IgE germline promoter reporter gene constructs carrying point mutations in the NF kappa B2 site were largely unresponsive to IL-4 stimulation in transient transfection experiments , while plasmids with similar mutations in the NF kappa B1 site responded to cytokine stimulation better than the wild-type promoter .

<dna>IgE germline promoter reporter gene constructs</dna> carrying point mutations in the <protein>NF kappa B2</protein> site were largely unresponsive to <protein>IL-4</protein> stimulation in transient transfection experiments , while plasmids with similar mutations in the <protein>NF kappa B1</protein> site responded to <protein>cytokine</protein> stimulation better than the <dna>wild-type promoter</dna> .


The NF kappa B2 effect was dependent on the presence of the STAT6 binding site , demonstrating that the NF kappa B2 motif is necessary but not sufficient for mediating cytokine up-regulation .

The <protein>NF kappa B2</protein> effect was dependent on the presence of the <dna>STAT6 binding site</dna> , demonstrating that the <protein>NF kappa B2 motif</protein> is necessary but not sufficient for mediating <protein>cytokine</protein> up-regulation .


In addition , the combination of a NF kappa B/rel binding site and the STAT6 response element conferred IL-4 inducibility to a heterologous minimal promoter , while the individual sites had no effect .

In addition , the combination of a <dna>NF kappa B/rel binding site</dna> and the <dna>STAT6 response element</dna> conferred <protein>IL-4</protein> inducibility to a <dna>heterologous minimal promoter</dna> , while the individual sites had no effect .


The available data suggest that the NF kappa B2 nucleoprotein complex may cooperate with DNA-bound STAT6 to achieve IL-4 -dependent activation of the human IgE germline gene .

The available data suggest that the <protein>NF kappa B2 nucleoprotein complex</protein> may cooperate with <protein>DNA-bound STAT6</protein> to achieve <protein>IL-4</protein> -dependent activation of the <dna>human IgE germline gene</dna> .


Alcohol-induced regulation of nuclear regulatory factor-kappa beta in human monocytes .

Alcohol-induced regulation of <protein>nuclear regulatory factor-kappa beta</protein> in <cell_type>human monocytes</cell_type> .


Acute ethanol exposure has the capacity to modulate immune functions , particularly , to down regulate monocyte production of inflammatory cytokines .

Acute ethanol exposure has the capacity to modulate immune functions , particularly , to down regulate monocyte production of <protein>inflammatory cytokines</protein> .


However , the intracellular mechanisms for these effects of ethanol are yet to be understood .


Considering that nuclear regulatory factor-kappa beta ( NF-kappa B ) /Rel is a common regulatory element of the promoter region of the inflammatory cytokine genes , herein , we tested the hypothesis that acute ethanol affects NF-kappa B activation in human monocytes .

Considering that <protein>nuclear regulatory factor-kappa beta</protein> <protein>( NF-kappa B ) /Rel</protein> is a common <protein>regulatory element</protein> of the <dna>promoter region</dna> of the <dna>inflammatory cytokine genes</dna> , herein , we tested the hypothesis that acute ethanol affects <protein>NF-kappa B</protein> activation in <cell_type>human monocytes</cell_type> .


Adherence-isolated monocytes showed constitutive DNA binding activity of NF-kappa B .

<cell_line>Adherence-isolated monocytes</cell_line> showed constitutive DNA binding activity of <protein>NF-kappa B</protein> .


A clinically relevant dose ( 25 mM ) of acute ethanol treatment in vitro increased NF-kappa B binding activity in monocytes with a preferential induction of the inhibitory , p50/p50 , NF-kappa B/Rel homodimer , and resulted in no induction of the p65/p50 heterodimer .

A clinically relevant dose ( 25 mM ) of acute ethanol treatment in vitro increased <protein>NF-kappa B</protein> binding activity in <cell_type>monocytes</cell_type> with a preferential induction of the inhibitory , <protein>p50/p50</protein> , <protein>NF-kappa B/Rel homodimer</protein> , and resulted in no induction of the <protein>p65/p50 heterodimer</protein> .


In contrast , lipopolysaccharide stimulation primarily induced the p65/p50 heterodimer that has been shown to result in gene activation .

In contrast , lipopolysaccharide stimulation primarily induced the <protein>p65/p50 heterodimer</protein> that has been shown to result in gene activation .


Thus , such unique activation of the inhibitory p50/p50 homodimer by acute ethanol treatment may result in inhibition rather than activation of NF-kappa B-regulated inflammatory cytokine genes .

Thus , such unique activation of the <protein>inhibitory p50/p50 homodimer</protein> by acute ethanol treatment may result in inhibition rather than activation of <dna>NF-kappa B-regulated inflammatory cytokine genes</dna> .


Consequently , these results suggest that physiologically relevant concentrations of ethanol may affect production of inflammatory cytokines , such as tumor necrosis factor-alpha , interleukin-1 beta , and interleukin-6 by disrupting NF-kappa B signaling in monocytes .

Consequently , these results suggest that physiologically relevant concentrations of ethanol may affect production of <protein>inflammatory cytokines</protein> , such as <protein>tumor necrosis factor-alpha</protein> , <protein>interleukin-1 beta</protein> , and <protein>interleukin-6</protein> by disrupting <protein>NF-kappa B</protein> signaling in <cell_type>monocytes</cell_type> .


LPS tolerance in monocytes/macrophages : three 3 ' cytosins are required in the DNA binding motif for detection of upregulated NF-kappa B p50 homodimers .

LPS tolerance in <cell_type>monocytes/macrophages</cell_type> : three 3 ' cytosins are required in the <dna>DNA binding motif</dna> for detection of upregulated <protein>NF-kappa B p50 homodimers</protein> .


When monocytes are stimulated with LPS ( lipopolysaccharide ) repeatedly then the initially high expression of the TNF ( tumor necrosis factor ) gene is only very low , i.e. the cells are tolerant to LPS .

When <cell_type>monocytes</cell_type> are stimulated with LPS ( lipopolysaccharide ) repeatedly then the initially high expression of the <dna>TNF ( tumor necrosis factor ) gene</dna> is only very low , i.e. the cells are tolerant to LPS .


Tolerant cells still express the CD14 receptor and they can still be activated to mobilize NF-kappa B into nucleus .

Tolerant cells still express the <protein>CD14 receptor</protein> and they can still be activated to mobilize <protein>NF-kappa B</protein> into nucleus .


Analysis of the binding proteins employing the -605 motif of the human TNF promoter ( GGGGCTGTCCC ) revealed that in tolerant cells of the human monocytic cell line Mono Mac 6 there is a predominance of p50p50 of NF-kappa B .

Analysis of the binding proteins employing the -605 motif of the <dna>human TNF promoter</dna> ( GGGGCTGTCCC ) revealed that in tolerant cells of the <cell_line>human monocytic cell line</cell_line> <cell_line>Mono Mac 6</cell_line> there is a predominance of <protein>p50p50</protein> of <protein>NF-kappa B</protein> .


We now show that a mutant motif that exchanges the terminal 3 ' C for a G fails to bind the p50 homodimer that is upregulated in LPS toler ant human Mono Mac 6 cells .

We now show that a <dna>mutant motif</dna> that exchanges the terminal 3 ' C for a G fails to bind the <protein>p50 homodimer</protein> that is upregulated in LPS toler ant <cell_line>human Mono Mac 6 cells</cell_line> .


The same is true for nuclear extracts taken from the murine P388D1 macrophage cell line when tested with the -516 motif of the murine TNF promoter ( GGGGGCTTTCCC ) .

The same is true for nuclear extracts taken from the <cell_line>murine P388D1 macrophage cell line</cell_line> when tested with the <dna>-516 motif</dna> of the <dna>murine TNF promoter</dna> ( GGGGGCTTTCCC ) .


Here the wild type motif gives efficient binding of p50p50 that again is upregulated in tolerant cells whereas a mutant with a 3 ' G shows hardly any binding of p50p50 .

Here the <dna>wild type motif</dna> gives efficient binding of <protein>p50p50</protein> that again is upregulated in <cell_type>tolerant cells</cell_type> whereas a mutant with a 3 ' G shows hardly any binding of <protein>p50p50</protein> .


Conversely , the murine kappa light chain enhancer motif ( GGGGACTTTCCG ) does not efficiently bind the nuclear p50p50 from tolerant murine P388 macrophages .

Conversely , the <dna>murine kappa light chain enhancer motif</dna> ( GGGGACTTTCCG ) does not efficiently bind the nuclear <protein>p50p50</protein> from <cell_line>tolerant murine P388 macrophages</cell_line> .


Binding is , however , readily detected when the 3 ' G is replaced by a C .


These data show that the detection of upregulated p50 homodimers in LPS tolerant cells is dependent on subtle differences in the sequence of the DNA binding motif .

These data show that the detection of upregulated <protein>p50 homodimers</protein> in LPS <cell_type>tolerant cells</cell_type> is dependent on subtle differences in the sequence of the <dna>DNA binding motif</dna> .


Cyclosporin A inhibits early mRNA expression of G0/G1 switch gene 2 ( G0S2 ) in cultured human blood mononuclear cells .

Cyclosporin A inhibits early mRNA expression of <dna>G0/G1 switch gene 2</dna> ( <dna>G0S2</dna> ) in <cell_line>cultured human blood mononuclear cells</cell_line> .


Cyclosporin A ( CsA ) may achieve its immunosuppressive effects by inhibiting the calcium- and calmodulin-dependent phosphatase calcineurin which is required for activation of target genes by members of the NFAT ( nuclear factor of activated T cells ) transcription factor family .

Cyclosporin A ( CsA ) may achieve its immunosuppressive effects by inhibiting the <protein>calcium- and calmodulin-dependent phosphatase</protein> <protein>calcineurin</protein> which is required for activation of <dna>target genes</dna> by members of the <protein>NFAT ( nuclear factor of activated T cells ) transcription factor family</protein> .


Among these target genes is the gene encoding interleukin-2 ( IL2 ) , a cytokine facilitating progression through the G1 phase of the cell cycle .

Among these <dna>target genes</dna> is the gene encoding <protein>interleukin-2</protein> ( <protein>IL2</protein> ) , a <protein>cytokine</protein> facilitating progression through the G1 phase of the cell cycle .


However , IL2 does not reverse CsA inhibition , suggesting that at least one other NFAT-sensitive gene may be involved .

However , <protein>IL2</protein> does not reverse CsA inhibition , suggesting that at least one other <dna>NFAT-sensitive gene</dna> may be involved .


The human G0/G1 switch gene , G0S2 , has potential NFAT-binding sites in the 5 ' flank and encodes a small basic potential phosphoprotein of unknown function .

The <dna>human G0/G1 switch gene</dna> , <dna>G0S2</dna> , has potential <dna>NFAT-binding sites</dna> in the <dna>5 ' flank</dna> and encodes a <protein>small basic potential phosphoprotein</protein> of unknown function .


Using a sensitive , reverse transcription-polymerase chain reaction ( RT-PCR ) assay , G0S2 mRNA levels were assayed in cultured blood mononuclear cells .

Using a sensitive , reverse transcription-polymerase chain reaction ( RT-PCR ) assay , <dna>G0S2</dna> mRNA levels were assayed in <cell_line>cultured blood mononuclear cells</cell_line> .


Freshly isolated cells contain high levels of G0S2 mRNA which rapidly decline .

Freshly isolated cells contain high levels of <rna>G0S2 mRNA</rna> which rapidly decline .


This `` spontaneous stimulation '' is also noted with some other G0S genes and has been attributed to some aspect of the isolation procedure .

This `` spontaneous stimulation '' is also noted with some other <dna>G0S genes</dna> and has been attributed to some aspect of the isolation procedure .


