We concluded from this , that one or more of the cell types present in synovial tissue contain a specific steroid receptor , but that this is lacking in synovial fluid polymorphonuclear leucocytes .

We concluded from this , that one or more of the cell types present in synovial tissue contain a <protein>specific steroid receptor</protein> , but that this is lacking in <cell_type>synovial fluid polymorphonuclear leucocytes</cell_type> .


Clinical implications of glucocorticoid receptors in human leukemia .

Clinical implications of <protein>glucocorticoid receptors</protein> in human leukemia .


Glucorticoid receptors were studied in various populations of normal human peripheral blood lymphocytes and leukemic lymphoblasts .

<protein>Glucorticoid receptors</protein> were studied in various populations of normal <cell_type>human peripheral blood lymphocytes</cell_type> and <cell_type>leukemic lymphoblasts</cell_type> .


Normal lymphocytes contain low levels of glucocorticoid receptor ( approximately 2 , 500 sites/cell ) which are identical in T- and non-T-fractions .

<cell_type>Normal lymphocytes</cell_type> contain low levels of <protein>glucocorticoid receptor</protein> ( approximately 2 , 500 sites/cell ) which are identical in T- and non-T-fractions .


Phytohemagglutinin treatment increases levels about 3-fold .

<protein>Phytohemagglutinin</protein> treatment increases levels about 3-fold .


Leukemic lymphoblasts contain larger numbers of receptor sites .

<cell_type>Leukemic lymphoblasts</cell_type> contain larger numbers of receptor sites .


Presence of receptor is correlated with in vitro sensitivitiy to glucocorticoids and in vivo response to therapy .


Quantity of receptor is also correlated with complete remission duration independently of leukemic cell type ( T or null ) , initial WBC , or age of patient .

Quantity of receptor is also correlated with complete remission duration independently of <cell_type>leukemic cell type</cell_type> ( T or null ) , initial <cell_type>WBC</cell_type> , or age of patient .


Quantitative determination of glucocorticoid receptor levels in acute lymphoblastic leukemia may be of value both as an independent prognostic variable and in suggesting which patients should receive glucocorticoid therapy .

Quantitative determination of <protein>glucocorticoid receptor</protein> levels in acute lymphoblastic leukemia may be of value both as an independent prognostic variable and in suggesting which patients should receive glucocorticoid therapy .


Functional and physical interaction of protein-tyrosine kinases Fyn and Csk in the T-cell signaling system .

Functional and physical interaction of <protein>protein-tyrosine kinases Fyn and Csk</protein> in the T-cell signaling system .


The Src-like protein-tyrosine kinase Fyn is associated with T-cell antigen receptor .

The <protein>Src-like protein-tyrosine kinase</protein> <protein>Fyn</protein> is associated with <protein>T-cell antigen receptor</protein> .


Transient expression of actively mutated Fyn , having Phe-528 instead of Tyr-528 or Thr-338 instead of Ile-338 , in Jurkat T-cells stimulated the serum response element ( SRE ) , 12-O-tetradecanoyl-phorbol-13-acetate response element , cyclic AMP response element , and c-fos promoter .

Transient expression of actively mutated <protein>Fyn</protein> , having Phe-528 instead of Tyr-528 or Thr-338 instead of Ile-338 , in <cell_type>Jurkat T-cells</cell_type> <dna>stimulated the serum response element</dna> ( <dna>SRE</dna> ) , <dna>12-O-tetradecanoyl-phorbol-13-acetate response element</dna> , <dna>cyclic AMP response element</dna> , and <dna>c-fos promoter</dna> .


The stimulation of SRE was particularly prominent not only with active Fyn but also with normal ( wild-type ) Fyn .

The stimulation of <dna>SRE</dna> was particularly prominent not only with active <protein>Fyn</protein> but also with <protein>normal ( wild-type ) Fyn</protein> .


SRE was also stimulated by both normal and active Lck .

<dna>SRE</dna> was also stimulated by both <protein>normal and active Lck</protein> .


Furthermore , normal and active Fyn stimulated transcription from the IL-2 gene promoter when transfected cells were stimulated by concanavalin A plus 12-O-tetradecanoylphorbol-13-acetate .

Furthermore , <protein>normal and active Fyn</protein> stimulated transcription from the <dna>IL-2 gene promoter</dna> when transfected cells were stimulated by <protein>concanavalin A</protein> plus 12-O-tetradecanoylphorbol-13-acetate .


Under the same conditions , Lck did not stimulate IL-2 promoter unless it was activated by mutation .

Under the same conditions , <protein>Lck</protein> did not stimulate <dna>IL-2 promoter</dna> unless it was activated by mutation .


Interestingly , a mutant Fyn , which has deletions within the SH2 region and so is able to transform chicken embryo fibroblasts , did not stimulate either the c-fos or IL-2 promoter , suggesting the importance of this region in T-cell signaling .

Interestingly , a <protein>mutant Fyn</protein> , which has deletions within the <protein>SH2 region</protein> and so is able to transform <cell_type>chicken embryo fibroblasts</cell_type> , did not stimulate either the <dna>c-fos or IL-2 promoter</dna> , suggesting the importance of this region in <cell_type>T-cell</cell_type> signaling .


Csk , which phosphorylates tyrosine residues in the negative regulatory sites of Src family kinases , down-regulated Fyn- and Lck-mediated stimulation of the serum response element and Fyn-mediated enhancement of IL-2 promoter activity .

<protein>Csk</protein> , which phosphorylates tyrosine residues in the negative regulatory sites of <protein>Src family kinases</protein> , down-regulated Fyn- and Lck-mediated stimulation of the <dna>serum response element</dna> and Fyn-mediated enhancement of <dna>IL-2 promoter</dna> activity .


These data suggest that Fyn and Lck , whose activities are regulated by Csk , are involved in different phases of T-cell activation .

These data suggest that <protein>Fyn</protein> and <protein>Lck</protein> , whose activities are regulated by <protein>Csk</protein> , are involved in different phases of <cell_type>T-cell</cell_type> activation .


A novel NF-kappa B complex containing p65 homodimers : implications for transcriptional control at the level of subunit dimerization .

A novel <protein>NF-kappa B complex</protein> containing <protein>p65 homodimers</protein> : implications for transcriptional control at the level of subunit dimerization .


The predominant inducible form of the NF-kappa B transcription factor is a heteromeric complex containing two Rel-related DNA-binding subunits , termed p65 and p50 .

The predominant inducible form of the <protein>NF-kappa B transcription factor</protein> is a <protein>heteromeric complex</protein> containing two <protein>Rel-related DNA-binding subunits</protein> , <protein>termed p65</protein> and <protein>p50</protein> .


Prior transfection studies have shown that when these p65 and p50 subunits are expressed independently as stable homodimers , p65 stimulates kappa B-directed transcription , whereas p50 functions as a kappa B-specific repressor .

Prior transfection studies have shown that when these <protein>p65 and p50 subunits</protein> are expressed independently as <protein>stable homodimers</protein> , <protein>p65</protein> stimulates kappa B-directed transcription , whereas <protein>p50</protein> functions as a kappa B-specific repressor .


While authentic p50 homodimers ( previously termed KBF1 ) have been detected in nuclear extracts from nontransfected cells , experimental evidence supporting the existence of p65 homodimers in vivo was lacking .

While authentic <protein>p50 homodimers</protein> ( previously termed <protein>KBF1</protein> ) have been detected in nuclear extracts from nontransfected cells , experimental evidence supporting the existence of <protein>p65 homodimers</protein> in vivo was lacking .


We now provide direct biochemical evidence for the presence of an endogenous pool of inducible p65 homodimers in intact human T cells .

We now provide direct biochemical evidence for the presence of an endogenous pool of inducible <protein>p65 homodimers</protein> in intact <cell_type>human T cells</cell_type> .


As with the prototypical NF-kappa B p50-p65 heterodimer , this novel p65 homodimeric form of NF-kappa B is functionally sequestered in the cytoplasm but rapidly appears in the nuclear compartment following cellular stimulation .

As with the prototypical <protein>NF-kappa B p50-p65 heterodimer</protein> , this novel <protein>p65 homodimeric form</protein> of <protein>NF-kappa B</protein> is functionally sequestered in the cytoplasm but rapidly appears in the nuclear compartment following cellular stimulation .


Site-directed mutagenesis studies indicate that the homodimerization function of p65 is dependent upon the presence of cysteine 216 and a conserved recognition motif for protein kinase A ( RRPS ; amino acids 273 to 276 ) , both of which reside within a 91-amino-acid segment of the Rel homology domain that mediates self-association .

Site-directed mutagenesis studies indicate that the homodimerization function of <protein>p65</protein> is dependent upon the presence of cysteine 216 and a conserved recognition motif for <protein>protein kinase A</protein> ( <protein>RRPS</protein> ; amino acids 273 to 276 ) , both of which reside within a 91-amino-acid segment of the <protein>Rel homology domain</protein> that mediates self-association .


In contrast , mutations at these two sites do not affect heterodimerization of p65 with p50 or its functional interaction with I kappa B alpha .

In contrast , mutations at these two sites do not affect heterodimerization of <protein>p65</protein> with <protein>p50</protein> or its functional interaction with I kappa B alpha .


These later findings indicate that neither homo- nor heterodimer formation is an absolute prerequisite for I kappa B alpha recognition of p65 .

These later findings indicate that neither homo- nor heterodimer formation is an absolute prerequisite for <protein>I kappa B alpha</protein> recognition of <protein>p65</protein> .


Taken together with prior in vivo transcription studies , these results suggest that the biological activities of p65 and p50 homodimers are independently regulated , thereby providing an integrated and flexible control mechanism for the rapid activation and repression of NF-kappa B / Rel -directed gene expression .

Taken together with prior in vivo transcription studies , these results suggest that the biological activities of <protein>p65 and p50 homodimers</protein> are independently regulated , thereby providing an integrated and flexible control mechanism for the rapid activation and repression of <protein>NF-kappa B</protein> / <protein>Rel</protein> -directed gene expression .


Carrier determination for X-linked agammaglobulinemia using X inactivation analysis of purified B cells .

Carrier determination for X-linked agammaglobulinemia using X inactivation analysis of <cell_type>purified B cells</cell_type> .


We report the development of a relatively quick and simple method for the assessment of X inactivation status for carrier determination in families affected by X-linked agammaglobulinemia ( XLA ) .


This method utilises an immunomagnetic separation technique for B cell purification and a polymerase chain reaction ( PCR ) based assay for the determination of methylation status at the androgen receptor ( AR ) gene locus to assess whether X inactivation is random or non-random at this locus .

This method utilises an immunomagnetic separation technique for <cell_type>B cell</cell_type> purification and a polymerase chain reaction ( PCR ) based assay for the determination of methylation status at the <dna>androgen receptor ( AR ) gene locus</dna> to assess whether X inactivation is random or non-random at this locus .


We report the results we have obtained using this assay to investigate females known to be carriers of various X-linked immunodeficiency disorders .


In addition , we investigated four females from different families affected by XLA , two of whom were of unknown carrier status , and we discuss the results obtained with this and other X-inactivation assays .


A similar assay has recently been described by Allen et al. ( 1992 ) and applied to members of one family affected by XLA .


Effects of IL-4 and Fc gamma receptor II engagement on Egr-1 expression during stimulation of B lymphocytes by membrane immunoglobulin crosslinking .

Effects of <protein>IL-4</protein> and <protein>Fc gamma receptor II</protein> engagement on <protein>Egr-1</protein> expression during stimulation of <cell_type>B lymphocytes</cell_type> by membrane immunoglobulin crosslinking .


