Moreover , using a heterologous promoter ( herpes thymidine kinase ) construct containing the NF-kappa B consensus sequence , it is shown that NF-kappa B binding by an IL-2-induced monocyte-derived nuclear protein confers reporter gene ( human growth hormone ) activity .

Moreover , using a <dna>heterologous promoter ( herpes thymidine kinase ) construct</dna> containing the <dna>NF-kappa B consensus sequence</dna> , it is shown that <protein>NF-kappa B</protein> binding by an <protein>IL-2-induced monocyte-derived nuclear protein</protein> confers reporter gene ( human growth hormone ) activity .


Taken together , our findings indicate that IL-2 induces gene expression of M-CSF in human blood-derived Mo and provide evidence for involvement of NF-kappa B in transcriptional regulation of this gene .

Taken together , our findings indicate that <protein>IL-2</protein> induces gene expression of <protein>M-CSF</protein> in <cell_type>human blood-derived Mo</cell_type> and provide evidence for involvement of <protein>NF-kappa B</protein> in transcriptional regulation of this gene .


Regulation of the interleukin-1 beta ( IL-1 beta ) gene by mycobacterial components and lipopolysaccharide is mediated by two nuclear factor-IL6 motifs .

Regulation of the <dna>interleukin-1 beta ( IL-1 beta ) gene</dna> by <protein>mycobacterial components</protein> and lipopolysaccharide is mediated by two <dna>nuclear factor-IL6 motifs</dna> .


The cytokines interleukin-1 beta ( IL-1 beta ) and tumor necrosis factor alpha ( TNF-alpha ) are released by mononuclear phagocytes in vitro after stimulation with mycobacteria and are considered to mediate pathophysiologic events , including granuloma formation and systemic symptoms .

The cytokines interleukin-1 beta ( <protein>IL-1 beta</protein> ) and tumor necrosis factor alpha ( <protein>TNF-alpha</protein> ) are released by <cell_type>mononuclear phagocytes</cell_type> in vitro after stimulation with mycobacteria and are considered to mediate pathophysiologic events , including granuloma formation and systemic symptoms .


We demonstrated that the Mycobacterium tuberculosis cell wall component lipoarabinomannan ( LAM ) is a very potent inducer of IL-1 beta gene expression in human monocytes and investigated the mechanism of this effect .

We demonstrated that the Mycobacterium tuberculosis cell wall component lipoarabinomannan ( LAM ) is a very potent inducer of <dna>IL-1 beta gene</dna> expression in <cell_type>human monocytes</cell_type> and investigated the mechanism of this effect .


We localized the LAM- , lipopolysaccharide ( LPS ) - , and TNF-alpha -inducible promoter activity to a -131/+15 ( positions -131 to +15 ) DNA fragment of the IL-1 beta gene by deletion analysis and chloramphenicol acetyltransferase assay .

We localized the LAM- , lipopolysaccharide ( LPS ) - , and <protein>TNF-alpha</protein> <dna>-inducible promoter</dna> activity to a -131/+15 ( positions -131 to +15 ) DNA fragment of the <dna>IL-1 beta gene</dna> by deletion analysis and <protein>chloramphenicol acetyltransferase</protein> assay .


Within this DNA fragment , there were two novel 9-bp motifs ( -90/-82 and -40/-32 ) with high homology to the nuclear factor-IL6 ( NF-IL6 ) binding site .

Within this DNA fragment , there were two novel 9-bp motifs ( -90/-82 and -40/-32 ) with high homology to the <dna>nuclear factor-IL6 ( NF-IL6 ) binding site</dna> .


Site-directed mutagenesis demonstrated that the two NF-IL-6 motifs could be independently activated by LAM , LPS , or TNF-alpha and that they acted in an orientation-independent manner .

Site-directed mutagenesis demonstrated that the two <dna>NF-IL-6 motifs</dna> could be independently activated by LAM , LPS , or <protein>TNF-alpha</protein> and that they acted in an orientation-independent manner .


DNA mobility shift assay revealed specific binding of nuclear protein ( s ) from LAM- , LPS- , or TNF-alpha- stimulated THP-1 cells to the NF-IL6 motifs .

DNA mobility shift assay revealed specific binding of <protein>nuclear protein</protein> ( s ) from <cell_line>LAM- , LPS- , or TNF-alpha- stimulated THP-1 cells</cell_line> to the NF-IL6 motifs .


We conclude that the two NF-IL6 sites mediate induction of IL-1 beta in response to the stimuli LAM , LPS , and TNF-alpha .

We conclude that the two <dna>NF-IL6 sites</dna> mediate induction of <protein>IL-1 beta</protein> in response to the stimuli LAM , LPS , and <protein>TNF-alpha</protein> .


HIV-1 Nef protein inhibits the recruitment of AP-1 DNA-binding activity in human T-cells .

<protein>HIV-1 Nef protein</protein> inhibits the recruitment of <protein>AP-1</protein> DNA-binding activity in <cell_type>human T-cells</cell_type> .


The human immunodeficiency virus type 1 long terminal repeat , HIV-1-LTR , contains binding sites for several cellular transcription factors which contribute to HIV-1 gene expression .

The <dna>human immunodeficiency virus type 1 long terminal repeat</dna> , <dna>HIV-1-LTR</dna> , contains binding sites for several cellular transcription factors which contribute to <dna>HIV-1 gene</dna> expression .


Our previous studies on the function of the HIV-1-encoded Nef protein suggested that Nef may be an inhibitor HIV-1 transcription .

Our previous studies on the function of the <protein>HIV-1-encoded Nef protein</protein> suggested that <protein>Nef</protein> may be an inhibitor HIV-1 transcription .


To determine whether Nef affects the binding of cellular factors implicated in HIV-1 regulation , 32P-labeled oligonucleotides corresponding to the binding sites were incubated with nuclear extracts prepared from Nef-expressing T-cell lines that were not stimulated or were stimulated with T-cell mitogens .

To determine whether <protein>Nef</protein> affects the binding of <protein>cellular factors</protein> implicated in HIV-1 regulation , 32P-labeled oligonucleotides corresponding to the binding sites were incubated with nuclear extracts prepared from <cell_line>Nef-expressing T-cell lines</cell_line> that were not stimulated or were stimulated with <protein>T-cell mitogens</protein> .


We found that Nef inhibited the recruitment of AP-1 DNA-binding activity in mitogen-stimulated human T-cells .

We found that <protein>Nef</protein> inhibited the recruitment of <protein>AP-1</protein> DNA-binding activity in mitogen-stimulated <cell_type>human T-cells</cell_type> .


Additionally , Nef expressing cells were transiently transfected with a plasmid in which HIV-1 AP-1 DNA recognition sequences were cloned downstream of the chloramphenicol acetyltransferase ( CAT ) gene .

Additionally , <cell_line>Nef expressing cells</cell_line> were transiently transfected with a plasmid in which <dna>HIV-1 AP-1 DNA recognition sequences</dna> were cloned downstream of the <protein>chloramphenicol acetyltransferase</protein> ( <protein>CAT</protein> ) gene .


Mitogen-mediated transcriptional activation of the CAT gene in this construct was inhibited in Nef-expressing cells but not in control cells .

Mitogen-mediated transcriptional activation of the <dna>CAT gene</dna> in this construct was inhibited in <cell_line>Nef-expressing cells</cell_line> but not in control cells .


These studies suggest that , by inhibiting AP-1 activation , Nef may play a role in regulating HIV-1 gene expression in infected T-cells .

These studies suggest that , by inhibiting <protein>AP-1</protein> activation , <protein>Nef</protein> may play a role in regulating <dna>HIV-1 gene</dna> expression in <cell_type>infected T-cells</cell_type> .


Regulation of the beta-globin locus .

Regulation of the <dna>beta-globin locus</dna> .


Transcription of the human beta-globin gene cluster depends upon upstream regulatory sequences , which are collectively termed the locus control region .

Transcription of the <dna>human beta-globin gene cluster</dna> depends upon <dna>upstream regulatory sequences</dna> , which are collectively termed the <dna>locus control region</dna> .


Recent studies have provided new insights into how the individual genes of the cluster are regulated through development .


The crux of transcriptional activation is how the locus control region communicates with the gene-proximal regulatory elements .

The crux of transcriptional activation is how the <dna>locus control region</dna> communicates with the <dna>gene-proximal regulatory elements</dna> .


Enhancing effect of 17 beta-estradiol on human NK cell activity .


The in vitro effect of 17 beta-estradiol on NK activity was studied .


The proliferation and NK activity of YT-N17 ( a human NK-like cell line ) were enhanced by 17 beta-estradiol ( E2 ) , and the enhancement was blocked by tamoxifen ( Tx ) , an antagonist of E2 .

The proliferation and NK activity of <cell_line>YT-N17</cell_line> ( a <cell_line>human NK-like cell line</cell_line> ) were enhanced by 17 beta-estradiol ( E2 ) , and the enhancement was blocked by tamoxifen ( Tx ) , an antagonist of E2 .


On the contrary , other steroid hormones such as Tx , progesterone , and testosterone had no effect .


YT-N17 contained 11.8 fmol/mg protein of estrogen receptor ( mean of two independent assays ) , a value which was 5-10-fold higher than that of other hematopoietic cell lines .

<cell_line>YT-N17</cell_line> contained 11.8 fmol/mg protein of <protein>estrogen receptor</protein> ( mean of two independent assays ) , a value which was 5-10-fold higher than that of other <cell_line>hematopoietic cell lines</cell_line> .


An enhancement of NK activity by E2 was also seen in large granular lymphocytes obtained from normal subjects , and it was again suppressed by Tx .

An enhancement of NK activity by E2 was also seen in <cell_type>large granular lymphocytes</cell_type> obtained from normal subjects , and it was again suppressed by Tx .


These data suggest that E2 is one of the activating factors for NK/LGL cells .

These data suggest that E2 is one of the activating factors for <cell_line>NK/LGL cells</cell_line> .


A serum response element and a binding site for NF-Y mediate the serum response of the human thrombospondin 1 gene .

A <dna>serum response element</dna> and a <dna>binding site for NF-Y</dna> mediate the serum response of the <dna>human thrombospondin 1 gene</dna> .


The expression of thrombospondin 1 ( TSP 1 ) , a member of the TSP gene family , is rapidly induced by growth factors .

The expression of <protein>thrombospondin 1</protein> ( <protein>TSP 1</protein> ) , a member of the <dna>TSP gene family</dna> , is rapidly induced by growth factors .


We tested the ability of human TSP 1-chloramphenicol acetyltransferase constructs to respond to serum in stably transfected NIH-3T3 cells .

We tested the ability of <dna>human TSP 1-chloramphenicol acetyltransferase constructs</dna> to respond to serum in stably transfected NIH-3T3 cells .


Two transcriptional elements in the TSP 1 promoter , a distal element at -1280 and a proximal element at -65 , were required for the response of the human TSP 1 gene to serum .

Two transcriptional elements in the <protein>TSP 1</protein> promoter , a distal element at -1280 and a proximal element at -65 , were required for the response of the human TSP 1 gene to serum .


The distal element contains the 5'-CC ( A + T ) 6GG-3 ' consensus sequence characteristic of a serum-response element ( SRE ) .


Deletions or mutations in this element reduced the serum response of the TSP 1 gene by 80-90 % .

Deletions or mutations in this element reduced the serum response of the <dna>TSP 1 gene</dna> by 80-90 % .


In gel-shift assays , the -1280 element and the c-fos SRE cross -competed , whereas their functional and binding mutants did not .

In gel-shift assays , the <dna>-1280 element</dna> and the <dna>c-fos SRE cross</dna> -competed , whereas their functional and <dna>binding mutants</dna> did not .