In cells that have been preincubated to lower mRNA levels , there is a transient increase in G0S2 mRNA , peaking between 1-2 h , in response to Concanavalin-A ( ConA ) , or to the combination of phorbol ester ( TPA ) , and the calcium ionophore , ionomycin .

In cells that have been preincubated to lower mRNA levels , there is a transient increase in <rna>G0S2 mRNA</rna> , peaking between 1-2 h , in response to Concanavalin-A ( ConA ) , or to the combination of phorbol ester ( TPA ) , and the calcium ionophore , ionomycin .


Both these responses are inhibited by CsA .


Our results suggest that G0S2 expression is required to commit cells to enter the G1 phase of the cell cycle , and that , while not excluding other possible targets , early inhibition of G0S2 expression by CsA may be important in achieving immunosuppression .

Our results suggest that <dna>G0S2</dna> expression is required to commit cells to enter the G1 phase of the cell cycle , and that , while not excluding other possible targets , early inhibition of <dna>G0S2</dna> expression by CsA may be important in achieving immunosuppression .


G0S2 may be of value as a reporter gene for analyzing the mechanism of action of CsA and its influence on the positive and negative selection of lymphocytes in response to self and not-self antigens .

<dna>G0S2</dna> may be of value as a reporter gene for analyzing the mechanism of action of CsA and its influence on the positive and negative selection of <cell_type>lymphocytes</cell_type> in response to self and not-self antigens .


c-Rel and p65 subunits bind to an upstream NF-kappaB site in human granulocyte macrophage-colony stimulating factor promoter involved in phorbol ester response in 5637 cells .

<protein>c-Rel and p65 subunits</protein> bind to an <dna>upstream NF-kappaB site</dna> in <dna>human granulocyte macrophage-colony stimulating factor promoter</dna> involved in phorbol ester response in <cell_line>5637 cells</cell_line> .


To further clarify the complex transcriptional regulation of the human GM-CSF gene , which was extensively investigated in activated T cells , we have studied the role of an upstream NF-kappaB like site in the 5637 non-lymphoid cell line , which derives from a bladder carcinoma and constitutively produces GM-CSF .

To further clarify the complex transcriptional regulation of the <dna>human GM-CSF gene</dna> , which was extensively investigated in <cell_type>activated T cells</cell_type> , we have studied the role of an upstream <dna>NF-kappaB like site</dna> in the <cell_line>5637 non-lymphoid cell line</cell_line> , which derives from a bladder carcinoma and constitutively produces <protein>GM-CSF</protein> .


This sequence , named the A element , has an active role on GM-CSF transcription and is responsive to the tumor promoter PMA in transient transfection experiments .

This sequence , named the A element , has an active role on <protein>GM-CSF</protein> transcription and is responsive to the <dna>tumor promoter PMA</dna> in transient transfection experiments .


We describe here a heterodimeric binding complex of NF-kappaB subunits ( c-Rel and p65 ) which is identical to the one obtained using the HIV-LTR-kappaB site as recognition sequence and different from the one ( c-Rel and p50 ) observed with nuclear extracts from Mo T-lymphoid HTLV-II infected cells .

We describe here a <protein>heterodimeric binding complex</protein> of <protein>NF-kappaB subunits</protein> ( <protein>c-Rel</protein> and <protein>p65</protein> ) which is identical to the one obtained using the <dna>HIV-LTR-kappaB site</dna> as recognition sequence and different from the one ( <protein>c-Rel</protein> and <protein>p50</protein> ) observed with nuclear extracts from <cell_line>Mo T-lymphoid HTLV-II infected cells</cell_line> .


Repression of human immunodeficiency virus type 1 through the novel cooperation of human factors YY1 and LSF [ published erratum appears in J Virol 1998 Feb ; 72 ( 2 ) : 1709 ]

Repression of human immunodeficiency virus type 1 through the novel cooperation of <protein>human factors</protein> <protein>YY1</protein> and <protein>LSF</protein> [ published erratum appears in J Virol 1998 Feb ; 72 ( 2 ) : 1709 ]


A subpopulation of stably infected CD4+ cells capable of producing virus upon stimulation has been identified in human immunodeficiency virus ( HIV ) -positive individuals ( T.-W.Chun , D.Finzi , J.Margolick , K.Chadwick , D.Schwartz , and R.F.Siliciano , Nat.Med.1 : 1284-1290 , 1995 ) .

A subpopulation of <cell_type>stably infected CD4+ cells</cell_type> capable of producing virus upon stimulation has been identified in human immunodeficiency virus ( HIV ) -positive individuals ( T.-W.Chun , D.Finzi , J.Margolick , K.Chadwick , D.Schwartz , and R.F.Siliciano , Nat.Med.1 : 1284-1290 , 1995 ) .


Few host factors that directly limit HIV-1 transcription and could support this state of nonproductive HIV-1 infection have been described .


YY1 , a widely distributed human transcription factor , is known to inhibit HIV-1 long terminal repeat ( LTR ) transcription and virus production .

<protein>YY1</protein> , a widely distributed <protein>human transcription factor</protein> , is known to inhibit <dna>HIV-1 long terminal repeat</dna> ( <dna>LTR</dna> ) transcription and virus production .


LSF ( also known as LBP-1 , UBP , and CP-2 ) has been shown to repress LTR transcription in vitro , but transient expression of LSF has no effect on LTR activity in vivo .

<protein>LSF</protein> ( also known as <protein>LBP-1</protein> , <protein>UBP</protein> , and <protein>CP-2</protein> ) has been shown to repress LTR transcription in vitro , but transient expression of <protein>LSF</protein> has no effect on <dna>LTR</dna> activity in vivo .


We report that both YY1 and LSF participate in the formation of a complex that recognizes the initiation region of the HIV-1 LTR .

We report that both <protein>YY1</protein> and <protein>LSF</protein> participate in the formation of a complex that recognizes the initiation region of the <dna>HIV-1 LTR</dna> .


Further , we have found that these factors cooperate in the repression of LTR expression and viral replication .

Further , we have found that these factors cooperate in the repression of <dna>LTR</dna> expression and viral replication .


This cooperative function may account for the divergent effects of LSF previously observed in vitro and in vivo .

This cooperative function may account for the divergent effects of <protein>LSF</protein> previously observed in vitro and in vivo .


Thus , the cooperation of two general cellular transcription factors may allow for the selective downregulation of HIV transcription .


Through this mechanism of gene regulation , YY1 and LSF could contribute to the establishment and maintenance of a population of cells stably but nonproductively infected with HIV-1 .

Through this mechanism of gene regulation , <protein>YY1</protein> and <protein>LSF</protein> could contribute to the establishment and maintenance of a population of cells stably but nonproductively infected with HIV-1 .


Analysis of interactions between huGATA-3 transcription factor and three GATA regulatory elements of HIV-1 long terminal repeat , by surface plasmon resonance .

Analysis of interactions between <protein>huGATA-3</protein> transcription factor and three <dna>GATA regulatory elements</dna> of <dna>HIV-1 long terminal repeat</dna> , by surface plasmon resonance .


Relative affinities of transcriptional regulatory elements for their respective factor have been essentially studied by bandshift analysis .

Relative affinities of <dna>transcriptional regulatory elements</dna> for their respective factor have been essentially studied by bandshift analysis .


Here we report a real-time study of factor/DNA interactions using a surface plasmon resonance approach and further characterization of recovered proteins involved in this interaction .

Here we report a real-time study of factor/DNA interactions using a surface plasmon resonance approach and further characterization of <protein>recovered proteins</protein> involved in this interaction .


For this purpose , human GATA-3 , either recombinant or in nuclear extracts , and three natural GATA elements of the HIV-1 long terminal repeat ( sites 1 , 2 , and 3 ) were chosen , in which only site 2 is a noncanonical GATA site .

For this purpose , <protein>human GATA-3</protein> , either recombinant or in nuclear extracts , and three <dna>natural GATA elements</dna> of the <dna>HIV-1 long terminal repeat</dna> ( <dna>sites 1 , 2 , and 3</dna> ) were chosen , in which only <dna>site 2</dna> is a noncanonical <dna>GATA site</dna> .


Direct analysis of sensorgrams , with recombinant huGATA-3 , allowed the comparison of association and dissociation profiles of the three DNA regions and their ranking according to their relative affinities .

Direct analysis of sensorgrams , with <protein>recombinant huGATA-3</protein> , allowed the comparison of association and dissociation profiles of the three <dna>DNA regions</dna> and their ranking according to their relative affinities .


This result , confirmed by competitions with each GATA site , demonstrated the higher relative affinity ( at least sevenfold ) of site 3 .

This result , confirmed by competitions with each <dna>GATA site</dna> , demonstrated the higher relative affinity ( at least sevenfold ) of site 3 .


Interactions between the canonical and unique GATA site 3 and nuclear extracts were also studied in real time and provided information on its association and dissociation rates for native huGATA-3 .

Interactions between the canonical and unique <dna>GATA site 3</dna> and nuclear extracts were also studied in real time and provided information on its association and dissociation rates for native <protein>huGATA-3</protein> .


Finally , recovered protein was identified as genuine huGATA-3 by SDS-PAGE , Western blotting , and bandshift assays .

Finally , recovered protein was identified as genuine <protein>huGATA-3</protein> by SDS-PAGE , Western blotting , and bandshift assays .


Copyright 1997 Academic Press .


Stable transfection of U937 cells with sense or antisense RXR-alpha cDNA suggests a role for RXR-alpha in the control of monoblastic differentiation induced by retinoic acid and vitamin D .

Stable transfection of <cell_line>U937 cells</cell_line> with sense or <dna>antisense RXR-alpha cDNA</dna> suggests a role for <protein>RXR-alpha</protein> in the control of monoblastic differentiation induced by retinoic acid and vitamin D .


Although retinoic acid ( RA ) has been known for many years to be a modulating agent that plays a role in generating both granulocytes and monocytes , the molecular mechanism underlying this role has not been defined in the monoblast lineage .

Although retinoic acid ( RA ) has been known for many years to be a modulating agent that plays a role in generating both <cell_type>granulocytes</cell_type> and <cell_type>monocytes</cell_type> , the molecular mechanism underlying this role has not been defined in the <cell_type>monoblast lineage</cell_type> .


In particular , the part played by the retinoid X receptors ( RXRs ) , which are members of the steroid/thyroid hormone nuclear receptor family , has not been explored .

In particular , the part played by the <protein>retinoid X receptors</protein> ( <protein>RXRs</protein> ) , which are members of the <protein>steroid/thyroid hormone nuclear receptor family</protein> , has not been explored .


In this study , therefore , the human monoblastic leukemia cell line U937 has been used as a model system to investigate the role of one of the RXRs , RXR-alpha , in monoblast differentiation .

In this study , therefore , the <cell_line>human monoblastic leukemia cell line U937</cell_line> has been used as a model system to investigate the role of one of the <protein>RXRs</protein> , <protein>RXR-alpha</protein> , in monoblast differentiation .


RXR-alpha mRNA was present in untreated U937 cells , and levels increased after induction of differentiation with phorbol ester .

<rna>RXR-alpha mRNA</rna> was present in untreated <cell_line>U937 cells</cell_line> , and levels increased after induction of differentiation with phorbol ester .


The same was found for RXR-beta mRNA .

The same was found for <rna>RXR-beta mRNA</rna> .


Using plasmids containing sense or antisense RXR-alpha sequences under the control of an inducible promoter , we generated stably transfected cell lines which expressed either increased or decreased levels of RXR-alpha , respectively .

Using plasmids containing sense or <dna>antisense RXR-alpha sequences</dna> under the control of an inducible promoter , we generated stably transfected cell lines which expressed either increased or decreased levels of <protein>RXR-alpha</protein> , respectively .