Egr-1 is an immediate early gene that is rapidly upregulated in response to mitogenic signals induced by antigen receptor crosslinking on murine B lymphocytes .

<dna>Egr-1</dna> is an <dna>immediate early gene</dna> that is rapidly upregulated in response to mitogenic signals induced by antigen receptor crosslinking on <cell_type>murine B lymphocytes</cell_type> .


It has been shown that levels of Egr-1 expression are closely correlated with B cell proliferation in several models of B cell activation and tolerance .

It has been shown that levels of <dna>Egr-1</dna> expression are closely correlated with <cell_type>B cell</cell_type> proliferation in several models of B cell activation and tolerance .


We compared the expression of Egr-1 during B cell stimulation with Fab'2 and IgG anti-immunoglobulin ( anti-Ig ) , since it is known that Fab'2 anti-Ig is mitogenic while IgG anti-Ig is not , owing to a dominant inhibitory effect of crosslinking the B cell Fc gamma RII to membrane Ig .

We compared the expression of <dna>Egr-1</dna> during B cell stimulation with <protein>Fab'2</protein> and <protein>IgG anti-immunoglobulin</protein> ( <protein>anti-Ig</protein> ) , since it is known that <protein>Fab'2 anti-Ig</protein> is mitogenic while <protein>IgG anti-Ig</protein> is not , owing to a dominant inhibitory effect of crosslinking the <protein>B cell Fc gamma RII</protein> to <protein>membrane Ig</protein> .


While mitogenic doses of Fab'2 anti-Ig induce large and rapid increases in Egr-1 expression , IgG anti-Ig results in smaller increases in Egr-1 mRNA , comparable to that seen with submitogenic concentrations of Fab'2 anti-Ig .

While mitogenic doses of <protein>Fab'2 anti-Ig</protein> induce large and rapid increases in Egr-1 expression , <protein>IgG anti-Ig</protein> results in smaller increases in <rna>Egr-1 mRNA</rna> , comparable to that seen with submitogenic concentrations of <protein>Fab'2 anti-Ig</protein> .


However , the correlation between Egr-1 expression and B cell proliferation breaks down when IL-4 is added as a co-mitogen to induce B cell proliferation with IgG anti-Ig or submitogenic concentrations of Fab'2 anti-Ig .

However , the correlation between Egr-1 expression and B cell proliferation breaks down when <protein>IL-4</protein> is added as a co-mitogen to induce B cell proliferation with <protein>IgG anti-Ig</protein> or submitogenic concentrations of <protein>Fab'2 anti-Ig</protein> .


No corresponding increases in Egr-1 mRNA levels are observed when IL-4 is added .

No corresponding increases in <rna>Egr-1 mRNA</rna> levels are observed when <protein>IL-4</protein> is added .


Therefore , IL-4 overcomes Fc receptor-mediated inhibition of B cell proliferation without affecting inhibition of Egr-1 mRNA induction , as demonstrated earlier for c-myc mRNA in this system .

Therefore , <protein>IL-4</protein> overcomes Fc receptor-mediated inhibition of <cell_type>B cell</cell_type> proliferation without affecting inhibition of <rna>Egr-1 mRNA</rna> induction , as demonstrated earlier for <rna>c-myc mRNA</rna> in this system .


Identification of a novel cyclosporin-sensitive element in the human tumor necrosis factor alpha gene promoter .

Identification of a novel <dna>cyclosporin-sensitive element</dna> in the <dna>human tumor necrosis factor alpha gene promoter</dna> .


Tumor necrosis factor alpha ( TNF-alpha ) , a cytokine with pleiotropic biological effects , is produced by a variety of cell types in response to induction by diverse stimuli .

<protein>Tumor necrosis factor alpha</protein> ( <protein>TNF-alpha</protein> ) , a <protein>cytokine</protein> with pleiotropic biological effects , is produced by a variety of cell types in response to induction by diverse stimuli .


In this paper , TNF-alpha mRNA is shown to be highly induced in a murine T cell clone by stimulation with T cell receptor ( TCR ) ligands or by calcium ionophores alone .

In this paper , <rna>TNF-alpha mRNA</rna> is shown to be highly induced in a <cell_line>murine T cell clone</cell_line> by stimulation with <protein>T cell receptor</protein> ( <protein>TCR</protein> ) ligands or by calcium ionophores alone .


Induction is rapid , does not require de novo protein synthesis , and is completely blocked by the immunosuppressant cyclosporin A ( CsA ) .


We have identified a human TNF-alpha promoter element , kappa 3 , which plays a key role in the calcium-mediated inducibility and CsA sensitivity of the gene .

We have identified a <dna>human TNF-alpha promoter element</dna> , <dna>kappa 3</dna> , which plays a key role in the calcium-mediated inducibility and CsA sensitivity of the gene .


In electrophoretic mobility shift assays , an oligonucleotide containing kappa 3 forms two DNA protein complexes with proteins that are present in extracts from unstimulated T cells .

In electrophoretic mobility shift assays , an oligonucleotide containing <dna>kappa 3</dna> forms two DNA protein complexes with proteins that are present in extracts from <cell_type>unstimulated T cells</cell_type> .


These complexes appear in nuclear extracts only after T cell stimulation .


Induction of the inducible nuclear complexes is rapid , independent of protein synthesis , and blocked by CsA , and thus , exactly parallels the induction of TNF-alpha mRNA by TCR ligands or by calcium ionophore .

Induction of the inducible <protein>nuclear complexes</protein> is rapid , independent of protein synthesis , and blocked by CsA , and thus , exactly parallels the induction of <rna>TNF-alpha mRNA</rna> by TCR ligands or by calcium ionophore .


Our studies indicate that the kappa 3 binding factor resembles the preexisting component of nuclear factor of activated T cells .

Our studies indicate that the <protein>kappa 3 binding factor</protein> resembles the preexisting component of <protein>nuclear factor</protein> of <cell_type>activated T cells</cell_type> .


Thus , the TNF-alpha gene is an immediate early gene in activated T cells and provides a new model system in which to study CsA-sensitive gene induction in activated T cells .

Thus , the <dna>TNF-alpha gene</dna> is an immediate early gene in activated <cell_type>T cells</cell_type> and provides a new model system in which to study <dna>CsA-sensitive gene</dna> induction in <cell_type>activated T cells</cell_type> .


Differences in expression of transcription factor AP-1 in human promyelocytic HL-60 cells during differentiation towards macrophages versus granulocytes .

Differences in expression of <protein>transcription factor AP-1</protein> in <cell_line>human promyelocytic HL-60 cells</cell_line> during differentiation towards <cell_type>macrophages</cell_type> versus <cell_type>granulocytes</cell_type> .


Commitment of HL-60 cells to macrophage or granulocytic differentiation was achieved by incubation with 4 beta-phorbol 12-myristate 13-acetate ( PMA ) for 30-60 min or with dimethyl sulphoxide ( DMSO ) for 24 h respectively .

Commitment of <cell_line>HL-60 cells</cell_line> to macrophage or granulocytic differentiation was achieved by incubation with 4 beta-phorbol 12-myristate 13-acetate ( PMA ) for 30-60 min or with dimethyl sulphoxide ( DMSO ) for 24 h respectively .


The commitment stage towards PMA-induced macrophage differentiation was associated with increases in jun B and c-fos mRNA levels , as well as with an increase in the binding activity of transcription factor AP-1 .

The commitment stage towards PMA-induced macrophage differentiation was associated with increases in jun B and c-fos mRNA levels , as well as with an increase in the binding activity of <protein>transcription factor AP-1</protein> .


Nevertheless , gel retardation analysis indicated that the AP-1 activity detected in untreated cells was drastically reduced during the commitment stage of DMSO-induced HL-60 differentiation towards granulocytes .

Nevertheless , gel retardation analysis indicated that the <protein>AP-1</protein> activity detected in untreated cells was drastically reduced during the commitment stage of DMSO-induced HL-60 differentiation towards <cell_type>granulocytes</cell_type> .


When HL-60 cells were treated with sodium butyrate , which induced monocytic differentiation , a remarkable increase in AP-1 binding activity was detected .

When <cell_line>HL-60 cells</cell_line> were treated with sodium butyrate , which induced monocytic differentiation , a remarkable increase in AP-1 binding activity was detected .


Treatment of HL-60 cells with 1 alpha , 25-dihydroxyvitamin D3 , another monocytic differentiation agent , induced a weak , but appreciable , increase in AP-1 activity .

Treatment of <cell_line>HL-60 cells</cell_line> with 1 alpha , 25-dihydroxyvitamin D3 , another monocytic differentiation agent , induced a weak , but appreciable , increase in <protein>AP-1</protein> activity .


Furthermore , addition of sodium butyrate or 1 alpha , 25-dihydroxyvitamin D3 to HL-60 cells induced the expression of c-fos , c-jun , jun B and jun D proto-oncogenes .

Furthermore , addition of sodium butyrate or 1 alpha , 25-dihydroxyvitamin D3 to <cell_line>HL-60 cells</cell_line> induced the expression of <dna>c-fos , c-jun , jun B and jun D proto-oncogenes</dna> .


In contrast , when HL-60 cells were treated with retinoic acid , a granulocytic differentiation inducer , no enhanced AP-1 binding activity was observed , and only a weak increase in jun D mRNA level was detected .

In contrast , when <cell_line>HL-60 cells</cell_line> were treated with retinoic acid , a granulocytic differentiation inducer , no enhanced AP-1 binding activity was observed , and only a weak increase in <rna>jun D mRNA</rna> level was detected .


These data indicate that formation of AP-1 is not required for the induction of HL-60 differentiation towards granulocytes , whereas induction of monocytic differentiation is correlated with an increase in AP-1 activity .

These data indicate that formation of <protein>AP-1</protein> is not required for the induction of <cell_line>HL-60</cell_line> differentiation towards <cell_type>granulocytes</cell_type> , whereas induction of monocytic differentiation is correlated with an increase in <protein>AP-1</protein> activity .


The differential expression of AP-1 activity may be critical in the differentiation of HL-60 cells towards monocytic or granulocytic lineages

The differential expression of <protein>AP-1</protein> activity may be critical in the differentiation of <cell_line>HL-60 cells</cell_line> towards <cell_type>monocytic or granulocytic lineages</cell_type>


Glucocorticoid receptors and sensitivity in leukemias .

<protein>Glucocorticoid receptors</protein> and sensitivity in leukemias .


In an attempt to investigate the utility of glucocorticoid receptor determination to predict clinical responsiveness in human leukemias we have studied glucocorticoid receptors in the leukemic cells from 46 patients and in the lymphocytes from 18 normal donors .

In an attempt to investigate the utility of <protein>glucocorticoid receptor</protein> determination to predict clinical responsiveness in human leukemias we have studied <protein>glucocorticoid receptors</protein> in the <cell_type>leukemic cells</cell_type> from 46 patients and in the <cell_type>lymphocytes</cell_type> from 18 normal donors .


In the normal lymphocytes there were 3 , 875 ( Median ) specific binding sites per cell .

In the <cell_type>normal lymphocytes</cell_type> there were 3 , 875 ( Median ) specific binding sites per cell .


The blasts from 17 patients with ANLL had on average higher levels of binding sites per cell ( Median = 7 , 250 , range : 0 to 15 , 295 ) than the other leukemias .

The <cell_type>blasts</cell_type> from 17 patients with ANLL had on average higher levels of <protein>binding sites</protein> per cell ( Median = 7 , 250 , range : 0 to 15 , 295 ) than the other leukemias .


Of the 15 patients with CLL , six had received glucocorticoid treatment for 3 to 5 years .