The proximal element contains the sequence 5'-GGCCAATGGG-3 ' , which closely resembles the consensus binding motif for the CCAAT-binding factor NF-Y ( CBF , CP1 , alpha CP1 ) .

The <dna>proximal element</dna> contains the sequence 5'-GGCCAATGGG-3 ' , which closely resembles the consensus binding motif for the <dna>CCAAT-binding factor</dna> <protein>NF-Y</protein> ( <protein>CBF</protein> , <protein>CP1</protein> , <protein>alpha CP1</protein> ) .


Deletions or mutations in this element also reduced the serum response by 80-90 % .


Methylation interference analysis of the -65 region identified a pattern of contacts with nuclear factors resembling that for NF-Y , and an NF-Y-binding site and the proximal TSP 1 element cross-competed in gel-shift assays , whereas their binding mutants did not .

Methylation interference analysis of the <dna>-65 region</dna> identified a pattern of contacts with <protein>nuclear factors</protein> resembling that for <protein>NF-Y</protein> , and an <dna>NF-Y-binding site</dna> and the <dna>proximal TSP 1 element</dna> cross-competed in gel-shift assays , whereas their <dna>binding mutants</dna> did not .


Finally , an abbreviated TSP 1 promoter/5'-flank , containing the SRE- and NF-Y-binding sites , mediated a serum response that was close in magnitude to that of the parent promoter .

Finally , an <dna>abbreviated TSP 1 promoter/5'-flank</dna> , containing the <dna>SRE- and NF-Y-binding sites</dna> , mediated a serum response that was close in magnitude to that of the <dna>parent promoter</dna> .


We conclude that the serum response of the human TSP 1 gene requires the coordinated function of an SRE- and NF-Y-binding site .

We conclude that the serum response of the human <dna>TSP 1 gene</dna> requires the coordinated function of an <dna>SRE- and NF-Y-binding site</dna> .


Cell-type-specific transactivation of the parathyroid hormone-related protein gene promoter by the human T-cell leukemia virus type I ( HTLV-I ) tax and HTLV-II tax proteins .

Cell-type-specific transactivation of the <dna>parathyroid hormone-related protein gene promoter</dna> by the <protein>human T-cell leukemia virus type I ( HTLV-I ) tax</protein> and <protein>HTLV-II tax proteins</protein> .


The human T-cell leukemia virus type I ( HTLV-I ) and HTLV-II Tax proteins are potent transactivators of viral and cellular gene expression .

The <protein>human T-cell leukemia virus type I ( HTLV-I ) and HTLV-II Tax proteins</protein> are potent <protein>transactivators</protein> of viral and cellular gene expression .


Using deletion mutants , the downstream parathyroid hormone-related protein ( PTHrP ) promoter is shown to be responsive to both HTLV-I and HTLV-II Tax as well as the AP1/c-jun proto-oncogene .

Using <dna>deletion mutants</dna> , the downstream <dna>parathyroid hormone-related protein ( PTHrP ) promoter</dna> is shown to be responsive to both <protein>HTLV-I and HTLV-II Tax</protein> as well as the <dna>AP1/c-jun proto-oncogene</dna> .


Transactivation of PTHrP by Tax was seen in T cells but not in B-cell lines or fibroblasts .

Transactivation of <protein>PTHrP</protein> by <protein>Tax</protein> was seen in <cell_type>T cells</cell_type> but not in <cell_line>B-cell lines</cell_line> or <cell_line>fibroblasts</cell_line> .


A carboxy terminal Tax deletion mutant was deficient in transactivation of both the PTHrP and IL2R alpha promoters but not the HTLV-I long terminal repeat ( LTR ) .

A <dna>carboxy terminal Tax deletion mutant</dna> was deficient in transactivation of both the <dna>PTHrP and IL2R alpha promoters</dna> but not the <dna>HTLV-I long terminal repeat</dna> ( <dna>LTR</dna> ) .


Exogenous provision of NFkB rescued IL2R alpha expression but not the PTHrP promoter .

Exogenous provision of <protein>NFkB</protein> rescued <protein>IL2R</protein> alpha expression but not the <dna>PTHrP promoter</dna> .


Thus , HTLV-I Tax , HTLV-II Tax , and c-jun transactivate PTHrP and may contribute to the pathogenesis of hypercalcemia in adult T-cell leukemia .

Thus , <protein>HTLV-I Tax</protein> , <protein>HTLV-II Tax</protein> , and <protein>c-jun</protein> transactivate <protein>PTHrP</protein> and may contribute to the pathogenesis of hypercalcemia in adult T-cell leukemia .


Involvement of Alu sequences in the cell-specific regulation of transcription of the gamma chain of Fc and T cell receptors .

Involvement of <dna>Alu sequences</dna> in the cell-specific regulation of transcription of the <protein>gamma chain</protein> of <protein>Fc</protein> and <protein>T cell receptors</protein> .


The Fc epsilon RI-gamma chains are expressed in a variety of hematopoietic cells where they play a critical role in signal transduction .

The <protein>Fc epsilon RI-gamma chains</protein> are expressed in a variety of <cell_type>hematopoietic cells</cell_type> where they play a critical role in signal transduction .


They are part of the high affinity IgE receptor in mast cells , basophils , Langerhans cells , and possibly other cells ; a component of the low affinity receptor for IgG ( Fc gamma RIIIA or CD16 ) in natural killer cells and macrophages ; and part of the T cell antigen receptor in subsets of T cells .

They are part of the <protein>high affinity IgE receptor</protein> in <cell_type>mast cells</cell_type> , <cell_type>basophils</cell_type> , <cell_type>Langerhans cells</cell_type> , and possibly other cells ; a component of the low affinity receptor for <protein>IgG</protein> ( <protein>Fc gamma RIIIA</protein> or <protein>CD16</protein> ) in natural <cell_type>killer cells</cell_type> and <cell_type>macrophages</cell_type> ; and part of the <protein>T cell antigen receptor</protein> in subsets of <cell_type>T cells</cell_type> .


Here we have investigated the transcriptional regulation of the gamma chain gene by analyzing the 2.5-kilobase sequence upstream of the transcription start site .

Here we have investigated the transcriptional regulation of the <protein>gamma chain</protein> gene by analyzing the <dna>2.5-kilobase sequence</dna> upstream of the transcription start site .


This sequence contains a promoter specific to cells of hematopoietic lineage .

This sequence contains a <dna>promoter</dna> specific to <cell_type>cells of hematopoietic lineage</cell_type> .


However , the tissue specificity of this promoter is only partial because it is active in all of the hematopoietic cells tested here , regardless of whether they constitutively express Fc epsilon RI- gamma chain transcripts .

However , the tissue specificity of this <dna>promoter</dna> is only partial because it is active in all of the <cell_type>hematopoietic cells</cell_type> tested here , regardless of whether they constitutively express Fc epsilon RI- <protein>gamma chain</protein> transcripts .


We have identified two adjacent cis-acting regulatory elements , both of which are part of an Alu repeat .


The first ( -445/-366 ) is a positive element active in both basophils and T cells .

The first ( -445/-366 ) is a positive element active in both <cell_type>basophils</cell_type> and <cell_type>T cells</cell_type> .


The second ( -365/-264 ) binds to nuclear factors , which appear to be different in basophils and T cells , and acts as a negative element in basophils and as a positive one in T cells .

The second ( -365/-264 ) binds to <protein>nuclear factors</protein> , which appear to be different in <cell_type>basophils</cell_type> and <cell_type>T cells</cell_type> , and acts as a negative element in <cell_type>basophils</cell_type> and as a positive one in <cell_type>T cells</cell_type> .


Thus , this Alu repeat ( 90 % identical to Alu consensus sequences ) has evolved to become both a positive and negative regulator .

Thus , this <dna>Alu repeat</dna> ( 90 % identical to <dna>Alu consensus sequences</dna> ) has evolved to become both a positive and negative regulator .


Chlorinated dibenzo-p-dioxins and dibenzofurans and the human immune system .


1 . Blood cell receptors in volunteers with moderately increased body burdens .

1 . <protein>Blood cell receptors</protein> in volunteers with moderately increased body burdens .


Using monoclonal antibodies ( mAbs ) and flow cytometry , we studied a variety of surface receptors on lymphocyte subpopulations of workers with moderately increased body burdens of 2 , 3 , 7 , 8-tetrachlorodibenzo-p-dioxin ( TCDD ) and of other polychlorinated dibenzo-p-dioxins and dibenzofurans ( PCDD/PCDF ) , expressed here as International-Toxicity Equivalencies ( I-TE ) .

Using <protein>monoclonal antibodies</protein> ( <protein>mAbs</protein> ) and flow cytometry , we studied a variety of <protein>surface receptors</protein> on <cell_type>lymphocyte subpopulations</cell_type> of workers with moderately increased body burdens of 2 , 3 , 7 , 8-tetrachlorodibenzo-p-dioxin ( TCDD ) and of other polychlorinated dibenzo-p-dioxins and dibenzofurans ( PCDD/PCDF ) , expressed here as International-Toxicity Equivalencies ( I-TE ) .


The hypothesis to be tested was whether or not humans exhibit a similar susceptibility to PCDDs/PCDFs with respect to the surface receptors found previously to respond to small doses of 2 , 3 , 7 , 8-tetrachlorodibenzo-p-dioxin ( TCDD ) in Callithrix jacchus .

The hypothesis to be tested was whether or not humans exhibit a similar susceptibility to PCDDs/PCDFs with respect to the <protein>surface receptors</protein> found previously to respond to small doses of 2 , 3 , 7 , 8-tetrachlorodibenzo-p-dioxin ( TCDD ) in Callithrix jacchus .


These are : helper-inducer ( memory ) T cells ( CD4+CD45R0+CD45RA-CD29highCD11a+ ) , CD20+ B cells , and cytotoxic T cells ( CD8+CD56+/CD57+ ) .

These are : <cell_type>helper-inducer ( memory ) T cells</cell_type> ( <cell_type>CD4+CD45R0+CD45RA-CD29highCD11a+</cell_type> ) , <cell_type>CD20+ B cells</cell_type> , and <cell_type>cytotoxic T cells</cell_type> ( <cell_type>CD8+CD56+/CD57+</cell_type> ) .


Furthermore , 68 triple-labellings with mAbs were performed on the cells of each volunteer to possibly generate further hypotheses .

Furthermore , 68 triple-labellings with <protein>mAbs</protein> were performed on the cells of each volunteer to possibly generate further hypotheses .


It was evaluated whether any of the variables might be used as a biomarker of effects for this class of compounds .


There were two main goals : ( 1 ) to evaluate whether workers with a moderately increased PCDD/PCDF-body burden [ 25-140 ppt TCDD or 104-522 ppt I-TE in blood fat ] exhibit changes in the surface receptors of white blood cells , as observed in previous studies in non-human primates , and ( 2 ) to clarify whether persons at the upper range [ 10-23 ppt TCDD or 30-90 ppt I-TE in blood fat ] of the body burden reference values of a not particularly exposed population show detectable deviations in these immunological variables , when compared with persons at the lower and medium range [ 1-3 ppt TCDD or 9-29 ppt I-TE ] of these body burden reference values .

There were two main goals : ( 1 ) to evaluate whether workers with a moderately increased PCDD/PCDF-body burden [ 25-140 ppt TCDD or 104-522 ppt I-TE in blood fat ] exhibit changes in the <protein>surface receptors</protein> of <cell_type>white blood cells</cell_type> , as observed in previous studies in non-human primates , and ( 2 ) to clarify whether persons at the upper range [ 10-23 ppt TCDD or 30-90 ppt I-TE in blood fat ] of the body burden reference values of a not particularly exposed population show detectable deviations in these immunological variables , when compared with persons at the lower and medium range [ 1-3 ppt TCDD or 9-29 ppt I-TE ] of these body burden reference values .