The sense cell lines ( U alpha S and its clonal derivative alpha G2S ) showed increased sensitivity to RA , while the antisense cell lines ( U alpha A and its clonal derivative alpha B5A ) showed decreased sensitivity to RA , as demonstrated by growth inhibition and by regulation of an RA-responsive reporter gene .

The <cell_line>sense cell lines</cell_line> ( <cell_line>U alpha S</cell_line> and its clonal derivative alpha <cell_line>G2S</cell_line> ) showed increased sensitivity to RA , while the <cell_line>antisense cell lines</cell_line> ( <cell_line>U alpha A</cell_line> and its clonal derivative alpha <cell_line>B5A</cell_line> ) showed decreased sensitivity to RA , as demonstrated by growth inhibition and by regulation of an <dna>RA-responsive reporter gene</dna> .


Both U alpha A and alpha B5A also failed to respond to another modulating agent , 1 alpha , 25-dihydroxycholecalciferol ( DHCC ) , but only U alpha S and not alpha G2S showed an enhanced response to DHCC .

Both <cell_line>U alpha A</cell_line> and alpha <cell_line>B5A</cell_line> also failed to respond to another modulating agent , 1 alpha , 25-dihydroxycholecalciferol ( DHCC ) , but only <cell_line>U alpha S</cell_line> and not <protein>alpha G2S</protein> showed an enhanced response to DHCC .


The combination of RA and DHCC together inhibited growth of both sense and antisense cell lines .

The combination of RA and DHCC together inhibited growth of both sense and <cell_line>antisense cell lines</cell_line> .


In addition , alpha G2S exhibited increased expression of CD11b and CD54 , while alpha B5A cells showed increased expression of CD102 , suggesting that RXR-alpha has a role in regulating expression of cell adhesion molecules in U937 cells .

In addition , <protein>alpha G2S</protein> exhibited increased expression of <protein>CD11b</protein> and <protein>CD54</protein> , while <cell_line>alpha B5A cells</cell_line> showed increased expression of <protein>CD102</protein> , suggesting that <protein>RXR-alpha</protein> has a role in regulating expression of <protein>cell adhesion molecules</protein> in <cell_line>U937 cells</cell_line> .


These results demonstrate that RXR-alpha has a role in mediating growth inhibition and cell adhesion during myelomonocytic differentiation , and suggest that different species of heterodimers involving RXR-alpha may control the acquisition of different features of mature monocyte/macrophage function .

These results demonstrate that <protein>RXR-alpha</protein> has a role in mediating growth inhibition and cell adhesion during myelomonocytic differentiation , and suggest that different species of <protein>heterodimers</protein> involving <protein>RXR-alpha</protein> may control the acquisition of different features of mature monocyte/macrophage function .


Transcriptional activation of the vascular cell adhesion molecule-1 gene in T lymphocytes expressing human T-cell leukemia virus type 1 Tax protein .

Transcriptional activation of the <dna>vascular cell adhesion molecule-1 gene</dna> in <cell_type>T lymphocytes</cell_type> expressing <protein>human T-cell leukemia virus type 1 Tax protein</protein> .


Recruitment and extravasation of T cells through the blood-brain barrier are favored by adhesion molecule -mediated interactions of circulating T cells with endothelial cells .

Recruitment and extravasation of <cell_type>T cells</cell_type> through the blood-brain barrier are favored by <protein>adhesion molecule</protein> -mediated interactions of <cell_type>circulating T cells</cell_type> with <cell_type>endothelial cells</cell_type> .


Since a common pathological finding in human T-cell leukemia virus type 1 ( HTLV-1 ) -associated diseases is the infiltration of HTLV-1-infected T lymphocytes into various organs , we have looked for the profile of adhesion molecules expressed by HTLV-1-transformed T cells .

Since a common pathological finding in human T-cell leukemia virus type 1 ( HTLV-1 ) -associated diseases is the infiltration of <cell_type>HTLV-1-infected T lymphocytes</cell_type> into various organs , we have looked for the profile of <protein>adhesion molecules</protein> expressed by <cell_type>HTLV-1-transformed T cells</cell_type> .


Flow cytometry analysis indicated that these cells were expressing high levels of vascular cell adhesion molecule 1 ( VCAM-1 [ CD106 ] ) , a 110-kDa member of the immunoglobulin gene superfamily , first identified on endothelial cells stimulated with inflammatory cytokines .

Flow cytometry analysis indicated that these cells were expressing high levels of <protein>vascular cell adhesion molecule 1</protein> ( <protein>VCAM-1</protein> [ <protein>CD106</protein> ] ) , a 110-kDa member of the <protein>immunoglobulin gene superfamily</protein> , first identified on <cell_type>endothelial cells</cell_type> stimulated with <protein>inflammatory cytokines</protein> .


This adhesion molecule was also expressed by T cells obtained from one patient with HTLV-1-associated myelopathy/tropical spastic paraparesis but not by activated T cells isolated from one normal blood donor .

This adhesion molecule was also expressed by <cell_type>T cells</cell_type> obtained from one patient with HTLV-1-associated myelopathy/tropical spastic paraparesis but not by <cell_type>activated T cells</cell_type> isolated from one normal blood donor .


The role of the viral trans-activator Tax protein in the induction of VCAM-1 was first indicated by the detection of this adhesion molecule on Jurkat T-cell clones stably expressing the tax gene .

The role of the <protein>viral trans-activator Tax protein</protein> in the induction of <protein>VCAM-1</protein> was first indicated by the detection of this <protein>adhesion molecule</protein> on <cell_line>Jurkat T-cell clones</cell_line> stably expressing the tax gene .


The effect of Tax on VCAM-1 gene transcription was next confirmed in JPX-9 cells , a subclone of Jurkat cells , carrying the tax sequences under the control of an inducible promoter .

The effect of <protein>Tax</protein> on <protein>VCAM-1</protein> gene transcription was next confirmed in <cell_line>JPX-9 cells</cell_line> , a subclone of <cell_line>Jurkat cells</cell_line> , carrying the <dna>tax sequences</dna> under the control of an <dna>inducible promoter</dna> .


Furthermore , deletion and mutation analyses of the VCAM-1 promoter performed with chloramphenicol acetyltransferase constructs revealed that Tax was trans activating the VCAM-1 promoter via two NF-kappaB sites present at bp -72 and -57 in the VCAM-1 gene promoter , with both of them being required for the Tax -induced expression of this adhesion molecule .

Furthermore , deletion and mutation analyses of the <dna>VCAM-1 promoter</dna> performed with <dna>chloramphenicol acetyltransferase constructs</dna> revealed that <protein>Tax</protein> was trans activating the <dna>VCAM-1 promoter</dna> via two <dna>NF-kappaB sites</dna> present at bp -72 and -57 in the <dna>VCAM-1 gene promoter</dna> , with both of them being required for the <protein>Tax</protein> -induced expression of this <protein>adhesion molecule</protein> .


Finally , gel mobility shift assays demonstrated the nuclear translocation of proteins specifically bound to these two NF-kappaB motifs , confirming that VCAM-1 was induced on Tax-expressing cells in a kappaB -dependent manner .

Finally , gel mobility shift assays demonstrated the nuclear translocation of proteins specifically bound to these two <dna>NF-kappaB motifs</dna> , confirming that <protein>VCAM-1</protein> was induced on <cell_line>Tax-expressing cells</cell_line> in a <protein>kappaB</protein> -dependent manner .


Collectively , these results therefore suggest that the exclusive Tax -induced expression of VCAM-1 on T cells may represent a pivotal event in the progression of HTLV-1-associated diseases .

Collectively , these results therefore suggest that the exclusive <protein>Tax</protein> -induced expression of <protein>VCAM-1</protein> on <cell_type>T cells</cell_type> may represent a pivotal event in the progression of HTLV-1-associated diseases .


HIV-1 Vpr suppresses immune activation and apoptosis through regulation of nuclear factor kappa B [ see comments ]

<protein>HIV-1 Vpr</protein> suppresses immune activation and apoptosis through regulation of <protein>nuclear factor kappa B</protein> [ see comments ]


The HIV-1 accessory gene product Vpr can influence viral pathogenesis by affecting viral replication as well as host cell transcription and proliferation .

The <protein>HIV-1 accessory gene product</protein> <protein>Vpr</protein> can influence viral pathogenesis by affecting viral replication as well as host cell transcription and proliferation .


We have investigated the effects of Vpr on host cell activation and confirm that it influences cellular proliferation .

We have investigated the effects of <protein>Vpr</protein> on host cell activation and confirm that it influences cellular proliferation .


However , we have also found that Vpr modulates T-cell receptor ( TCR ) -triggered apoptosis in a manner similar to that of glucocorticoids .

However , we have also found that <protein>Vpr</protein> modulates T-cell receptor ( <protein>TCR</protein> ) -triggered apoptosis in a manner similar to that of glucocorticoids .


In the absence of TCR -mediated activation , Vpr induces apoptosis whereas in its presence , Vpr interrupts the expected induction of apoptosis .

In the absence of <protein>TCR</protein> -mediated activation , <protein>Vpr</protein> induces apoptosis whereas in its presence , <protein>Vpr</protein> interrupts the expected induction of apoptosis .


This regulation of apoptosis is linked to Vpr suppression of NF-kappa B activity via the induction of I kappa B , an inhibitor of NF-kappa B .

This regulation of apoptosis is linked to <protein>Vpr</protein> suppression of <protein>NF-kappa B</protein> activity via the induction of <protein>I kappa B</protein> , an inhibitor of <protein>NF-kappa B</protein> .


Further , Vpr suppresses expression of IL-2 , IL-10 , IL-12 , TNF alpha and IL-4 , all of which are NF-kappa B -dependent .

Further , <protein>Vpr</protein> suppresses expression of <protein>IL-2</protein> , <protein>IL-10</protein> , <protein>IL-12</protein> , <protein>TNF alpha</protein> and <protein>IL-4</protein> , all of which are <protein>NF-kappa B</protein> -dependent .


The effects of Vpr could be reversed by RU486 .

The effects of <protein>Vpr</protein> could be reversed by RU486 .


Our finding that Vpr can regulate NF-kappa B supports the hypothesis that some aspects of viral pathogenesis are the consequence of cell dysregulation by Vpr .

Our finding that <protein>Vpr</protein> can regulate <protein>NF-kappa B</protein> supports the hypothesis that some aspects of viral pathogenesis are the consequence of cell dysregulation by <protein>Vpr</protein> .


Tissue transglutaminase-dependent posttranslational modification of the retinoblastoma gene product in promonocytic cells undergoing apoptosis .

Tissue transglutaminase-dependent posttranslational modification of the <protein>retinoblastoma gene product</protein> in <cell_line>promonocytic cells</cell_line> undergoing apoptosis .


The retinoblastoma gene product ( pRB ) plays an important role in controlling both cell release from the G1 phase and apoptosis .

The <protein>retinoblastoma gene product</protein> ( <protein>pRB</protein> ) plays an important role in controlling both cell release from the G1 phase and apoptosis .


We show here that in the early phases of apoptosis , pRB is posttranslationally modified by a tissue transglutaminase ( tTG ) -catalyzed reaction .

We show here that in the early phases of apoptosis , <protein>pRB</protein> is posttranslationally modified by a tissue transglutaminase ( <protein>tTG</protein> ) -catalyzed reaction .


In fact , by employing a novel haptenized lysis synthetic substrate which allows the isolation of glutaminyl-tTG substrates in vivo , we identified pRB as a potential tTG substrate in U937 cells undergoing apoptosis .