Their lymphocytes had lower number of receptors ( Median = 2 , 000 ) than the other cases which were newly diagnosed ( Median = 4 , 500 ) .

Their <cell_type>lymphocytes</cell_type> had lower number of receptors ( Median = 2 , 000 ) than the other cases which were newly diagnosed ( Median = 4 , 500 ) .


Four patients had ALL/AUL , three patients had blast crisis as terminal phase of CML , and seven had leukemic Non-Hodgkin lymphomas ( Median = 3 , 500 sites/cell ) .


In 24 patients we have also studied the in vitro sensitivity of the leukemic cells to dexamethasone .

In 24 patients we have also studied the in vitro sensitivity of the <cell_type>leukemic cells</cell_type> to dexamethasone .


There was no marked correlation between glucocorticoid receptor levels and in vitro sensitivity .

There was no marked correlation between <protein>glucocorticoid receptor</protein> levels and in vitro sensitivity .


An attempt to correlate receptor levels with clinical responsiveness demonstrated that glucocorticoid receptor determination might be of value in patients with lymphoid malignancies but probably not in patients with other leukemias .

An attempt to correlate receptor levels with clinical responsiveness demonstrated that <protein>glucocorticoid receptor</protein> determination might be of value in patients with lymphoid malignancies but probably not in patients with other leukemias .


'Activation-labile ' glucocorticoid-receptor complexes of a steroid-resistant variant of CEM-C7 human lymphoid cells .

<protein>'Activation-labile ' glucocorticoid-receptor complexes</protein> of a steroid-resistant variant of <cell_line>CEM-C7 human lymphoid cells</cell_line> .


For cytoplasmic glucocorticoid-receptor complexes to enter and accumulate in the nucleus a temperature-dependent event , 'activation ' is required .

For <protein>cytoplasmic glucocorticoid-receptor complexes</protein> to enter and accumulate in the nucleus a temperature-dependent event , 'activation ' is required .


Activation can be achieved in vitro by increased ionic strength , dilution or gel filtration and is manifested by an increased affinity of steroid-receptor complex for DNA and an altered elution profile from ion-exchange resins .

Activation can be achieved in vitro by increased ionic strength , dilution or gel filtration and is manifested by an increased affinity of <protein>steroid-receptor complex</protein> for <dna>DNA</dna> and an altered elution profile from ion-exchange resins .


Munck and Foley have shown that activated complexes isolated from thymocytes elute from DEAE-cellulose in a manner identical to complexes activated in vitro .

Munck and Foley have shown that <protein>activated complexes</protein> isolated from thymocytes elute from DEAE-cellulose in a manner identical to complexes activated in vitro .


We report here that DEAE-cellulose chromatography of steroid-receptor complexes from CEM-C7 , a cloned human leukaemic T-cell line sensitive to the cytolytic action of glucocorticoids , and its steroid-resistant subclone 4R4 demonstrated that steroid receptors of clone 4R4 can not form stable activated complexes .

We report here that DEAE-cellulose chromatography of <protein>steroid-receptor complexes</protein> from <cell_line>CEM-C7</cell_line> , a <cell_line>cloned human leukaemic T-cell line</cell_line> sensitive to the cytolytic action of glucocorticoids , and its <cell_line>steroid-resistant subclone 4R4</cell_line> demonstrated that <protein>steroid receptors</protein> of <cell_line>clone 4R4</cell_line> can not form stable activated complexes .


This defines a new defect in receptor action , activation lability ( r+act1 ) , which is unlike either the r- , r+nt- , or r+nti phenotypes previously described for mouse lymphoid variants .

This defines a new defect in receptor action , activation lability ( r+act1 ) , which is unlike either the <cell_line>r- , r+nt- , or r+nti phenotypes</cell_line> previously described for <cell_line>mouse lymphoid variants</cell_line> .


Granulocytes in the endometrium of post-partum women .

<cell_type>Granulocytes</cell_type> in the endometrium of post-partum women .


Endometrial samples of women at various stages of gonadal activity after parturition were examined for the presence and numbers of endometrial granulocytes .

Endometrial samples of women at various stages of gonadal activity after parturition were examined for the presence and numbers of <cell_type>endometrial granulocytes</cell_type> .


Although samples at all the stages contained significant numbers of the granulocytes ( i.e. greater than 7/high-power field ) , the 100 % values for late-proliferative and adaptation hyperplasia were significantly higher than the values for the resting ( 81.8 % ) , early ( 82.4 % ) and mid- ( 87.9 % ) proliferative and secretory ( 83.3 % ) phases .

Although samples at all the stages contained significant numbers of the <cell_type>granulocytes</cell_type> ( i.e. greater than 7/high-power field ) , the 100 % values for late-proliferative and adaptation hyperplasia were significantly higher than the values for the resting ( 81.8 % ) , early ( 82.4 % ) and mid- ( 87.9 % ) proliferative and secretory ( 83.3 % ) phases .


We suggest that this correlates with the suggestion that the granulocytes constitute a receptor system for oestrogens .

We suggest that this correlates with the suggestion that the <cell_type>granulocytes</cell_type> constitute a receptor system for oestrogens .


Interaction of glucocorticoids with macrophages .

Interaction of glucocorticoids with <cell_type>macrophages</cell_type> .


Identification of glucocorticoid receptors in monocytes and macrophages .

Identification of <protein>glucocorticoid receptors</protein> in <cell_type>monocytes</cell_type> and <cell_type>macrophages</cell_type> .


Glucocorticoid binding was measured in resident and thioglycollate-elicited mouse peritoneal macrophages , rabbit alveolar macrophages , and human monocytes .

Glucocorticoid binding was measured in resident and <cell_type>thioglycollate-elicited mouse peritoneal macrophages</cell_type> , <cell_type>rabbit alveolar macrophages</cell_type> , and <cell_type>human monocytes</cell_type> .


Two assays of binding were used -- an assay with intact cells in suspension or monolayers , and an assay of cytosol and nuclear forms of glucocorticoid receptors .

Two assays of binding were used -- an assay with <cell_type>intact cells</cell_type> in suspension or monolayers , and an assay of cytosol and nuclear forms of <protein>glucocorticoid receptors</protein> .


The mononuclear phagocytes contained approximately equal to 4 -- 10 X 10 ( 3 ) high affinity receptor sites per cell , with dissociation constants of approximately equal to 2 -- 8 nM dexamethasone .

The <cell_type>mononuclear phagocytes</cell_type> contained approximately equal to 4 -- 10 X 10 ( 3 ) <protein>high affinity receptor sites</protein> per cell , with dissociation constants of approximately equal to 2 -- 8 nM dexamethasone .


The binding to the saturable sites was specific for steroids with glucocorticoid or antiglucocorticoid activity .

The binding to the <protein>saturable sites</protein> was specific for steroids with glucocorticoid or antiglucocorticoid activity .


Cortisol , corticosterone , and progesterone competed with dexamethasone for binding , whereas estradiol , dihydrotestosterone , and 11-epicortisol competed very little .


Binding of dexamethasone to cytosol and nuclear forms of the receptor complex and temperature-sensitive translocation of cytosol forms to nuclear forms were shown .

Binding of dexamethasone to cytosol and nuclear forms of the <protein>receptor complex</protein> and temperature-sensitive translocation of cytosol forms to nuclear forms were shown .


At 37 degrees C the predominant form of the hormone-receptor complex was nuclear .

At 37 degrees C the predominant form of the <protein>hormone-receptor complex</protein> was nuclear .


These results demonstrate that corticosteroids interact with macrophages at physiological concentrations .

These results demonstrate that corticosteroids interact with <cell_type>macrophages</cell_type> at physiological concentrations .


Nitric oxide signaling : a possible role for G proteins .

Nitric oxide signaling : a possible role for <protein>G proteins</protein> .


We have previously reported various inductive effects of nitric oxide on human PBMC .

We have previously reported various inductive effects of nitric oxide on <cell_type>human PBMC</cell_type> .


We describe a novel and potentially important mechanism of nitric oxide signaling-through direct activation of guanine nucleotide-binding proteins ( G proteins ) .

We describe a novel and potentially important mechanism of nitric oxide signaling-through direct activation of <protein>guanine nucleotide-binding proteins</protein> ( <protein>G proteins</protein> ) .


We have found that nitric oxide treatment of membranes isolated from fresh human PBMC enhances the ability of these membranes to hydrolyze [ gamma-32P ] GTP and bind [ gamma-35S ] GTP .

We have found that nitric oxide treatment of membranes isolated from <cell_type>fresh human PBMC</cell_type> enhances the ability of these membranes to hydrolyze [ gamma-32P ] GTP and bind [ gamma-35S ] GTP .


In addition , treatment of whole cells with nitric oxide yielded membranes with enhanced GTPase activity .


Furthermore , the GTPase activity of pure , recombinant Gs alpha , Gi alpha 1 , and p21ras was greatly enhanced by nitric oxide .

Furthermore , the GTPase activity of pure , recombinant <protein>Gs alpha</protein> , <protein>Gi alpha 1</protein> , and <protein>p21ras</protein> was greatly enhanced by nitric oxide .


In support of the existence of this pathway in whole cells , we found that the G protein inhibitor , GDP-beta-S , blocked NF-kappa B translocation induced by nitric oxide or LPS in permeabilized cells .

In support of the existence of this pathway in whole cells , we found that the G protein inhibitor , GDP-beta-S , blocked <protein>NF-kappa B</protein> translocation induced by nitric oxide or LPS in <cell_type>permeabilized cells</cell_type> .


In addition , nitric oxide greatly reduced the pertussis toxin-mediated ADP-ribosylation of 45- and 41-kDa proteins in membranes of these cells .

In addition , nitric oxide greatly reduced the pertussis toxin-mediated ADP-ribosylation of <protein>45- and 41-kDa proteins</protein> in membranes of these cells .


Because G proteins play a central role in many diverse signaling systems , activation by an endogenous and inducible oxidant may represent a novel signaling pathway .

Because <protein>G proteins</protein> play a central role in many diverse signaling systems , activation by an endogenous and inducible oxidant may represent a novel signaling pathway .


The granulocyte-macrophage colony-stimulating factor promoter cis-acting element CLE0 mediates induction signals in T cells and is recognized by factors related to AP1 and NFAT .

The <dna>granulocyte-macrophage colony-stimulating factor promoter cis-acting element</dna> <dna>CLE0</dna> mediates induction signals in <cell_type>T cells</cell_type> and is recognized by factors related to <protein>AP1</protein> and <protein>NFAT</protein> .


Expression of the granulocyte-macrophage colony-stimulating factor ( GM-CSF ) gene in T cells is activated by the combination of phorbol ester ( phorbol myristate acetate ) and calcium ionophore ( A23187 ) , which mimic antigen stimulation through the T-cell receptor .

Expression of the <dna>granulocyte-macrophage colony-stimulating factor ( GM-CSF ) gene</dna> in T cells is activated by the combination of phorbol ester ( phorbol myristate acetate ) and calcium ionophore ( A23187 ) , which mimic antigen stimulation through the <protein>T-cell receptor</protein> .


We have previously shown that a fragment containing bp -95 to +27 of the mouse GM-CSF promoter can confer inducibility to reporter genes in the human Jurkat T-cell line .

We have previously shown that a fragment containing <dna>bp -95 to +27</dna> of the <dna>mouse GM-CSF promoter</dna> can confer inducibility to <dna>reporter genes</dna> in the <cell_line>human Jurkat T-cell line</cell_line> .