Regression analysis of our data revealed slight trends for some of the biomarkers ( e.g. CD45R0+ ) .


With one exception , these were all increases .


of the alterations observed are of medical relevance .


The slight increase in the percentage of CD4+CD45R0+ cells remained significant even after covariant analysis taking age-related changes into account .

The slight increase in the percentage of <cell_type>CD4+CD45R0+ cells</cell_type> remained significant even after covariant analysis taking age-related changes into account .


Altogether , the data do not provide any evidence to support an assumption that moderately increased body burdens of PCDDs/PCDFs in adults induce decreases in the cellular components of the human immune system .


Adult humans certainly are less susceptible to this action of PCDDs/PCDFs than adolescent Callithrix jacchus


Inhibition of HIV-1 latency reactivation by dehydroepiandrosterone ( DHEA ) and an analog of DHEA .


The initial infection with human immunodeficiency virus type 1 ( HIV-1 ) in most individuals usually results in the establishment of a latent or chronic infection before eventual progression toward acquired immunodeficiency syndrome .


HIV-1 can also establish a latent or persistent infection in some T cell lines that show minimal constitutive virus expression .

HIV-1 can also establish a latent or persistent infection in some <cell_line>T cell lines</cell_line> that show minimal constitutive virus expression .


However , activation of the T cell lines leading to enhanced HIV-1 replication can be induced by antigens , mitogens , and cytokines ( tumor necrosis factor alpha [ TNF-alpha ] , interleukin 1 , and interleukin-2 ) .

However , activation of the <cell_line>T cell lines</cell_line> leading to enhanced HIV-1 replication can be induced by <protein>antigens</protein> , mitogens , and <protein>cytokines</protein> ( <protein>tumor necrosis factor alpha</protein> [ <protein>TNF-alpha</protein> ] , <protein>interleukin 1</protein> , and <protein>interleukin-2</protein> ) .


Various gene products from other viruses ( HTLV-1 , HSV , EBV , CMV , HBV , and HHV-6 ) can also enhance HIV-1 long terminal repeat ( LTR ) -driven reporter gene activity .

Various gene products from other viruses ( HTLV-1 , HSV , EBV , CMV , HBV , and HHV-6 ) can also enhance HIV-1 <dna>long terminal repeat</dna> ( <dna>LTR</dna> ) -driven <dna>reporter gene</dna> activity .


On the basis of these observations , it has been proposed that reactivation of latent HIV-1 harbored in chronically infected T lymphocytes , monocytes , or macrophages plays an important role in the pathogenesis of AIDS .

On the basis of these observations , it has been proposed that reactivation of latent HIV-1 harbored in <cell_type>chronically infected T lymphocytes</cell_type> , <cell_type>monocytes</cell_type> , or <cell_type>macrophages</cell_type> plays an important role in the pathogenesis of AIDS .


So far , there are no drugs or therapy available that can provide protection against HIV-1 latency reactivation .


ACH-2 , derived from a human T cell line ( CEM ) , is chronically infected with HIV-1 , with low levels of constitutive virus expression .

<cell_line>ACH-2</cell_line> , derived from a <cell_line>human T cell line</cell_line> ( <cell_line>CEM</cell_line> ) , is chronically infected with HIV-1 , with low levels of constitutive virus expression .


ACH-2 can be converted to productive infection by stimulation of the cells with 12-O-tetradecanoylphorbol-13-acetate ( TPA ) , mitogen or cytokines ( TNF-alpha ) , or infection with HSV .

<cell_line>ACH-2</cell_line> can be converted to productive infection by stimulation of the cells with 12-O-tetradecanoylphorbol-13-acetate ( TPA ) , mitogen or <protein>cytokines</protein> ( <protein>TNF-alpha</protein> ) , or infection with HSV .


Therefore the ACH-2 cell line is a good candidate for studying the effects of drugs on HIV-1 activation .

Therefore the <cell_line>ACH-2</cell_line> cell line is a good candidate for studying the effects of drugs on HIV-1 activation .


Previously , we have reported that DHEA and synthetic analogs of DHEA can be modest inhibitors of HIV-1 IIIB replication in phytohemagglutinin-stimulated peripheral blood lymphocyte cultures .

Previously , we have reported that DHEA and synthetic analogs of DHEA can be modest inhibitors of HIV-1 IIIB replication in <cell_line>phytohemagglutinin-stimulated peripheral blood lymphocyte cultures</cell_line> .


( ABSTRACT TRUNCATED AT 250 WORDS )


Occurrence of a silencer of the interleukin-2 gene in naive but not in memory resting T helper lymphocytes .

Occurrence of a <dna>silencer</dna> of the <dna>interleukin-2 gene</dna> in naive but not in <cell_type>memory resting T helper lymphocytes</cell_type> .


In the immune system the first activation of a naive T cell by antigen is a key step in the shaping of the peripheral T cell specificity repertoire and maintenance of self-tolerance .

In the immune system the first activation of a <cell_type>naive T cell</cell_type> by antigen is a key step in the shaping of the peripheral T cell specificity repertoire and maintenance of self-tolerance .


In the present study , analysis of the interleukin-2 ( IL-2 ) gene activation shows that naive human helper T cells ( cord blood CD4+ T cells , adult CD4+CD45RO- T cells ) regulate IL-2 transcription by a mechanism involving both a silencer and an activator acting on the purine-rich IL-2 promoter elements ( NF-AT binding sites ) .

In the present study , analysis of the <dna>interleukin-2 ( IL-2 ) gene</dna> activation shows that <cell_type>naive human helper T cells</cell_type> ( <cell_type>cord blood CD4+ T cells</cell_type> , <cell_type>adult CD4+CD45RO- T cells</cell_type> ) regulate <protein>IL-2</protein> transcription by a mechanism involving both a <dna>silencer</dna> and an <dna>activator</dna> acting on the <dna>purine-rich IL-2 promoter elements</dna> ( <dna>NF-AT binding sites</dna> ) .


By contrast , memory cells , either in vitro activated helper T cells reverting to a resting state , or CD4+ T ( memory ) clones , or CD4+CD45RO+ T cells isolated ex vivo , no longer have a silencer .

By contrast , memory cells , either in <cell_line>vitro activated helper T cells</cell_line> reverting to a resting state , or <cell_line>CD4+ T ( memory ) clones</cell_line> , or <cell_line>CD4+CD45RO+ T cells</cell_line> isolated ex vivo , no longer have a <dna>silencer</dna> .


Their IL-2 transcription seems to be controlled solely by the transition from inactive to active functional state of a positive transcription factor binding to these promoter elements as well as its cytoplasmic or nuclear location : in resting memory T cells the activator is located in the cytoplasm and is inactive , whereas in stimulated cells it is functional in promoting transcription and now resides in the nucleus .

Their <protein>IL-2</protein> transcription seems to be controlled solely by the transition from inactive to active functional state of a <protein>positive transcription factor</protein> binding to these promoter elements as well as its cytoplasmic or nuclear location : in <cell_type>resting memory T cells</cell_type> the <dna>activator</dna> is located in the cytoplasm and is inactive , whereas in <cell_line>stimulated cells</cell_line> it is functional in promoting transcription and now resides in the nucleus .


Thus , the regulation of the gene coding for the main T cell growth factor changes irreversibly after the first encounter of T cells with antigen .

Thus , the regulation of the gene coding for the main <protein>T cell growth factor</protein> changes irreversibly after the first encounter of T cells with antigen .


It is most likely that the presence of a silencer contributes to the more stringent activation requirements of naive CD4+ T cells .

It is most likely that the presence of a <dna>silencer</dna> contributes to the more stringent activation requirements of <cell_type>naive CD4+ T cells</cell_type> .


Expression of mRNA for the GATA-binding proteins in human eosinophils and basophils : potential role in gene transcription .

Expression of <rna>mRNA</rna> for the <protein>GATA-binding proteins</protein> in <cell_type>human eosinophils</cell_type> and <cell_type>basophils</cell_type> : potential role in gene transcription .


The expression of the hematopoietic transcription factors GATA-1 , GATA-2 , and GATA-3 was studied in eosinophils and basophils .

The expression of the <protein>hematopoietic transcription factors</protein> <protein>GATA-1</protein> , <protein>GATA-2</protein> , and <protein>GATA-3</protein> was studied in <cell_type>eosinophils</cell_type> and <cell_type>basophils</cell_type> .


Eosinophils express mRNA for GATA-1 , GATA-2 , and GATA-3 .

<cell_type>Eosinophils</cell_type> express <rna>mRNA</rna> for <protein>GATA-1</protein> , <protein>GATA-2</protein> , and <protein>GATA-3</protein> .


Basophils express GATA-2 and GATA-3 .

<cell_type>Basophils</cell_type> express <protein>GATA-2</protein> and <protein>GATA-3</protein> .


Treatment of HL-60 eosinophilic sublines with either interleukin-5 or butyric acid increased the expression of GATA-1 mRNA concomitant with the expression of eosinophil-specific genes , whereas levels of GATA-2 mRNA remained relatively constant .

Treatment of <cell_line>HL-60 eosinophilic sublines</cell_line> with either <protein>interleukin-5</protein> or butyric acid increased the expression of <rna>GATA-1 mRNA</rna> concomitant with the expression of <dna>eosinophil-specific genes</dna> , whereas levels of <rna>GATA-2 mRNA</rna> remained relatively constant .


The presence of mRNA for these proteins in eosinophils and basophils suggests that gene transcription in these lineages may be regulated by GATA-binding proteins .

The presence of <rna>mRNA</rna> for these proteins in <cell_type>eosinophils</cell_type> and <cell_type>basophils</cell_type> suggests that gene transcription in these lineages may be regulated by <protein>GATA-binding proteins</protein> .


Oxidoreductive regulation of nuclear factor kappa B .

Oxidoreductive regulation of <protein>nuclear factor kappa B</protein> .


Involvement of a cellular reducing catalyst thioredoxin .

Involvement of a cellular reducing catalyst <protein>thioredoxin</protein> .


We have investigated an oxidoreductive regulatory pathway for the DNA binding activity of a pleiotropic cellular transcription factor , nuclear factor kappa B ( NF kappa B ) , has been investigated by using NF kappa B prepared from the nucleus and the cytosol of the primary human T lymphocytes .

We have investigated an oxidoreductive regulatory pathway for the DNA binding activity of a <protein>pleiotropic cellular transcription factor</protein> , <protein>nuclear factor kappa B</protein> ( <protein>NF kappa B</protein> ) , has been investigated by using <protein>NF kappa B</protein> prepared from the nucleus and the cytosol of the <cell_type>primary human T lymphocytes</cell_type> .


We show that a cellular reducing catalyst thioredoxin ( Trx ) plays a major role in activation of the DNA binding of NF kappa B in vitro and stimulation of transcription from the NF kappa B -dependent gene expression .

We show that a cellular reducing catalyst <protein>thioredoxin</protein> ( <protein>Trx</protein> ) plays a major role in activation of the DNA binding of <protein>NF kappa B</protein> in vitro and stimulation of transcription from the <protein>NF kappa B</protein> -dependent gene expression .


We demonstrate evidence suggesting that redox regulation of NF kappa B by Trx might be exerted at a step after dissociation of the inhibitory molecule I kappa B , a cytosolic-anchoring protein for NF kappa B .

We demonstrate evidence suggesting that redox regulation of <protein>NF kappa B</protein> by <protein>Trx</protein> might be exerted at a step after dissociation of the inhibitory molecule <protein>I kappa B</protein> , a <protein>cytosolic-anchoring protein</protein> for <protein>NF kappa B</protein> .