In fact , by employing a novel haptenized lysis synthetic substrate which allows the isolation of glutaminyl-tTG substrates in vivo , we identified <protein>pRB</protein> as a potential tTG substrate in <cell_line>U937 cells</cell_line> undergoing apoptosis .


In keeping with this finding , we showed that apoptosis of U937 cells is characterized by the rapid disappearance of the 105 , 000- to 110 , 000-molecular-weight pRB forms concomitantly with the appearance of a smear of immunoreactive products with a molecular weight of greater than 250 , 000 .

In keeping with this finding , we showed that apoptosis of <cell_line>U937 cells</cell_line> is characterized by the rapid disappearance of the 105 , 000- to 110 , 000-molecular-weight <protein>pRB</protein> forms concomitantly with the appearance of a smear of immunoreactive products with a molecular weight of greater than 250 , 000 .


The shift in pRB molecular weight was reproduced by adding exogenous purified tTG to extracts obtained from viable U937 cells and was prevented by dansylcadaverine , a potent enzyme inhibitor .

The shift in <protein>pRB</protein> molecular weight was reproduced by adding exogenous purified tTG to extracts obtained from viable <cell_line>U937 cells</cell_line> and was prevented by dansylcadaverine , a potent enzyme inhibitor .


The effect of the pRB posttranslational modification during apoptosis was investigated by determining the E2F-1 levels and by isolating and characterizing pRB-null clones from U937 cells .

The effect of the <protein>pRB</protein> posttranslational modification during apoptosis was investigated by determining the <protein>E2F-1</protein> levels and by isolating and characterizing <cell_line>pRB-null clones</cell_line> from <cell_line>U937 cells</cell_line> .


Notably , the lack of pRB in these U937-derived clones renders these p53-null cells highly resistant to apoptosis induced by serum withdrawal , calphostin C , and ceramide .

Notably , the lack of <protein>pRB</protein> in these <cell_line>U937-derived clones</cell_line> renders these <cell_line>p53-null cells</cell_line> highly resistant to apoptosis induced by serum withdrawal , calphostin C , and ceramide .


Taken together , these data suggest that tTG , acting on the pRB protein , might play an important role in the cell progression through the death program .

Taken together , these data suggest that <protein>tTG</protein> , acting on the <protein>pRB protein</protein> , might play an important role in the cell progression through the death program .


STAT3 is a serine kinase target in T lymphocytes .

<protein>STAT3</protein> is a <protein>serine kinase target</protein> in <cell_type>T lymphocytes</cell_type> .


Interleukin 2 and T cell antigen receptor signals converge upon serine 727 .

<protein>Interleukin 2</protein> and <protein>T cell antigen receptor</protein> signals converge upon serine 727 .


Interleukin 2 ( IL-2 ) induces tyrosine phosphorylation of STATs 3 and 5 ( signal transducer and activator of transcription ) .

<protein>Interleukin 2</protein> ( <protein>IL-2</protein> ) induces tyrosine phosphorylation of <protein>STATs 3 and 5</protein> ( signal transducer and activator of transcription ) .


We now show that IL-2 regulation of STAT3 proteins in T cells is a complex response involving activation of two forms of STAT3 : 90-kDa STAT3alpha and an 83-kDa carboxyl-terminal truncated STAT3beta .

We now show that <protein>IL-2</protein> regulation of <protein>STAT3 proteins</protein> in <cell_type>T cells</cell_type> is a complex response involving activation of two forms of <protein>STAT3</protein> : 90-kDa <protein>STAT3alpha</protein> and an <protein>83-kDa carboxyl-terminal truncated STAT3beta</protein> .


The phosphorylation of STAT proteins on serine residues is also required for competent STAT transcription .

The phosphorylation of <protein>STAT proteins</protein> on serine residues is also required for competent <protein>STAT</protein> transcription .


A critical serine phosphorylation site in STAT3alpha is at position 727 .

A critical <protein>serine phosphorylation site</protein> in <protein>STAT3alpha</protein> is at position 727 .


In this study we have produced an antisera specific for STAT3alpha proteins phosphorylated on serine 727 and used this to monitor the phosphorylation of this residue during T lymphocyte activation .

In this study we have produced an antisera specific for <protein>STAT3alpha proteins</protein> phosphorylated on serine 727 and used this to monitor the phosphorylation of this residue during T lymphocyte activation .


Our results show that phosphorylation of STAT3alpha on serine 727 is not constitutive in quiescent T cells but can be induced by the cytokine IL-2 .

Our results show that phosphorylation of <protein>STAT3alpha</protein> on serine 727 is not constitutive in <cell_type>quiescent T cells</cell_type> but can be induced by the <protein>cytokine IL-2</protein> .


Interestingly , triggering of the T cell antigen receptor complex or activation of protein kinase C with phorbol esters also induces phosphorylation of serine 727 but without simultaneously inducing STAT3 tyrosine phosphorylation or DNA binding .

Interestingly , triggering of the <protein>T cell antigen receptor complex</protein> or activation of <protein>protein kinase C</protein> with phorbol esters also induces phosphorylation of serine 727 but without simultaneously inducing <protein>STAT3</protein> tyrosine phosphorylation or DNA binding .


Hence , the present results show that STAT3 serine phosphorylation can be regulated independently of the tyrosine phosphorylation of this molecule .

Hence , the present results show that <protein>STAT3</protein> serine phosphorylation can be regulated independently of the tyrosine phosphorylation of this molecule .


IL-2 and T cell antigen receptor complex induction of STAT3alpha serine 727 phosphorylation is dependent on the activity of the MEK /ERK pathway .

<protein>IL-2</protein> and <protein>T cell antigen receptor complex</protein> induction of <protein>STAT3alpha</protein> serine 727 phosphorylation is dependent on the activity of the <protein>MEK</protein> <protein>/ERK</protein> pathway .


Previous studies have identified H-7-sensitive kinase pathways that regulate STAT3 DNA binding .

Previous studies have identified H-7-sensitive kinase pathways that regulate <protein>STAT3</protein> DNA binding .


We show that H-7-sensitive pathways regulate STAT3 DNA binding in T cells .

We show that H-7-sensitive pathways regulate <protein>STAT3</protein> DNA binding in <cell_type>T cells</cell_type> .


Nevertheless , we show that H-7-sensitive kinases do not regulate STAT3 tyrosine phosphorylation or phosphorylation of serine 727 .

Nevertheless , we show that <protein>H-7-sensitive kinases</protein> do not regulate <protein>STAT3</protein> tyrosine phosphorylation or phosphorylation of serine 727 .


These results thus show that STAT3 proteins are targets for multiple kinase pathways in T cells and can integrate signals from both cytokine receptors and antigen receptors .

These results thus show that <protein>STAT3 proteins</protein> are targets for multiple kinase pathways in <cell_type>T cells</cell_type> and can integrate signals from both <protein>cytokine receptors</protein> and <protein>antigen receptors</protein> .


NF-kappa B/Rel family members regulating the ICAM-1 promoter in monocytic THP-1 cells .

<protein>NF-kappa B/Rel family members</protein> regulating the <dna>ICAM-1 promoter</dna> in <cell_line>monocytic THP-1 cells</cell_line> .


A kappa B-site was identified in the promoter of the intercellular adhesion molecule-1 ( ICAM-1 ) gene , which is involved in regulation of ICAM-1 expression by tumor necrosis factor alpha ( TNF-alpha ) and glucocorticoids .

A <dna>kappa B-site</dna> was identified in the promoter of the <dna>intercellular adhesion molecule-1 ( ICAM-1 ) gene</dna> , which is involved in regulation of <protein>ICAM-1</protein> expression by <protein>tumor necrosis factor alpha</protein> ( <protein>TNF-alpha</protein> ) and glucocorticoids .


We now report on the transcription factors which bind and transactivate this enhancer sequence .

We now report on the <protein>transcription factors</protein> which bind and transactivate this <dna>enhancer sequence</dna> .


In vitro , the ICAM-1 kappa B site appeared to bind RelA and c-Rel homodimers as well as heterodimers with NF-kappa B1 , but weakly NF-kappa B1 homodimers .

In vitro , the <dna>ICAM-1 kappa B site</dna> appeared to bind <protein>RelA</protein> and <protein>c-Rel homodimers</protein> as well as heterodimers with <protein>NF-kappa B1</protein> , but weakly <protein>NF-kappa B1 homodimers</protein> .


In addition , both RelA and c-Rel , but not NF-kappa B1 , were shown to transactivate an ICAM-1 kappa B-reporter construct .

In addition , both <protein>RelA</protein> and <protein>c-Rel</protein> , but not <protein>NF-kappa B1</protein> , were shown to transactivate an <dna>ICAM-1 kappa B-reporter construct</dna> .


In monocytic THP-1 cells TNF-alpha induced two nuclear complexes which in vitro bound to the ICAM-1 kappa B site .

In <cell_line>monocytic THP-1 cells</cell_line> <protein>TNF-alpha</protein> induced two <protein>nuclear complexes</protein> which in vitro bound to the <dna>ICAM-1 kappa B site</dna> .


Using antibodies in an electrophoretic mobility supershift assay , one of these complexes was shown to contain NF-kappa B1 and RelA , and to bind with higher affinity to the consensus kappa B site in the HIV long terminal repeat .

Using antibodies in an electrophoretic mobility supershift assay , one of these complexes was shown to contain <protein>NF-kappa B1</protein> and <protein>RelA</protein> , and to bind with higher affinity to the <dna>consensus kappa B site</dna> in the <dna>HIV long terminal repeat</dna> .


The second complex contained RelA , and exhibited higher affinity towards the ICAM-1 kappa B than to the HIV kappa B site .

The second complex contained <protein>RelA</protein> , and exhibited higher affinity towards the <dna>ICAM-1 kappa B</dna> than to the <dna>HIV kappa B site</dna> .


The glucocorticoid receptor was shown to repress activity of both the RelA homodimer and the NF-kappa B1/RelA heterodimer .

The <protein>glucocorticoid receptor</protein> was shown to repress activity of both the <protein>RelA homodimer</protein> and the <protein>NF-kappa B1/RelA heterodimer</protein> .


We argue that in vivo RelA homodimers are likely to play a dominant role in TNF-alpha -induced ICAM-1 transcription in monocytic cells .

We argue that in vivo <protein>RelA homodimers</protein> are likely to play a dominant role in <protein>TNF-alpha</protein> -induced <protein>ICAM-1</protein> transcription in <cell_type>monocytic cells</cell_type> .


Competent transcription initiation by RNA polymerase II in cell-free extracts from xeroderma pigmentosum groups B and D in an optimized RNA transcription assay .

Competent transcription initiation by <protein>RNA polymerase II</protein> in <cell_type>cell-free extracts</cell_type> from xeroderma pigmentosum groups B and D in an optimized RNA transcription assay .


The human autosomal recessive disease , xeroderma pigmentosum ( XP ) , can result from mutations in any one of seven genes , designated XPA through XPG .

The human autosomal recessive disease , xeroderma pigmentosum ( XP ) , can result from mutations in any one of seven genes , designated <dna>XPA</dna> through <dna>XPG</dna> .


Of these , the XPB and XPD genes encode proteins that are subunits of a general transcription factor , TFIIH , involved in both nucleotide excision repair ( NER ) and initiation of mRNA transcription by RNA polymerase II .

Of these , the <dna>XPB and XPD genes</dna> encode proteins that are subunits of a <protein>general transcription factor</protein> , <protein>TFIIH</protein> , involved in both nucleotide excision repair ( NER ) and initiation of mRNA transcription by <protein>RNA polymerase II</protein> .


In humans , mutation of the XPB or XPD gene impairs NER , resulting in hyper-sensitivity to sunlight and greatly increased skin tumor formation .