Here we use an in vitro transcription system to demonstrate that a cis-acting element ( positions -54 to -40 ) , referred to as CLE0 , is a target for the induction signals .

Here we use an in vitro transcription system to demonstrate that a <dna>cis-acting element</dna> ( <dna>positions -54 to -40</dna> ) , referred to as <dna>CLE0</dna> , is a target for the induction signals .


We observed induction with templates containing intact CLE0 but not with templates with deleted or mutated CLE0 .

We observed induction with templates containing intact <dna>CLE0</dna> but not with templates with <dna>deleted or mutated CLE0</dna> .


We also observed that two distinct signals were required for the stimulation through CLE0 , since only extracts from cells treated with both phorbol myristate acetate and A23187 supported optimal induction .

We also observed that two distinct signals were required for the stimulation through <dna>CLE0</dna> , since only extracts from cells treated with both phorbol myristate acetate and A23187 supported optimal induction .


Stimulation probably was mediated by CLE0-binding proteins because depletion of these proteins specifically reduced GM-CSF transcription .

Stimulation probably was mediated by <protein>CLE0-binding proteins</protein> because depletion of these proteins specifically reduced <protein>GM-CSF</protein> transcription .


One of the binding factors possessed biochemical and immunological features identical to those of the transcription factor AP1 .

One of the <protein>binding factors</protein> possessed biochemical and immunological features identical to those of the <protein>transcription factor AP1</protein> .


Another factor resembled the T-cell-specific factor NFAT .

Another factor resembled the <protein>T-cell-specific factor NFAT</protein> .


The characteristics of these two factors are consistent with their involvement in GM-CSF induction .


The presence of CLE0-like elements in the promoters of interleukin-3 ( IL-3 ) , IL-4 , IL-5 , GM-CSF , and NFAT sites in the IL-2 promoter suggests that the factors we detected , or related factors that recognize these sites , may account for the coordinate induction of these genes during T-cell activation .

The presence of <dna>CLE0-like elements</dna> in the <dna>promoters</dna> of <dna>interleukin-3 ( IL-3 ) , IL-4 , IL-5 , GM-CSF , and NFAT sites</dna> in the <dna>IL-2 promoter</dna> suggests that the factors we detected , or related factors that recognize these sites , may account for the coordinate induction of these genes during <cell_type>T-cell</cell_type> activation .


Identification and characterization of an Alu-containing , T-cell-specific enhancer located in the last intron of the human CD8 alpha gene .

Identification and characterization of an <dna>Alu-containing , T-cell-specific enhancer</dna> located in the last <dna>intron</dna> of the <dna>human CD8 alpha gene</dna> .


Expression of the human CD8 alpha gene is restricted to cells of the lymphoid lineage and developmentally regulated during thymopoiesis .

Expression of the <dna>human CD8 alpha gene</dna> is restricted to cells of the <cell_type>lymphoid lineage</cell_type> and developmentally regulated during thymopoiesis .


As an initial step towards understanding the molecular basis for tissue-specific expression of this gene , we surveyed the surrounding chromatin structure for potential cis-acting regulatory regions by DNase I hypersensitivity mapping and found four hypersensitive sites , three of which were T cell restricted .

As an initial step towards understanding the molecular basis for tissue-specific expression of this gene , we surveyed the surrounding chromatin structure for potential <dna>cis-acting regulatory regions</dna> by <protein>DNase I</protein> hypersensitivity mapping and found four hypersensitive sites , three of which were T cell restricted .


By using a reporter-based expression approach , a T-cell-specific enhancer was identified by its close association with a prominent T-cell-restricted hypersensitive sites in the last intron of the CD8 alpha gene .

By using a reporter-based expression approach , a <dna>T-cell-specific enhancer</dna> was identified by its close association with a prominent <dna>T-cell-restricted hypersensitive sites</dna> in the last intron of the <dna>CD8 alpha gene</dna> .


Deletion studies demonstrated that the minimal enhancer is adjacent to a negative regulatory element .


DNA sequence analysis of the minimal enhancer revealed a striking cluster of consensus binding sites for Ets-1 , TCF-1 , CRE , GATA-3 , LyF-1 , and bHLH proteins which were verified by electrophoretic mobility shift assays .

DNA sequence analysis of the <dna>minimal enhancer</dna> revealed a striking cluster of <dna>consensus binding sites</dna> for <protein>Ets-1 , TCF-1 , CRE , GATA-3 , LyF-1 , and bHLH proteins</protein> which were verified by electrophoretic mobility shift assays .


In addition , the 5 ' end of the enhancer was composed of an Alu repeat which contained the GATA-3 , bHLH , and LyF-1 binding sites .

In addition , the <dna>5 ' end</dna> of the <dna>enhancer</dna> was composed of an Alu repeat which contained the <dna>GATA-3 , bHLH , and LyF-1 binding sites</dna> .


Site-directed mutation of the Ets-1 and GATA-3 sites dramatically reduced enhancer activity .

Site-directed mutation of the <dna>Ets-1 and GATA-3 sites</dna> dramatically reduced enhancer activity .


The functional importance of the other binding sites only became apparent when combinations of mutations were analyzed .


Taken together , these results suggest that the human CD8 alpha gene is regulated by the interaction of multiple T-cell nuclear proteins with a transcriptional enhancer located in the last intron of the gene .

Taken together , these results suggest that the <dna>human CD8 alpha gene</dna> is regulated by the interaction of multiple <protein>T-cell nuclear proteins</protein> with a transcriptional enhancer located in the last <dna>intron</dna> of the gene .


Comparison of the CD8 alpha enhancer with other recently identified T-cell-specific regulatory elements suggests that a common set of transcription factors regulates several T-cell genes .

Comparison of the <dna>CD8 alpha enhancer</dna> with other recently identified <dna>T-cell-specific regulatory elements</dna> suggests that a common set of <protein>transcription factors</protein> regulates several <dna>T-cell genes</dna> .


Molecular regulation of the human IL-3 gene : inducible T cell-restricted expression requires intact AP-1 and Elf-1 nuclear protein binding sites .

Molecular regulation of the <dna>human IL-3 gene</dna> : inducible T cell-restricted expression requires intact <dna>AP-1 and Elf-1 nuclear protein binding sites</dna> .


Interleukin 3 ( IL-3 ) is a hematopoietic stem-cell growth and differentiation factor that is expressed solely in activated T and NK cells .

<protein>Interleukin 3</protein> ( <protein>IL-3</protein> ) is a <protein>hematopoietic stem-cell growth and differentiation factor</protein> that is expressed solely in activated <cell_type>T and NK cells</cell_type> .


Studies to date have identified elements 5 ' to the IL-3 coding sequences that regulate its transcription , but the sequences that confer T cell-specific expression remain to be clearly defined .

Studies to date have identified elements 5 ' to the <dna>IL-3 coding sequences</dna> that regulate its transcription , but the sequences that confer T cell-specific expression remain to be clearly defined .


We have now identified DNA sequences that are required for T cell-restricted IL-3 gene transcription .

We have now identified <dna>DNA sequences</dna> that are required for T cell-restricted <dna>IL-3 gene</dna> transcription .


A series of transient transfections performed with human IL-3-chloramphenicol acetyltransferase ( CAT ) reporter plasmids in T and non-T cells revealed that a plasmid containing 319 bp of 5 ' flanking sequences was active exclusively in T cells .

A series of transient transfections performed with <dna>human IL-3-chloramphenicol acetyltransferase ( CAT ) reporter plasmids</dna> in <cell_type>T and non-T cells</cell_type> revealed that a <dna>plasmid</dna> containing 319 bp of <dna>5 ' flanking sequences</dna> was active exclusively in <cell_type>T cells</cell_type> .


Deletion analysis revealed that T cell specificity was conferred by a 49-bp fragment ( bp -319 to -270 ) that included a potential binding site for AP-1 transcription factors 6 bp upstream of a binding site for Elf-1 , a member of the Ets family of transcription factors .

Deletion analysis revealed that <cell_type>T cell</cell_type> specificity was conferred by a <dna>49-bp fragment</dna> ( <dna>bp -319 to -270</dna> ) that included a <dna>potential binding site</dna> for <protein>AP-1 transcription factors</protein> 6 bp upstream of a <dna>binding site for Elf-1</dna> , a member of the <protein>Ets family</protein> of <protein>transcription factors</protein> .


DNaseI footprint and electrophoretic mobility shift assay analyses performed with MLA-144 T cell nuclear extracts demonstrated that this 49-bp region contains a nuclear protein binding region that includes consensus AP-1 and Elf-1 binding sites .

<protein>DNaseI</protein> footprint and electrophoretic mobility shift assay analyses performed with <cell_line>MLA-144 T cell</cell_line> nuclear extracts demonstrated that this <dna>49-bp region</dna> contains a <dna>nuclear protein binding region</dna> that includes <dna>consensus AP-1 and Elf-1 binding sites</dna> .


In addition , extracts prepared from purified human T cells contained proteins that bound to synthetic oligonucleotides corresponding to the AP-1 and Elf-1 binding sites .

In addition , extracts prepared from <cell_type>purified human T cells</cell_type> contained proteins that bound to synthetic oligonucleotides corresponding to the <dna>AP-1 and Elf-1 binding sites</dna> .


In vitro-transcribed and -translated Elf-1 protein bound specifically to the Elf-1 site , and Elf-1 antisera competed and super shifted nuclear protein complexes present in MLA-144 nuclear extracts .

In vitro-transcribed and -translated <protein>Elf-1 protein</protein> bound specifically to the <dna>Elf-1 site</dna> , and Elf-1 antisera competed and super shifted nuclear protein complexes present in <cell_line>MLA-144</cell_line> nuclear extracts .


Moreover , addition of anti-Jun family antiserum in electrophoretic mobility shift assay reactions completely blocked formation of the AP-1-related complexes .

Moreover , addition of anti-Jun family antiserum in electrophoretic mobility shift assay reactions completely blocked formation of the <protein>AP-1-related complexes</protein> .


Transient transfection studies in MLA-144 T cells revealed that constructs containing mutations in the AP-1 site almost completely abolished CAT activity while mutation of the Elf-1 site or the NF-IL-3 site , a previously described nuclear protein binding site ( bp. -155 to -148 ) in the IL-3 promoter , reduced CAT activity to < 25 % of the activity given by wild-type constructs .

Transient transfection studies in <cell_line>MLA-144 T cells</cell_line> revealed that constructs containing mutations in the <dna>AP-1 site</dna> almost completely abolished CAT activity while mutation of the <dna>Elf-1 site</dna> or the <dna>NF-IL-3 site</dna> , a previously described <dna>nuclear protein binding site</dna> ( <dna>bp. -155 to -148</dna> ) in the <dna>IL-3 promoter</dna> , reduced CAT activity to < 25 % of the activity given by wild-type constructs .


We conclude that expression of the human IL-3 gene requires the AP-1 and Elf-1 binding sites ; however , unlike other previously characterized cytokine genes such as IL-2 , the AP-1 and Elf-1 factors can bind independently in the IL-3 gene .

We conclude that expression of the <dna>human IL-3 gene</dna> requires the <dna>AP-1 and Elf-1 binding sites</dna> ; however , unlike other previously characterized <dna>cytokine genes</dna> such as <protein>IL-2</protein> , the <protein>AP-1 and Elf-1 factors</protein> can bind independently in the <dna>IL-3 gene</dna> .


( ABSTRACT TRUNCATED AT 400 WORDS )


Combination IL-2 and IL-4 reduces glucocorticoid receptor-binding affinity and T cell response to glucocorticoids .

