These results suggest that the SRG3 protein is involved in the glucocorticoid-induced apoptosis in the thymoma cell line .

These results suggest that the <protein>SRG3 protein</protein> is involved in the glucocorticoid-induced apoptosis in the <cell_line>thymoma cell line</cell_line> .


This implicates that the SRG3 may play an important regulatory role during T cell development in thymus .

This implicates that the <dna>SRG3</dna> may play an important regulatory role during T cell development in thymus .


Transcriptional induction of collagenase-1 in differentiated monocyte-like ( U937 ) cells is regulated by AP-1 and an upstream C/EBP-beta site .

Transcriptional induction of <protein>collagenase-1</protein> in <cell_line>differentiated monocyte-like ( U937 ) cells</cell_line> is regulated by <protein>AP-1</protein> and an <dna>upstream C/EBP-beta site</dna> .


In this report , we demonstrate that the AP-1 site and a distal promoter element regulate transcriptional induction of collagenase-1 during monocytic differentiation .

In this report , we demonstrate that the <protein>AP-1</protein> site and a <dna>distal promoter element</dna> regulate transcriptional induction of <protein>collagenase-1</protein> during monocytic differentiation .


Chloramphenicol acetyltransferase expression constructs containing regions of the human collagenase-1 promoter were stably or transiently transfected into U937 cells , and reporter activity was assessed at various times after the onset of phorbol 12-myristate 13-acetate ( PMA ) -mediated differentiation .

<dna>Chloramphenicol acetyltransferase expression constructs</dna> containing regions of the <dna>human collagenase-1 promoter</dna> were stably or transiently transfected into <cell_line>U937 cells</cell_line> , and reporter activity was assessed at various times after the onset of phorbol 12-myristate 13-acetate ( PMA ) -mediated differentiation .


Rapid and strong induction of promoter activity was lost in constructs with a mutant AP-1 element ; however , at 16-96 h post-PMA , the mutant collagenase-1 promoter displayed AP-1 independent PMA-mediated transactivation .

Rapid and strong induction of promoter activity was lost in constructs with a <dna>mutant AP-1 element</dna> ; however , at 16-96 h post-PMA , the <dna>mutant collagenase-1 promoter</dna> displayed <protein>AP-1</protein> independent PMA-mediated transactivation .


The AP-1 mutant constructs also showed delayed transcriptional activation in PMA-treated fibroblasts .

The <dna>AP-1 mutant constructs</dna> also showed delayed transcriptional activation in <cell_line>PMA-treated fibroblasts</cell_line> .


Western and supershift analyses indicated that functional Jun and Fos proteins were present in nuclear extracts of PMA-differentiated U937 cells .

Western and supershift analyses indicated that <protein>functional Jun and Fos proteins</protein> were present in nuclear extracts of <cell_line>PMA-differentiated U937 cells</cell_line> .


Promoter deletion constructs demonstrated the potential role of distal promoter sequences in regulating collagenase-1 transcription .

Promoter deletion constructs demonstrated the potential role of <dna>distal promoter sequences</dna> in regulating <protein>collagenase-1</protein> transcription .


In particular , Western , supershift , and promoter deletion analyses suggested a role for CCAAT/enhancer-binding protein-beta ( C/EBP-beta ) binding site between -2010 and -1954 in regulating transcription of collagenase-1 in monocytic cells .

In particular , Western , supershift , and promoter deletion analyses suggested a role for <dna>CCAAT/enhancer-binding protein-beta ( C/EBP-beta ) binding site</dna> between <dna>-2010 and -1954</dna> in regulating transcription of <protein>collagenase-1</protein> in <cell_type>monocytic cells</cell_type> .


Our findings suggest that distinct regulatory elements , acting somewhat independently of each other , control expression of collagenase-1 .

Our findings suggest that distinct <dna>regulatory elements</dna> , acting somewhat independently of each other , control expression of <protein>collagenase-1</protein> .


In addition , our data suggests that the rapid PMA-mediated induction of collagenase-1 transcription is controlled by a mechanism distinct from that regulating the sustained expression of this proteinase in activated macrophages .

In addition , our data suggests that the rapid PMA-mediated induction of <protein>collagenase-1</protein> transcription is controlled by a mechanism distinct from that regulating the sustained expression of this <protein>proteinase</protein> in <cell_type>activated macrophages</cell_type> .


Quantification of vitamin D receptor mRNA by competitive polymerase chain reaction in PBMC : lack of correspondence with common allelic variants .

Quantification of <rna>vitamin D receptor mRNA</rna> by competitive polymerase chain reaction in <cell_type>PBMC</cell_type> : lack of correspondence with <dna>common allelic variants</dna> .


It has been recently claimed that polymorphism for the vitamin D receptor ( VDR ) influences several aspects of calcium and bone metabolism .

It has been recently claimed that polymorphism for the <protein>vitamin D receptor</protein> ( <protein>VDR</protein> ) influences several aspects of calcium and bone metabolism .


To evaluate the physiologic plausibility of these claims , we compared the abundance of the VDR mRNA in peripheral blood mononuclear cells ( PBMCs ) between different VDR genotypes using a quantitative reverse transcribed polymerase chain reaction-based method .

To evaluate the physiologic plausibility of these claims , we compared the abundance of the <rna>VDR mRNA</rna> in <cell_type>peripheral blood mononuclear cells</cell_type> ( <cell_type>PBMCs</cell_type> ) between different <protein>VDR genotypes</protein> using a quantitative reverse transcribed polymerase chain reaction-based method .


The method is based on the coamplification of VDR cDNA and an internal standard consisting of known concentrations of a human VDR CDNA mutated at a BglII restriction site ; the interassay coefficient of variation is 11 % .

The method is based on the coamplification of <dna>VDR cDNA</dna> and an internal standard consisting of known concentrations of a <dna>human VDR CDNA</dna> mutated at a <dna>BglII restriction site</dna> ; the interassay coefficient of variation is 11 % .


To validate the method , we made use of earlier receptor binding studies indicating that normal human monocytes and activated , but not resting , lymphocytes expressed the VDR .

To validate the method , we made use of earlier receptor binding studies indicating that <cell_type>normal human monocytes</cell_type> and activated , but not resting , <cell_type>lymphocytes</cell_type> expressed the <protein>VDR</protein> .


The concentration of the VDR mRNA was 10 ( -8 ) to 10 ( -7 ) g/g of total RNA in cell-sorted monocytes and in in vitro activated lymphocytes , but only 10 ( -12 ) g/g of total mRNA in resting lymphocytes , establishing that the VDR mRNA determined by our method in PBMCs is due to constitutive expression in monocytes .

The concentration of the <rna>VDR mRNA</rna> was 10 ( -8 ) to 10 ( -7 ) g/g of total RNA in <cell_type>cell-sorted monocytes</cell_type> and in <cell_type>in vitro activated lymphocytes</cell_type> , but only 10 ( -12 ) g/g of total mRNA in resting <cell_type>lymphocytes</cell_type> , establishing that the <rna>VDR mRNA</rna> determined by our method in <cell_type>PBMCs</cell_type> is due to constitutive expression in <cell_type>monocytes</cell_type> .


Following an initial genotype screening of 85 normal volunteers by polymerase chain reaction or restriction fragment length polymorphism analysis , 14 individuals with the Bb genotype , 12 with the bb genotype , and 12 with the BB genotype were selected .

Following an initial genotype screening of 85 normal volunteers by polymerase chain reaction or <dna>restriction fragment length polymorphism</dna> analysis , 14 individuals with the Bb genotype , 12 with the bb genotype , and 12 with the BB genotype were selected .


The concentration of the VDR mRNA , corrected for the number of monocytes , was similar among the three genotype groups , as were the other variables examined : serum calcitriol , serum osteocalcin , and vertebral and hip bone density .

The concentration of the <rna>VDR mRNA</rna> , corrected for the number of <cell_type>monocytes</cell_type> , was similar among the three genotype groups , as were the other variables examined : <protein>serum calcitriol</protein> , <protein>serum osteocalcin</protein> , and vertebral and hip bone density .


We conclude that VDR polymorphism does not affect the abundance of the VDR mRNA .

We conclude that <protein>VDR</protein> polymorphism does not affect the abundance of the <rna>VDR mRNA</rna> .


Oncogenic forms of NOTCH1 lacking either the primary binding site for RBP-Jkappa or nuclear localization sequences retain the ability to associate with RBP-Jkappa and activate transcription .

Oncogenic forms of <protein>NOTCH1</protein> lacking either the <dna>primary binding site</dna> for <protein>RBP-Jkappa</protein> or <protein>nuclear localization sequences</protein> retain the ability to associate with <protein>RBP-Jkappa</protein> and activate transcription .


Truncated forms of the NOTCH1 transmembrane receptor engineered to resemble mutant forms of NOTCH1 found in certain cases of human T cell leukemia/lymphoma ( T-ALL ) efficiently induce T-ALL when expressed in the bone marrow of mice .

Truncated forms of the <protein>NOTCH1 transmembrane receptor</protein> engineered to resemble mutant forms of <protein>NOTCH1</protein> found in certain cases of human T cell leukemia/lymphoma ( T-ALL ) efficiently induce T-ALL when expressed in the bone marrow of mice .


Unlike full-sized NOTCH1 , two such truncated forms of the protein either lacking a major portion of the extracellular domain ( DeltaE ) or consisting only of the intracellular domain ( ICN ) were found to activate transcription in cultured cells , presumably through RBP-Jkappa response elements within DNA .

Unlike full-sized <protein>NOTCH1</protein> , two such truncated forms of the protein either lacking a major portion of the <protein>extracellular domain</protein> ( <protein>DeltaE</protein> ) or consisting only of the <protein>intracellular domain</protein> ( <protein>ICN</protein> ) were found to activate transcription in cultured cells , presumably through <protein>RBP-Jkappa</protein> response elements within DNA .


Both truncated forms also bound to the transcription factor RBP-Jkappa in extracts prepared from human and murine T-ALL cell lines .

