mobility-shift assays of <cell_line>HUVEC</cell_line> <protein>nuclear proteins</protein> revealed a decrease in <protein>TNF-alpha-stimulated nuclear factor-kappa B</protein> ( <protein>NF-kappa B</protein> ) activation after pretreatment of <cell_line>HUVEC</cell_line> with TCP succinate but not with TCP , TCP acetate , or succinate alone .


In conclusion , we demonstrate that the vitamin E derivative TCP succinate prevents monocytic cell adhesion to cytokine-stimulated endothelial cells by inhibiting the activation of NF-kappa B , further emphasizing the antiatherosclerotic potential of lipid soluble antioxidants .

In conclusion , we demonstrate that the vitamin E derivative TCP succinate prevents monocytic cell adhesion to cytokine-stimulated <cell_type>endothelial cells</cell_type> by inhibiting the activation of <protein>NF-kappa B</protein> , further emphasizing the antiatherosclerotic potential of lipid soluble antioxidants .


The product of the murine homolog of the Drosophila extra sex combs gene displays transcriptional repressor activity .

The product of the <dna>murine homolog</dna> of the <dna>Drosophila extra sex combs gene</dna> displays <protein>transcriptional repressor</protein> activity .


The heterogeneous nuclear ribonucleoprotein K protein represents a novel class of proteins that may act as docking platforms that orchestrate cross-talk among molecules involved in signal transduction and gene expression .

The <protein>heterogeneous nuclear ribonucleoprotein K protein</protein> represents a novel class of proteins that may act as docking platforms that orchestrate cross-talk among molecules involved in signal transduction and gene expression .


Using a fragment of K protein as bait in the yeast two-hybrid screen , we isolated a cDNA that encodes a protein whose primary structure has extensive similarity to the Drosophila melanogaster extra sex combs ( esc ) gene product , Esc , a putative silencer of homeotic genes .

Using a fragment of <protein>K protein</protein> as bait in the yeast two-hybrid screen , we isolated a <dna>cDNA</dna> that encodes a protein whose primary structure has extensive similarity to the <protein>Drosophila melanogaster extra sex combs ( esc ) gene product</protein> , <protein>Esc</protein> , a putative silencer of <dna>homeotic genes</dna> .


The cDNA that we isolated is identical to the cDNA of the recently positionally cloned mouse embryonic ectoderm development gene , eed .

The <dna>cDNA</dna> that we isolated is identical to the <dna>cDNA</dna> of the recently positionally cloned <dna>mouse embryonic ectoderm development gene</dna> , eed .


Like Esc , Eed contains six WD-40 repeats in the C-terminal half of the protein and is thought to repress homeotic gene expression during mouse embryogenesis .

Like <protein>Esc</protein> , <protein>Eed</protein> contains six <protein>WD-40 repeats</protein> in the <protein>C-terminal half</protein> of the protein and is thought to repress homeotic gene expression during mouse embryogenesis .