To examine the effect of Trx in intact cells , we performed transient assay with a chloramphenicol acetyltransferase-expressing plasmid under the control of human immunodeficiency virus ( HIV ) long terminal repeat and an effector plasmid expressing human Trx .

To examine the effect of <protein>Trx</protein> in intact cells , we performed transient assay with a <dna>chloramphenicol acetyltransferase-expressing plasmid</dna> under the control of <dna>human immunodeficiency virus ( HIV ) long terminal repeat</dna> and an <dna>effector plasmid</dna> expressing <protein>human Trx</protein> .


The promoter activity from HIV long terminal repeat was greatly augmented by co-transfecting the Trx-expressing plasmid , whose effect was dependent on the NF kappa B -binding sites .

The promoter activity from <dna>HIV long terminal repeat</dna> was greatly augmented by co-transfecting the <dna>Trx-expressing plasmid</dna> , whose effect was dependent on the <protein>NF kappa B</protein> -binding sites .


These findings have suggested that cysteine residue ( s ) of NF kappa B might be involved in the DNA-recognition by NF kappa B and that the redox control mechanism mediated by Trx might have a regulatory role in the NF kappa B -mediated gene expression .

These findings have suggested that cysteine residue ( s ) of <protein>NF kappa B</protein> might be involved in the DNA-recognition by <protein>NF kappa B</protein> and that the redox control mechanism mediated by <protein>Trx</protein> might have a regulatory role in the <protein>NF kappa B</protein> -mediated gene expression .


These results may also provide a clue to understanding of the molecular process of AIDS pathogenesis and its possible biochemical intervention .


Synergism between the CD3 antigen- and CD2 antigen-derived signals .


Exploration at the level of induction of DNA-binding proteins and characterization of the inhibitory activity of cyclosporine .

Exploration at the level of induction of <protein>DNA-binding proteins</protein> and characterization of the inhibitory activity of cyclosporine .


We have demonstrated earlier that the crosslinkage of the CD3/TCR complex with the CD2 antigen results in the proliferation of normal human T cells .

We have demonstrated earlier that the crosslinkage of the <protein>CD3/TCR complex</protein> with the <protein>CD2 antigen</protein> results in the proliferation of <cell_type>normal human T cells</cell_type> .


The effect of this synergism was perceptible at the level of induction of the IL-2 gene , a process critical for T cell growth .

The effect of this synergism was perceptible at the level of induction of the <dna>IL-2 gene</dna> , a process critical for T cell growth .


To further understand the molecular and nuclear basis for this synergism , we have explored the induction of DNA-binding proteins in highly purified normal human T cells signaled via the CD3 and/or CD2 proteins .

To further understand the molecular and nuclear basis for this synergism , we have explored the induction of <protein>DNA-binding proteins</protein> in highly purified <cell_type>normal human T cells</cell_type> signaled via the <protein>CD3</protein> and/or <protein>CD2 proteins</protein> .


The effect of transmembrane signaling of T cells with ionomycin , and/or sn-1 , 2 dioctanoyl glycerol , was also determined .

The effect of transmembrane signaling of <cell_type>T cells</cell_type> with ionomycin , and/or sn-1 , 2 dioctanoyl glycerol , was also determined .


The emergence of nuclear binding proteins was investigated using interleukin-2 sequence specific oligonucleotide probes in the electrophoretic mobility shift assay .

The emergence of <protein>nuclear binding proteins</protein> was investigated using <dna>interleukin-2 sequence specific oligonucleotide probes</dna> in the electrophoretic mobility shift assay .


Our studies demonstrate for the first time that CD3 antigen-derived signals and CD2 antigen -derived signals are synergistic in inducing the emergence of transcription factors that bind to the NF-AT1 , AP-1 , and NF-kB sites located in the promoter/enhancer region of the IL-2 gene .

Our studies demonstrate for the first time that <protein>CD3</protein> antigen-derived signals and <protein>CD2 antigen</protein> -derived signals are synergistic in inducing the emergence of <protein>transcription factors</protein> that bind to the <dna>NF-AT1 , AP-1 , and NF-kB sites</dna> located in the <dna>promoter/enhancer region</dna> of the <protein>IL-2</protein> gene .


Moreover , cyclosporine , at concentrations readily accomplished in clinical practice , was found to inhibit the emergence of these DNA-binding proteins in normal human T cells signaled via cell surface proteins implicated in antigen-dependent T cell activation and in T cells stimulated by mobilization of cellular calcium and activation of protein kinase C .

Moreover , cyclosporine , at concentrations readily accomplished in clinical practice , was found to inhibit the emergence of these <protein>DNA-binding proteins</protein> in <cell_type>normal human T cells</cell_type> signaled via cell surface proteins implicated in antigen-dependent T cell activation and in <cell_type>T cells</cell_type> stimulated by mobilization of cellular calcium and activation of <protein>protein kinase C</protein> .


Induced myeloid differentiation of K562 cells with downregulation of erythroid and megakaryocytic transcription factors : a novel experimental model for hemopoietic lineage restriction .

Induced myeloid differentiation of <cell_line>K562 cells</cell_line> with downregulation of <protein>erythroid and megakaryocytic transcription factors</protein> : a novel experimental model for hemopoietic lineage restriction .


The human erythroleukemia cell line K562 can be induced to differentiate along the erythroid and megakaryocytic lineages .

The <cell_line>human erythroleukemia cell line K562</cell_line> can be induced to differentiate along the erythroid and megakaryocytic lineages .


Here we demonstrate that hexamethylene bisacetamide ( HMBA ) induced K562 cells to differentiate along a third pathway .

Here we demonstrate that hexamethylene bisacetamide ( HMBA ) induced <cell_line>K562 cells</cell_line> to differentiate along a third pathway .


This was accompanied by downregulation of two transcription factors normally expressed in erythroid , mast and megakaryocyte lineages .

This was accompanied by downregulation of two <protein>transcription factors</protein> normally expressed in <cell_type>erythroid , mast and megakaryocyte lineages</cell_type> .


Northern analysis demonstrated coordinate downregulation of alpha globin and gamma globin in addition to the two lineage-restricted transcription factors , SCL and GATA-1 .

Northern analysis demonstrated coordinate downregulation of <protein>alpha globin</protein> and <protein>gamma globin</protein> in addition to the two lineage-restricted <protein>transcription factors</protein> , <protein>SCL</protein> and <protein>GATA-1</protein> .


Proliferation of the K562 cells was also suppressed .

Proliferation of the <cell_line>K562 cells</cell_line> was also suppressed .


Clonal assay showed that the suppression was irreversible and appeared analogous to the commitment of murine erythroleukemia ( MEL ) cells to terminal differentiation .

Clonal assay showed that the suppression was irreversible and appeared analogous to the commitment of <cell_line>murine erythroleukemia ( MEL ) cells</cell_line> to terminal differentiation .


In contrast to MEL cells , however , K562 cells acquired a macrophage-like morphology and exhibited a complete failure to generate benzidine-positive cells .

In contrast to <cell_line>MEL cells</cell_line> , however , <cell_line>K562 cells</cell_line> acquired a macrophage-like morphology and exhibited a complete failure to generate <cell_line>benzidine-positive cells</cell_line> .


Electron microscopy revealed a marked increase in granules resembling those specific for eosinophils .

Electron microscopy revealed a marked increase in granules resembling those specific for <cell_type>eosinophils</cell_type> .


Surface marker analysis showed that HMBA-induced cells expressed reduced levels of glycophorin A , CD5 , CD7 and CD11b .

Surface marker analysis showed that <cell_line>HMBA-induced cells</cell_line> expressed reduced levels of <protein>glycophorin A</protein> , <protein>CD5</protein> , <protein>CD7</protein> and <protein>CD11b</protein> .


No upregulation of megakaryocyte or lymphoid markers occurred .

No upregulation of <protein>megakaryocyte or lymphoid markers</protein> occurred .


Thus the response of K562 cells to HMBA may provide a useful experimental system for studying the molecular mechanisms responsible for downmodulation of lineage-restricted transcription factors during hemopoietic lineage commitment .

Thus the response of <cell_line>K562 cells</cell_line> to HMBA may provide a useful experimental system for studying the molecular mechanisms responsible for downmodulation of lineage-restricted <protein>transcription factors</protein> during hemopoietic lineage commitment .


Activation of primary human T-lymphocytes through CD2 plus CD28 adhesion molecules induces long-term nuclear expression of NF-kappa B .

Activation of <cell_type>primary human T-lymphocytes</cell_type> through <protein>CD2 plus CD28 adhesion molecules</protein> induces long-term nuclear expression of <protein>NF-kappa B</protein> .


Stimulation of highly purified human T-cells via CD2 and CD28 adhesion molecules induces and maintains proliferation for more than 3 weeks .

Stimulation of <cell_type>highly purified human T-cells</cell_type> via <protein>CD2 and CD28 adhesion molecules</protein> induces and maintains proliferation for more than 3 weeks .


This potent interleukin 2 ( IL-2 ) -dependent activation does not require monocytes or accessory cells .

This potent <protein>interleukin 2</protein> ( <protein>IL-2</protein> ) -dependent activation does not require <cell_type>monocytes</cell_type> or <cell_type>accessory cells</cell_type> .


Long-lasting IL-2 receptivity is associated with high-level expression of the inducible IL-2 receptor alpha chain ( IL-2R alpha ) gene that is regulated at both transcriptional and posttranscriptional levels .

Long-lasting <protein>IL-2</protein> receptivity is associated with high-level expression of the inducible <dna>IL-2 receptor alpha chain ( IL-2R alpha ) gene</dna> that is regulated at both transcriptional and posttranscriptional levels .


Increase of IL-2R alpha gene transcription involves the enhanced binding of the transcription factor NF-kappa B to its consensus sequence in the 5'-regulatory region of the IL-2R alpha gene .

Increase of <dna>IL-2R alpha gene</dna> transcription involves the enhanced binding of the transcription factor <protein>NF-kappa B</protein> to its <dna>consensus sequence</dna> in the <dna>5'-regulatory region</dna> of the <dna>IL-2R alpha gene</dna> .


To dissect the molecular basis for the unusually persistent transcription of the IL-2R alpha gene , we analyzed nuclear NF-kappa B binding to a radiolabeled IL-2R alpha kappa B-specific oligonucleotide probe during the time course of CD2 + CD28 activation .

To dissect the molecular basis for the unusually persistent transcription of the <dna>IL-2R alpha gene</dna> , we analyzed nuclear <protein>NF-kappa B</protein> binding to a radiolabeled <protein>IL-2R alpha</protein> kappa B-specific oligonucleotide probe during the time course of <protein>CD2</protein> + <protein>CD28</protein> activation .


Resting T-cell nuclear extracts contained KBF1/p50 homodimer .

Resting T-cell nuclear extracts contained <protein>KBF1/p50 homodimer</protein> .


After stimulation , two new kappa B-specific complexes were identified as NF-kappa B p50-p65 heterodimer and putative c-Rel homodimer or c-Rel-p65 heterodimer .

After stimulation , two new <protein>kappa B-specific complexes</protein> were identified as <protein>NF-kappa B</protein> <protein>p50-p65 heterodimer</protein> and putative <protein>c-Rel homodimer</protein> or <protein>c-Rel-p65 heterodimer</protein> .


Both inducible complexes persisted for at least 3 weeks .


Their relative levels were very similar for the duration of proliferation .


In parallel , CD2 + CD28 activation triggered a significant intracellular thiol decrease , suggesting that oxygen radicals are involved in the signaling pathway of adhesion molecules .

In parallel , <protein>CD2</protein> + <protein>CD28</protein> activation triggered a significant intracellular thiol decrease , suggesting that oxygen radicals are involved in the signaling pathway of <protein>adhesion molecules</protein> .