Both truncated forms also bound to the <protein>transcription factor RBP-Jkappa</protein> in extracts prepared from <cell_line>human and murine T-ALL cell lines</cell_line> .


Transcriptional activation required the presence of a weak RBP-Jkappa-binding site within the NOTCH1 ankyrin repeat region of the intracellular domain .

Transcriptional activation required the presence of a weak <protein>RBP-Jkappa-binding site</protein> within the <protein>NOTCH1 ankyrin repeat region</protein> of the <protein>intracellular domain</protein> .


Unexpectedly , a second , stronger RBP-Jkappa-binding site , which lies within the intracellular domain close to the transmembrane region and significantly augments association with RBP-Jkappa , was not needed for oncogenesis or for transcriptional activation .

Unexpectedly , a second , stronger <protein>RBP-Jkappa-binding site</protein> , which lies within the <protein>intracellular domain</protein> close to the transmembrane region and significantly augments association with <protein>RBP-Jkappa</protein> , was not needed for oncogenesis or for transcriptional activation .


While ICN appeared primarily in the nucleus , DeltaE localized to cytoplasmic and nuclear membranes , suggesting that intranuclear localization is not essential for oncogenesis or transcriptional activation .

While <protein>ICN</protein> appeared primarily in the nucleus , <protein>DeltaE</protein> localized to cytoplasmic and nuclear membranes , suggesting that intranuclear localization is not essential for oncogenesis or transcriptional activation .


In support of this interpretation , mutation of putative nuclear localization sequences decreased nuclear localization and increased transcriptional activation by membrane-bound DeltaE .

In support of this interpretation , mutation of putative <protein>nuclear localization sequences</protein> decreased nuclear localization and increased transcriptional activation by <protein>membrane-bound DeltaE</protein> .


Transcriptional activation by this mutant form of membrane-bound DeltaE was approximately equivalent to that produced by intranuclear ICN .

Transcriptional activation by this mutant form of <protein>membrane-bound DeltaE</protein> was approximately equivalent to that produced by <protein>intranuclear ICN</protein> .


These data are most consistent with NOTCH1 oncogenesis and transcriptional activation being independent of association with RBP-Jkappa at promoter sites .

These data are most consistent with <protein>NOTCH1</protein> oncogenesis and transcriptional activation being independent of association with <protein>RBP-Jkappa</protein> at <dna>promoter sites</dna> .


Sp family members preferentially interact with the promoter proximal repeat within the HTLV-I enhancer .

<protein>Sp family members</protein> preferentially interact with the <dna>promoter proximal repeat</dna> within the <dna>HTLV-I enhancer</dna> .


Human T cell lymphotropic virus type I ( HTLV-I ) encodes the transactivator protein , Tax , which facilitates viral transcription from three 21 bp repeated elements within the U3 region of the long terminal repeat ( LTR ) .

Human T cell lymphotropic virus type I ( HTLV-I ) encodes the <protein>transactivator protein</protein> , <protein>Tax</protein> , which facilitates viral transcription from three <dna>21 bp repeated elements</dna> within the <dna>U3 region</dna> of the <dna>long terminal repeat</dna> ( <dna>LTR</dna> ) .


Examination of the basal factors interacting with the 21 bp repeat elements through electrophoretic mobility shift ( EMS ) analyses has demonstrated the formation of DNA-protein complexes common to each of the 21 bp repeats ( C1-C3 ) as well as three DNA-protein complexes specific to the promoter proximal ( pp ) repeat ( U1 ( U1A/U1B ) and U2 ; 1-4 ) .

Examination of the <protein>basal factors</protein> interacting with the <dna>21 bp repeat elements</dna> through electrophoretic mobility shift ( EMS ) analyses has demonstrated the formation of <protein>DNA-protein complexes</protein> common to each of the <dna>21 bp repeats</dna> ( <dna>C1-C3</dna> ) as well as three <protein>DNA-protein complexes</protein> specific to the <dna>promoter proximal ( pp ) repeat</dna> ( <dna>U1</dna> ( <dna>U1A/U1B</dna> ) and <dna>U2</dna> ; 1-4 ) .


These studies have indicated that the individual repeats are not identical with respect to the cellular factors with which they interact .

These studies have indicated that the individual repeats are not identical with respect to the <protein>cellular factors</protein> with which they interact .


EMS analyses utilizing a series of mutated pp repeat elements demonstrate that the nucleotide sequence requirements for U1 ( U1A/U1B ) and U2 formation are separable from those required for C1-C3 formation .

EMS analyses utilizing a series of <dna>mutated pp repeat elements</dna> demonstrate that the nucleotide sequence requirements for <dna>U1</dna> ( <dna>U1A/U1B</dna> ) and <dna>U2</dna> formation are separable from those required for <dna>C1-C3</dna> formation .


Competition EMS analyses utilizing Sp1 and CREB binding site oligonucleotides demonstrate that Sp family members are critical components of U1 ( U1A/U1B ) and U2 and that ATF/CREB family members are critical components of C1-C3 .

Competition EMS analyses utilizing Sp1 and CREB binding site oligonucleotides demonstrate that <protein>Sp family members</protein> are critical components of <dna>U1</dna> ( <dna>U1A/U1B</dna> ) and <dna>U2</dna> and that <protein>ATF/CREB family members</protein> are critical components of <dna>C1-C3</dna> .


EMS supershift analyses have demonstrated that Sp1 is involved in U1A formation while Sp3 is involved in U1B and U2 formation .

EMS supershift analyses have demonstrated that <protein>Sp1</protein> is involved in U1A formation while <protein>Sp3</protein> is involved in U1B and U2 formation .


EMS analyses performed with nuclear extracts from Tax-expressing Jurkat cells and HTLV-I-transformed peripheral blood mononuclear cells demonstrate that Tax prevents the formation of U1 ( U1A/U1B ) and U2 DNA-protein complexes .

EMS analyses performed with nuclear extracts from <cell_line>Tax-expressing Jurkat cells</cell_line> and <cell_type>HTLV-I-transformed peripheral blood mononuclear cells</cell_type> demonstrate that <protein>Tax</protein> prevents the formation of <dna>U1</dna> ( <dna>U1A/U1B</dna> ) and <dna>U2</dna> <protein>DNA-protein complexes</protein> .


Therefore , Tax appears to inhibit the interaction of Sp family members with the pp repeat .

Therefore , <protein>Tax</protein> appears to inhibit the interaction of <protein>Sp family members</protein> with the <dna>pp repeat</dna> .


Based on these observations , it is possible that the interaction of Sp and ATF/CREB family members with the pp repeat during basal and Tax -mediated transcription may play a critical role in viral gene expression during the initial stages of virus infection or during activation of a latent infection .

Based on these observations , it is possible that the interaction of <protein>Sp</protein> and <protein>ATF/CREB family members</protein> with the <dna>pp repeat</dna> during basal and <protein>Tax</protein> -mediated transcription may play a critical role in viral gene expression during the initial stages of virus infection or during activation of a latent infection .


Lineage- and stage-specific expression of runt box polypeptides in primitive and definitive hematopoiesis .

Lineage- and stage-specific expression of <protein>runt box polypeptides</protein> in primitive and definitive hematopoiesis .


Translocations involving the human CBFA2 locus have been associated with leukemia .

Translocations involving the <dna>human CBFA2 locus</dna> have been associated with leukemia .


This gene , originally named AML1 , is a human homologue of the Drosophila gene runt that controls early events in fly embryogenesis .

This gene , originally named <dna>AML1</dna> , is a human homologue of the <dna>Drosophila gene runt</dna> that controls early events in fly embryogenesis .


To clarify the role of mammalian runt products in normal and leukemic hematopoiesis , we have studied their pattern of expression in mouse hematopoietic tissues in the adult and during ontogeny using an anti-runt box antiserum .

To clarify the role of <protein>mammalian runt products</protein> in normal and leukemic hematopoiesis , we have studied their pattern of expression in mouse hematopoietic tissues in the adult and during ontogeny using an anti-runt box antiserum .


In the adult bone marrow , we found expression of runt polypeptides in differentiating myeloid cells and in B lymphocytes .

In the adult bone marrow , we found expression of runt polypeptides in <cell_type>differentiating myeloid cells</cell_type> and in <cell_type>B lymphocytes</cell_type> .


Within the erythroid lineage , runt expression is biphasic , clearly present in the erythroblasts of early blood islands and of the fetal liver , but absent in the adult .

Within the <cell_type>erythroid lineage</cell_type> , runt expression is biphasic , clearly present in the <cell_type>erythroblasts</cell_type> of early blood islands and of the fetal liver , but absent in the adult .


Biochemical analysis by Western blotting of fetal and adult hematopoietic populations shows several runt isoforms .

Biochemical analysis by Western blotting of <cell_type>fetal and adult hematopoietic populations</cell_type> shows several <protein>runt isoforms</protein> .


At least one of them appears to be myeloid specific .


Signal transducer and activator of transcription-3 ( STAT3 ) is constitutively activated in normal , self-renewing B-1 cells but only inducibly expressed in conventional B lymphocytes [ see comments ]

<protein>Signal transducer and activator of transcription-3</protein> ( <protein>STAT3</protein> ) is constitutively activated in normal , <cell_type>self-renewing B-1 cells</cell_type> but only inducibly expressed in conventional <cell_type>B lymphocytes</cell_type> [ see comments ]


Cytokine and growth factor receptor engagement leads to the rapid phosphorylation and activation of latent , cytosolic signal transducers and activators of transcription ( STAT ) proteins , which then translocate to the nucleus where they regulate transcriptional events from specific promoter sequences .

<protein>Cytokine</protein> and <protein>growth factor receptor</protein> engagement leads to the rapid phosphorylation and activation of latent , <protein>cytosolic signal transducers and activators of transcription ( STAT ) proteins</protein> , which then translocate to the nucleus where they regulate transcriptional events from <dna>specific promoter sequences</dna> .


STAT3 expression in particular has been associated with Abl , Src , and HTLV-1 transformation of normal cells .