Combination <protein>IL-2</protein> and <protein>IL-4</protein> reduces glucocorticoid receptor-binding affinity and <cell_type>T cell</cell_type> response to glucocorticoids .


The mechanisms contributing to persistent T cell activation and poor response to glucocorticoids in chronic inflammatory illnesses such as steroid resistant ( SR ) asthma are poorly defined .

The mechanisms contributing to persistent <cell_type>T cell</cell_type> activation and poor response to glucocorticoids in chronic inflammatory illnesses such as steroid resistant ( SR ) asthma are poorly defined .


We examined the possibility that certain cytokines , specifically IL-2 and IL-4 , could affect T cell response to glucocorticoids .

We examined the possibility that certain cytokines , specifically <protein>IL-2</protein> and <protein>IL-4</protein> , could affect T cell response to glucocorticoids .


A [ 3H ] dexamethasone radioligand-binding assay was used to measure the number of glucocorticoid receptors ( GR ) and dissociation constant ( Kd ) in PBMC from normal donors and patients with SR asthma , cultured in the absence and presence of these cytokines .

A [ 3H ] dexamethasone radioligand-binding assay was used to measure the number of <protein>glucocorticoid receptors</protein> ( <protein>GR</protein> ) and dissociation constant ( Kd ) in <cell_type>PBMC</cell_type> from normal donors and patients with SR asthma , cultured in the absence and presence of these <protein>cytokines</protein> .


PBMC from normal donors incubated for 48 h in the presence of combination IL-2 + IL-4 had nuclear GR with significantly reduced binding affinity ( GR Kd = 36.1 +/- 1.63 nM , mean +/- SEM ; p = 0.0001 ) as compared with PBMC incubated with medium alone ( GR Kd = 6.74 +/- 0.46 nM ) .

PBMC from normal donors incubated for 48 h in the presence of combination <protein>IL-2</protein> + <protein>IL-4</protein> had nuclear GR with significantly reduced binding affinity ( GR Kd = 36.1 +/- 1.63 nM , mean +/- SEM ; p = 0.0001 ) as compared with <cell_type>PBMC</cell_type> incubated with medium alone ( GR Kd = 6.74 +/- 0.46 nM ) .


The cytosolic GR Kd remained unchanged .


However , when PBMC were incubated with IL-2 alone or IL-4 alone , no change in GR-binding affinity was observed .

However , when <cell_type>PBMC</cell_type> were incubated with <protein>IL-2</protein> alone or <protein>IL-4</protein> alone , no change in GR-binding affinity was observed .


Furthermore , when T cells and non-T cells were individually stimulated with combination IL-2 + IL-4 , a significant reduction in GR-binding affinity was observed only in the T cell population ( p = 0.0001 ) .

Furthermore , when <cell_type>T cells</cell_type> and <cell_type>non-T cells</cell_type> were individually stimulated with combination IL-2 + IL-4 , a significant reduction in GR-binding affinity was observed only in the <cell_type>T cell population</cell_type> ( p = 0.0001 ) .


The IL-2 + IL-4-induced alteration in PBMC GR Kd was associated with an increase in GR number ( 8348 +/- 964 vs 1710 +/- 228 sites/cell ; p = 0.0003 ) .


More importantly , the alteration in PBMC GR-binding affinity with IL-2 + IL-4 was associated with a functional change in T cell response to methylprednisolone MPN , i.e. , a reduced inhibitory effect of MPN on PMA/ionomycin-induced T cell proliferation .

More importantly , the alteration in PBMC GR-binding affinity with <protein>IL-2</protein> + <protein>IL-4</protein> was associated with a functional change in T cell response to methylprednisolone MPN , i.e. , a reduced inhibitory effect of MPN on PMA/ionomycin-induced <cell_type>T cell</cell_type> proliferation .


These effects of IL-2 + IL-4 on PBMC GR affinity and response to MPN were blocked by co-incubation with IFN-gamma .

These effects of <protein>IL-2</protein> + <protein>IL-4</protein> on <cell_type>PBMC</cell_type> <protein>GR</protein> affinity and response to MPN were blocked by co-incubation with <protein>IFN-gamma</protein> .


Freshly isolated PBMC from four patients with SR asthma had a significantly reduced GR-binding affinity ( Kd = 40.0 +/- 2.68 nM ; p = 0.0001 ) when compared with seven normal subjects ( 7.15 +/- 0.41 nM ) .

Freshly isolated <cell_type>PBMC</cell_type> from four patients with SR asthma had a significantly reduced GR-binding affinity ( Kd = 40.0 +/- 2.68 nM ; p = 0.0001 ) when compared with seven normal subjects ( 7.15 +/- 0.41 nM ) .


The altered PBMC GR binding from patients with SR asthma reversed to normal when incubated with medium alone , but was sustained with IL-2 + IL-4 .

The altered PBMC GR binding from patients with SR asthma reversed to normal when incubated with medium alone , but was sustained with <protein>IL-2</protein> + <protein>IL-4</protein> .


These observations suggest that with persistent inflammation certain cytokines may contribute to an impaired response to glucocorticoids .

These observations suggest that with persistent inflammation certain <protein>cytokines</protein> may contribute to an impaired response to glucocorticoids .


Furthermore , the effects of IL-2 and IL-4 were blocked by IFN-gamma .

Furthermore , the effects of <protein>IL-2</protein> and <protein>IL-4</protein> were blocked by <protein>IFN-gamma</protein> .


Characterization of the human CD4 gene promoter : transcription from the CD4 gene core promoter is tissue-specific and is activated by Ets proteins .

Characterization of the <dna>human CD4 gene promoter</dna> : transcription from the <dna>CD4 gene core promoter</dna> is tissue-specific and is activated by <protein>Ets proteins</protein> .


We analyzed the 5 ' transcription control sequences of the human CD4 gene .

We analyzed the <dna>5 ' transcription control sequences</dna> of the <dna>human CD4 gene</dna> .


We located the transcription initiation site and showed that the CD4 core promoter ( positions -40 to +16 ) lacks a classical `` TATA '' or initiator positioning consensus sequence but directs precise and efficient transcription when coupled to the ubiquitously active simian virus 40 enhancer .

We located the <dna>transcription initiation site</dna> and showed that the <dna>CD4 core promoter</dna> ( <dna>positions -40 to +16</dna> ) lacks a classical `` TATA '' or <dna>initiator positioning consensus sequence</dna> but directs precise and efficient transcription when coupled to the ubiquitously active <dna>simian virus 40 enhancer</dna> .


The transcriptional activity of the CD4 gene promoter correlated with CD4 expression in various cell types .

The transcriptional activity of the <dna>CD4 gene promoter</dna> correlated with CD4 expression in various cell types .


Interestingly , the CD4 core promoter also displayed a tissue-specific transcriptional activity .

Interestingly , the <dna>CD4 core promoter</dna> also displayed a tissue-specific transcriptional activity .


Within this fragment , three nucleic acid sequences are completely conserved in the murine CD4 gene .

Within this fragment , three nucleic acid sequences are completely conserved in the <dna>murine CD4 gene</dna> .


One of these sequences contains a perfect ETS consensus sequence .

One of these sequences contains a <dna>perfect ETS consensus sequence</dna> .


Another ETS consensus sequence is located 1060 nt upstream .

Another <dna>ETS consensus sequence</dna> is located 1060 nt upstream .


Electrophoretic-mobility-shift assays showed that the core promoter ETS motif binds an Ets-related protein specifically expressed at high levels in CD4+ cells .

Electrophoretic-mobility-shift assays showed that the core promoter ETS motif binds an <protein>Ets-related protein</protein> specifically expressed at high levels in <cell_type>CD4+ cells</cell_type> .


Moreover , in CD4- cells , overexpression of Ets-1 or Ets-2 efficiently and specifically activated transcription from the CD4 promoter and core promoter .

Moreover , in <cell_type>CD4- cells</cell_type> , overexpression of <protein>Ets-1</protein> or <protein>Ets-2</protein> efficiently and specifically activated transcription from the <dna>CD4 promoter</dna> and <dna>core promoter</dna> .


These data indicate that Ets transcription factors play a central role in controlling CD4 gene expression , by binding to both a classical remote site and an unusual proximal activator sequence

These data indicate that <protein>Ets transcription factors</protein> play a central role in controlling CD4 gene expression , by binding to both a <dna>classical remote site</dna> and an unusual <dna>proximal activator sequence</dna>


Glucocorticoid receptor activation and inactivation in c ultured human lymphocytes .

Glucocorticoid receptor activation and inactivation in c <cell_line>ultured human lymphocytes</cell_line> .


Although glucocorticoids are not cytolytic for and do not inhibit the growth of the IM-9 line of cultured human lymphoblasts , these cells have a high steroid-binding capacity .

Although glucocorticoids are not cytolytic for and do not inhibit the growth of the <cell_line>IM-9 line</cell_line> of <cell_line>cultured human lymphoblasts</cell_line> , these cells have a high steroid-binding capacity .


We have used IM-9 cells in order to examine whether unoccupied glucocorticoid receptors are inactivated and activated in intact cells .

We have used <cell_type>IM-9 cells</cell_type> in order to examine whether <protein>unoccupied glucocorticoid receptors</protein> are inactivated and activated in <cell_type>intact cells</cell_type> .


when IM-9 cells are incubated in glucose-free medium in a nitrogen atmosphere , both their ability to bind triamcinolone acetonide and their ATP levels decline and , when glucose and oxygen are reintroduced , ATP levels and receptor activity return .

when <cell_type>IM-9 cells</cell_type> are incubated in glucose-free medium in a nitrogen atmosphere , both their ability to bind triamcinolone acetonide and their ATP levels decline and , when glucose and oxygen are reintroduced , ATP levels and receptor activity return .


The specific glucocorticoid-binding activity of cytosol prepared from cells exposed to various degrees of energy limitation is directly correlated with the ATP content .


Receptor activation in intact cells is rapid and independent of protein synthesis .

Receptor activation in <cell_type>intact cells</cell_type> is rapid and independent of protein synthesis .


Cytosol prepared from inactivated cells can not be activated by addition of ATP .


The inactivation of glucocorticoid receptors that occurs when cytosol from normal IM-9 cells is incubated at 25 degrees C is inhibited by molybdate , vanadate , fluoride , ATP , and several other nucleotides .

The inactivation of <protein>glucocorticoid receptors</protein> that occurs when cytosol from normal <cell_line>IM-9 cells</cell_line> is incubated at 25 degrees C is inhibited by molybdate , vanadate , fluoride , ATP , and several other nucleotides .


The experiments with intact human lymphoblasts suggest that assays of specific glucocorticoid-binding capacity do not necessarily reflect the cellular content of receptor protein .

The experiments with <cell_type>intact human lymphoblasts</cell_type> suggest that assays of specific glucocorticoid-binding capacity do not necessarily reflect the cellular content of <protein>receptor protein</protein> .


Glucocorticoids and lymphocytes .

Glucocorticoids and <cell_type>lymphocytes</cell_type> .


II .


Cell cycle-dependent changes in glucocorticoid receptor content .

Cell cycle-dependent changes in <protein>glucocorticoid receptor</protein> content .


To study variations in glucocorticoid receptor levels during the cell cycle , we have separated mitogen-stimulated human peripheral lymphocytes and rat lymph node cells by unit gravity sedimentation and measured glucocorticoid binding in the resultant fractions .

To study variations in <protein>glucocorticoid receptor</protein> levels during the cell cycle , we have separated <cell_line>mitogen-stimulated human peripheral lymphocytes</cell_line> and <cell_type>rat lymph node cells</cell_type> by unit gravity sedimentation and measured glucocorticoid binding in the resultant fractions .