Finally , micromolar amounts of pyrrolidine dithiocarbamate , an oxygen radical scavenger that efficiently blocked the nuclear appearance of NF-kappa B in T-lymphocytes , also inhibited IL-2 secretion , IL-2R alpha cell surface expression , and T-cell proliferation .

Finally , micromolar amounts of pyrrolidine dithiocarbamate , an oxygen radical scavenger that efficiently blocked the nuclear appearance of <protein>NF-kappa B</protein> in <cell_type>T-lymphocytes</cell_type> , also inhibited <protein>IL-2</protein> secretion , <protein>IL-2R alpha</protein> cell surface expression , and T-cell proliferation .


Together , these results suggest that NF-kappa B plays an important role in long-term activation of human primary T-lymphocytes via CD2 + CD28 .

Together , these results suggest that <protein>NF-kappa B</protein> plays an important role in long-term activation of <cell_type>human primary T-lymphocytes</cell_type> via <protein>CD2</protein> + <protein>CD28</protein> .


Immobilization and recovery of fusion proteins and B-lymphocyte cells using magnetic separation .

Immobilization and recovery of <protein>fusion proteins</protein> and <cell_type>B-lymphocyte cells</cell_type> using magnetic separation .


A new approach to facilitate immobilization and affinity purification of recombinant proteins and selected human B lymphocytes has been developed .

A new approach to facilitate immobilization and affinity purification of <protein>recombinant proteins</protein> and selected <cell_type>human B lymphocytes</cell_type> has been developed .


Using magnetic beads with attached DNA containing the Escherichia coli lac operator , fusion proteins comprising the DNA-binding lac repressor could be affinity-purified and recovered by gentle elution conditions , such as with a lactose analogue or by enzymatic means using either deoxyribonuclease ( DNase ) or restriction endonucleases .

Using magnetic beads with attached DNA containing the <dna>Escherichia coli lac operator</dna> , <protein>fusion proteins</protein> comprising the <dna>DNA-binding lac repressor</dna> could be affinity-purified and recovered by gentle elution conditions , such as with a lactose analogue or by enzymatic means using either <protein>deoxyribonuclease</protein> ( <protein>DNase</protein> ) or <protein>restriction endonucleases</protein> .


The results show for the first time that a DNA-binding protein can be used for affinity purification of fusion proteins as exemplified by the specific and gentle recovery of beta-galactosidase and alkaline phosphatase from bacterial lysates using immunomagnetic separation .

The results show for the first time that a <protein>DNA-binding protein</protein> can be used for affinity purification of <protein>fusion proteins</protein> as exemplified by the specific and gentle recovery of <protein>beta-galactosidase</protein> and <protein>alkaline phosphatase</protein> from bacterial lysates using immunomagnetic separation .


The approach was further extended to cell separation by the efficient recovery and elution of human CD37 B lymphocytes from peripheral blood .

The approach was further extended to cell separation by the efficient recovery and elution of <cell_line>human CD37 B lymphocytes</cell_line> from peripheral blood .


ras protein activity is essential for T-cell antigen receptor signal transduction .

<protein>ras protein</protein> activity is essential for <protein>T-cell antigen receptor</protein> signal transduction .


In a Jurkat cell model of T-cell activation an interleukin-2 promoter/reporter gene construct was activated by antigen receptor agonism in combination with the lymphokine interleukin-1 .

In a <cell_line>Jurkat cell model</cell_line> of T-cell activation an <dna>interleukin-2 promoter/reporter gene construct</dna> was activated by <protein>antigen receptor</protein> agonism in combination with the <protein>lymphokine</protein> <protein>interleukin-1</protein> .


Antigen receptor signals could be mimicked by suboptimal activation of protein kinase C ( PKC ) with phorbol esters in combination with calcium mobilization by an ionophore .

Antigen receptor signals could be mimicked by suboptimal activation of <protein>protein kinase C</protein> ( <protein>PKC</protein> ) with phorbol esters in combination with calcium mobilization by an ionophore .


In cotransfection experiments , oncogenic rats obviated the need for PKC stimulation but did not replace either the calcium signal or interleukin-1 .

In cotransfection experiments , oncogenic rats obviated the need for <protein>PKC</protein> stimulation but did not replace either the calcium signal or <protein>interleukin-1</protein> .


Activated ras expression also replaced the requirement for PKC stimulation in activation of the T-cell transcription factor NF-AT .

Activated <protein>ras</protein> expression also replaced the requirement for <protein>PKC</protein> stimulation in activation of the <protein>T-cell transcription factor</protein> <protein>NF-AT</protein> .


A dominant inhibitory ras mutant specifically blocked antigen receptor agonism , indicating that ras activity is required for antigen receptor signaling .

A <protein>dominant inhibitory ras mutant</protein> specifically blocked antigen receptor agonism , indicating that <protein>ras</protein> activity is required for <protein>antigen receptor</protein> signaling .


In addition , an inhibitor of PKC blocked both activated ras and phorbol ester stimulation , suggesting a role for ras upstream of PKC .

In addition , an inhibitor of <protein>PKC</protein> blocked both activated <protein>ras</protein> and phorbol ester stimulation , suggesting a role for <protein>ras</protein> upstream of <protein>PKC</protein> .


Expression of the Tat protein of HIV1 in human promonocytic U937 cells .

Expression of the <protein>Tat protein</protein> of HIV1 in <cell_line>human promonocytic U937 cells</cell_line> .


Numerous studies have shown that , upon HIV1 infection , human promonocytic U937 cells were induced to differentiate , as indicated , for example , by increased expression of adhesion molecules .

Numerous studies have shown that , upon HIV1 infection , <cell_line>human promonocytic U937 cells</cell_line> were induced to differentiate , as indicated , for example , by increased expression of <protein>adhesion molecules</protein> .


One of the viral proteins involved in this process might be the Tat protein .

One of the viral proteins involved in this process might be the <protein>Tat protein</protein> .


Indeed , this viral protein , which is essential for productive infection , has also been shown to display growth-stimulating properties and immunomodulatory activities .

Indeed , this <protein>viral protein</protein> , which is essential for productive infection , has also been shown to display growth-stimulating properties and immunomodulatory activities .


In order to apprehend the role of the HIV1 tat gene in inducing the differentiation of HIV1-infected U937 cells , we have successfully introduced this gene into U937 cells by infecting them with retroviral particles transducing tat .

In order to apprehend the role of the <dna>HIV1 tat gene</dna> in inducing the differentiation of <cell_line>HIV1-infected U937 cells</cell_line> , we have successfully introduced this gene into <cell_line>U937 cells</cell_line> by infecting them with retroviral particles transducing <dna>tat</dna> .


The effect of the Tat protein constitutively expressed by these cells upon their differentiation was then evaluated by looking for the expression of the c-fos and of the c-fms proto-oncogenes which are linked to the differentiation of myelomonoblastic cells .

The effect of the <protein>Tat protein</protein> constitutively expressed by these cells upon their differentiation was then evaluated by looking for the expression of the <dna>c-fos</dna> and of the <dna>c-fms proto-oncogenes</dna> which are linked to the differentiation of <cell_type>myelomonoblastic cells</cell_type> .


Northern blot analysis revealed in these cells , an increase in the transcription of these two proto-oncogenes , and this increase was amplified after treatment with phorbol myristate acetate .

Northern blot analysis revealed in these cells , an increase in the transcription of these two <dna>proto-oncogenes</dna> , and this increase was amplified after treatment with phorbol myristate acetate .


No such increase was observed in control U937 cells .

No such increase was observed in control <cell_line>U937 cells</cell_line> .


These results indicate that , among HIV1 gene products , the Tat protein appears to trigger monocytic differentiation , and suggests that this viral protein directs progenitors of the monocyte/macrophage lineage towards a differentiation stage in which production of viral antigens and virions might be more efficient .

These results indicate that , among <protein>HIV1 gene products</protein> , the <protein>Tat protein</protein> appears to trigger monocytic differentiation , and suggests that this <protein>viral protein</protein> directs progenitors of the <cell_type>monocyte/macrophage lineage</cell_type> towards a differentiation stage in which production of <protein>viral antigens</protein> and virions might be more efficient .


Analysis of the preexisting and nuclear forms of nuclear factor of activated T cells .

Analysis of the preexisting and nuclear forms of <protein>nuclear factor of activated T cells</protein> .


The nuclear factor of activated T cells ( NF-AT ) 3 is an inducible DNA-binding protein that is essential for transcriptional induction of the IL-2 gene during T cell activation .

The <protein>nuclear factor of activated T cells</protein> ( <protein>NF-AT</protein> ) 3 is an inducible <protein>DNA-binding protein</protein> that is essential for transcriptional induction of the <dna>IL-2 gene</dna> during T cell activation .


NF-AT is thought to consist of two components : a ubiquitous , inducible nuclear component that we have identified as Fos and Jun proteins , and a preexisting , T cell-specific component ( NF-ATp ) which is the target for the immunosuppressive agents cyclosporin A ( CsA ) and FK506 .

<protein>NF-AT</protein> is thought to consist of two components : a <protein>ubiquitous , inducible nuclear component</protein> that we have identified as <protein>Fos</protein> and <protein>Jun proteins</protein> , and a preexisting , <protein>T cell-specific component</protein> ( <protein>NF-ATp</protein> ) which is the target for the immunosuppressive agents cyclosporin A ( CsA ) and FK506 .


We have previously shown that nuclear extracts from activated T cells form two inducible NF-AT complexes with an oligonucleotide corresponding to the distal NF-AT site of the murine IL-2 promoter , although hypotonic extracts of unstimulated T cells form a single complex containing NF-ATp .

We have previously shown that nuclear extracts from <cell_type>activated T cells</cell_type> form two inducible <protein>NF-AT complexes</protein> with an oligonucleotide corresponding to the <dna>distal NF-AT site</dna> of the <dna>murine IL-2 promoter</dna> , although hypotonic extracts of unstimulated <cell_type>T cells</cell_type> form a single complex containing <protein>NF-ATp</protein> .


We show that the ability to detect NF-ATp in a gel shift assay , which is essential for purification and biochemical studies of this protein , is strikingly dependent on the precise sequence of the NF-AT oligonucleotide used as the labeled probe .

We show that the ability to detect <protein>NF-ATp</protein> in a gel shift assay , which is essential for purification and biochemical studies of this protein , is strikingly dependent on the precise sequence of the <protein>NF-AT</protein> oligonucleotide used as the labeled probe .


Moreover we present evidence that the component that forms the faster-migrating ( `` lower '' ) nuclear NF-AT complex is derived by a calcium-dependent , cyclosporin-sensitive , posttranslational modification of NF-ATp , and that Fos and Jun proteins stabilize its interaction with DNA .

Moreover we present evidence that the component that forms the faster-migrating ( `` lower '' ) <protein>nuclear NF-AT complex</protein> is derived by a calcium-dependent , cyclosporin-sensitive , posttranslational modification of <protein>NF-ATp</protein> , and that <protein>Fos</protein> and <protein>Jun</protein> proteins stabilize its interaction with DNA .


The results are discussed in the context of a model relating the two nuclear NF-AT complexes to NF-ATp .

The results are discussed in the context of a model relating the two nuclear <protein>NF-AT complexes</protein> to <protein>NF-ATp</protein> .


Cell cycle analysis of E2F in primary human T cells reveals novel E2F complexes and biochemically distinct forms of free E2F .

Cell cycle analysis of <protein>E2F</protein> in <cell_type>primary human T cells</cell_type> reveals <protein>novel E2F complexes</protein> and biochemically distinct forms of free <protein>E2F</protein> .


The transcription factor E2F activates the expression of multiple genes involved in cell proliferation , such as c-myc and the dihydrofolate reductase gene .