In humans , mutation of the <dna>XPB or XPD gene</dna> impairs NER , resulting in hyper-sensitivity to sunlight and greatly increased skin tumor formation .


However , no transcription deficiency has been demonstrated in either XP-B or XP-D .


We have employed an optimized cell-free RNA transcription assay to analyze transcription activity of XP-B and XP-D .


Although the growth rate was normal , the XP-B and XP-D cells contained reduced amounts of TFIIH .

Although the growth rate was normal , the <cell_line>XP-B and XP-D cells</cell_line> contained reduced amounts of <protein>TFIIH</protein> .


Extracts prepared from XP-B and XP-D lymphoblastoid cells exhibited similar transcription activity from the adenovirus major late promoter when compared to that in extracts from normal cells .

Extracts prepared from <cell_line>XP-B and XP-D lymphoblastoid cells</cell_line> exhibited similar transcription activity from the <dna>adenovirus major late promoter</dna> when compared to that in extracts from normal cells .


Thus , we conclude that the XP-B and XP-D lymphoblastoid cells do not have impaired RNA transcription activity .

Thus , we conclude that the <cell_line>XP-B and XP-D lymphoblastoid cells</cell_line> do not have impaired RNA transcription activity .


We consider the possible consequences of the reduced cellular content of TFIIH for the clinical symptoms in XP-B or XP-D patients , and discuss a 'conditional phenotype ' that may involve an impairment of cellular function only under certain growth conditions .

We consider the possible consequences of the reduced cellular content of <protein>TFIIH</protein> for the clinical symptoms in XP-B or XP-D patients , and discuss a 'conditional phenotype ' that may involve an impairment of cellular function only under certain growth conditions .


A novel form of the myeloid-specific zinc finger protein ( MZF-2 ) .

A novel form of the <protein>myeloid-specific zinc finger protein</protein> ( <protein>MZF-2</protein> ) .


BACKGROUND : Myeloid cell development is controlled by tissue-specific transcription factors .

BACKGROUND : Myeloid cell development is controlled by tissue-specific <protein>transcription factors</protein> .


Human myeloid zinc finger protein ( MZF-1 ) is a putative transcription factor containing 13 zinc fingers , and has been suggested that it regulates the development of neutrophilic granulocytes .

<protein>Human myeloid zinc finger protein</protein> ( <protein>MZF-1</protein> ) is a putative transcription factor containing <protein>13 zinc fingers</protein> , and has been suggested that it regulates the development of <cell_type>neutrophilic granulocytes</cell_type> .


RESULTS : Here , we have isolated the murine and human cDNAs which encode a novel form of MZF protein ( MZF-2 ) .

RESULTS : Here , we have isolated the <dna>murine and human cDNAs</dna> which encode a novel form of <protein>MZF protein</protein> ( <protein>MZF-2</protein> ) .


Murine and human MZF-2 proteins consisted of 814 and 775 amino acids , respectively , and have identity of 75.3 % between them .

<protein>Murine and human MZF-2 proteins</protein> consisted of 814 and 775 amino acids , respectively , and have identity of 75.3 % between them .


The C-terminal half of human MZF-2 , carrying the zinc finger domains , was completely identical with that of human MZF-1 , whereas the N-terminal half of MZF-2 was different from the corresponding region of human MZF-1 , and was coded by distinct exons .

The <protein>C-terminal half</protein> of <protein>human MZF-2</protein> , carrying the <protein>zinc finger domains</protein> , was completely identical with that of <protein>human MZF-1</protein> , whereas the <protein>N-terminal half</protein> of <protein>MZF-2</protein> was different from the corresponding region of human <protein>MZF-1</protein> , and was coded by distinct <dna>exons</dna> .


MZF-2 mRNA was expressed in myeloid cells , particularly in the cells committed to the neutrophilic lineage , and down-regulated by G-CSF .

<rna>MZF-2 mRNA</rna> was expressed in myeloid cells , particularly in the cells committed to the <cell_type>neutrophilic lineage</cell_type> , and down-regulated by <protein>G-CSF</protein> .


CONCLUSIONS : MZF-1 and MZF-2 mRNAs seem to be produced by the alternative use of two different transcription initiation sites .

CONCLUSIONS : <rna>MZF-1 and MZF-2 mRNAs</rna> seem to be produced by the alternative use of two different <dna>transcription initiation sites</dna> .


The distinct N-terminal half of MZF-2 carries two characteristic domains , a leucine-rich domain called LeR and an acidic domain , which suggests a unique function of MZF-2 in neutrophil development .

The distinct <protein>N-terminal half</protein> of <protein>MZF-2</protein> carries two characteristic domains , a <protein>leucine-rich domain</protein> called <protein>LeR</protein> and an <protein>acidic domain</protein> , which suggests a unique function of <protein>MZF-2</protein> in neutrophil development .


Impaired cortisol binding to glucocorticoid receptors in hypertensive patients .

Impaired cortisol binding to <protein>glucocorticoid receptors</protein> in hypertensive patients .


We compared glucocorticoid receptor binding characteristics and glucocorticoid responsiveness of human mononuclear leukocytes ( HML ) from hypertensive patients and matched normotensive volunteers .

We compared glucocorticoid receptor binding characteristics and glucocorticoid responsiveness of <cell_type>human mononuclear leukocytes</cell_type> ( <cell_type>HML</cell_type> ) from hypertensive patients and matched normotensive volunteers .


We also considered associations of these variables with plasma renin activity , aldosterone , cortisol , corticotropin , and electrolyte concentrations .

We also considered associations of these variables with plasma <protein>renin</protein> activity , aldosterone , cortisol , corticotropin , and electrolyte concentrations .


We calculated binding affinity ( Kd ; nmol/L ) and capacity ( Bmax ; sites/cell ) for dexamethasone and cortisol from homologous and heterologous competition curves for specific [ 3H ] dexamethasone binding sites on HML isolated from the blood of normotensive volunteers and subjects with essential hypertension .

We calculated binding affinity ( Kd ; nmol/L ) and capacity ( Bmax ; sites/cell ) for dexamethasone and cortisol from homologous and heterologous competition curves for specific [ 3H ] dexamethasone binding sites on <cell_type>HML</cell_type> isolated from the blood of normotensive volunteers and subjects with essential hypertension .


Glucocorticoid responsiveness of HML was evaluated as IC50 values ( nmol/L ) for dexamethasone and cortisol for the inhibition of lysozyme release .

Glucocorticoid responsiveness of <cell_type>HML</cell_type> was evaluated as IC50 values ( nmol/L ) for dexamethasone and cortisol for the inhibition of <protein>lysozyme</protein> release .


We measured plasma hormones by radioimmunoassay .


Kd values ( mean+/-SE ) for cortisol in HML of hypertensive patients were higher than in control subjects ( 24.6+/-2.4 versus 17.5+/-1.7 nmol/L , P < .04 ) .

Kd values ( mean+/-SE ) for cortisol in <cell_type>HML</cell_type> of hypertensive patients were higher than in control subjects ( 24.6+/-2.4 versus 17.5+/-1.7 nmol/L , P < .04 ) .


Binding capacity ( 4978+/-391 versus 4131+/-321 sites/cell ) , Kd values for dexamethasone ( 6.7+/-0.5 versus 5.7+/-0.3 nmol/L ) , and IC50 values for dexamethasone ( 3.4+/-0.3 versus 3.1+/-0.2 nmol/L ) and cortisol ( 12.2+/-1.6 versus 9.5+/-0.3 nmol/L ) were not significantly different .


Patients with renin values less than 0.13 ng angiotensin I/L per second were markedly less sensitive to cortisol than those with higher values .

Patients with <protein>renin</protein> values less than 0.13 ng angiotensin I/L per second were markedly less sensitive to cortisol than those with higher values .


Both Kd ( 30.3+/-2.5 versus 19.2+/-2.4 nmol/L ) and IC50 values ( 15.5+/-1.8 versus 8.9+/-1.2 nmol/L ) for cortisol were significantly higher in patients with lower renin values ( P < .03 ) .

Both Kd ( 30.3+/-2.5 versus 19.2+/-2.4 nmol/L ) and IC50 values ( 15.5+/-1.8 versus 8.9+/-1.2 nmol/L ) for cortisol were significantly higher in patients with lower <protein>renin</protein> values ( P < .03 ) .


Other variables , including plasma hormone and electrolyte values and binding characteristics for dexamethasone , were not different .


These data suggest that cortisol binding to glucocorticoid receptor is slightly impaired in patients with essential hypertension .

These data suggest that cortisol binding to <protein>glucocorticoid receptor</protein> is slightly impaired in patients with essential hypertension .


In vivo , this could lead to inappropriate binding of cortisol to mineralocorticoid receptors .


Hence , decreased sensitivity to cortisol is associated with renin suppression .

Hence , decreased sensitivity to cortisol is associated with <protein>renin</protein> suppression .


This hypothesis is supported by evidence of hypertension and low renin activity , which others have described in patients with primary glucocorticoid resistance due to mutations of the glucocorticoid receptor .

This hypothesis is supported by evidence of hypertension and low <protein>renin</protein> activity , which others have described in patients with primary glucocorticoid resistance due to mutations of the <protein>glucocorticoid receptor</protein> .


PU.1/Pip and basic helix loop helix zipper transcription factors interact with binding sites in the CD20 promoter to help confer lineage- and stage-specific expression of CD20 in B lymphocytes .

<protein>PU.1/Pip</protein> and <protein>basic helix loop helix zipper transcription factors</protein> interact with <dna>binding sites</dna> in the <dna>CD20 promoter</dna> to help confer lineage- and stage-specific expression of <protein>CD20</protein> in <cell_type>B lymphocytes</cell_type> .


CD20 is a B-lineage-specific gene expressed at the pre-B-cell stage of B-cell development that disappears on differentiation to plasma cells .

<protein>CD20</protein> is a <dna>B-lineage-specific gene</dna> expressed at the pre-B-cell stage of B-cell development that disappears on differentiation to <cell_type>plasma cells</cell_type> .


As such , it serves as an excellent paradigm for the study of lineage and developmental stage-specific gene expression .


Using in vivo footprinting we identified two sites in the promoter at -45 and -160 that were occupied only in CD20 + B cells .

Using in vivo footprinting we identified two sites in the promoter at <dna>-45 and -160</dna> that were occupied only in <protein>CD20</protein> + B cells .


The -45 site is an E box that binds basic helix-loop-helix-zipper proteins whereas the -160 site is a composite PU.1 and Pip binding site .

The <dna>-45 site</dna> is an <dna>E box</dna> that binds <protein>basic helix-loop-helix-zipper proteins</protein> whereas the <dna>-160 site</dna> is a <dna>composite PU.1 and Pip binding site</dna> .


Transfection studies with reporter constructs and various expression vectors verified the importance of these sites .

Transfection studies with <dna>reporter constructs</dna> and various expression vectors verified the importance of these sites .


The composite PU.1 and Pip site likely accounts for both lineage and stage-specific expression of CD20 whereas the CD20 E box binding proteins enhance overall promoter activity and may link the promoter to a distant enhancer .

The <dna>composite PU.1 and Pip site</dna> likely accounts for both lineage and stage-specific expression of <protein>CD20</protein> whereas the <protein>CD20 E box binding proteins</protein> enhance overall promoter activity and may link the promoter to a <dna>distant enhancer</dna> .


Glucocorticoids and the immune function in the human immunodeficiency virus infection : a study in hypercortisolemic and cortisol-resistant patients .


Immunological studies in human immunodeficiency virus ( HIV ) -positive patients suggest that the disease progression is accompanied by a defective production of type 1 cytokines ( interleukin-2 ( IL-2 ) and IL-12 ] , an increased production of type 2 cytokines ( IL-4 , IL-6 , and IL-10 ) , and an increased production of IgE .