By morphologic criteria and thymidine incorporation , the fractions were separated into populations of G0 and G1 phase and S and post-S phase cells .

By morphologic criteria and thymidine incorporation , the fractions were separated into populations of <cell_type>G0 and G1 phase and S and post-S phase cells</cell_type> .


A 2- to 3-fold increase in glucocorticoid receptor sites per cell , for cells in the S and post-S phase over those in G0 and G1 , was observed with both nonstimulated rat lymph node cell suspensions and concanavalin A-stimulated human peripheral lymphocytes .

A 2- to 3-fold increase in glucocorticoid receptor sites per cell , for cells in the S and post-S phase over those in G0 and G1 , was observed with both <cell_type>nonstimulated rat lymph node cell suspensions</cell_type> and <cell_line>concanavalin A-stimulated human peripheral lymphocytes</cell_line> .


These observations together with those from other studies indicate that formation of new glucocorticoid receptors near the S phase may be a general phenomenon in proliferating cells .

These observations together with those from other studies indicate that formation of new <protein>glucocorticoid receptors</protein> near the S phase may be a general phenomenon in <cell_type>proliferating cells</cell_type> .


We propose that this increase in glucocorticoid receptors during the cell cycle may explain the increase in glucocorticoid receptors in mitogen-stimulated lymphocytes .

We propose that this increase in <protein>glucocorticoid receptors</protein> during the cell cycle may explain the increase in <protein>glucocorticoid receptors</protein> in <cell_type>mitogen-stimulated lymphocytes</cell_type> .


The leukocyte migration inhibition response to certain breast cancer-related antigens ( MCF-7 and MuMTV ) : their potential as discriminants .

The <cell_type>leukocyte</cell_type> migration inhibition response to certain <protein>breast cancer-related antigens</protein> ( <protein>MCF-7</protein> and <protein>MuMTV</protein> ) : their potential as discriminants .


Certain oncogenic viruses have been implicated in human breast cancer , including the murine mammary tumor virus ( MuMTV ) and the Mason-Pfizer monkey virus ( MPMV ) .


We have used the leukocyte migration inhibition ( LMI ) response to assay the response to several potential breast cancer-related antigens , including MuMTV , MPMV , and a breast cancer cultured cell line , MCF-7 , in 96 breast cancer patients , in 32 women with benign breast disease , and in 67 normal women .

We have used the leukocyte migration inhibition ( LMI ) response to assay the response to several potential breast cancer-related antigens , including <protein>MuMTV</protein> , <protein>MPMV</protein> , and a <cell_line>breast cancer cultured cell line</cell_line> , <cell_line>MCF-7</cell_line> , in 96 breast cancer patients , in 32 women with benign breast disease , and in 67 normal women .


The lowest tenth percentile of control ( LMI ) responses was used as the cutoff point to designate responders .


Breast cancer patients showed significant responses to MuMTV ( 49 % and to MCF-7 ( 50 % ) , but not to MPMV ( 29 % ) .

Breast cancer patients showed significant responses to <protein>MuMTV</protein> ( 49 % and to MCF-7 ( 50 % ) , but not to <protein>MPMV</protein> ( 29 % ) .


In a paired-antigen study using MuMTV and MCF-7 , 75 % of the breast cancer patients responded , versus 18 % of the normal women ( P less than 0.0050 ) .

In a paired-antigen study using <protein>MuMTV</protein> and <protein>MCF-7</protein> , 75 % of the breast cancer patients responded , versus 18 % of the normal women ( P less than 0.0050 ) .


The potential for this assay to distinguish `` normal '' from `` breast cancer '' was analyzed using a migration index derived from discriminant analysis .


The ability of the assay to discriminate `` normal '' from `` cancer '' was significant ( P less than 0.001 ) and showed a sensitivity of detecting `` cancer '' of 75 % .


The overall responses to MuMTV and MCF-7 were analyzed with reference to certain prognostic factors , but showed no relation to age , menstrual status , estrogen receptor status , or stage of disease .

The overall responses to <protein>MuMTV</protein> and <protein>MCF-7</protein> were analyzed with reference to certain prognostic factors , but showed no relation to age , menstrual status , estrogen receptor status , or stage of disease .


The above reactions suggest that a large proportion of breast cancer patients exhibit presensitization to antigenfs found in MuMTV and MCF-7 , which may be cross-reactive with antigens in the primary cancer .

The above reactions suggest that a large proportion of breast cancer patients exhibit presensitization to antigenfs found in <protein>MuMTV</protein> and <protein>MCF-7</protein> , which may be cross-reactive with antigens in the primary cancer .


These responses appear to be independent of major prognostic variables .


Further refinement of this assay may yield one which is more highly discriminating for breast cancer .


Chronic lymphatic leukaemia : cellular effects of glucocorticoids in vitro .


Glucocorticoid receptor levels and steroid induced inhibition of nucleic acid precursors have been examined in lymphocytes from 27 patients at different stages of chronic lymphatic leukaemia .

<protein>Glucocorticoid receptor</protein> levels and steroid induced inhibition of nucleic acid precursors have been examined in <cell_type>lymphocytes</cell_type> from 27 patients at different stages of chronic lymphatic leukaemia .


No correlation can be found between the level of glucocorticoid receptors and the stage of the disease .

No correlation can be found between the level of <protein>glucocorticoid receptors</protein> and the stage of the disease .


On the other hand , a significant difference ( P less than 0.02 ) was found between stage O and stage III/IV patients , in terms of the in vitro effect of dexamethasone on [ 3H ] uridine incorporation .


Pyrrolidine dithiocarbamate inhibits NF-kappa B mobilization and TNF production in human monocytes .

Pyrrolidine dithiocarbamate inhibits <protein>NF-kappa B</protein> mobilization and TNF production in <cell_type>human monocytes</cell_type> .


The human TNF promoter contains four potential nuclear factor-kappa B ( NF-kappa B ) -binding sites , with the strongest binding seen for the -605 motif .

The <dna>human TNF promoter</dna> contains four potential <dna>nuclear factor-kappa B ( NF-kappa B ) -binding sites</dna> , with the strongest binding seen for the -605 motif .


Nuclear extracts from unstimulated cells of the human monocytic cell line , Mono Mac 6 , contain one specific binding protein ( complex II ) , consistent with a constitutive p50 homodimer .

Nuclear extracts from <cell_line>unstimulated cells</cell_line> of the <cell_line>human monocytic cell line</cell_line> , <cell_line>Mono Mac 6</cell_line> , contain one <protein>specific binding protein</protein> ( <protein>complex II</protein> ) , consistent with a <protein>constitutive p50 homodimer</protein> .


Stimulation of Mono Mac 6 cells with LPS will increase complex II and will strongly induce a second specific complex ( complex I ) , which represents the p50/65 heterodimer .

Stimulation of <cell_line>Mono Mac 6 cells</cell_line> with LPS will increase <protein>complex II</protein> and will strongly induce a second specific complex ( <protein>complex I</protein> ) , which represents the <protein>p50/65 heterodimer</protein> .


Treatment of Mono Mac 6 cells with pyrrolidine-dithiocarbamate ( PDTC ) at 300 microM will block the LPS-induced complex I almost completely and will reduce complex II to the constitutive level .

Treatment of <cell_type>Mono Mac 6 cells</cell_type> with pyrrolidine-dithiocarbamate ( PDTC ) at 300 microM will block the <protein>LPS-induced complex I</protein> almost completely and will reduce <protein>complex II</protein> to the constitutive level .


Binding activity of other nuclear factors that recognize the SP-1 and c/EBP motifs of the human TNF promoter is not affected by such treatment .

Binding activity of other <protein>nuclear factors</protein> that recognize the SP-1 and c/EBP motifs of the <dna>human TNF promoter</dna> is not affected by such treatment .


Northern blot analysis demonstrates that PDTC treatment will strongly reduce LPS-induced TNF transcripts .

Northern blot analysis demonstrates that PDTC treatment will strongly reduce <protein>LPS-induced TNF transcripts</protein> .


Secreted TNF protein as detected in the Wehi 164S/ActD bioassay and in a sandwich immunoassay was similarly reduced by PDTC .

Secreted <protein>TNF protein</protein> as detected in the Wehi 164S/ActD bioassay and in a sandwich immunoassay was similarly reduced by PDTC .


Kinetic analyses show that after LPS stimulation , NF-kappa B will peak at 1 h , TNF transcript prevalence at 2 h , and TNF protein at 4 h .

Kinetic analyses show that after LPS stimulation , <protein>NF-kappa B</protein> will peak at 1 h , TNF transcript prevalence at 2 h , and <protein>TNF protein</protein> at 4 h .


PDTC did not shift this response to LPS to a later time , but suppressed NF-kappa B mobilization , TNF transcripts , and TNF protein over the entire 8-h observation period .

PDTC did not shift this response to LPS to a later time , but suppressed NF-kappa B mobilization , <protein>TNF transcripts</protein> , and <protein>TNF protein</protein> over the entire 8-h observation period .


Analysis of freshly isolated , LPS-stimulated blood monocytes showed a similar blockade of NF-kappa B .

Analysis of freshly isolated , <cell_type>LPS-stimulated blood monocytes</cell_type> showed a similar blockade of <protein>NF-kappa B</protein> .


Furthermore , in these primary cells , induction of TNF transcripts , as determined by Northern blot analysis and by quantitative polymerase chain reaction , was prevented by PDTC as was TNF protein production .

Furthermore , in these primary cells , induction of <protein>TNF transcripts</protein> , as determined by Northern blot analysis and by quantitative polymerase chain reaction , was prevented by PDTC as was <protein>TNF protein</protein> production .


These data show that dithiocarbamates can profoundly affect cytokine expression and suggest that NF-kappa B is involved in LPS-induced TNF gene expression in human monocytes .

These data show that dithiocarbamates can profoundly affect cytokine expression and suggest that <protein>NF-kappa B</protein> is involved in LPS-induced TNF gene expression in <cell_type>human monocytes</cell_type> .


Comparing regions of the Epstein-Barr virus ZEBRA protein which function as transcriptional activating sequences in Saccharomyces cerevisiae and in B cells .

Comparing regions of the <protein>Epstein-Barr virus ZEBRA protein</protein> which function as <dna>transcriptional activating sequences</dna> in Saccharomyces cerevisiae and in <cell_type>B cells</cell_type> .


The ZEBRA protein activates expression of Epstein-Barr virus early-lytic-cycle genes in human B lymphocytes .

The <protein>ZEBRA protein</protein> activates expression of Epstein-Barr virus <dna>early-lytic-cycle genes</dna> in <cell_type>human B lymphocytes</cell_type> .


Here it is shown that ZEBRA also behaves as a sequence-specific transcriptional activator in Saccharomyces cerevisiae .

Here it is shown that <protein>ZEBRA</protein> also behaves as a <protein>sequence-specific transcriptional activator</protein> in Saccharomyces cerevisiae .


Deletional mutagenesis defined three regions of ZEBRA that participate in activation in S. cerevisiae .

Deletional mutagenesis defined three regions of <protein>ZEBRA</protein> that participate in activation in S. cerevisiae .


These regions are designated YI ( amino acids [ aa ] 1 to 25 ) , YII ( aa 51 to 102 ) , and YIII ( aa 228 to 245 ) .

These regions are designated <protein>YI</protein> ( <protein>amino acids [ aa ] 1 to 25</protein> ) , <protein>YII</protein> ( <protein>aa 51 to 102</protein> ) , and <protein>YIII</protein> ( <protein>aa 228 to 245</protein> ) .