The <protein>transcription factor</protein> <protein>E2F</protein> activates the expression of multiple <dna>genes</dna> involved in cell proliferation , such as <dna>c-myc</dna> and the <dna>dihydrofolate reductase gene</dna> .


Regulation of E2F involves its interactions with other cellular proteins , including the retinoblastoma protein ( Rb ) , the Rb-related protein p107 , cyclin A , and cdk2 .

Regulation of <protein>E2F</protein> involves its interactions with other <protein>cellular proteins</protein> , including the <protein>retinoblastoma protein</protein> ( <protein>Rb</protein> ) , the <protein>Rb-related protein</protein> <protein>p107</protein> , <protein>cyclin A</protein> , and <protein>cdk2</protein> .


We undertook a detailed analysis of E2F DNA-binding activities and their cell cycle behavior in primary human T cells .

We undertook a detailed analysis of <protein>E2F</protein> DNA-binding activities and their cell cycle behavior in primary human <cell_type>T cells</cell_type> .


Three E2F DNA-binding activities were identified in resting ( G0 ) T cells with mobilities in gel shift assays distinct from those of previously defined E2F complexes .

Three <protein>E2F</protein> DNA-binding activities were identified in resting ( G0 ) <cell_type>T cells</cell_type> with mobilities in gel shift assays distinct from those of previously defined <protein>E2F</protein> complexes .


One of these activities was found to be a novel , less abundant , Rb- E2F complex .

One of these activities was found to be a novel , less abundant , Rb- <protein>E2F</protein> complex .


The most prominent E2F activity in resting T cells ( termed complex X ) was abundant in both G0 and G1 but disappeared as cells entered S phase , suggesting a possible role in negatively regulating E2F function .

The most prominent <protein>E2F</protein> activity in resting <cell_type>T cells</cell_type> ( termed complex X ) was abundant in both G0 and G1 but disappeared as cells entered S phase , suggesting a possible role in negatively regulating <protein>E2F</protein> function .


Complex X could be dissociated by adenovirus E1A with a requirement for an intact E1A conserved region 2 .

<protein>Complex X</protein> could be dissociated by adenovirus E1A with a requirement for an intact <dna>E1A conserved region 2</dna> .


However , X failed to react with a variety of antibodies against Rb or p107 , implicating the involvement of an E1A-binding protein other than Rb or p107 .

However , <protein>X</protein> failed to react with a variety of antibodies against <protein>Rb</protein> or <protein>p107</protein> , implicating the involvement of an <protein>E1A-binding protein</protein> other than <protein>Rb</protein> or <protein>p107</protein> .


In addition to these novel E2F complexes , three distinct forms of unbound ( free ) E2F were resolved in gel shift experiments .

In addition to these <protein>novel E2F complexes</protein> , three distinct forms of unbound ( free ) <protein>E2F</protein> were resolved in gel shift experiments .


These species showed different cell cycle kinetics .


UV cross-linking experiments suggested that a distinct E2F DNA-binding protein is uniquely associated with the S-phase p107 complex and is not associated with Rb .

UV cross-linking experiments suggested that a distinct <protein>E2F</protein> DNA-binding protein is uniquely associated with the <protein>S-phase p107 complex</protein> and is not associated with <protein>Rb</protein> .


Together , these results suggest that E2F consists of multiple , biochemically distinct DNA-binding proteins which function at different points in the cell cycle .

Together , these results suggest that <protein>E2F</protein> consists of multiple , biochemically distinct <protein>DNA-binding proteins</protein> which function at different points in the cell cycle .


Defective translocation of protein kinase C in multidrug-resistant HL-60 cells confers a reversible loss of phorbol ester-induced monocytic differentiation .

Defective translocation of <protein>protein kinase C</protein> in <cell_line>multidrug-resistant HL-60 cells</cell_line> confers a reversible loss of phorbol ester-induced monocytic differentiation .


Previous studies have demonstrated that human HL-60 myeloid leukemia cells differentiate in response to phorbol esters .

Previous studies have demonstrated that <cell_line>human HL-60 myeloid leukemia cells</cell_line> differentiate in response to phorbol esters .


This event is associated with induction of the c-jun early response gene and appearance of a monocytic phenotype .

This event is associated with induction of the <dna>c-jun early response gene</dna> and appearance of a monocytic phenotype .


The present studies have examined the effects of vincristine-selected , multidrug resistance on 12-O-tetradecanoylphorbol-13-acetate ( TPA ) -induced HL-60 cell differentiation .


The results demonstrate that multidrug-resistant HL-60 cells , designated HL-60/vinc , fail to respond to TPA with an increase in c-jun transcripts or other phenotypic characteristics of monocytic differentiation .

The results demonstrate that <cell_line>multidrug-resistant HL-60 cells</cell_line> , designated HL-60/vinc , fail to respond to TPA with an increase in <rna>c-jun transcripts</rna> or other phenotypic characteristics of monocytic differentiation .


By contrast , treatment of HL-60/vinc cells with okadaic acid , an inhibitor of serine/threonine protein phosphatases , induces c-jun transcription , growth arrest , and expression of the c-fms gene .

By contrast , treatment of <cell_line>HL-60/vinc cells</cell_line> with okadaic acid , an inhibitor of <protein>serine/threonine protein phosphatases</protein> , induces <dna>c-jun</dna> transcription , growth arrest , and expression of the <dna>c-fms gene</dna> .


Studies were also performed with an HL-60/vinc revertant ( HL-60/vinc/R ) line that has regained partial sensitivity to vincristine .

Studies were also performed with an <cell_line>HL-60/vinc revertant</cell_line> ( <cell_line>HL-60/vinc/R</cell_line> ) line that has regained partial sensitivity to vincristine .


The finding that HL-60/vinc/R cells respond to TPA with induction of a monocytic phenotype , but not c-jun expression , suggests that c-jun induction is not obligatory for monocytic differentiation .

The finding that <cell_line>HL-60/vinc/R cells</cell_line> respond to TPA with induction of a monocytic phenotype , but not <dna>c-jun</dna> expression , suggests that <dna>c-jun</dna> induction is not obligatory for monocytic differentiation .


Other studies further demonstrate that the jun-B and fra-1 genes are induced by TPA in both HL-60/vinc and HL-60/vinc/R cells , whereas c-fos expression is attenuated in the HL-60/vinc line .

Other studies further demonstrate that the <dna>jun-B</dna> and <dna>fra-1</dna> <dna>genes</dna> are induced by TPA in both <cell_line>HL-60/vinc</cell_line> and <cell_line>HL-60/vinc/R cells</cell_line> , whereas <dna>c-fos</dna> expression is attenuated in the HL-60/vinc line .


Since TPA activates protein kinase C ( PKC ) , we examined translocation of PKC from the cytosol to the membrane fraction .

Since TPA activates <protein>protein kinase C</protein> ( <protein>PKC</protein> ) , we examined translocation of <protein>PKC</protein> from the cytosol to the membrane fraction .


Although HL-60 and HL-60/vinc/R cells demonstrated translocation of PKC activity , this subcellular redistribution was undetectable in HL-60/vinc cells .

Although <cell_line>HL-60</cell_line> and <cell_line>HL-60/vinc/R cells</cell_line> demonstrated translocation of <protein>PKC</protein> activity , this subcellular redistribution was undetectable in <cell_line>HL-60/vinc cells</cell_line> .


Activity of the mitogen-activated protein kinase family with associated phosphorylation of c-Jun Y-peptide was markedly diminished in TPA-treated HL-60/vinc cells , but not in response to okadaic acid .

Activity of the <protein>mitogen-activated protein kinase family</protein> with associated phosphorylation of <protein>c-Jun Y-peptide</protein> was markedly diminished in <cell_line>TPA-treated HL-60/vinc cells</cell_line> , but not in response to okadaic acid .


Taken together , these findings suggest that vincristine resistance confers insensitivity to TPA-induced differentiation and can include defects in PKC -mediated signaling events and induction of jun/fos early response gene expression .

Taken together , these findings suggest that vincristine resistance confers insensitivity to TPA-induced differentiation and can include defects in <protein>PKC</protein> -mediated signaling events and induction of <dna>jun/fos early response gene</dna> expression .


Molecular basis of a multiple lymphokine deficiency in a patient with severe combined immunodeficiency .

Molecular basis of a multiple <protein>lymphokine</protein> deficiency in a patient with severe combined immunodeficiency .


We have previously reported that the T lymphocytes of a child with severe combined immunodeficiency are defective in the transcription of several lymphokine genes that include IL2 , IL3 , IL4 , and IL5 , which encode interleukins 2 , 3 , 4 , and 5 ( IL-2 , -3 , -4 , and -5 ) .

We have previously reported that the <cell_type>T lymphocytes</cell_type> of a child with severe combined immunodeficiency are defective in the transcription of several <dna>lymphokine genes</dna> that include <dna>IL2</dna> , <dna>IL3</dna> , <dna>IL4</dna> , and <dna>IL5</dna> , which encode <protein>interleukins</protein> 2 , 3 , 4 , and 5 ( <protein>IL-2</protein> , <protein>-3</protein> , <protein>-4</protein> , and <protein>-5</protein> ) .


To determine whether the defect in the patient 's T lymphocytes involved a trans-acting factor common to the affected lymphokine genes , we examined the ability of nuclear factors from the patient 's T lymphocytes to bind response elements present in the regulatory region of IL2 .

To determine whether the defect in the patient 's <cell_type>T lymphocytes</cell_type> involved a <protein>trans-acting factor</protein> common to the affected <dna>lymphokine genes</dna> , we examined the ability of <protein>nuclear factors</protein> from the patient 's <cell_type>T lymphocytes</cell_type> to bind response elements present in the <dna>regulatory region</dna> of <dna>IL2</dna> .


Nuclear factor NF-kB , activation protein 1 ( AP-1 ) , OCT-1 , and NF-IL-2B binding activity were normal .

<protein>Nuclear factor NF-kB</protein> , <protein>activation protein 1</protein> ( <protein>AP-1</protein> ) , <protein>OCT-1</protein> , and <protein>NF-IL-2B</protein> binding activity were normal .


In contrast , the binding of the nuclear factor of activated T cells ( NF-AT ) to its response element in the IL2 enhancer and to an NF-AT-like response element present in the IL4 enhancer was abnormal .

In contrast , the binding of the <protein>nuclear factor of activated T cells</protein> ( <protein>NF-AT</protein> ) to its response element in the <dna>IL2 enhancer</dna> and to an <dna>NF-AT-like response element</dna> present in the <dna>IL4 enhancer</dna> was abnormal .


To ascertain whether the abnormal NF-AT binding activity was related to an impaired function , we transfected patient and control T lymphocytes with constructs containing the reporter gene encoding chloramphenicol acetyl transferase ( CAT ) under the control of the entire IL2 regulatory region or of multimers of individual enhancer sequences .

To ascertain whether the abnormal <protein>NF-AT</protein> binding activity was related to an impaired function , we transfected patient and control <cell_type>T lymphocytes</cell_type> with constructs containing the reporter gene encoding <protein>chloramphenicol acetyl transferase</protein> ( <protein>CAT</protein> ) under the control of the entire <dna>IL2 regulatory region</dna> or of multimers of individual <dna>enhancer sequences</dna> .


CAT expression directed by the IL2 regulatory region or by a multimer of the NF-AT-binding site was markedly lower in the patient relative to controls .

CAT expression directed by the <dna>IL2 regulatory region</dna> or by a multimer of the <dna>NF-AT-binding site</dna> was markedly lower in the patient relative to controls .


In contrast , CAT gene expression directed by a multimer of the OCT-1 proximal ( OCT-1p ) -binding site was equivalent in patient and controls .