<protein>STAT3</protein> expression in particular has been associated with <protein>Abl</protein> , <protein>Src</protein> , and HTLV-1 transformation of <cell_type>normal cells</cell_type> .


B-1 lymphocytes are self-renewing , CD5+ B cells that display a propensity for malignant transformation and are the normal counterpart to human chronic lymphocytic leukemias .

<cell_type>B-1 lymphocytes</cell_type> are self-renewing , <cell_type>CD5+ B cells</cell_type> that display a propensity for malignant transformation and are the normal counterpart to human chronic lymphocytic leukemias .


Further , B-1 cells are characterized by aberrant intracellular signaling , including hyperresponsiveness to phorbol ester PKC agonists .

Further , <cell_type>B-1 cells</cell_type> are characterized by aberrant intracellular signaling , including hyperresponsiveness to phorbol ester PKC agonists .


Here we demonstrate that B-1 lymphocytes constitutively express nuclear activated STAT3 , which is not expressed by unmanipulated conventional ( B-2 ) lymphocytes .

Here we demonstrate that <cell_type>B-1 lymphocytes</cell_type> constitutively express <protein>nuclear activated STAT3</protein> , which is not expressed by <cell_type>unmanipulated conventional ( B-2 ) lymphocytes</cell_type> .


In contrast , STAT3 activation is induced in B-2 cells after antigen receptor engagement in a delayed fashion ( after 3 h ) .

In contrast , <protein>STAT3</protein> activation is induced in <cell_type>B-2 cells</cell_type> after antigen receptor engagement in a delayed fashion ( after 3 h ) .


Induction of STAT3 is inhibited by both the serine/threonine protein kinase inhibitor H-7 and the immunosuppressive drug rapamycin and requires de novo protein synthesis , demonstrating novel coupling between sIg and STAT proteins that differs from the classical paradigm for STAT induction by cytokine receptors .

Induction of <protein>STAT3</protein> is inhibited by both the serine/threonine protein kinase inhibitor H-7 and the immunosuppressive drug rapamycin and requires de novo protein synthesis , demonstrating novel coupling between <protein>sIg</protein> and <protein>STAT proteins</protein> that differs from the classical paradigm for <protein>STAT</protein> induction by <protein>cytokine receptors</protein> .


The inability of prolonged stimulation of conventional B-2 cells with anti-Ig , a treatment sufficient to induce CD5 expression , to result in sustained STAT3 activation suggests that STAT3 is a specific nuclear marker for B-1 cells .

The inability of prolonged stimulation of conventional <cell_type>B-2 cells</cell_type> with <protein>anti-Ig</protein> , a treatment sufficient to induce <protein>CD5</protein> expression , to result in sustained <protein>STAT3</protein> activation suggests that <protein>STAT3</protein> is a specific nuclear marker for <cell_type>B-1 cells</cell_type> .


Thus , STAT3 may play a role in B cell antigen-specific signaling responses , and its constitutive activation is associated with a normal cell population exhibiting intrinsic proliferative behavior .

Thus , <protein>STAT3</protein> may play a role in B cell antigen-specific signaling responses , and its constitutive activation is associated with a normal cell population exhibiting intrinsic proliferative behavior .


Significance of quantitative analysis of AML1/ETO transcripts in peripheral blood stem cells from t ( 8 ; 21 ) acute myelogenous leukemia .

Significance of quantitative analysis of <rna>AML1/ETO transcripts</rna> in <cell_type>peripheral blood stem cells</cell_type> from t ( 8 ; 21 ) acute myelogenous leukemia .


Autologous peripheral blood stem cell transplantation ( PBSCT ) is replacing autologous bone marrow transplantation ( BMT ) in the treatment of leukemia .

Autologous <cell_type>peripheral blood stem cell</cell_type> transplantation ( PBSCT ) is replacing autologous bone marrow transplantation ( BMT ) in the treatment of leukemia .


One of the potential advantages of autologous PBSCT is the possibility that peripheral blood stem cells ( PBSC ) are less likely to be contaminated by leukemic cells than bone marrow grafts .

One of the potential advantages of autologous PBSCT is the possibility that <cell_type>peripheral blood stem cells</cell_type> ( <cell_type>PBSC</cell_type> ) are less likely to be contaminated by <cell_type>leukemic cells</cell_type> than bone marrow grafts .


However , the major problem still remains the high incidence of leukemic relapse following autologous PBSCT , which may be caused by the reinfusion of PBSC contaminated by leukemic cells .

However , the major problem still remains the high incidence of leukemic relapse following autologous PBSCT , which may be caused by the reinfusion of <cell_type>PBSC</cell_type> contaminated by <cell_type>leukemic cells</cell_type> .


Recently , we have developed a quantitative assay using competitive reverse transcriptase polymerase chain reaction that estimates the number of AML1/ETO transcripts in t ( 8 ; 21 ) acute myelogenous leukemia ( AML ) , in order to determine the degree of leukemic cell contamination in PBSC harvests , and to monitor minimal residual disease ( MRD ) quantitatively in patients with t ( 8 ; 21 ) AML .

Recently , we have developed a quantitative assay using competitive <protein>reverse transcriptase polymerase</protein> chain reaction that estimates the number of <rna>AML1/ETO transcripts</rna> in t ( 8 ; 21 ) acute myelogenous leukemia ( AML ) , in order to determine the degree of leukemic cell contamination in <cell_type>PBSC</cell_type> harvests , and to monitor minimal residual disease ( MRD ) quantitatively in patients with t ( 8 ; 21 ) AML .


Our data indicate that although PBSC harvests collected after consolidation chemotherapy are contaminated by leukemic cells , the degree of leukemic cell contamination decreases with repeated cycles of chemotherapy .

Our data indicate that although <cell_type>PBSC</cell_type> harvests collected after consolidation chemotherapy are contaminated by <cell_type>leukemic cells</cell_type> , the degree of leukemic cell contamination decreases with repeated cycles of chemotherapy .


Furthermore , the MRD in PBSC harvests is less than in the corresponding bone marrow obtained on the day of the PBSC collection .

Furthermore , the MRD in <cell_type>PBSC</cell_type> harvests is less than in the corresponding bone marrow obtained on the day of the <cell_type>PBSC</cell_type> collection .


There appears to be no relationship between the number of AML1/ETO transcripts found in the infused PBSC harvests and the incidence of leukemic relapse following autologous PBSCT in our study .

There appears to be no relationship between the number of <rna>AML1/ETO transcripts</rna> found in the infused <cell_type>PBSC</cell_type> harvests and the incidence of leukemic relapse following autologous PBSCT in our study .


However , a substantial decrease of AML1/ETO transcripts was seen following autologous PBSCT .

However , a substantial decrease of <rna>AML1/ETO transcripts</rna> was seen following autologous PBSCT .


Thus , the quantitative analysis of AML1/ETO transcripts may be clinically useful in patients with t ( 8 ; 21 ) AML .

Thus , the quantitative analysis of <rna>AML1/ETO transcripts</rna> may be clinically useful in patients with t ( 8 ; 21 ) AML .


Upregulation of c-Fos in activated T lymphoid and monocytic cells by human immunodeficiency virus-1 Tat protein .

Upregulation of <protein>c-Fos</protein> in <cell_type>activated T lymphoid and monocytic cells</cell_type> by <protein>human immunodeficiency virus-1 Tat protein</protein> .


The regulatory Tat protein of the human immunodeficiency virus type-1 ( HIV-1 ) is essential for viral replication and also shows pleiotropic activities on various cell functions .

The <protein>regulatory Tat protein</protein> of the human immunodeficiency virus type-1 ( HIV-1 ) is essential for viral replication and also shows pleiotropic activities on various cell functions .


To get further insights into the molecular mechanisms underlying the biological activity of Tat , we investigated the effect of endogenous and exogenous Tat protein on c-fos gene expression in T lymphoblastoid ( Jurkat ) and monocytic ( U937 ) cell lines , as well as in primary peripheral blood mononuclear cells ( PBMC ) .

To get further insights into the molecular mechanisms underlying the biological activity of <protein>Tat</protein> , we investigated the effect of endogenous and exogenous <protein>Tat protein</protein> on c-fos gene expression in <cell_line>T lymphoblastoid ( Jurkat ) and monocytic ( U937 ) cell lines</cell_line> , as well as in <cell_type>primary peripheral blood mononuclear cells</cell_type> ( <cell_type>PBMC</cell_type> ) .


Transient cotransfection of tat cDNA in sense orientation ( tat/S ) , together with a plasmid containing the c-fos promoter ( FC3 , from- 711 to +42 ) in front of the bacterial chloramphenicol acetyltransferase ( CAT ) gene significantly enhanced CAT activity in Jurkat cells activated by the addition of 15 % fetal calf serum ( FCS ) or 5 micrograms/mL phytohemagglutinin plus 10 ( -7 ) mol/L phorbol myristate acetate ( PMA ) and U937 cells activated by 15 % FCS or 10 ( -7 ) mol/L PMA .

Transient cotransfection of <dna>tat cDNA</dna> in sense orientation ( <dna>tat/S</dna> ) , together with a <dna>plasmid</dna> containing the <dna>c-fos promoter</dna> ( <dna>FC3</dna> , from- <dna>711 to +42</dna> ) in front of the <dna>bacterial chloramphenicol acetyltransferase ( CAT ) gene</dna> significantly enhanced <protein>CAT</protein> activity in <cell_line>Jurkat cells</cell_line> activated by the addition of 15 % fetal calf serum ( FCS ) or 5 micrograms/mL <protein>phytohemagglutinin</protein> plus 10 ( -7 ) mol/L phorbol myristate acetate ( PMA ) and <cell_line>U937 cells</cell_line> activated by 15 % FCS or 10 ( -7 ) mol/L PMA .


This effect was specifically due to Tat , since Jurkat and U937 cells cotransfected either with tat cDNA in antisense orientation ( tat/AS ) , tat carrying a mutation in the aminoacid cys22-gly22 ( tat 22/S ) or with the backbone vector alone ( pRPneo-SL3 ) did not show any significant difference in c-fos promoter activity as compared to cells transfected with FC3 plasmid alone .