Two of the three regions of the native ZEBRA protein act together to mediate activation when assayed on ZEBRA binding sites .

Two of the three regions of the native <protein>ZEBRA protein</protein> act together to mediate activation when assayed on <protein>ZEBRA binding sites</protein> .


However , when fused to the DNA binding domain of GAL 4 and assayed on GAL4 binding sites , regions YII and YIII were each sufficient to confer activation in S. cerevisiae .

However , when fused to the <protein>DNA binding domain</protein> of <protein>GAL</protein> 4 and assayed on <protein>GAL4 binding sites</protein> , <protein>regions YII and YIII</protein> were each sufficient to confer activation in S. cerevisiae .


Regions of ZEBRA which affected activation in S. cerevisiae were also required in human B lymphocytes .

Regions of <protein>ZEBRA</protein> which affected activation in S. cerevisiae were also required in <cell_type>human B lymphocytes</cell_type> .


The amino-terminal region of ZEBRA ( aa 1 to 98 ) was required for activation both in S. cerevisiae and in human B cells ; deletion of the carboxy-terminal 18 aa also significantly reduced activation in both cell types .

The <protein>amino-terminal region</protein> of <protein>ZEBRA</protein> ( <protein>aa 1 to 98</protein> ) was required for activation both in S. cerevisiae and in <cell_type>human B cells</cell_type> ; deletion of the <protein>carboxy-terminal 18 aa</protein> also significantly reduced activation in both cell types .


Thus , the behavior of ZEBRA in human B cells and S. cerevisiae suggests that the protein contains universal activation motifs that interact with conserved components of the transcription machinery .

Thus , the behavior of <protein>ZEBRA</protein> in <cell_type>human B cells</cell_type> and S. cerevisiae suggests that the protein contains <protein>universal activation motifs</protein> that interact with conserved components of the transcription machinery .


However , certain deletion mutants of ZEBRA containing mutations in the N-terminal region exhibited discordant behaviors in S. cerevisiae and in B cells .

However , certain <protein>deletion mutants</protein> of <protein>ZEBRA</protein> containing mutations in the <protein>N-terminal region</protein> exhibited discordant behaviors in S. cerevisiae and in B cells .


For example , deletion of ZEBRA aa 26 to 51 impaired activation to a great extent in B cells but had little or no effect in S. cerevisiae .

For example , deletion of <protein>ZEBRA aa 26 to 51</protein> impaired activation to a great extent in <cell_type>B cells</cell_type> but had little or no effect in S. cerevisiae .


The discordant mutants may reflect interactions with a variable domain of a conserved component or unique interactions with specialized components of the basal transcription apparatus in different cells .

The discordant mutants may reflect interactions with a <protein>variable domain</protein> of a conserved component or unique interactions with specialized components of the basal transcription apparatus in different cells .


Functional interaction of the v-Rel and c-Rel oncoproteins with the TATA-binding protein and association with transcription factor IIB .

Functional interaction of the <protein>v-Rel and c-Rel oncoproteins</protein> with the <protein>TATA-binding protein</protein> and association with <protein>transcription factor IIB</protein> .


Rel family proteins regulate the expression of genes linked to kappa B-binding motifs .

<protein>Rel family proteins</protein> regulate the expression of genes linked to kappa B-binding motifs .


Little is known , however , of the mechanism by which they enhance transcription .


We have investigated the ability of the v-Rel and c-Rel oncoproteins to interact with components of the basal transcription machinery .

We have investigated the ability of the <protein>v-Rel and c-Rel oncoproteins</protein> to interact with components of the basal transcription machinery .


Here we report that both the acidic transcription activation domain mapping to the unique C terminus of chicken c-Rel and the F9 cell-specific activation region common to both v-Rel and c-Rel interact with the TATA-binding protein ( TBP ) and transcription factor IIB ( TFIIB ) in vitro and in vivo .

Here we report that both the acidic transcription activation domain mapping to the unique <protein>C terminus</protein> of <protein>chicken c-Rel</protein> and the <protein>F9 cell-specific activation region</protein> common to both <protein>v-Rel</protein> and <protein>c-Rel</protein> interact with the <protein>TATA-binding protein</protein> ( <protein>TBP</protein> ) and <protein>transcription factor IIB</protein> ( <protein>TFIIB</protein> ) in vitro and in vivo .


We also demonstrate that TPB interaction with Rel activation regions leads to synergistic activation of transcription of a kappa B-linked reporter gene .

We also demonstrate that <protein>TPB</protein> interaction with <protein>Rel activation regions</protein> leads to synergistic activation of transcription of a <dna>kappa B-linked reporter gene</dna> .


Combined with the observation that the mouse c-Rel and human RelA proteins also interact with TBP and TFIIB in vitro , these results suggest that association with basal transcription factors is important for the transcriptional activities of Rel family proteins .

Combined with the observation that the <protein>mouse c-Rel and human RelA proteins</protein> also interact with <protein>TBP</protein> and <protein>TFIIB</protein> in vitro , these results suggest that association with <protein>basal transcription factors</protein> is important for the transcriptional activities of <protein>Rel family proteins</protein> .


p21ras and calcineurin synergize to regulate the nuclear factor of activated T cells .

<protein>p21ras</protein> and calcineurin synergize to regulate the <protein>nuclear factor</protein> of activated <cell_type>T cells</cell_type> .


In T lymphocytes , triggering of the T cell receptor ( TCR ) induces several signaling cascades which ultimately synergize to induce the activity of the nuclear factor of activated T cells ( NFAT ) , a DNA binding complex critical to the inducibility and T cell specificity of the T cell growth factor interleukin 2 .

In <cell_type>T lymphocytes</cell_type> , triggering of the <protein>T cell receptor</protein> ( <protein>TCR</protein> ) induces several signaling cascades which ultimately synergize to induce the activity of the <protein>nuclear factor</protein> of activated <cell_type>T cells</cell_type> ( <cell_line>NFAT</cell_line> ) , a <protein>DNA binding complex</protein> critical to the inducibility and T cell specificity of the <protein>T cell growth factor</protein> <protein>interleukin 2</protein> .


One immediate consequence of T cell activation via the TCR is an increase in cytosolic calcium .

One immediate consequence of T cell activation via the <protein>TCR</protein> is an increase in cytosolic calcium .


Calcium signals are important for NFAT induction , and recent studies have identified calcineurin , a calcium-calmodulin dependent serine-threonine phosphatase , as a prominent component of the calcium signaling pathway in T cells .

Calcium signals are important for <cell_line>NFAT</cell_line> induction , and recent studies have identified <protein>calcineurin</protein> , a <protein>calcium-calmodulin dependent serine-threonine phosphatase</protein> , as a prominent component of the calcium signaling pathway in <cell_type>T cells</cell_type> .


A second important molecule in TCR signal transduction is the guanine nucleotide binding protein , p21ras , which is coupled to the TCR by a protein tyrosine kinase dependent mechanism .

A second important molecule in TCR signal transduction is the <protein>guanine nucleotide binding protein</protein> , <protein>p21ras</protein> , which is coupled to the <protein>TCR</protein> by a <protein>protein tyrosine kinase</protein> dependent mechanism .


The experiments presented here show that expression by transfection of mutationally activated calcineurin or activated p21ras alone is insufficient for NFAT transactivation .

The experiments presented here show that expression by transfection of mutationally activated <protein>calcineurin</protein> or activated <protein>p21ras</protein> alone is insufficient for <cell_type>NFAT</cell_type> transactivation .


However , coexpression of the activated calcineurin with activated p21ras could mimic TCR signals in NFAT induction .

However , coexpression of the activated calcineurin with activated p21ras could mimic TCR signals in <cell_type>NFAT</cell_type> induction .


These data identify calcineurin and p21ras as cooperative partners in T cell activation .

These data identify <protein>calcineurin</protein> and <protein>p21ras</protein> as cooperative partners in <cell_type>T cell</cell_type> activation .


Expression of the chicken GATA factor family during early erythroid development and differentiation .

Expression of the <protein>chicken GATA factor family</protein> during early erythroid development and differentiation .


The DNA motif WGATAR has been identified within transcriptional regulatory domains of globin and other erythroid-specific genes and the activator proteins that bind to this regulatory element , the GATA factors , belong to a multi-gene family that is expressed in chicken erythroid cells .

The DNA motif WGATAR has been identified within <dna>transcriptional regulatory domains</dna> of <dna>globin</dna> and other <dna>erythroid-specific genes</dna> and the <protein>activator proteins</protein> that bind to this <dna>regulatory element</dna> , the <protein>GATA factors</protein> , belong to a <protein>multi-gene family</protein> that is expressed in <cell_type>chicken erythroid cells</cell_type> .


Here we show that , as in chickens , multiple members of the GATA factor family are expressed in human and murine erythroid cells .

Here we show that , as in chickens , multiple members of the <protein>GATA factor family</protein> are expressed in <cell_type>human and murine erythroid cells</cell_type> .


During the early stages of chicken embryogenesis ( well before blood island formation ) , each of the GATA family members is transcribed with a unique temporal and spatial pattern .

During the early stages of chicken embryogenesis ( well before blood island formation ) , each of the <protein>GATA family members</protein> is transcribed with a unique temporal and spatial pattern .


In the primitive erythroid lineage , transcription of the embryonic epsilon-globin gene parallels GATA-1 expression while the switch to beta-globin transcription in definitive erythroid cells is directly preceded by a pronounced increase in GATA-3 accumulation .

In the <cell_type>primitive erythroid lineage</cell_type> , transcription of the <dna>embryonic epsilon-globin gene</dna> parallels GATA-1 expression while the switch to beta-globin transcription in <cell_type>definitive erythroid cells</cell_type> is directly preceded by a pronounced increase in <protein>GATA-3</protein> accumulation .


The timing and pattern of expression of these different mRNAs during avian erythroid development and differentiation suggests that temporally regulated changes in GATA factor expression are required for vertebrate hematopoiesis .

The timing and pattern of expression of these different <rna>mRNAs</rna> during avian erythroid development and differentiation suggests that temporally regulated changes in <protein>GATA factor</protein> expression are required for vertebrate hematopoiesis .


Molecular regulation of human interleukin 2 and T-cell function by interleukin 4 .

Molecular regulation of <protein>human interleukin 2</protein> and <cell_type>T-cell</cell_type> function by <protein>interleukin 4</protein> .


Distinct functional T-cell subsets , differing in the patterns of lymphokines produced , regulate cell-mediated and humoral immune responses .

Distinct functional <cell_type>T-cell subsets</cell_type> , differing in the patterns of <protein>lymphokines</protein> produced , regulate cell-mediated and humoral immune responses .


The two major types and their principal products , interleukin 4 and interferon gamma ( IL-4 and IFN-gamma ) , are reciprocally negatively interactive .

The two major types and their principal products , <protein>interleukin 4</protein> and <protein>interferon gamma</protein> ( <protein>IL-4</protein> and <protein>IFN-gamma</protein> ) , are reciprocally negatively interactive .


To analyze the molecular mechanism of IL-4-mediated suppression of cell-mediated immunity we studied its effects on expression of interleukin 2 ( IL-2 ) and IFN-gamma .

To analyze the molecular mechanism of IL-4-mediated suppression of cell-mediated immunity we studied its effects on expression of <protein>interleukin 2</protein> ( <protein>IL-2</protein> ) and <protein>IFN-gamma</protein> .


IL-4 pretreatment of Jurkat cells prior to stimulation resulted in a decrease in transcription of the IL2 gene .

<protein>IL-4</protein> pretreatment of <cell_type>Jurkat cells</cell_type> prior to stimulation resulted in a decrease in transcription of the <dna>IL2 gene</dna> .