In contrast , <dna>CAT gene</dna> expression directed by a multimer of the <dna>OCT-1 proximal ( OCT-1p ) -binding site</dna> was equivalent in patient and controls .


These results indicate that an abnormality of/or influencing NF-AT may underlie the multiple lymphokine deficiency in this patient .

These results indicate that an abnormality of/or influencing <protein>NF-AT</protein> may underlie the multiple <protein>lymphokine</protein> deficiency in this patient .


FK506 and ciclosporin : molecular probes for studying intracellular signal transduction .


The immunosuppressants ciclosporin and FK506 block the Ca ( 2+ ) -dependent signal-transduction pathway emanating from the T-cell receptor , thereby inhibiting the activation of helper T cells .

The immunosuppressants ciclosporin and FK506 block the Ca ( 2+ ) -dependent signal-transduction pathway emanating from the <protein>T-cell receptor</protein> , thereby inhibiting the activation of <cell_type>helper T cells</cell_type> .


Using these drugs as probes , chemists and biologists have uncovered several intracellular signalling molecules bridging the generation of second-messenger Ca2+ ions and the transcriptional activation of IL-2 , among which are calmodulin , calcineurin and the nuclear factor of activated T cells ( NF-AT ) .

Using these drugs as probes , chemists and biologists have uncovered several intracellular signalling molecules bridging the generation of second-messenger Ca2+ ions and the transcriptional activation of <protein>IL-2</protein> , among which are <protein>calmodulin</protein> , <protein>calcineurin</protein> and the <protein>nuclear factor of activated T cells</protein> ( <protein>NF-AT</protein> ) .


Hence , Ca2+ binds to calmodulin , leading to the binding of calmodulin to calcineurin ; the activated calcineurin , in turn , may dephosphorylate the cytoplasmic subunit of NF-AT , resulting in its translocation from the cytoplasm into the nucleus to form a competent transcriptional activator .

Hence , Ca2+ binds to <protein>calmodulin</protein> , leading to the binding of <protein>calmodulin</protein> to <protein>calcineurin</protein> ; the activated <protein>calcineurin</protein> , in turn , may dephosphorylate the <protein>cytoplasmic subunit of NF-AT</protein> , resulting in its translocation from the cytoplasm into the nucleus to form a <protein>competent transcriptional activator</protein> .


As described by Jun Liu , these drugs manifest their effects in an unprecedented fashion .

As described by <protein>Jun</protein> Liu , these drugs manifest their effects in an unprecedented fashion .


They do not directly intercept intracellular signalling molecules .


Instead , they form tight complexes with two different classes of abundant cytosolic receptors called immunophilins upon entering the cell , and consequently inhibit their peptidyl prolyl cis-trans isomerase activities .

Instead , they form tight complexes with two different classes of abundant <protein>cytosolic receptors</protein> called <protein>immunophilins</protein> upon entering the cell , and consequently inhibit their <protein>peptidyl prolyl cis-trans isomerase</protein> activities .


The two structurally distinct immunophilin-drug complexes bind to , and inhibit , the phosphatase activity of calcineurin .

The two structurally distinct <protein>immunophilin-drug complexes</protein> bind to , and inhibit , the <protein>phosphatase</protein> activity of <protein>calcineurin</protein> .


p105 and p98 precursor proteins play an active role in NF-kappa B -mediated signal transduction .

<protein>p105</protein> and <protein>p98</protein> <protein>precursor proteins</protein> play an active role in <protein>NF-kappa B</protein> -mediated signal transduction .


The Rel/NF-kappa B family of transcription factors is composed of two distinct subgroups , proteins that undergo proteolytic processing and contain SWI6/ankyrin repeats in their carboxyl termini ( p105 , p98 ) , and those without such repeats that do not require processing ( p65 , c-Rel , RelB , and Dorsal ) .

The <protein>Rel/NF-kappa B family</protein> of <protein>transcription factors</protein> is composed of two distinct subgroups , proteins that undergo proteolytic processing and contain <protein>SWI6/ankyrin repeats</protein> in their <protein>carboxyl termini</protein> ( <protein>p105</protein> , <protein>p98</protein> ) , and those without such repeats that do not require processing ( <protein>p65</protein> , <protein>c-Rel</protein> , <protein>RelB</protein> , and <protein>Dorsal</protein> ) .


We demonstrate that the p105 and p98 precursors share functional properties with the I kappa B proteins , which also contain SWI6/ankyrin repeats .

We demonstrate that the <protein>p105</protein> and <protein>p98</protein> precursors share functional properties with the <protein>I kappa B proteins</protein> , which also contain <protein>SWI6/ankyrin repeats</protein> .


Both p105 and p98 were found to form stable complexes with other Rel/NF-kappa B family members , including p65 and c-Rel .

Both <protein>p105</protein> and <protein>p98</protein> were found to form stable complexes with other <protein>Rel/NF-kappa B family members</protein> , including <protein>p65</protein> and <protein>c-Rel</protein> .


Association with the precursors is sufficient for cytoplasmic retention of either p65 or c-Rel , both of which are otherwise nuclear .

Association with the precursors is sufficient for cytoplasmic retention of either <protein>p65</protein> or <protein>c-Rel</protein> , both of which are otherwise nuclear .


These complexes undergo stimulus-responsive processing to produce active p50/c-Rel and p55/c-Rel complexes .

These complexes undergo stimulus-responsive processing to produce active <protein>p50/c-Rel</protein> and <protein>p55/c-Rel</protein> complexes .


These observations suggest a second pathway leading to NF-kappa B induction , in which processing of the precursors rather than phosphorylation of I kappa B plays a major role .

These observations suggest a second pathway leading to <protein>NF-kappa B</protein> induction , in which processing of the precursors rather than phosphorylation of <protein>I kappa B</protein> plays a major role .


NF-kappa B controls expression of inhibitor I kappa B alpha : evidence for an inducible autoregulatory pathway .

<protein>NF-kappa B</protein> controls expression of inhibitor <protein>I kappa B alpha</protein> : evidence for an inducible autoregulatory pathway .


The eukaryotic transcription factor nuclear factor-kappa B ( NF-kappa B ) participates in many parts of the genetic program mediating T lymphocyte activation and growth .

The eukaryotic <protein>transcription factor</protein> <protein>nuclear factor-kappa B</protein> ( <protein>NF-kappa B</protein> ) participates in many parts of the genetic program mediating <cell_type>T lymphocyte</cell_type> activation and growth .


Nuclear expression of NF-kappa B occurs after its induced dissociation from its cytoplasmic inhibitor I kappa B alpha .

Nuclear expression of <protein>NF-kappa B</protein> occurs after its induced dissociation from its cytoplasmic inhibitor <protein>I kappa B alpha</protein> .


Phorbol ester and tumor necrosis factor-alpha induction of nuclear NF-kappa B is associated with both the degradation of performed I kappa B alpha and the activation of I kappa B alpha gene expression .

Phorbol ester and <protein>tumor necrosis factor-alpha</protein> induction of <protein>nuclear NF-kappa B</protein> is associated with both the degradation of performed <protein>I kappa B alpha</protein> and the activation of <protein>I kappa B</protein> alpha gene expression .


Transfection studies indicate that the I kappa B alpha gene is specifically induced by the 65-kilodalton transactivating subunit of NF-kappa B .

Transfection studies indicate that the <dna>I kappa B alpha gene</dna> is specifically induced by the <protein>65-kilodalton transactivating subunit</protein> of <protein>NF-kappa B</protein> .


Association of the newly synthesized I kappa B alpha with p65 restores intracellular inhibition of NF-kappa B DNA binding activity and prolongs the survival of this labile inhibitor .

Association of the newly synthesized <protein>I kappa B alpha</protein> with <protein>p65</protein> restores intracellular inhibition of <protein>NF-kappa B</protein> DNA binding activity and prolongs the survival of this labile inhibitor .


Together , these results show that NF-kappa B controls the expression of I kappa B alpha by means of an inducible autoregulatory pathway .

Together , these results show that <protein>NF-kappa B</protein> controls the expression of <protein>I kappa B</protein> alpha by means of an inducible autoregulatory pathway .


The human prointerleukin 1 beta gene requires DNA sequences both proximal and distal to the transcription start site for tissue-specific induction .

The <dna>human prointerleukin 1 beta gene</dna> requires DNA sequences both proximal and distal to the <dna>transcription start site</dna> for tissue-specific induction .


In these studies , we have identified DNA sequences and specific protein interactions necessary for transcriptional regulation of the human prointerleukin 1 beta ( proIL-1 beta ) gene .

In these studies , we have identified <dna>DNA sequences</dna> and specific protein interactions necessary for transcriptional regulation of the <dna>human prointerleukin 1 beta ( proIL-1 beta ) gene</dna> .


A cell-type-independent lipopolysaccharide ( LPS ) -responsive enhancer element located between -3757 and -2729 bp upstream from the transcription start site ( cap site ) consisted of at least six discrete subregions which were essential to the maximal induction by LPS in transfected monocytes .

A <dna>cell-type-independent lipopolysaccharide ( LPS ) -responsive enhancer element</dna> located between <dna>-3757 and -2729 bp upstream</dna> from the <dna>transcription start site</dna> ( <dna>cap site</dna> ) consisted of at least six discrete subregions which were essential to the maximal induction by LPS in <cell_line>transfected monocytes</cell_line> .


The enhancer also appeared to mediate phorbol myristate acetate induction in monocytes and IL-1 responsiveness in fibroblasts .

The enhancer also appeared to mediate phorbol myristate acetate induction in <cell_type>monocytes</cell_type> and <protein>IL-1</protein> responsiveness in <cell_type>fibroblasts</cell_type> .


Deletion and base substitution mutations along with DNA binding studies demonstrated that the enhancer contained a minimum of three functional protein binding sequences , two of which appeared to be important for gene induction .

Deletion and base substitution mutations along with DNA binding studies demonstrated that the enhancer contained a minimum of three functional <dna>protein binding sequences</dna> , two of which appeared to be important for gene induction .


One of the essential proteins which bound to the enhancer was similar or identical to members of the C/EBP family of transcription factors required for both IL-1 -and LPS-specific induction of the IL-6 gene ( i.e. , the NF-IL6 proteins ) .

One of the essential proteins which bound to the enhancer was similar or identical to members of the <protein>C/EBP family</protein> of <protein>transcription factors</protein> required for both <protein>IL-1</protein> -and LPS-specific induction of the <dna>IL-6 gene</dna> ( i.e. , the <dna>NF-IL6 proteins</dna> ) .


When ligated to the proIL-1 beta cap site -proximal region ( located between -131 to +12 ) , both the proIL-1 beta and the simian virus 40 enhancer elements functioned more efficiently in monocytes than in HeLa cells , which are not normally competent for IL-1 beta expression .

When ligated to the <dna>proIL-1 beta cap site</dna> <dna>-proximal region</dna> ( located between -131 to +12 ) , both the <dna>proIL-1 beta and the simian virus 40 enhancer elements</dna> functioned more efficiently in <cell_type>monocytes</cell_type> than in <cell_type>HeLa cells</cell_type> , which are not normally competent for <protein>IL-1 beta</protein> expression .


When ligated to the murine c-fos promoter , however , the proIL-1 beta enhancer was inducible in phorbol myristate acetate-stimulated HeLa cells , suggesting the existence of a proIL-1 beta promoter-proximal requirement for tissue specificity .

When ligated to the <dna>murine c-fos promoter</dna> , however , the <dna>proIL-1 beta enhancer</dna> was inducible in <cell_line>phorbol myristate acetate-stimulated HeLa cells</cell_line> , suggesting the existence of a <protein>proIL-1 beta</protein> promoter-proximal requirement for tissue specificity .