This effect was specifically due to <protein>Tat</protein> , since <cell_line>Jurkat</cell_line> and <cell_line>U937 cells</cell_line> cotransfected either with <dna>tat cDNA</dna> in antisense orientation ( <dna>tat/AS</dna> ) , <dna>tat</dna> carrying a mutation in the <protein>aminoacid cys22-gly22</protein> ( <dna>tat 22/S</dna> ) or with the <dna>backbone vector alone</dna> ( <dna>pRPneo-SL3</dna> ) did not show any significant difference in <dna>c-fos promoter</dna> activity as compared to cells transfected with <dna>FC3 plasmid</dna> alone .


By using deletion mutants of the c-fos promoter , we found that the minimal DNA sequence required for Tat activity was located between nucleotides -404/-220 and that the serum responsive element ( SRE , -317/-288 ) , present within this region , was still responsive to Tat .

By using deletion mutants of the <dna>c-fos promoter</dna> , we found that the <dna>minimal DNA sequence</dna> required for <protein>Tat</protein> activity was located between <dna>nucleotides -404/-220</dna> and that the <dna>serum responsive element</dna> ( <dna>SRE</dna> , <dna>-317/-288</dna> ) , present within this region , was still responsive to <protein>Tat</protein> .


A single point mutation in the SRE completely abrogated the responsiveness to tat/S .

A <dna>single point mutation</dna> in the <dna>SRE</dna> completely abrogated the responsiveness to <dna>tat/S</dna> .


Exogenous recombinant Tat protein was also able to upregulate c-fos promoter activity in serum-activated Jurkat and U937 cells , as well as endogenous c-fos mRNA expression and c-Fos protein synthesis in both serum-activated cell lines and primary PBMC .

Exogenous <protein>recombinant Tat protein</protein> was also able to upregulate <dna>c-fos promoter</dna> activity in serum-activated Jurkat and <cell_line>U937 cells</cell_line> , as well as endogenous <rna>c-fos mRNA</rna> expression and <protein>c-Fos</protein> protein synthesis in both <cell_line>serum-activated cell lines</cell_line> and <cell_type>primary PBMC</cell_type> .


c-Fos protein was shown essential for an optimal transactivation of the HIV-1 long terminal repeat ( LTR ) by Tat : incubation of Jurkat cells with antisense , but not sense , c-fos oligonucleotides significantly reduced either the Tat -enhanced expression of an LTR-CAT reporter construct or the levels of gag p24 in the culture supernatants of Jurkat cells and PBMC acutely infected with HIV-1 .

<protein>c-Fos</protein> protein was shown essential for an optimal transactivation of the HIV-1 <dna>long terminal repeat</dna> ( <dna>LTR</dna> ) by <protein>Tat</protein> : incubation of <cell_line>Jurkat cells</cell_line> with antisense , but not sense , c-fos oligonucleotides significantly reduced either the <protein>Tat</protein> -enhanced expression of an <dna>LTR-CAT reporter construct</dna> or the levels of <protein>gag p24</protein> in the culture supernatants of <cell_line>Jurkat cells</cell_line> and <cell_type>PBMC</cell_type> acutely infected with HIV-1 .


Our data suggest that the c-fos upregulation mediated by Tat might play a significant role in the control of viral gene transactivation .

Our data suggest that the <dna>c-fos</dna> upregulation mediated by <protein>Tat</protein> might play a significant role in the control of viral gene transactivation .


Cytokine signal networks and a new era in biomedical research .

<protein>Cytokine signal networks</protein> and a new era in biomedical research .


Elucidation of the biochemical nature of the signal transduction pathway that regulate transcription and replication is the focus of attention in molecular biology .


This research may make feasible manipulation of growth and differentiation of mammalian cells , which in turn would have profound implication in biomedical research on cell and gene therapy , and development of pharmaceutical products .

This research may make feasible manipulation of growth and differentiation of <cell_type>mammalian cells</cell_type> , which in turn would have profound implication in biomedical research on cell and gene therapy , and development of pharmaceutical products .


Cytokines control growth , differentiation , death , and function of cells of lymphocytic , hemopoietic systems , and together with nerve cells provide a pertinent model to study intercellular communications and intercellular signal networks .

<protein>Cytokines</protein> control growth , differentiation , death , and function of cells of <cell_type>lymphocytic , hemopoietic systems</cell_type> , and together with <cell_type>nerve cells</cell_type> provide a pertinent model to study intercellular communications and intercellular signal networks .


This review outlines general features of signal transduction and several aspects of cytokine networks are discussed with emphasis on : transcriptional regulation of Th1 and Th2-specific cytokine genes in T cells , the roles of cytokines and their receptors in growth and differentiation of hemopoietic cells , and the manipulation of cytokine networks .

This review outlines general features of signal transduction and several aspects of <protein>cytokine networks</protein> are discussed with emphasis on : transcriptional regulation of Th1 and Th2-specific <dna>cytokine genes</dna> in <cell_type>T cells</cell_type> , the roles of <protein>cytokines</protein> and their receptors in growth and differentiation of <cell_type>hemopoietic cells</cell_type> , and the manipulation of <protein>cytokine networks</protein> .


Regulation of the tissue factor gene in human monocytic cells .

Regulation of the <dna>tissue factor gene</dna> in <cell_type>human monocytic cells</cell_type> .


Role of AP-1 , NF-kappa B/Rel , and Sp1 proteins in uninduced and lipopolysaccharide-induced expression .

Role of <protein>AP-1</protein> , <protein>NF-kappa B/Rel</protein> , and <protein>Sp1</protein> proteins in uninduced and lipopolysaccharide-induced expression .


Tissue factor ( TF ) expression by peripheral blood monocytes during sepsis initiates intravascular thrombosis .

<protein>Tissue factor</protein> ( TF ) expression by <cell_type>peripheral blood monocytes</cell_type> during sepsis initiates intravascular thrombosis .


Bacterial lipopolysaccharide ( LPS ) rapidly induces TF gene transcription in monocytes .

Bacterial lipopolysaccharide ( LPS ) rapidly induces <dna>TF gene</dna> transcription in <cell_type>monocytes</cell_type> .


The human TF promoter contains binding sites for the transcription factors AP-1 , c-Rel/p65 , Egr-1 , and Sp1 .

The <dna>human TF promoter</dna> contains binding sites for the transcription factors <protein>AP-1</protein> , <protein>c-Rel/p65</protein> , <protein>Egr-1</protein> , and <protein>Sp1</protein> .


NF-kappa B/Rel proteins have been shown to physically interact with both AP-1 and Sp1 proteins .

<protein>NF-kappa B/Rel proteins</protein> have been shown to physically interact with both <protein>AP-1 and Sp1 proteins</protein> .


In this study , we investigated the role of these transcription factors in uninduced and LPS-induced TF gene expression in human monocytic THP-1 cells .

In this study , we investigated the role of these <protein>transcription factors</protein> in uninduced and LPS-induced <dna>TF gene</dna> expression in <cell_type>human monocytic THP-1 cells</cell_type> .


Deletional analysis indicated that five Sp1 sites mediated basal expression in uninduced cells .

Deletional analysis indicated that five <dna>Sp1 sites</dna> mediated basal expression in <cell_type>uninduced cells</cell_type> .


The two AP-1 sites bound c-Fos/c-Jun heterodimers in both unstimulated and LPS-stimulated cells .

The two <dna>AP-1 sites</dna> bound <protein>c-Fos/c-Jun heterodimers</protein> in both <cell_type>unstimulated and LPS-stimulated cells</cell_type> .


Maximal LPS induction of the TF promoter required the two AP-1 sites and the kappa B site within the LPS response element .

Maximal LPS induction of the <dna>TF promoter</dna> required the two <protein>AP-1</protein> sites and the <dna>kappa B site</dna> within the <dna>LPS response element</dna> .


Disruption of the conserved spacing between the proximal AP-1 site and the kappa B site abolished LPS induction .

Disruption of the conserved spacing between the <dna>proximal AP-1 site</dna> and the <dna>kappa B site</dna> abolished LPS induction .


Replacement of the two AP-1 sites with intrinsically bent DNA partially restored LPS induction , suggesting an additional structural role for the AP-1 sites .

Replacement of the two <dna>AP-1 sites</dna> with intrinsically bent DNA partially restored LPS induction , suggesting an additional structural role for the <dna>AP-1 sites</dna> .


Synergistic transactivation of the LPS response element in Drosophila Schneider cells by coexpression of c-Fos , c-Jun , c-Rel , and p65 or c-Jun and p65 required the transactivation domains of c-Jun and p65 .

Synergistic transactivation of the <dna>LPS response element</dna> in <cell_line>Drosophila Schneider cells</cell_line> by coexpression of <protein>c-Fos</protein> , <protein>c-Jun</protein> , <protein>c-Rel</protein> , and <protein>p65</protein> or <protein>c-Jun</protein> and <protein>p65</protein> required the <protein>transactivation domains</protein> of <protein>c-Jun</protein> and <protein>p65</protein> .


These data indicated that c-Fos /c-Jun , c-Rel /p65 , and Sp1 regulate TF gene expression in human monocytic cells .

These data indicated that <protein>c-Fos</protein> <protein>/c-Jun</protein> , <protein>c-Rel</protein> <protein>/p65</protein> , and <protein>Sp1</protein> regulate <dna>TF gene</dna> expression in <cell_type>human monocytic cells</cell_type> .


Inhibitory effect of growth hormone on TNF-alpha secretion and nuclear factor-kappaB translocation in lipopolysaccharide-stimulated human monocytes .

Inhibitory effect of growth hormone on <protein>TNF-alpha</protein> secretion and <protein>nuclear factor-kappaB</protein> translocation in <cell_type>lipopolysaccharide-stimulated human monocytes</cell_type> .


Several studies have pointed to a link between immune and endocrine systems , including a regulatory function of GH on monocyte activation .