IL-4 suppressed IL-2 and IFN-gamma mRNA levels in primary human T cells , and addition of anti-CD28 antibodies relieved this suppression .

<protein>IL-4</protein> suppressed IL-2 and IFN-gamma mRNA levels in <cell_type>primary human T cells</cell_type> , and addition of <protein>anti-CD28 antibodies</protein> relieved this suppression .


Using enhancer-reporter constructs , IL-4 specifically down-regulated the NFIL-2B element .

Using <dna>enhancer-reporter constructs</dna> , <protein>IL-4</protein> specifically down-regulated the <dna>NFIL-2B element</dna> .


Electrophoretic mobility shift assays using a DNA oligomer containing the NFIL-2B binding site indicated that IL-4 inhibited the NFIL-2B complex and that the NFIL-2B DNA binding factor is distinct from AP- 1 .

Electrophoretic mobility shift assays using a DNA oligomer containing the <dna>NFIL-2B binding site</dna> indicated that <protein>IL-4</protein> inhibited the <protein>NFIL-2B complex</protein> and that the <protein>NFIL-2B DNA binding factor</protein> is distinct from <protein>AP-</protein> 1 .


These results suggest that IL-4 may regulate development and function of T-cell subsets involved in cell-mediated immunity in part by inhibiting factors required for transcription of the IL2 gene

These results suggest that <protein>IL-4</protein> may regulate development and function of <cell_type>T-cell subsets</cell_type> involved in cell-mediated immunity in part by <protein>inhibiting factors</protein> required for transcription of the <dna>IL2 gene</dna>


Immunochemical differences between glucocorticoid receptors from corticoid-sensitive and -resistant malignant lymphocytes .

Immunochemical differences between <protein>glucocorticoid receptors</protein> from <cell_type>corticoid-sensitive and -resistant malignant lymphocytes</cell_type> .


We have explored the possibility of using antibodies against purified rat liver glucocorticoid receptors to study the immunochemical properties of glucocorticoid receptors from murine and human malignant lymphocytes .

We have explored the possibility of using <protein>antibodies</protein> against purified rat liver <protein>glucocorticoid receptors</protein> to study the immunochemical properties of <protein>glucocorticoid receptors</protein> from <cell_type>murine and human malignant lymphocytes</cell_type> .


For this purpose , purified immune immunoglobulin G was covalently linked to Sepharose CL-4B .

For this purpose , <protein>purified immune immunoglobulin G</protein> was covalently linked to <protein>Sepharose CL-4B</protein> .


We then examined the ability of the affinity gel to recognize cytosolic [ 3H ] triamcinolone acetonide-receptor complexes from the corticoid-sensitive ( CS ) and -resistant strains of mouse lymphoma P1798 , from CS lymphocytes of patients with chronic lymphatic leukemia , and from a CS clone of human leukemic lymphoblasts in tissue culture ( CH6 ) .

We then examined the ability of the affinity gel to recognize <protein>cytosolic [ 3H ] triamcinolone acetonide-receptor complexes</protein> from the <cell_line>corticoid-sensitive ( CS ) and -resistant strains of mouse lymphoma P1798</cell_line> , from <cell_type>CS lymphocytes</cell_type> of patients with chronic lymphatic leukemia , and from a <cell_line>CS clone</cell_line> of <cell_type>human leukemic lymphoblasts</cell_type> in tissue culture ( <cell_line>CH6</cell_line> ) .


Mouse thymus was used as a source of glucocorticoid receptor from normal CS lymphocytes .

Mouse thymus was used as a source of <protein>glucocorticoid receptor</protein> from <cell_type>normal CS lymphocytes</cell_type> .


Whereas the immunoaffinity column retained 70 to 84 % of the 58- to 62-A ( Stokes radius ) [ 3H ] triamcinolone acetonide-receptor complexes characteristic of the CS mouse and human lymphocytes , it failed to recognize the 27- to 28-A ( Stokes radius ) glucocorticoid receptor present in corticoid-resistant mouse lymphoma P1798 cells .

Whereas the immunoaffinity column retained 70 to 84 % of the 58- to 62-A ( Stokes radius ) <protein>[ 3H ] triamcinolone acetonide-receptor complexes</protein> characteristic of the <cell_line>CS mouse and human lymphocytes</cell_line> , it failed to recognize the 27- to 28-A ( Stokes radius ) <protein>glucocorticoid receptor</protein> present in corticoid-resistant <cell_line>mouse lymphoma P1798 cells</cell_line> .


Therefore , under appropriate experimental conditions , it was possible to demonstrate cross-reactivity between the antiserum against rat liver glucocorticoid receptor and the 58- to 62-A ( Stokes radius ) glucocorticoid receptor from species as diverse as mouse and humans .

Therefore , under appropriate experimental conditions , it was possible to demonstrate cross-reactivity between the antiserum against <protein>rat liver glucocorticoid receptor</protein> and the 58- to 62-A ( Stokes radius ) <protein>glucocorticoid receptor</protein> from species as diverse as mouse and humans .


Heterogeneity of the in vitro responses to glucocorticoids in acute leukemia .


In leukocyte population freshly isolated from the blood of 26 patients with acute leukemia , we have measured several parameters including glucocorticoid receptors , nucleoside incorporation , percentage of cells in S phase , and steroid-induced cell lysis .

In <cell_type>leukocyte population</cell_type> freshly isolated from the blood of 26 patients with acute leukemia , we have measured several parameters including <protein>glucocorticoid receptors</protein> , nucleoside incorporation , percentage of cells in S phase , and steroid-induced cell lysis .


In addition , in some cases , the short-term response to steroid therapy was determined .


Although , in all the patients studied , leukocytes were found to contain glucocorticoid receptors , we failed to demonstrate any correlation between the level of binding sites and the in vitro or in vivo response to glucocorticoids .

Although , in all the patients studied , <cell_type>leukocytes</cell_type> were found to contain <protein>glucocorticoid receptors</protein> , we failed to demonstrate any correlation between the level of binding sites and the in vitro or in vivo response to glucocorticoids .


This absence of correlation could be in part explained by the marked heterogeneity of the steroid response demonstrated in leukocyte subpopulations .


It appears , however , that the degree of steroid action in vitro as well as the extent of spontaneous and dexamethasone-induced cell death may be related to the number of cells in the S phase of the cell cycle .


Glucocorticoid receptors in cytosol and nuclear extract of human leukocytes .

<protein>Glucocorticoid receptors</protein> in cytosol and nuclear extract of <cell_type>human leukocytes</cell_type> .


Cortisol binding by cytosol and 0.4 M KCl extract of the nuclear fraction of human leukocytes were studied by gel chromatography and ion exchange filtration on DEAE cellulose .

Cortisol binding by cytosol and 0.4 M KCl extract of the nuclear fraction of <cell_type>human leukocytes</cell_type> were studied by gel chromatography and ion exchange filtration on DEAE cellulose .


The cytoplasmic cortisol binding protein has a molecular weight 95 000 and the soluble nuclear binding protein 50 000 .

The <protein>cytoplasmic cortisol binding protein</protein> has a molecular weight 95 000 and the <protein>soluble nuclear binding protein</protein> 50 000 .


The absence of the uptake of radioactive cortisol by isolated nuclei and the apparent requirement of the cytosol for glucocorticoid specific binding in nuclear receptor sites was observed .


The association constant characterising the binding of cortisol to cytosol was KA = 3.5 . 10 ( 9 ) l/mol .


Corticosteroid-induced lymphopenia , immunosuppression , and body defense .


The apparent paradox of heightened adrenal corticosteroid levels associated with reduction in the competence of the body 's defensive apparatus to cope with exposure to new microbial antigens is considered .

The apparent paradox of heightened adrenal corticosteroid levels associated with reduction in the competence of the body 's defensive apparatus to cope with exposure to new <protein>microbial antigens</protein> is considered .


The question is asked how this lowered defensive capability , which occurs in the face of a threat to body integrity , is consistent with Cannon 's principals of the `` wisdom of the body. ''


The suggestion is offered that the immunologic response to self-antigens exposed by disease or trauma may be suppressed by corticosteroid to offset the likelihood of autoimmune attack .

The suggestion is offered that the immunologic response to <protein>self-antigens</protein> exposed by disease or trauma may be suppressed by corticosteroid to offset the likelihood of autoimmune attack .


Protein tyrosine kinase activation is required for lipopolysaccharide induction of cytokines in human blood monocytes .

<protein>Protein tyrosine kinase</protein> activation is required for lipopolysaccharide induction of <protein>cytokines</protein> in <cell_type>human blood monocytes</cell_type> .


Bacterial LPS induce production of cytokines such as IL-1 , IL-6 , and TNF in mononuclear phagocytes , and this represents a central component in the pathogenesis of septic shock syndrome .

Bacterial LPS induce production of <protein>cytokines</protein> such as <protein>IL-1</protein> , <protein>IL-6</protein> , and <protein>TNF</protein> in <cell_type>mononuclear phagocytes</cell_type> , and this represents a central component in the pathogenesis of septic shock syndrome .


However , the mechanisms by which LPS activates these cells to express cytokines are not completely characterized .

However , the mechanisms by which LPS activates these cells to express <protein>cytokines</protein> are not completely characterized .


The present study addressed the role of different protein kinases in the LPS induction of cytokines .

The present study addressed the role of different <protein>protein kinases</protein> in the LPS induction of <protein>cytokines</protein> .


It is shown that LPS induced a 12- to 16-fold increase in IL-1 beta , IL-6 , and TNF-alpha mRNA levels , and this was completely or more than 80 % blocked by the protein tyrosine kinase specific inhibitors herbimycin A and genistein at the concentrations of 1.7 and 37 microM , respectively .


Protein kinase C inhibition by staurosporine reduced LPS induction of TNF-alpha , whereas it had no effects on IL-6 and IL-1 beta .

<protein>Protein kinase C</protein> inhibition by staurosporine reduced LPS induction of <protein>TNF-alpha</protein> , whereas it had no effects on <protein>IL-6</protein> and IL-1 beta .


Inhibition of protein kinase A by H89 reduced IL-6 mRNA levels but did not detectably change IL-1 beta or TNF-alpha mRNA levels .

Inhibition of <protein>protein kinase A</protein> by H89 reduced <rna>IL-6 mRNA</rna> levels but did not detectably change IL-1 beta or TNF-alpha mRNA levels .


In contrast , LPS did not increase leukemia inhibitory factor mRNA , which was constitutively expressed and not significantly reduced by these inhibitors .

In contrast , LPS did not increase <rna>leukemia inhibitory factor mRNA</rna> , which was constitutively expressed and not significantly reduced by these inhibitors .


In addition to cytokine mRNA levels , LPS-induced IL-6 protein synthesis and IL-6 bioactivity were also reduced to baseline levels by the PTK inhibitors herbimycin A and genistein .

In addition to <rna>cytokine mRNA</rna> levels , LPS-induced IL-6 protein synthesis and IL-6 bioactivity were also reduced to baseline levels by the PTK inhibitors herbimycin A and genistein .


Both PTK inhibitors also reduced the LPS activation of nuclear factor-kappa B ( NF-kappa B ) , which is a transcription factor involved in the expression of cytokine genes such as IL-6 and TNF-alpha .

Both PTK inhibitors also reduced the LPS activation of <protein>nuclear factor-kappa B</protein> ( <protein>NF-kappa B</protein> ) , which is a <protein>transcription factor</protein> involved in the expression of <rna>cytokine genes</rna> such as <dna>IL-6</dna> and <dna>TNF-alpha</dna> .