Immunological studies in human immunodeficiency virus ( HIV ) -positive patients suggest that the disease progression is accompanied by a defective production of type <protein>1 cytokines</protein> ( <protein>interleukin-2</protein> ( <protein>IL-2</protein> ) and <protein>IL-12</protein> ] , an increased production of <protein>type 2 cytokines</protein> ( <protein>IL-4</protein> , <protein>IL-6</protein> , and <protein>IL-10</protein> ) , and an increased production of <protein>IgE</protein> .


HIV infection is also associated with activation of the hypothalamo-pituitary-adrenal axis function and increased plasma and urinary cortisol concentrations .


As cortisol is involved in the physiological regulation of cytokines , a study was conducted to examine cytokine patterns in two groups of hypercortisolemic patients , one with normal sensitivity to glucocorticoids and the other with glucocorticoid resistance .

As cortisol is involved in the physiological regulation of <protein>cytokines</protein> , a study was conducted to examine <protein>cytokine</protein> patterns in two groups of hypercortisolemic patients , one with normal sensitivity to glucocorticoids and the other with glucocorticoid resistance .


Ten HIV-infected patients with normal receptor affinity to glucocorticoids ( AIDS-C ) , 10 HIV-infected patients with low receptor affinity to glucocorticoids ( AIDS-GR ) , and 20 healthy subjects were studied .


Receptor characteristics of peripheral blood mononuclear cells were evaluated by [ 3H ] dexamethasone binding .

Receptor characteristics of <cell_type>peripheral blood mononuclear cells</cell_type> were evaluated by [ 3H ] dexamethasone binding .


Serum cortisol and urinary free cortisol were measured by RIA .


Serum ACTH and IgE were measured by immunoradiometric assay , and IL-2 , IL-4 , and IL-10 cytokines and interferon-gamma were measured by enzyme-linked immunosorbent assay .

Serum ACTH and <protein>IgE</protein> were measured by immunoradiometric assay , and <protein>IL-2</protein> , <protein>IL-4</protein> , and <protein>IL-10</protein> <protein>cytokines</protein> and <protein>interferon-gamma</protein> were measured by enzyme-linked immunosorbent assay .


AIDS-C patients showed low IL-2 and high IL-4 , IL-10 , and IgE concentratios ; conversely , AIDS-GR patients showed high IL-2 and low IL-4 and IgE concentrations .

AIDS-C patients showed low <protein>IL-2</protein> and high <protein>IL-4</protein> , <protein>IL-10</protein> , and <protein>IgE</protein> concentratios ; conversely , AIDS-GR patients showed high <protein>IL-2</protein> and low <protein>IL-4</protein> and <protein>IgE</protein> concentrations .


Thus , in HIV infection , elevated cortisol levels suppress cell-mediated immunity and stimulate humoral immunity , whereas this response is not detected in cortisol-resistant patients .


These findings indicate that cortisol and its receptors are critically involved in the regulation of immune function in HIV infection .


Monocytic differentiation of HL-60 promyelocytic leukemia cells correlates with the induction of Bcl-xL .

Monocytic differentiation of <cell_line>HL-60 promyelocytic leukemia cells</cell_line> correlates with the induction of <protein>Bcl-xL</protein> .


Treatment of human promyelocytic leukemia HL-60 cells with phorbol esters ultimately induces the differentiation of these cells along the monocyte/macrophage lineage , whereas treatment with retinoic acid or DMSO induces granulocytic/neutrophillic differentiation .

Treatment of <cell_line>human promyelocytic leukemia HL-60 cells</cell_line> with phorbol esters ultimately induces the differentiation of these cells along the <cell_type>monocyte/macrophage lineage</cell_type> , whereas treatment with retinoic acid or DMSO induces granulocytic/neutrophillic differentiation .


In this study , we demonstrate the disparate fates of HL-60 cells treated with the phorbol ester 12 , 13-phorbol dibutyric acid ( PDBu ) or DMSO .

In this study , we demonstrate the disparate fates of <cell_line>HL-60 cells</cell_line> treated with the phorbol ester 12 , 13-phorbol dibutyric acid ( PDBu ) or DMSO .


After DMSO treatment , HL-60 cells eventually died via apoptosis , whereas the viability of PDBu-treated cells was not affected during the same interval .

After DMSO treatment , <cell_line>HL-60 cells</cell_line> eventually died via apoptosis , whereas the viability of <cell_line>PDBu-treated cells</cell_line> was not affected during the same interval .


The levels of the apoptosis effector proteins Bak and Bad were enhanced , whereas there was a slight down-regulation of the apoptosis suppressor protein Bcl-2 after treatment of the cells with PDBu and DMSO .

The levels of the apoptosis effector proteins <protein>Bak</protein> and <protein>Bad</protein> were enhanced , whereas there was a slight down-regulation of the <protein>apoptosis suppressor protein</protein> <protein>Bcl-2</protein> after treatment of the cells with PDBu and DMSO .


Treatment with DMSO resulted in the elevation of the apoptosis effector Bax , whereas treatment with PDBu did not significantly alter the levels of this protein .

Treatment with DMSO resulted in the elevation of the <protein>apoptosis effector</protein> <protein>Bax</protein> , whereas treatment with PDBu did not significantly alter the levels of this protein .


However , treatment of HL-60 cells with PDBu induced the rapid expression of the apoptosis suppressor protein Bcl-xL , whereas the expression of this protein remained unaltered in DMSO-treated cells .

However , treatment of <cell_line>HL-60 cells</cell_line> with PDBu induced the rapid expression of the <protein>apoptosis suppressor protein</protein> <protein>Bcl-xL</protein> , whereas the expression of this protein remained unaltered in DMSO-treated cells .


The generality of this finding was confirmed by the induction of Bcl-xL in human myeloid U-937 cells , human peripheral blood monocytes exposed to phorbol ester , and mouse thioglycollate-activated and resident peritoneal macrophages .

The generality of this finding was confirmed by the induction of <protein>Bcl-xL</protein> in <cell_line>human myeloid U-937 cells</cell_line> , <cell_type>human peripheral blood monocytes</cell_type> exposed to phorbol ester , and <cell_line>mouse thioglycollate-activated and resident peritoneal macrophages</cell_line> .


PDBu-treated HL-60 cells remained viable for 7 days and thereafter began to die via apoptosis , with a concomitant down-regulation of Bcl-xL .

<cell_line>PDBu-treated HL-60 cells</cell_line> remained viable for 7 days and thereafter began to die via apoptosis , with a concomitant down-regulation of <protein>Bcl-xL</protein> .


In conclusion , we propose that Bcl-xL expression is associated with differentiation and survival of hematopoietic cells along the monocyte/macrophage lineage .

In conclusion , we propose that <protein>Bcl-xL</protein> expression is associated with differentiation and survival of <cell_type>hematopoietic cells</cell_type> along the <cell_type>monocyte/macrophage lineage</cell_type> .


Monochloramine inhibits phorbol ester-inducible neutrophil respiratory burst activation and T cell interleukin-2 receptor expression by inhibiting inducible protein kinase C activity .

Monochloramine inhibits phorbol ester-inducible neutrophil respiratory burst activation and T cell <protein>interleukin-2</protein> receptor expression by inhibiting inducible <protein>protein kinase C</protein> activity .


Monochloramine derivatives are long lived physiological oxidants produced by neutrophils during the respiratory burst .

Monochloramine derivatives are long lived physiological oxidants produced by <cell_type>neutrophils</cell_type> during the respiratory burst .


The effects of chemically prepared monochloramine ( NH2Cl ) on protein kinase C ( PKC ) and PKC -mediated cellular responses were studied in elicited rat peritoneal neutrophils and human Jurkat T cells .

The effects of chemically prepared monochloramine ( NH2Cl ) on <protein>protein kinase C</protein> ( <protein>PKC</protein> ) and <protein>PKC</protein> -mediated cellular responses were studied in <cell_type>elicited rat peritoneal neutrophils</cell_type> and <cell_line>human Jurkat T cells</cell_line> .


Neutrophils pretreated with NH2Cl ( 30-50 microM ) showed a marked decrease in the respiratory burst activity induced by phorbol 12-myristate 13-acetate ( PMA ) , which is a potent PKC activator .

Neutrophils pretreated with NH2Cl ( 30-50 microM ) showed a marked decrease in the respiratory burst activity induced by phorbol 12-myristate 13-acetate ( PMA ) , which is a potent <protein>PKC</protein> activator .


These cells , however , were viable and showed a complete respiratory burst upon arachidonic acid stimulation , which induces the respiratory burst by a PKC -independent mechanism .

These cells , however , were viable and showed a complete respiratory burst upon arachidonic acid stimulation , which induces the respiratory burst by a <protein>PKC</protein> -independent mechanism .


The NH2Cl-treated neutrophils showed a decrease in both PKC activity and PMA-induced phosphorylation of a 47-kDa protein , which corresponds to the cytosolic factor of NADPH oxidase , p47 ( phox ) .

The <cell_line>NH2Cl-treated neutrophils</cell_line> showed a decrease in both <protein>PKC</protein> activity and PMA-induced phosphorylation of a <protein>47-kDa protein</protein> , which corresponds to the <protein>cytosolic factor</protein> of <protein>NADPH oxidase</protein> , <protein>p47 ( phox )</protein> .


Jurkat T cells pretreated with NH2Cl ( 20-70 microM ) showed a decrease in the expression of the interleukin-2 receptor alpha chain following PMA stimulation .

<cell_line>Jurkat T cells</cell_line> pretreated with NH2Cl ( 20-70 microM ) showed a decrease in the expression of the <protein>interleukin-2</protein> <protein>receptor alpha chain</protein> following PMA stimulation .


This was also accompanied by a decrease in both PKC activity and nuclear transcription factor- kappaB activation , also without loss of cell viability .

This was also accompanied by a decrease in both <protein>PKC</protein> activity and nuclear transcription factor- <protein>kappaB</protein> activation , also without loss of cell viability .


These results show that NH2Cl inhibits PKC -mediated cellular responses through inhibition of the inducible PKC activity .

These results show that NH2Cl inhibits <protein>PKC</protein> -mediated cellular responses through inhibition of the inducible <protein>PKC</protein> activity .


Evidence that calcineurin is rate-limiting for primary human lymphocyte activation .


Cyclosporine ( CsA ) is both a clinical immunosuppressive drug and a probe to dissect intracellular signaling pathways .


In vitro , CsA inhibits lymphocyte gene activation by inhibiting the phosphatase activity of calcineurin ( CN ) .

In vitro , CsA inhibits <dna>lymphocyte gene</dna> activation by inhibiting the <protein>phosphatase</protein> activity of <protein>calcineurin</protein> ( <protein>CN</protein> ) .


In clinical use , CsA treatment inhibits 50-75 % of CN activity in circulating leukocytes .

In clinical use , CsA treatment inhibits 50-75 % of <protein>CN</protein> activity in <cell_type>circulating leukocytes</cell_type> .


We modeled this degree of CN inhibition in primary human leukocytes in vitro in order to study the effect of partial CN inhibition on the downstream signaling events that lead to gene activation .

We modeled this degree of <protein>CN</protein> inhibition in <cell_type>primary human leukocytes</cell_type> in vitro in order to study the effect of partial <protein>CN</protein> inhibition on the downstream signaling events that lead to gene activation .


In CsA-treated leukocytes stimulated by calcium ionophore , the degree of reduction in CN activity was accompanied by a similar degree of inhibition of each event tested : dephosphorylation of nuclear factor of activated T cell proteins , nuclear DNA binding , activation of a transfected reporter gene construct , IFN-gamma and IL-2 mRNA accumulation , and IFN-gamma production .