The present study demonstrates that human THP-1 promonocytic cells , engineered by gene transfer to constitutively produce human growth hormone ( hGH ) , secreted depressed amounts of TNF-alpha in response to challenge by LPS .

The present study demonstrates that human <cell_line>THP-1 promonocytic cells</cell_line> , engineered by gene transfer to constitutively produce human growth hormone ( hGH ) , secreted depressed amounts of <protein>TNF-alpha</protein> in response to challenge by LPS .


The effect of GH appears to occur in an autocrine fashion , since the inhibitory effect on TNF-alpha secretion by constitutive GH production could be abolished in the presence of anti-hGH mAb .

The effect of GH appears to occur in an autocrine fashion , since the inhibitory effect on <protein>TNF-alpha</protein> secretion by constitutive GH production could be abolished in the presence of <protein>anti-hGH mAb</protein> .


The GH-induced inhibitory effect was also observed using normal human monocytes and monocyte-derived macrophages .

The GH-induced inhibitory effect was also observed using <cell_type>normal human monocytes</cell_type> and <cell_type>monocyte-derived macrophages</cell_type> .


Inhibition of TNF-alpha production by THP-1-hGH-transfected cells cultured in the presence of LPS is dependent on a selective pathway , since no inhibition of TNF-alpha production was observed when cells were cultured in the presence of PMA .

Inhibition of <protein>TNF-alpha</protein> production by <cell_line>THP-1-hGH-transfected cells</cell_line> cultured in the presence of LPS is dependent on a selective pathway , since no inhibition of <protein>TNF-alpha</protein> production was observed when cells were cultured in the presence of PMA .


Inhibition of TNF-alpha secretion by LPS-stimulated THP-1-hGH cells was associated with a decrease in nuclear translocation of nuclear factor-kappaB .

Inhibition of <protein>TNF-alpha</protein> secretion by <cell_line>LPS-stimulated THP-1-hGH cells</cell_line> was associated with a decrease in nuclear translocation of <protein>nuclear factor-kappaB</protein> .


The capacity of GH to inhibit LPS-induced TNF-alpha production by monocytes without altering other pathways leading to TNF-alpha production may be of potential relevance in septic shock , since GH is available for clinical use .

The capacity of GH to inhibit LPS-induced <protein>TNF-alpha</protein> production by <cell_type>monocytes</cell_type> without altering other pathways leading to <protein>TNF-alpha</protein> production may be of potential relevance in septic shock , since GH is available for clinical use .


Nuclear factor-kappaB activation in human monocytes stimulated with lipopolysaccharide is inhibited by fibroblast conditioned medium and exogenous PGE2 .

<protein>Nuclear factor-kappaB</protein> activation in <cell_type>human monocytes</cell_type> stimulated with lipopolysaccharide is inhibited by fibroblast conditioned medium and exogenous PGE2 .


The nuclear factor kappaB ( NF-kappaB ) is thought to be crucially involved in the gene activation of several cytokines , including tumor necrosis factor alpha ( TNF ) .

The <protein>nuclear factor kappaB</protein> ( <protein>NF-kappaB</protein> ) is thought to be crucially involved in the gene activation of several <protein>cytokines</protein> , including <protein>tumor necrosis factor alpha</protein> ( <protein>TNF</protein> ) .


Previously , we showed that fibroblast conditioned medium ( FCM ) is able to inhibit both TNF mRNA accumulation and protein release in peripheral blood-derived human monocytes ( PBM ) stimulated with lipopolysaccharide ( LPS ) .

Previously , we showed that fibroblast conditioned medium ( FCM ) is able to inhibit both <rna>TNF mRNA</rna> accumulation and protein release in <cell_line>peripheral blood-derived human monocytes</cell_line> ( <cell_line>PBM</cell_line> ) stimulated with lipopolysaccharide ( LPS ) .


In this study we have investigated the effect of FCM on the LPS-induced DNA-binding activity of NF-kappaB , by means of electrophoretic shift assay ( EMSA ) .

In this study we have investigated the effect of FCM on the LPS-induced DNA-binding activity of <protein>NF-kappaB</protein> , by means of electrophoretic shift assay ( EMSA ) .


We provide evidence that FCM strongly inhibits the LPS-induced NF-kappaB activation in PBM .

We provide evidence that FCM strongly inhibits the LPS-induced <protein>NF-kappaB</protein> activation in <cell_line>PBM</cell_line> .


Furthermore , we show that exogenous PGE2 mimics the NF-kappaB inhibitory effect of FCM .

Furthermore , we show that exogenous PGE2 mimics the <protein>NF-kappaB</protein> inhibitory effect of FCM .


On the other hand , FCM produced in the presence of indomethacin does not inhibit NF-kappaB activation by LPS .

On the other hand , FCM produced in the presence of indomethacin does not inhibit <protein>NF-kappaB</protein> activation by LPS .


Our results lend further support to the hypothesis that inflammatory and immune responses of monocytes/macrophages may be modulated at the molecular level by signals originating from tissue structural cells such as fibroblasts .

Our results lend further support to the hypothesis that inflammatory and immune responses of <cell_type>monocytes/macrophages</cell_type> may be modulated at the molecular level by signals originating from <cell_type>tissue structural cells</cell_type> such as <cell_type>fibroblasts</cell_type> .


Human neutrophil elastase proteolytically activates the platelet integrin alphaIIbbeta3 through cleavage of the carboxyl terminus of the alphaIIb subunit heavy chain .

<protein>Human neutrophil elastase</protein> proteolytically activates the <protein>platelet integrin</protein> <protein>alphaIIbbeta3</protein> through cleavage of the <protein>carboxyl terminus</protein> of the <protein>alphaIIb subunit heavy chain</protein> .


Involvement in the potentiation of platelet aggregation .


Neutrophil elastase ( NE ) and cathepsin G are two serine proteinases released concomitantly by stimulated polymorphonuclear neutrophils .

<protein>Neutrophil elastase</protein> ( <protein>NE</protein> ) and <protein>cathepsin G</protein> are two <protein>serine proteinases</protein> released concomitantly by stimulated <cell_type>polymorphonuclear neutrophils</cell_type> .


We previously demonstrated that while NE by itself does not activate human platelets , it strongly enhances the weak aggregation induced by a threshold concentration of cathepsin G ( threshold of cathepsin G ) ( Renesto , P. , and Chignard , M. ( 1993 ) Blood 82 , 139-144 ) .

We previously demonstrated that while <protein>NE</protein> by itself does not activate <cell_type>human platelets</cell_type> , it strongly enhances the weak aggregation induced by a threshold concentration of <protein>cathepsin G</protein> ( threshold of <protein>cathepsin G</protein> ) ( Renesto , P. , and Chignard , M. ( 1993 ) Blood 82 , 139-144 ) .


The aim of this study was to delineate the molecular mechanisms involved in this potentiation process .


Two main pieces of data prompted us to focus on the activation of the platelet fibrinogen receptor , the alphaIIbbeta3 integrin .

Two main pieces of data prompted us to focus on the activation of the <protein>platelet fibrinogen receptor</protein> , the <protein>alphaIIbbeta3 integrin</protein> .


First , previous studies have shown this integrin to be particularly prone to proteolytic regulation of its function .

First , previous studies have shown this <protein>integrin</protein> to be particularly prone to proteolytic regulation of its function .


Second , we found that the potentiating activity of NE on the threshold of cathepsin G -induced platelet aggregation was strictly dependent on the presence of exogenous fibrinogen .

Second , we found that the potentiating activity of <protein>NE</protein> on the threshold of <protein>cathepsin G</protein> -induced platelet aggregation was strictly dependent on the presence of <protein>exogenous fibrinogen</protein> .


Using flow cytometry analysis , NE was shown to trigger a time-dependent binding of PAC-1 and AP-5 , two monoclonal antibodies specific for the activated and ligand-occupied conformers of alphaIIbbeta3 .

Using flow cytometry analysis , <protein>NE</protein> was shown to trigger a time-dependent binding of <protein>PAC-1</protein> and <protein>AP-5</protein> , two <protein>monoclonal antibodies</protein> specific for the activated and ligand-occupied conformers of <protein>alphaIIbbeta3</protein> .


Furthermore , the potentiated aggregation was shown to result from an increased capacity of platelets to bind fibrinogen .


Indeed , the combination of NE and threshold of cathepsin G increased the binding of PAC-1 approximately 5.5-fold over basal values measured on nontreated platelets , whereas this binding raised only by approximately 3-fold in threshold of cathepsin G-stimulated platelets ( p < 0.05 ) .

Indeed , the combination of <protein>NE</protein> and threshold of <protein>cathepsin G</protein> increased the binding of <protein>PAC-1</protein> approximately 5.5-fold over basal values measured on <cell_type>nontreated platelets</cell_type> , whereas this binding raised only by approximately 3-fold in threshold of <cell_type>cathepsin G-stimulated platelets</cell_type> ( p < 0.05 ) .


By contrast , phosphatidic acid accumulation , pleckstrin phosphorylation , and calcium mobilization produced by the combination of NE and threshold of cathepsin G were not significantly different from those measured with threshold of cathepsin G alone ( p > 0.05 ) , indicating that the phospholipase C /protein kinase C pathway is not involved in the potentiation of aggregation .

By contrast , phosphatidic acid accumulation , pleckstrin phosphorylation , and calcium mobilization produced by the combination of <protein>NE</protein> and threshold of <protein>cathepsin G</protein> were not significantly different from those measured with threshold of <protein>cathepsin G</protein> alone ( p > 0.05 ) , indicating that the <protein>phospholipase C</protein> <protein>/protein kinase C</protein> pathway is not involved in the potentiation of aggregation .


The foregoing data , as well as the requirement of catalytically active NE to trigger alphaIIbbeta3 activation and potentiate threshold of cathepsin G -initiated platelet aggregation , led us to examine whether the structure of this integrin was affected by NE .