Lymphocytes from the site of disease suggest adenovirus is one cause of persistent or recurrent inflammatory arthritis .

<cell_type>Lymphocytes</cell_type> from the site of disease suggest adenovirus is one cause of persistent or recurrent inflammatory arthritis .


The assessment of synovial lymphocyte reactivity to adenovirus antigen stimulation was undertaken in patients with persistent or recurrent inflammatory arthritis .


The 3H-thymidine uptake procedure was employed , incorporating multiple microbiological antigens .

The 3H-thymidine uptake procedure was employed , incorporating multiple <protein>microbiological antigens</protein> .


Five patients were found with repeated maximal responses to adenovirus antigen ; in one of these adenovirus nucleotide sequences were present in a synovial biopsy specimen .

Five patients were found with repeated maximal responses to <protein>adenovirus antigen</protein> ; in one of these <dna>adenovirus nucleotide sequences</dna> were present in a synovial biopsy specimen .


It is concluded that adenovirus may be one cause of persistent or recurrent inflammatory arthritis .


Expression of PILOT , a putative transcription factor , requires two signals and is cyclosporin A sensitive in T cells .

Expression of <protein>PILOT</protein> , a putative <protein>transcription factor</protein> , requires two signals and is cyclosporin A sensitive in <cell_type>T cells</cell_type> .


Few known genes ( IL-2 , members of the IL-8 family , interferon-gamma ) are induced in T cells only through the combined effect of phorbol myristic acetate ( PMA ) and a Ca ( 2+ ) -ionophore , and expression of only these genes can be fully suppressed by Cyclosporin A ( CyA ) .

Few known <dna>genes</dna> ( <protein>IL-2</protein> , members of the <dna>IL-8 family</dna> , <dna>interferon-gamma</dna> ) are induced in <cell_type>T cells</cell_type> only through the combined effect of phorbol myristic acetate ( PMA ) and a Ca ( 2+ ) -ionophore , and expression of only these <dna>genes</dna> can be fully suppressed by Cyclosporin A ( CyA ) .


We have identified a putative transcription factor , designated PILOT , with an identical dual signal requirement for expression .

We have identified a putative <protein>transcription factor</protein> , designated <protein>PILOT</protein> , with an identical dual signal requirement for expression .


Induction of the PILOT gene is detectable in human T cells 20 min following activation in the presence of cycloheximide and is fully suppressed by CyA .

Induction of the <protein>PILOT</protein> gene is detectable in human <cell_type>T cells</cell_type> 20 min following activation in the presence of cycloheximide and is fully suppressed by CyA .


The PILOT protein has a calculated M ( r ) of 42.6 kDa and contains three zinc fingers of the C2H2-type at the carboxyl-terminus which are highly homologous to the zinc finger regions of the transcription factors EGR1 , EGR2 , and pAT 133 .

The <protein>PILOT</protein> protein has a calculated M ( r ) of 42.6 kDa and contains three <protein>zinc fingers</protein> of the <protein>C2H2-type</protein> at the <protein>carboxyl-terminus</protein> which are highly homologous to the <protein>zinc finger regions</protein> of the <protein>transcription factors</protein> <protein>EGR1</protein> , <protein>EGR2</protein> , and <protein>pAT 133</protein> .


In contrast to T cells , in fibroblasts PILOT gene expression requires only one signal ( PMA ) and is not affected by CyA .

In contrast to <cell_type>T cells</cell_type> , in <cell_type>fibroblasts</cell_type> <protein>PILOT</protein> gene expression requires only one signal ( PMA ) and is not affected by CyA .


This observation directly demonstrates the existence of a Ca2+ signal-dependent regulatory element obligatory for expression of some genes in T cells but not in fibroblasts .

This observation directly demonstrates the existence of a <protein>Ca2+ signal-dependent regulatory element</protein> obligatory for expression of some <dna>genes</dna> in <cell_type>T cells</cell_type> but not in <cell_type>fibroblasts</cell_type> .


This differential expression model will be valuable in the dissection of the dual signal pathway in T cells and the effects of CyA upon it .

This differential expression model will be valuable in the dissection of the dual signal pathway in <cell_type>T cells</cell_type> and the effects of CyA upon it .


Antisense oligonucleotides to the p65 subunit of NF-kappa B block CD11b expression and alter adhesion properties of differentiated HL-60 granulocytes .

Antisense oligonucleotides to the <protein>p65 subunit</protein> of <protein>NF-kappa B</protein> block <protein>CD11b</protein> expression and alter adhesion properties of differentiated <cell_line>HL-60 granulocytes</cell_line> .


NF-kappa B is a pleiotropic regulator of a variety of genes implicated in the cellular response to injury .

<protein>NF-kappa B</protein> is a <protein>pleiotropic regulator</protein> of a variety of genes implicated in the cellular response to injury .


This function has been attributed to the coordinated binding of subunits of NF-kappa B to distinct regions of the promoter elements of numerous genes , including cytokines , growth factor receptors , and adhesion molecules .

This function has been attributed to the coordinated binding of subunits of <protein>NF-kappa B</protein> to distinct regions of the <dna>promoter elements</dna> of numerous genes , including <protein>cytokines</protein> , <protein>growth factor receptors</protein> , and <protein>adhesion molecules</protein> .


Antisense phosphorothioate oligonucleotides to the p50 and p65 subunits of the NF-kappa B complex were used to define the physiologic role of this transcription factor in resting and stimulated granulocytes .

Antisense phosphorothioate oligonucleotides to the <protein>p50</protein> and <protein>p65 subunits</protein> of the <protein>NF-kappa B complex</protein> were used to define the physiologic role of this <protein>transcription factor</protein> in <cell_line>resting and stimulated granulocytes</cell_line> .


A reduction in the expression of p65 was produced by treatment with the phosphorothioate antisense oligodeoxynucleotide .

A reduction in the expression of <protein>p65</protein> was produced by treatment with the phosphorothioate antisense oligodeoxynucleotide .


This reduction was accompanied by rapid changes in the cellular adhesion of dimethyl sulfoxide-differentiated HL-60 leukemia cells stimulated by 12-O-tetradecanoylphorbol 13-acetate ( TPA ) .

This reduction was accompanied by rapid changes in the cellular adhesion of <cell_line>dimethyl sulfoxide-differentiated HL-60 leukemia cells</cell_line> stimulated by 12-O-tetradecanoylphorbol 13-acetate ( TPA ) .


These effects were characterized by a marked reduction in CD11b integrin expression on the surface of treated cells .

These effects were characterized by a marked reduction in <protein>CD11b</protein> <protein>integrin</protein> expression on the surface of <cell_line>treated cells</cell_line> .


Furthermore , the p65 antisense oligomer effectively abolished an upregulation of CD11b that was produced by formyl-met-leu-phe and TPA .

Furthermore , the <protein>p65</protein> antisense oligomer effectively abolished an upregulation of <protein>CD11b</protein> that was produced by formyl-met-leu-phe and TPA .


However , the p65 antisense phosphorothioate oligodeoxynucleotide had no significant effect on the production of reactive oxygen intermediates or on phagocytosis by these cells .

However , the <protein>p65</protein> antisense phosphorothioate oligodeoxynucleotide had no significant effect on the production of reactive oxygen intermediates or on phagocytosis by these cells .


These findings indicate that antisense oligomers to p65 can be used to define the role of NF-kappa B in the activation pathways of neutrophils .

These findings indicate that antisense oligomers to <protein>p65</protein> can be used to define the role of <protein>NF-kappa B</protein> in the activation pathways of <cell_type>neutrophils</cell_type> .


The p65 subunit of NF-kappa B regulates I kappa B by two distinct mechanisms .

The <protein>p65 subunit</protein> of <protein>NF-kappa B</protein> regulates <protein>I kappa B</protein> by two distinct mechanisms .


Transcription factor NF-kappa B ( p50/p65 ) is generally localized to the cytoplasm by its inhibitor I kappa B .

Transcription factor <protein>NF-kappa B</protein> ( <protein>p50/p65</protein> ) is generally localized to the cytoplasm by its inhibitor <protein>I kappa B</protein> .


Overproduced I kappa B , free from NF-kappa B , is rapidly degraded .

Overproduced <protein>I kappa B</protein> , free from <protein>NF-kappa B</protein> , is rapidly degraded .


Overexpression of p65 increases endogenous I kappa B protein in both carcinoma and lymphoid cells by two mechanisms : protein stabilization and increased transcription of I kappa B mRNA .

Overexpression of <protein>p65</protein> increases endogenous <protein>I kappa B</protein> protein in both <cell_type>carcinoma and lymphoid cells</cell_type> by two mechanisms : protein stabilization and increased transcription of <rna>I kappa B mRNA</rna> .


In contrast , p65 delta , a naturally occurring splice variant , fails to markedly augment I kappa B protein levels .

In contrast , <protein>p65 delta</protein> , a naturally occurring splice variant , fails to markedly augment <protein>I kappa B</protein> protein levels .


Both overexpressed p65 and coexpressed p50 are cytoplasmic , whereas p65 delta is partly nuclear , indicating that the I kappa B induced by p65 can maintain NF-kappa B in the cytoplasm .

Both overexpressed <protein>p65</protein> and coexpressed <protein>p50</protein> are cytoplasmic , whereas <protein>p65 delta</protein> is partly nuclear , indicating that the <protein>I kappa B</protein> induced by <protein>p65</protein> can maintain <protein>NF-kappa B</protein> in the cytoplasm .


Thus , p65 and I kappa B are linked in an autoregulatory loop , ensuring that NF-kappa B is held in the cytoplasm until cells are specifically induced to translocate it to the nucleus .

Thus , <protein>p65</protein> and <protein>I kappa B</protein> are linked in an autoregulatory loop , ensuring that <protein>NF-kappa B</protein> is held in the cytoplasm until cells are specifically induced to translocate it to the nucleus .


Lipopolysaccharide induces phosphorylation of MAD3 and activation of c-Rel and related NF-kappa B proteins in human monocytic THP-1 cells .

Lipopolysaccharide induces phosphorylation of <protein>MAD3</protein> and activation of <protein>c-Rel</protein> and related <protein>NF-kappa B</protein> proteins in <cell_line>human monocytic THP-1 cells</cell_line> .


Many effects of lipopolysaccharide ( LPS ) on gene expression , including that of human immunodeficiency virus ( HIV ) , in monocytic cells are mediated by activation of kappa B DNA-binding proteins .

Many effects of lipopolysaccharide ( LPS ) on gene expression , including that of human immunodeficiency virus ( HIV ) , in <cell_type>monocytic cells</cell_type> are mediated by activation of kappa B DNA-binding proteins .


However , the specific members of the NF-kappa B/Rel transcription factor family involved in the LPS response , and the mechanisms through which LPS-generated signals are transduced remain unclear .

However , the specific members of the <protein>NF-kappa B/Rel transcription factor family</protein> involved in the LPS response , and the mechanisms through which LPS-generated signals are transduced remain unclear .


Here we show that LPS induces nuclear expression of c-Rel/p50 heterodimers as well as p50/p65 ( NF-kappa B ) kappa B DNA-binding complexes in human monocytic THP-1 cells .

Here we show that LPS induces nuclear expression of <protein>c-Rel/p50 heterodimers</protein> as well as <protein>p50/p65</protein> ( <protein>NF-kappa B</protein> ) kappa B DNA-binding complexes in <cell_line>human monocytic THP-1 cells</cell_line> .


Nuclear localization of these proteins occurred concomitantly with a rapid decrease in their cytosolic levels and was independent of phorbol ester-sensitive protein kinase C .

Nuclear localization of these proteins occurred concomitantly with a rapid decrease in their cytosolic levels and was independent of <protein>phorbol ester-sensitive protein kinase C</protein> .