In <cell_line>CsA-treated leukocytes</cell_line> stimulated by calcium ionophore , the degree of reduction in <protein>CN</protein> activity was accompanied by a similar degree of inhibition of each event tested : dephosphorylation of <protein>nuclear factor of activated T cell proteins</protein> , nuclear DNA binding , activation of a <dna>transfected reporter gene construct</dna> , IFN-gamma and IL-2 mRNA accumulation , and <protein>IFN-gamma</protein> production .


Furthermore , the degree of CN inhibition was reflected by a similar degree of reduction in lymphocyte proliferation and IFN-gamma production in the allogeneic mixed lymphocyte cultures .

Furthermore , the degree of <protein>CN</protein> inhibition was reflected by a similar degree of reduction in lymphocyte proliferation and <protein>IFN-gamma</protein> production in the <cell_line>allogeneic mixed lymphocyte cultures</cell_line> .


These data support the conclusion that CN activity is rate-limiting for the activation of primary human T lymphocytes .

These data support the conclusion that <protein>CN</protein> activity is rate-limiting for the activation of <cell_type>primary human T lymphocytes</cell_type> .


Thus , the reduction of CN activity observed in CsA-treated patients is accompanied by a similar degree of reduction in lymphocyte gene activation , and accounts for the immunosuppression observed .

Thus , the reduction of <protein>CN</protein> activity observed in CsA-treated patients is accompanied by a similar degree of reduction in <dna>lymphocyte gene</dna> activation , and accounts for the immunosuppression observed .


Selection of down-regulated sequences along the monocytic differentiation of leukemic HL60 cells .

Selection of <dna>down-regulated sequences</dna> along the monocytic differentiation of <cell_line>leukemic HL60 cells</cell_line> .


In order to dissect the molecular mechanisms of monocytic differentiation we have developed a subtractive hybridisation method based on a simplified 'representational difference analysis' .


We have selected 16 sequences and confirmed their down-regulation along the TPA-induced monocytic differentiation of HL60 cells .

We have selected <dna>16 sequences</dna> and confirmed their down-regulation along the TPA-induced monocytic differentiation of <cell_line>HL60 cells</cell_line> .


Among these sequences we have identified the alpha-tubulin , the TaxREB protein and two ribosomal protein sequences which had not been previously described as differentially expressed .

Among these sequences we have identified the <protein>alpha-tubulin</protein> , the <protein>TaxREB protein</protein> and two <dna>ribosomal protein sequences</dna> which had not been previously described as differentially expressed .


These results add to our knowledge about the molecules implicated along the monocytic differentiation and growth arrest of leukemic cells and provide a first step in the study of their respective roles .

These results add to our knowledge about the molecules implicated along the monocytic differentiation and growth arrest of <cell_type>leukemic cells</cell_type> and provide a first step in the study of their respective roles .


Control of NF-kappa B activity by the I kappa B beta inhibitor .

Control of <protein>NF-kappa B</protein> activity by the <protein>I kappa B beta inhibitor</protein> .


The transcription factor NF-kappa B is maintained in an inactive cytoplasmic state by I kappa B inhibitors .

The <protein>transcription factor</protein> <protein>NF-kappa B</protein> is maintained in an inactive cytoplasmic state by <protein>I kappa B inhibitors</protein> .


In mammalian cells , I kappa B alpha and I kappa B beta proteins have been purified and shown to be the inhibitors of NF-kappa B through their association with the p65 or c-Rel subunits .

In <cell_type>mammalian cells</cell_type> , <protein>I kappa B alpha</protein> and <protein>I kappa B beta proteins</protein> have been purified and shown to be the inhibitors of <protein>NF-kappa B</protein> through their association with the <protein>p65 or c-Rel subunits</protein> .


In addition , we have isolated a third NF-kappa B inhibitor , I kappa B epsilon ( 1 ) .

In addition , we have isolated a third <protein>NF-kappa B inhibitor</protein> , <protein>I kappa B epsilon</protein> ( 1 ) .


Upon treatment with a large variety of inducers , I kappa B alpha , I kappa B beta are proteolytically degraded , resulting in NF-kappa B translocation into the nucleus .

Upon treatment with a large variety of inducers , <protein>I kappa B alpha</protein> , <protein>I kappa B beta</protein> are proteolytically degraded , resulting in <protein>NF-kappa B</protein> translocation into the nucleus .


Here we show that in E29.1 T cell hybridoma I kappa B alpha and I kappa B beta are equally associated with p65 and that I kappa B beta is degraded in response to TNF alpha in contrast to what has been originally published .

Here we show that in <cell_line>E29.1 T cell hybridoma</cell_line> <protein>I kappa B alpha</protein> and <protein>I kappa B beta</protein> are equally associated with <protein>p65</protein> and that <protein>I kappa B beta</protein> is degraded in response to <protein>TNF alpha</protein> in contrast to what has been originally published .


Our data also suggest that , unlike I kappa B alpha , I kappa B beta is constitutively phosphorylated and resynthesized as a hypophosphorylated form .

Our data also suggest that , unlike <protein>I kappa B alpha</protein> , <protein>I kappa B beta</protein> is constitutively phosphorylated and resynthesized as a <protein>hypophosphorylated form</protein> .


The absence of slow migrating forms of I kappa B beta following stimulation suggests that the phosphorylation does not necessarily constitute the signal-induced event which targets the molecule for proteolysis .

The absence of slow migrating forms of <protein>I kappa B beta</protein> following stimulation suggests that the phosphorylation does not necessarily constitute the signal-induced event which targets the molecule for proteolysis .


Agonistic activity of a CD40-specific single-chain Fv constructed from the variable regions of mAb G28-5 .

Agonistic activity of a <protein>CD40-specific single-chain Fv</protein> constructed from the <protein>variable regions</protein> of <protein>mAb G28-5</protein> .


A single-chain Fv ( sFv ) was expressed from the variable regions of the CD40-specific mAb G28-5 .

A <protein>single-chain Fv</protein> ( <protein>sFv</protein> ) was expressed from the <protein>variable regions</protein> of the CD40-specific <protein>mAb G28-5</protein> .


The molecule bound CD40 with a high affinity ( 2.2 nM ) and was a monomer in solution .

The molecule bound <protein>CD40</protein> with a high affinity ( 2.2 nM ) and was a monomer in solution .


Surprisingly , G28-5 sFv was a potent CD40 agonist that rapidly crosslinked CD40 on the cell surface but did not crosslink CD40-Ig in solution .

Surprisingly , <protein>G28-5 sFv</protein> was a potent <protein>CD40 agonist</protein> that rapidly crosslinked <protein>CD40</protein> on the cell surface but did not crosslink <protein>CD40-Ig</protein> in solution .


G28-5 sFv was a more potent agonist than G28-5 IgG and was able to stimulate CD40 responses by B cells and monocytes .

<protein>G28-5 sFv</protein> was a more potent agonist than <protein>G28-5 IgG</protein> and was able to stimulate <protein>CD40</protein> responses by <cell_type>B cells</cell_type> and monocytes .


G28-5 IgG partially blocked , whereas G28-5 sFv augmented CD40 responses during stimulation with natural ligand ( gp39-CD8 fusion protein ) .

<protein>G28-5 IgG</protein> partially blocked , whereas <protein>G28-5 sFv</protein> augmented <protein>CD40</protein> responses during stimulation with <protein>natural ligand</protein> ( <protein>gp39-CD8 fusion protein</protein> ) .


These results indicate that the functional activity of ligands built from the binding site of G28-5 is highly dependent upon the size and physical properties of the molecule both in solution and on the cell surfaces .

These results indicate that the functional activity of ligands built from the <protein>binding site</protein> of <protein>G28-5</protein> is highly dependent upon the size and physical properties of the molecule both in solution and on the cell surfaces .


C/EBP activates the human corticotropin-releasing hormone gene promoter .

<protein>C/EBP</protein> activates the <dna>human corticotropin-releasing hormone gene promoter</dna> .


The purpose of these studies was to identify whether transcription factors , associated with cytokine signalling , affected promoter activity of the corticotropin releasing hormone ( CRH ) gene .

The purpose of these studies was to identify whether transcription factors , associated with cytokine signalling , affected promoter activity of the <dna>corticotropin releasing hormone ( CRH ) gene</dna> .


Fragments of a 3.6 kb sequence of the human CRH gene promoter were amplified by PCR and ligated upstream of a CAT reporter .

Fragments of a 3.6 kb sequence of the <dna>human CRH gene promoter</dna> were amplified by PCR and ligated upstream of a <dna>CAT reporter</dna> .


These constructs were transfected into a variety of cell lines , either alone or together , with transcription factor expression vectors .

These constructs were transfected into a variety of cell lines , either alone or together , with <protein>transcription factor</protein> expression vectors .


Basal activity of a 3070 bp CRH promoter fragment was only seen in neuronal and lymphoblastoid cell lines .

Basal activity of a <dna>3070 bp CRH promoter fragment</dna> was only seen in neuronal and <cell_line>lymphoblastoid cell lines</cell_line> .


Promoter activity was increased by the transcription factors C/EBPbeta ( NF-IL6 ) and more strongly , by C/EBPdelta ( NF-IL6beta ) .

Promoter activity was increased by the <protein>transcription factors</protein> <protein>C/EBPbeta</protein> ( <protein>NF-IL6</protein> ) and more strongly , by <protein>C/EBPdelta</protein> ( <protein>NF-IL6beta</protein> ) .


Increased CRH promoter activity following phorbol ester treatment was inhibited by a dominant negative NF-IL6 mutant , showing that the effects of phorbol ester were principally mediated by C/EBP .

Increased <protein>CRH</protein> promoter activity following phorbol ester treatment was inhibited by a dominant negative <protein>NF-IL6 mutant</protein> , showing that the effects of phorbol ester were principally mediated by <protein>C/EBP</protein> .


Moreover , the inverse changes in the expression of CRH in the hypothalamus and spleens of arthritic rats were paralleled by similar inverse changes in NF-IL6beta expression in these organs .

Moreover , the inverse changes in the expression of <protein>CRH</protein> in the hypothalamus and spleens of arthritic rats were paralleled by similar inverse changes in <protein>NF-IL6beta</protein> expression in these organs .


These data show that some transcription factors associated with cytokine signalling can also activate the CRH promoter .

These data show that some <protein>transcription factors</protein> associated with <protein>cytokine</protein> signalling can also activate the <dna>CRH promoter</dna> .


Aberrant splicing of the TSG101 and FHIT genes occurs frequently in multiple malignancies and in normal tissues and mimics alterations previously described in tumours .

Aberrant splicing of the <dna>TSG101 and FHIT genes</dna> occurs frequently in multiple malignancies and in normal tissues and mimics alterations previously described in tumours .


Intragenic deletions of TSG101 , the human homolog of a mouse gene ( tsg101 ) that acts to suppress malignant cell growth , were reported in human breast tumours .

Intragenic deletions of <dna>TSG101</dna> , the human homolog of a <dna>mouse gene</dna> ( <dna>tsg101</dna> ) that acts to suppress malignant cell growth , were reported in <cell_type>human breast tumours</cell_type> .


We screened TSG101 for somatic mutations in DNA and RNA samples isolated from a variety of common human malignancies , EBV-immortalised B-cells , and normal lung parenchyma .

We screened <dna>TSG101</dna> for somatic mutations in DNA and RNA samples isolated from a variety of common human malignancies , <cell_type>EBV-immortalised B-cells</cell_type> , and normal lung parenchyma .


Intragenic TSG101 deletions in RNA transcripts were frequently found in all types of samples .