The foregoing data , as well as the requirement of catalytically active <protein>NE</protein> to trigger <protein>alphaIIbbeta3</protein> activation and potentiate threshold of <protein>cathepsin G</protein> -initiated platelet aggregation , led us to examine whether the structure of this <protein>integrin</protein> was affected by <protein>NE</protein> .


Immunoblot and flow cytometry analysis revealed a limited proteolysis of the carboxyl terminus of the alphaIIb subunit heavy chain ( alphaIIbH ) , as judged by the disappearance of the epitope for the monoclonal antibody PMI-1 .

Immunoblot and flow cytometry analysis revealed a limited proteolysis of the <protein>carboxyl terminus</protein> of the <protein>alphaIIb subunit heavy chain</protein> ( <protein>alphaIIbH</protein> ) , as judged by the disappearance of the epitope for the <protein>monoclonal antibody PMI-1</protein> .


Mass spectrometry studies performed on a synthetic peptide mapping over the cleavage domain of alphaIIbH predicted the site of proteolysis as located between Val837 and Asp838 .

Mass spectrometry studies performed on a synthetic peptide mapping over the <protein>cleavage domain</protein> of <protein>alphaIIbH</protein> predicted the site of proteolysis as located between Val837 and Asp838 .


Treatment by NE of ATP-depleted platelets or Chinese hamster ovary cells expressing human recombinant alphaIIbbeta3 clearly established that activation of the integrin was independent of signal transduction events and was concomitant with the proteolysis of alphaIIbH .

Treatment by <protein>NE</protein> of <cell_type>ATP-depleted platelets</cell_type> or <cell_line>Chinese hamster ovary cells</cell_line> expressing <protein>human recombinant alphaIIbbeta3</protein> clearly established that activation of the <protein>integrin</protein> was independent of signal transduction events and was concomitant with the proteolysis of <protein>alphaIIbH</protein> .


AP-1 derived from mature monocytes and astrocytes preferentially interacts with the HTLV-I promoter central 21 bp repeat .

<protein>AP-1</protein> derived from <cell_type>mature monocytes</cell_type> and <cell_type>astrocytes</cell_type> preferentially interacts with the <dna>HTLV-I promoter</dna> central <dna>21 bp repeat</dna> .


Characterization of the cellular transcription factors interacting with the human T cell lymphotropic virus type I ( HTLV-I ) long terminal repeat ( LTR ) is essential to dissecting the mechanisms involved in viral transcription that may be pertinent to the oncogenic and neuropathogenic processes associated with HTLV-I infection in both the immune and nervous systems .

Characterization of the cellular <protein>transcription factors</protein> interacting with the <dna>human T cell lymphotropic virus type I ( HTLV-I ) long terminal repeat</dna> ( <dna>LTR</dna> ) is essential to dissecting the mechanisms involved in viral transcription that may be pertinent to the oncogenic and neuropathogenic processes associated with HTLV-I infection in both the immune and nervous systems .


Electrophoretic mobility shift ( EMS ) analyses utilizing oligonucleotides homologous to each of the 21 bp repeat elements reacted with nuclear extracts derived from cell lines of lymphocytic , monocytic , neuronal , and glial cell origin have demonstrated differential binding of cellular factors to the three 21 bp repeats ( 1-4 ) .

Electrophoretic mobility shift ( EMS ) analyses utilizing oligonucleotides homologous to each of the <dna>21 bp repeat elements</dna> reacted with nuclear extracts derived from <cell_line>cell lines</cell_line> of <cell_type>lymphocytic , monocytic , neuronal , and glial cell origin</cell_type> have demonstrated differential binding of <protein>cellular factors</protein> to the three <dna>21 bp repeats</dna> ( 1-4 ) .


ATF/CREB and Sp family members interacted with the 21 bp repeats to form DNA-protein complexes common to all cell types examined .

<protein>ATF/CREB</protein> and <protein>Sp family members</protein> interacted with the <dna>21 bp repeats</dna> to form <protein>DNA-protein complexes</protein> common to all cell types examined .


However , a unique DNA-protein complex was detected when the promoter central 21 bp repeat was reacted with nuclear extracts derived from either the U-373 MG glioblastoma cell line or the THP-1 mature monocytic cell line .

However , a unique <protein>DNA-protein complex</protein> was detected when the promoter central <dna>21 bp repeat</dna> was reacted with nuclear extracts derived from either the <cell_line>U-373 MG glioblastoma cell line</cell_line> or the <cell_line>THP-1 mature monocytic cell line</cell_line> .


Based on nucleotide sequence requirements and immunoreactivity , we demonstrate that this DNA-protein complex is comprised of the AP-1 components , Fos and Jun .

Based on nucleotide sequence requirements and immunoreactivity , we demonstrate that this <protein>DNA-protein complex</protein> is comprised of the <protein>AP-1</protein> components , <protein>Fos</protein> and <protein>Jun</protein> .


HIV does not replicate in naive CD4 T cells stimulated with CD3/CD28 .

HIV does not replicate in <cell_type>naive CD4 T cells</cell_type> stimulated with <protein>CD3/CD28</protein> .


In this report , we demonstrate that the T cell tropic strain of HIV , LAI , does not replicate in naive CD4 T cells stimulated by cross-linking CD3 and CD28 .

In this report , we demonstrate that the T cell tropic strain of HIV , LAI , does not replicate in <cell_type>naive CD4 T cells</cell_type> stimulated by cross-linking <protein>CD3</protein> and <protein>CD28</protein> .


In contrast , LAI replicates well in memory CD4 T cells stimulated in the same way .

In contrast , LAI replicates well in <cell_type>memory CD4 T cells</cell_type> stimulated in the same way .


Unlike this physiologically relevant stimulation , PHA stimulates productive LAI replication in both naive and memory T cells .

Unlike this physiologically relevant stimulation , <protein>PHA</protein> stimulates productive LAI replication in both <cell_type>naive and memory T cells</cell_type> .


These studies were conducted with highly purified ( FACS-isolated ) subsets of CD4 T cells identified by expression of both CD45RA and CD62L .

These studies were conducted with highly purified ( FACS-isolated ) subsets of <cell_type>CD4 T cells</cell_type> identified by expression of both <protein>CD45RA</protein> and <protein>CD62L</protein> .


Remixing of purified T cells showed that naive T cells do not suppress LAI replication in memory T cells and that memory T cells do not restore LAI expression in naive T cells .

Remixing of <cell_type>purified T cells</cell_type> showed that <cell_type>naive T cells</cell_type> do not suppress LAI replication in <cell_type>memory T cells</cell_type> and that <cell_type>memory T cells</cell_type> do not restore LAI expression in <cell_type>naive T cells</cell_type> .


The suppression of productive LAI replication in naive T cells is not due to differential expression of viral coreceptors , nor is it due to inhibition of activation of the important HIV transcription factors , nuclear factor-kappaB and activator protein-1 .

The suppression of productive LAI replication in <cell_type>naive T cells</cell_type> is not due to differential expression of <protein>viral coreceptors</protein> , nor is it due to inhibition of activation of the important HIV <protein>transcription factors</protein> , <protein>nuclear factor-kappaB</protein> and <protein>activator protein-1</protein> .


The inherent resistance of naive T cells to productive HIV infection , coupled with their proliferative advantage as demonstrated here , provides a sound basis for proposed clinical therapies using ex vivo expansion and reinfusion of CD4 T cells from HIV-infected adults .

The inherent resistance of <cell_type>naive T cells</cell_type> to productive HIV infection , coupled with their proliferative advantage as demonstrated here , provides a sound basis for proposed clinical therapies using ex vivo expansion and reinfusion of <cell_type>CD4 T cells</cell_type> from HIV-infected adults .


A PMLRARalpha transgene initiates murine acute promyelocytic leukemia .

A <dna>PMLRARalpha transgene</dna> initiates murine acute promyelocytic leukemia .


The malignant cells of acute promyelocytic leukemia ( APL ) contain a reciprocal chromosomal translocation that fuses the promyelocytic leukemia gene ( PML ) with the retinoic acid receptor alpha gene ( RAR alpha ) .

The <cell_type>malignant cells</cell_type> of acute promyelocytic leukemia ( APL ) contain a reciprocal chromosomal translocation that fuses the <dna>promyelocytic leukemia gene</dna> ( PML ) with the <dna>retinoic acid receptor alpha gene</dna> ( <protein>RAR alpha</protein> ) .


To test the hypothesis that the chimera PMLRAR alpha plays a role in leukemogenesis , we expressed a PMLRAR alpha cDNA in myeloid cells of transgenic mice .

To test the hypothesis that the <protein>chimera PMLRAR alpha</protein> plays a role in leukemogenesis , we expressed a <dna>PMLRAR alpha cDNA</dna> in <cell_type>myeloid cells</cell_type> of transgenic mice .


PMLRAR alpha transgenic mice exhibited impaired neutrophil maturation early in life , which progressed at a low frequency over the course of several months to overt APL .


Both the preleukemic state and the leukemia could be transplanted to nontransgenic mice , and the transplanted preleukemia could progress to APL .


The APL recapitulated features of the human disease , including a response to retinoic acid .


Retinoic acid caused the leukemic cells to differentiate in vitro and in vivo , eliciting remissions of both the preleukemic state and APL in mice .

Retinoic acid caused the <cell_type>leukemic cells</cell_type> to differentiate in vitro and in vivo , eliciting remissions of both the preleukemic state and APL in mice .


Our results demonstrate that PMLRAR alpha impairs neutrophil differentiation and initiates the development of APL .

Our results demonstrate that <protein>PMLRAR alpha</protein> impairs neutrophil differentiation and initiates the development of APL .


The transgenic mice described here provide an apparently accurate model for human APL that includes clear evidence of tumor progression .


The model should be useful for exploring the molecular pathogenesis of APL and the mechanisms of the therapeutic response to retinoic acid , as well as for preclinical studies of therapeutic regimens .


DNA methylation changes in hematologic malignancies : biologic and clinical implications .


DNA methylation changes are among the most common detectable abnormalities in human neoplasia .


Hypermethylation within the promoters of selected genes appears to be especially common in all types of human hematopoietic neoplasms , and is usually associated with inactivation of the involved gene ( s ) .

Hypermethylation within the <dna>promoters</dna> of selected genes appears to be especially common in all types of <cell_type>human hematopoietic neoplasms</cell_type> , and is usually associated with inactivation of the involved gene ( s ) .


Such hypermethylation-associated silencing of gene expression has been shown for several genes regulating the growth and differentiation of hematopoietic cells , including the estrogen receptor ( ER ) gene , P15 , P16 and others .

Such hypermethylation-associated silencing of gene expression has been shown for several genes regulating the growth and differentiation of <cell_type>hematopoietic cells</cell_type> , including the <dna>estrogen receptor ( ER ) gene</dna> , P15 , P16 and others .


Hypermethylation within the promoters of some genes appear to be an early event in the pathogenesis of neoplasia ( ER , P15 ) , while other genes seem to become methylated during the progression of leukemias ( HIC1 , c-abl ) .

Hypermethylation within the <dna>promoters</dna> of some genes appear to be an early event in the pathogenesis of neoplasia ( <dna>ER</dna> , <dna>P15</dna> ) , while other genes seem to become methylated during the progression of leukemias ( HIC1 , c-abl ) .


The high prevalence of promoter methylation suggests that this molecular abnormality can be used to monitor disease activity during therapy .


In addition , new technology allows the sensitive identification of gene hypermethylation in a background of normal cells , suggesting possible new strategies for the detection of minimal residual disease .

In addition , new technology allows the sensitive identification of gene hypermethylation in a background of <cell_type>normal cells</cell_type> , suggesting possible new strategies for the detection of minimal residual disease .


Finally , reactivation of tumor-suppressor gene expression through pharmacologic inhibition of DNA methyltransferase and resultant DNA demethylation appears to be a promising new avenue of therapy in acute leukemia .

Finally , reactivation of tumor-suppressor gene expression through pharmacologic inhibition of <protein>DNA methyltransferase</protein> and resultant DNA demethylation appears to be a promising new avenue of therapy in acute leukemia .


Redox regulation of the mitogen-activated protein kinase pathway during lymphocyte activation .

Redox regulation of the <protein>mitogen-activated protein kinase</protein> pathway during lymphocyte activation .


We have previously demonstrated an obligatory requirement for intracellular reactive oxygen species generation during T lymphocyte activation , and have proposed that intracellular reactive oxygen species may act as signalling agents in the regulation of certain cellular processes , for example , during cell cycle entry .


To test this hypothesis , we have been interested to determine which , if any , cell cycle entry events are affected by oxidative signalling .


In earlier studies , we have identified the transcription factors NF-kappa B and AP-1 as molecular targets for oxidative signalling processes during cell cycle entry , and have shown that oxidative signalling is involved in the regulation of early changes in gene expression during the G0 to G1 phase transition .

In earlier studies , we have identified the <protein>transcription factors</protein> <protein>NF-kappa B</protein> and <protein>AP-1</protein> as molecular targets for oxidative signalling processes during cell cycle entry , and have shown that oxidative signalling is involved in the regulation of early changes in gene expression during the G0 to G1 phase transition .


To extend these initial observations , we have examined the effect of antioxidant treatment on the activity of the mitogen-activated protein kinases erk1 and erk2 , as members of a signal transduction pathway known to directly regulate transcription factor function .

To extend these initial observations , we have examined the effect of antioxidant treatment on the activity of the <protein>mitogen-activated protein kinases</protein> <protein>erk1</protein> and <protein>erk2</protein> , as members of a signal transduction pathway known to directly regulate transcription factor function .


Using as a probe cysteamine , an aminothiol compound with both antioxidant and antiproliferative activity , we have identified erk2 , a key element of the MAP kinase pathway , as being responsive to oxidative signalling during lymphocyte activation .

Using as a probe cysteamine , an aminothiol compound with both antioxidant and antiproliferative activity , we have identified <protein>erk2</protein> , a key element of the MAP kinase pathway , as being responsive to oxidative signalling during lymphocyte activation .


These observations provide further evidence to suggest a role for intracellular oxidant generation as a regulatory mechanism during cell cycle entry , and establish a link between oxidative signalling and other aspects of the intracellular signalling network that is activated in response to mitogenic stimulation .


Physical and functional interaction between the human T-cell lymphotropic virus type 1 Tax1 protein and the CCAAT binding protein NF-Y .

Physical and functional interaction between the <protein>human T-cell lymphotropic virus type 1 Tax1 protein</protein> and the <protein>CCAAT binding protein</protein> <protein>NF-Y</protein> .


Tax1 , a potent activator of human T-cell lymphotropic virus type 1 ( HTLV-1 ) transcription , has been shown to modulate expression of many cellular genes .

<protein>Tax1</protein> , a potent activator of human T-cell lymphotropic virus type 1 ( HTLV-1 ) transcription , has been shown to modulate expression of many <dna>cellular genes</dna> .


Tax1 does not bind DNA directly but regulates transcription through protein-protein interactions with sequence-specific transcription factors .

<protein>Tax1</protein> does not bind DNA directly but regulates transcription through protein-protein interactions with <protein>sequence-specific transcription factors</protein> .


Using the yeast two-hybrid system to screen for proteins which interact with Tax1 , we isolated the B subunit of the CCAAT binding protein NF-Y from a HeLa cDNA library .

Using the yeast two-hybrid system to screen for proteins which interact with <protein>Tax1</protein> , we isolated the <protein>B subunit</protein> of the <protein>CCAAT binding protein</protein> <protein>NF-Y</protein> from a <dna>HeLa cDNA library</dna> .


The interaction of Tax1 with NF-YB was specific in that NF-YB did not interact with a variety of other transcription factors , including human immunodeficiency virus Tat , human papillomavirus E6 , and Bicoid , or with the M7 ( amino acids 29CP-AS ) Tax1 mutant .

The interaction of <protein>Tax1</protein> with <protein>NF-YB</protein> was specific in that <protein>NF-YB</protein> did not interact with a variety of other <protein>transcription factors</protein> , including <protein>human immunodeficiency virus Tat</protein> , <protein>human papillomavirus E6</protein> , and <protein>Bicoid</protein> , or with the <protein>M7 ( amino acids 29CP-AS ) Tax1 mutant</protein> .


However , NF-YB did interact with the C-terminal Tax1 mutants M22 ( 130TL-AS ) and M47 ( 319LL-RS ) .

However , <protein>NF-YB</protein> did interact with the <protein>C-terminal Tax1 mutants</protein> <protein>M22</protein> ( <protein>130TL-AS</protein> ) and <protein>M47</protein> ( <protein>319LL-RS</protein> ) .


We also show that in vitro-translated NF-YB specifically bound to a glutathione S-transferase-Tax1 fusion protein .

We also show that in vitro-translated <protein>NF-YB</protein> specifically bound to a <protein>glutathione S-transferase-Tax1 fusion protein</protein> .


Further , Tax1 coimmunoprecipitated with NF-Y from nuclear extracts of HTLV-1-transformed cells , providing evidence for in vivo interaction of Tax1 and NF-YB .

Further , <protein>Tax1</protein> coimmunoprecipitated with <protein>NF-Y</protein> from nuclear extracts of <cell_type>HTLV-1-transformed cells</cell_type> , providing evidence for in vivo interaction of <protein>Tax1</protein> and <protein>NF-YB</protein> .


We further demonstrate that Tax1 specifically activated the NF-Y-responsive DQbeta promoter , as well as a minimal promoter which contains only the Y-box element .

We further demonstrate that <protein>Tax1</protein> specifically activated the <dna>NF-Y-responsive DQbeta promoter</dna> , as well as a <dna>minimal promoter</dna> which contains only the <dna>Y-box element</dna> .


In addition , mutation of the Y-box element alone abrogated Tax1 -mediated activation .

In addition , mutation of the <dna>Y-box element</dna> alone abrogated <protein>Tax1</protein> -mediated activation .


Taken together , these data indicate that Tax1 interacts with NF-Y through the B subunit and that this interaction results in activation of the major histocompatibility complex class II promoter .

Taken together , these data indicate that <protein>Tax1</protein> interacts with <protein>NF-Y</protein> through the <protein>B subunit</protein> and that this interaction results in activation of the <dna>major histocompatibility complex class II promoter</dna> .


Through activation of this and other NF-Y driven promoters , the Tax1 -NF-Y interaction may play a critical role in causing cellular transformation and HTLV-1 pathogenesis .

Through activation of this and other <protein>NF-Y</protein> driven <dna>promoters</dna> , the <protein>Tax1</protein> <protein>-NF-Y</protein> interaction may play a critical role in causing cellular transformation and HTLV-1 pathogenesis .


High levels of the DNA-binding activity of E2F in adult T-cell leukemia and human T-cell leukemia virus type I-infected cells : possible enhancement of DNA-binding of E2F by the human T-cell leukemia virus I transactivating protein , Tax .

High levels of the DNA-binding activity of <protein>E2F</protein> in adult T-cell leukemia and <cell_type>human T-cell leukemia virus type I-infected cells</cell_type> : possible enhancement of DNA-binding of <protein>E2F</protein> by the <protein>human T-cell leukemia virus I transactivating protein</protein> , <protein>Tax</protein> .


Transcription factor E2F binds to cellular promoters of certain growth- and cell cycle-controlling genes and forms distinct heteromeric complexes with other nuclear proteins .

<protein>Transcription factor</protein> <protein>E2F</protein> binds to <dna>cellular promoters</dna> of certain <dna>growth- and cell cycle-controlling genes</dna> and forms distinct <protein>heteromeric complexes</protein> with other <protein>nuclear proteins</protein> .


It has therefore been proposed that E2F is involved in cellular proliferation control .

It has therefore been proposed that <protein>E2F</protein> is involved in cellular proliferation control .


Human T-cell leukemia virus type I ( HTLV-I ) is an etiological agent of adult T-cell leukemia ( ATL ) .