Intragenic <dna>TSG101</dna> deletions in <rna>RNA transcripts</rna> were frequently found in all types of samples .


Analysis of DNA failed to show genomic rearrangements corresponding to transcripts containing deletions in the same samples .


The breakpoints of most transcript deletions coincide with genuine or cryptic splice site sequences , suggesting that they result from alternative or aberrant splicing .

The breakpoints of most transcript deletions coincide with genuine or cryptic <dna>splice site sequences</dna> , suggesting that they result from alternative or aberrant splicing .


A similar spectrum of transcript deletions has previously been described in the putative tumour suppressor gene FHIT .

A similar spectrum of transcript deletions has previously been described in the <dna>putative tumour suppressor gene</dna> <dna>FHIT</dna> .


We analysed FHIT in the same series of RNA samples and detected truncated FHIT transcripts frequently in both tumour and normal tissues .

We analysed <dna>FHIT</dna> in the same series of <rna>RNA samples</rna> and detected truncated <rna>FHIT transcripts</rna> frequently in both tumour and normal tissues .


In addition , transcripts from TSG101 , FHIT and seven other genes were analysed in RNA isolated from normal peripheral blood lymphocytes .

In addition , transcripts from <dna>TSG101</dna> , <dna>FHIT</dna> and seven other genes were analysed in RNA isolated from <cell_type>normal peripheral blood lymphocytes</cell_type> .


Large TSG101 and FHIT intragenic transcript deletions were detected and these appeared to be the predominant transcript in 'aged ' lymphocytes .

Large <dna>TSG101</dna> and <dna>FHIT</dna> intragenic transcript deletions were detected and these appeared to be the predominant transcript in <cell_type>'aged ' lymphocytes</cell_type> .


Similar alterations were not detected in transcripts of the other genes which were analysed .


Our findings demonstrate that truncated TSG101 and FHIT transcripts are commonly detected in both normal and malignant tissues and that a significant fraction of these are likely to be the result of aberrant splicing .

Our findings demonstrate that <rna>truncated TSG101 and FHIT transcripts</rna> are commonly detected in both normal and malignant tissues and that a significant fraction of these are likely to be the result of aberrant splicing .


While we can not exclude that alterations in TSG101 and FHIT occur during cancer development , our data indicate that in this context the commonly observed transcript abnormalities are misleading .

While we can not exclude that alterations in <dna>TSG101</dna> and <dna>FHIT</dna> occur during cancer development , our data indicate that in this context the commonly observed transcript abnormalities are misleading .


Inhibition of proliferation and apoptosis of human and rat T lymphocytes by curcumin , a curry pigment .

Inhibition of proliferation and apoptosis of <cell_type>human and rat T lymphocytes</cell_type> by curcumin , a curry pigment .


Curcumin ( diferuoylmethane ) , the yellow pigment in the rhizome of tumeric ( Curcuma longa ) , an ingredient of curry spice , is known to exhibit a variety of pharmacological effects including antitumor , antiinflammatory , and antiinfectious activities .


Although its precise mode of action remains elusive , curcumin has been shown to suppress the activity of the AP-1 transcription factor in cells stimulated to proliferate .

Although its precise mode of action remains elusive , curcumin has been shown to suppress the activity of the <protein>AP-1</protein> <protein>transcription factor</protein> in cells stimulated to proliferate .


In this study , we observed that curcumin ( 50 microM ) inhibited proliferation of rat thymocytes stimulated with concanavalin A ( Con A ) as well as that of human Jurkat lymphoblastoid cells in the logarithmic growth phase .

In this study , we observed that curcumin ( 50 microM ) inhibited proliferation of <cell_type>rat thymocytes</cell_type> stimulated with concanavalin A ( Con A ) as well as that of <cell_type>human Jurkat lymphoblastoid cells</cell_type> in the logarithmic growth phase .


The pigment also inhibited apoptosis in dexamethasone-treated rat thymocytes and in UV-irradiated Jurkat cells as judged by DNA ladder formation , cellular morphological changes , and flow cytometry analysis .

The pigment also inhibited apoptosis in <cell_line>dexamethasone-treated rat thymocytes</cell_line> and in <cell_line>UV-irradiated Jurkat cells</cell_line> as judged by DNA ladder formation , cellular morphological changes , and flow cytometry analysis .


The inhibition of apoptosis by curcumin in rat thymocytes was accompanied by partial suppression of AP-1 activity .

The inhibition of apoptosis by curcumin in <cell_type>rat thymocytes</cell_type> was accompanied by partial suppression of <protein>AP-1</protein> activity .


Complete suppression of AP-1 activity was observed in Con A-treated , proliferating thymocytes .

Complete suppression of <protein>AP-1</protein> activity was observed in <cell_line>Con A-treated , proliferating thymocytes</cell_line> .


The capacity of curcumin to inhibit both cell growth and death strongly implies that these two biological processes share a common pathway at some point and that curcumin affects a common step , presumably involving a modulation of the AP-1 transcription factor .

The capacity of curcumin to inhibit both cell growth and death strongly implies that these two biological processes share a common pathway at some point and that curcumin affects a common step , presumably involving a modulation of the <protein>AP-1</protein> <protein>transcription factor</protein> .


Interleukin-7 upregulates the interleukin-2-gene expression in activated human T lymphocytes at the transcriptional level by enhancing the DNA binding activities of both nuclear factor of activated T cells and activator protein-1 .

<protein>Interleukin-7</protein> upregulates the <dna>interleukin-2-gene</dna> expression in <cell_type>activated human T lymphocytes</cell_type> at the transcriptional level by enhancing the DNA binding activities of both nuclear factor of <cell_type>activated T cells</cell_type> and <protein>activator protein-1</protein> .


In the present report , we studied the role of the stromal-derived cytokine interleukin-7 ( IL-7 ) in the IL-2-gene regulation in activated T lymphocytes .

In the present report , we studied the role of the <protein>stromal-derived cytokine interleukin-7</protein> ( <protein>IL-7</protein> ) in the <dna>IL-2-gene</dna> regulation in <cell_type>activated T lymphocytes</cell_type> .


Production of IL-2 requires the formation of transcription factors involved in the IL-2-gene regulation .

Production of <protein>IL-2</protein> requires the formation of <protein>transcription factors</protein> involved in the <dna>IL-2-gene</dna> regulation .


T-cell receptor ( TCR ) /CD3 engagement results in the activation of nuclear factor of activated T cells ( NFAT ) , activator protein-1 ( AP-1 ) , and nuclear factor kappaB ( NFkappaB ) , whereas the CD28 responsive complex ( CD28RC ) is activated in response to the CD28 signal .

<protein>T-cell receptor</protein> <protein>( TCR ) /CD3</protein> engagement results in the activation of nuclear factor of <cell_type>activated T cells</cell_type> ( <protein>NFAT</protein> ) , <protein>activator protein-1</protein> ( <protein>AP-1</protein> ) , and <protein>nuclear factor kappaB</protein> ( <protein>NFkappaB</protein> ) , whereas the <protein>CD28 responsive complex</protein> ( <protein>CD28RC</protein> ) is activated in response to the <protein>CD28</protein> signal .


Costimulation of phytohemagglutinin/anti-CD28 activated T lymphocytes with IL-7 induces a fivefold enhanced IL-2 -mRNA accumulation and a 2.5-fold enhanced protein secretion .

Costimulation of <cell_line>phytohemagglutinin/anti-CD28 activated T lymphocytes</cell_line> with <protein>IL-7</protein> induces a fivefold enhanced <protein>IL-2</protein> -mRNA accumulation and a 2.5-fold enhanced protein secretion .


The IL-2-gene transcription rate is increased 3.4-fold , indicating that the effect of IL-7 is in part mediated at the transcriptional level .

The <dna>IL-2-gene</dna> transcription rate is increased 3.4-fold , indicating that the effect of <protein>IL-7</protein> is in part mediated at the transcriptional level .


The molecular mechanisms underlying the IL-7 effect involve the upregulation of the DNA binding activity of NFAT ( 60 % ) and AP-1 ( 120 % ) , without affecting the activities of NFkappaB and CD28RC , which was confirmed by transfection assays .

The molecular mechanisms underlying the <protein>IL-7</protein> effect involve the upregulation of the DNA binding activity of <protein>NFAT</protein> ( 60 % ) and <protein>AP-1</protein> ( 120 % ) , without affecting the activities of <protein>NFkappaB</protein> and <protein>CD28RC</protein> , which was confirmed by transfection assays .


We also show that the IL-7 -induced enhancement of the AP-1 -DNA binding activity is not cyclosporin A-sensitive .

We also show that the <protein>IL-7</protein> -induced enhancement of the <protein>AP-1</protein> -DNA binding activity is not cyclosporin A-sensitive .


Since AP-1 is part of the NFAT complex , we conclude that the IL-7 -signaling pathway is involved in the activation of the fos and jun proteins of which AP-1 consists .

Since <protein>AP-1</protein> is part of the <protein>NFAT complex</protein> , we conclude that the <protein>IL-7</protein> -signaling pathway is involved in the activation of the <protein>fos and jun proteins</protein> of which <protein>AP-1</protein> consists .


The spatial distribution of human immunoglobulin genes within the nucleus : evidence for gene topography independent of cell type and transcriptional activity .

The spatial distribution of <dna>human immunoglobulin genes</dna> within the nucleus : evidence for gene topography independent of cell type and transcriptional activity .


The three-dimensional positioning of immunoglobulin ( Ig ) genes within the nucleus of human cells was investigated using in situ hybridization and confocal microscopy .

The three-dimensional positioning of <dna>immunoglobulin ( Ig ) genes</dna> within the nucleus of <cell_type>human cells</cell_type> was investigated using in situ hybridization and confocal microscopy .


The visualization of heavy and light chain genes in B-lymphoid cells showed that the three Ig genes are differentially and nonrandomly distributed in different nuclear subvolumes : the kappa genes were found to be preferentially confined to an outer nuclear volume , whereas the gamma and lambda genes consistently occupied more central positions within the nucleus , the lambda genes being more interior when compared with the gamma genes .

The visualization of <dna>heavy and light chain genes</dna> in <cell_type>B-lymphoid cells</cell_type> showed that the three <dna>Ig genes</dna> are differentially and nonrandomly distributed in different nuclear subvolumes : the <dna>kappa genes</dna> were found to be preferentially confined to an outer nuclear volume , whereas the <dna>gamma and lambda genes</dna> consistently occupied more central positions within the nucleus , the <dna>lambda genes</dna> being more interior when compared with the <dna>gamma genes</dna> .


The data further show that these overall topographical distributions are independent of gene transcriptional activity and are conserved in different cell types .

The data further show that these overall topographical distributions are independent of gene transcriptional activity and are conserved in <cell_type>different cell types</cell_type> .


Although subtle gene movements within those defined topographical regions can not be excluded by this study , the results indicate that tissue specificity of gene expression is not accompanied by drastic changes in gene nuclear topography , rather suggesting that gene organization within the nucleus may be primarily dependent on structural constraints imposed on the respective chromosomes .

Although subtle gene movements within those defined topographical regions can not be excluded by this study , the results indicate that tissue specificity of gene expression is not accompanied by drastic changes in gene nuclear topography , rather suggesting that gene organization within the nucleus may be primarily dependent on structural constraints imposed on the respective <dna>chromosomes</dna> .


Dominant cytotoxic T lymphocyte response to the immediate-early trans-activator protein , BZLF1 , in persistent type A or B Epstein-Barr virus infection .

Dominant <cell_type>cytotoxic T lymphocyte</cell_type> response to the <protein>immediate-early trans-activator protein</protein> , <protein>BZLF1</protein> , in persistent type A or B Epstein-Barr virus infection .