We show here by mobility-shift assay that E2F-containing DNA-binding complexes were detected in HTLV-I-infected T-cell lines and leukemic cells obtained from ATL patients but not in an uninfected T-cell line , Jurkat , and normal peripheral blood mononuclear cells .

We show here by mobility-shift assay that <protein>E2F-containing DNA-binding complexes</protein> were detected in <cell_line>HTLV-I-infected T-cell lines</cell_line> and <cell_type>leukemic cells</cell_type> obtained from ATL patients but not in an <cell_line>uninfected T-cell line</cell_line> , <cell_line>Jurkat</cell_line> , and <cell_type>normal peripheral blood mononuclear cells</cell_type> .


The Tax protein , encoded by HTLV-I , is a potent transcription activator of viral and several cellular genes .

The <protein>Tax protein</protein> , encoded by HTLV-I , is a potent transcription activator of viral and several <dna>cellular genes</dna> .


We demonstrate that expression of Tax can induce the E2F-containing DNA-binding complexes in Jurkat T cells .

We demonstrate that expression of <protein>Tax</protein> can induce the <protein>E2F-containing DNA-binding complexes</protein> in <cell_line>Jurkat T cells</cell_line> .


Thus , Tax , through enhancement of the DNA-binding activity of E2F , may be capable of regulating cellular gene expression implicated in the proliferation and transformation of T cells in ATL .

Thus , <protein>Tax</protein> , through enhancement of the DNA-binding activity of <protein>E2F</protein> , may be capable of regulating cellular gene expression implicated in the proliferation and transformation of <cell_type>T cells</cell_type> in ATL .


This activity may be relevant to the mechanisms whereby HTLV-I which does not contain oncogenes induces neoplasia .

This activity may be relevant to the mechanisms whereby HTLV-I which does not contain <dna>oncogenes</dna> induces neoplasia .


Dissociation of the Jak kinase pathway from G-CSF receptor signaling in neutrophils .

Dissociation of the <protein>Jak kinase</protein> pathway from <protein>G-CSF receptor</protein> signaling in neutrophils .


Activation of the granulocyte colony-stimulating factor receptor ( G-CSFR ) induces rapid tyrosine phosphorylation of multiple intracellular substrates in proliferating cells and nonproliferating , terminally differentiated neutrophils .

Activation of the <protein>granulocyte colony-stimulating factor receptor</protein> ( <protein>G-CSFR</protein> ) induces rapid tyrosine phosphorylation of multiple <protein>intracellular substrates</protein> in <cell_type>proliferating cells</cell_type> and <cell_type>nonproliferating , terminally differentiated neutrophils</cell_type> .


The kinases that couple ligand binding to tyrosine phosphorylation of cellular substrates by the G-CSFR with activation of specific functional programs are largely unknown .

The kinases that couple ligand binding to tyrosine phosphorylation of <protein>cellular substrates</protein> by the G-CSFR with activation of specific functional programs are largely unknown .


In this study , we examined early signaling events in proliferating and terminally differentiated cells following G-CSF stimulation to determine whether identical signaling cascades are activated .

In this study , we examined early signaling events in <cell_type>proliferating</cell_type> and <cell_type>terminally differentiated cells</cell_type> following <protein>G-CSF</protein> stimulation to determine whether identical signaling cascades are activated .


In murine Ba/F3 cells transfected with the human G-CSFR and NFS-60 cells constitutively expressing the murine G-CSFR , G-CSF induced tyrosine phosphorylation and activation of Jak1 , Jak2 , and Tyk2 .

In <cell_line>murine Ba/F3 cells</cell_line> transfected with the <protein>human G-CSFR</protein> and <cell_line>NFS-60 cells</cell_line> constitutively expressing the <protein>murine G-CSFR</protein> , <protein>G-CSF</protein> induced tyrosine phosphorylation and activation of <protein>Jak1</protein> , <protein>Jak2</protein> , and <protein>Tyk2</protein> .


Tyrosine phosphorylation of Stat3 and , to a lesser extent , Stat1 was also detected following G-CSF stimulation .

Tyrosine phosphorylation of <protein>Stat3</protein> and , to a lesser extent , <protein>Stat1</protein> was also detected following <protein>G-CSF</protein> stimulation .


Using a mitogenically incompetent human G-CSFR mutant in which Pro639 and Pro641 were substituted by alanine , the box 1 PDP motif was found to be required for activation of Jak kinases , tyrosine phosphorylation of the G-CSFR , and recruitment of Stat proteins .

Using a <protein>mitogenically incompetent human G-CSFR mutant</protein> in which Pro639 and Pro641 were substituted by alanine , the <protein>box 1 PDP motif</protein> was found to be required for activation of <protein>Jak kinases</protein> , tyrosine phosphorylation of the <protein>G-CSFR</protein> , and recruitment of <protein>Stat proteins</protein> .


Notably , no activation of Jak1 , Jak2 , Tyk2 , Stat1 , or Stat3 was observed in neutrophils following G-CSF stimulation .

Notably , no activation of <protein>Jak1</protein> , <protein>Jak2</protein> , <protein>Tyk2</protein> , <protein>Stat1</protein> , or <protein>Stat3</protein> was observed in <cell_type>neutrophils</cell_type> following <protein>G-CSF</protein> stimulation .


In addition , there was no detectable activation in neutrophils of the recently cloned Jak3 kinase , which has been reported to be expressed at high levels as myeloid cells undergo terminal neutrophilic maturation .

In addition , there was no detectable activation in <cell_type>neutrophils</cell_type> of the recently cloned <protein>Jak3 kinase</protein> , which has been reported to be expressed at high levels as <cell_type>myeloid cells</cell_type> undergo terminal neutrophilic maturation .


These results indicate a lack of involvement of Jak kinases in signaling by the G-CSFR in neutrophils , and suggest utilization of alternative signal transduction pathways distinct from those in proliferating cells .

These results indicate a lack of involvement of <protein>Jak kinases</protein> in signaling by the <protein>G-CSFR</protein> in <cell_type>neutrophils</cell_type> , and suggest utilization of alternative signal transduction pathways distinct from those in <cell_type>proliferating cells</cell_type> .


Activation of the Jak -Stat pathway correlates with proliferative signaling by the G-CSFR and requires the membrane-proximal box 1 PXP motif , which is conserved in members of the cytokine receptor superfamily .

Activation of the <protein>Jak</protein> <protein>-Stat</protein> pathway correlates with proliferative signaling by the <protein>G-CSFR</protein> and requires the <protein>membrane-proximal box 1 PXP motif</protein> , which is conserved in members of the <protein>cytokine receptor superfamily</protein> .


Impaired induction of c-fos/c-jun genes and of transcriptional regulatory proteins binding distinct c-fos/c-jun promoter elements in activated human T cells during aging .

Impaired induction of <dna>c-fos/c-jun genes</dna> and of <protein>transcriptional regulatory proteins</protein> binding distinct <dna>c-fos/c-jun promoter elements</dna> in <cell_type>activated human T cells</cell_type> during aging .


The activation of transcriptional factor c-Fos/c-Jun AP-1 is essential for normal T cell responsiveness and is often impaired in T cells during aging .

The activation of <protein>transcriptional factor</protein> c-Fos/c-Jun <protein>AP-1</protein> is essential for normal <cell_type>T cell</cell_type> responsiveness and is often impaired in <cell_type>T cells</cell_type> during aging .


In the present study , we investigated whether aberrancies in the regulation of c-fos/c-jun at the mRNA or protein level might underlie the age-associated impairments of AP-1 in human T cells .

In the present study , we investigated whether aberrancies in the regulation of <dna>c-fos/c-jun</dna> at the mRNA or protein level might underlie the age-associated impairments of <protein>AP-1</protein> in <cell_type>human T cells</cell_type> .


Whereas T cells from young subjects stimulated with cross-linked anti-CD3epsilon mAb OKT3 plus PMA or with the lectin PHA plus PMA demonstrated considerable increases in c-Fos protein expression , the expression of c-Fos but not c-Jun was markedly reduced in stimulated T cells from certain elderly subjects .

Whereas <cell_type>T cells</cell_type> from young subjects stimulated with <protein>cross-linked anti-CD3epsilon mAb</protein> <protein>OKT3</protein> plus PMA or with the <protein>lectin</protein> <protein>PHA</protein> plus PMA demonstrated considerable increases in <protein>c-Fos</protein> protein expression , the expression of <protein>c-Fos</protein> but not <protein>c-Jun</protein> was markedly reduced in stimulated <cell_type>T cells</cell_type> from certain elderly subjects .


In addition , RNase protection assays revealed that anti-CD3/ PMA-stimulated T cells from a substantial proportion of elderly subjects exhibited decreased levels of c-fos and/or c-jun mRNA compared to T cells from young subjects .

In addition , <protein>RNase</protein> protection assays revealed that anti-CD3/ <cell_type>PMA-stimulated T cells</cell_type> from a substantial proportion of elderly subjects exhibited decreased levels of <rna>c-fos and/or c-jun mRNA</rna> compared to <cell_type>T cells</cell_type> from young subjects .


Using electrophoretic mobility shift assays , the levels of nuclear regulatory proteins recognizing the AP-1 consensus TRE motif , the proximal c-jun TRE-like promoter element , and the c-fos serum response element ( SRE ) were determined in resting and stimulated T cells .

Using electrophoretic mobility shift assays , the levels of <protein>nuclear regulatory proteins</protein> recognizing the <dna>AP-1 consensus TRE motif</dna> , the <dna>proximal c-jun TRE-like promoter element</dna> , and the <dna>c-fos serum response element</dna> ( <dna>SRE</dna> ) were determined in <cell_type>resting and stimulated T cells</cell_type> .


Although the stimulation of T cells from young subjects resulted in coordinated increases of nuclear protein complexes binding the AP-1 TRE , c-jun TRE , and c-fos SRE DNA sequence motifs , age-related reductions in the activation of AP-1 were accompanied by decreased levels of c-jun TRE and c-fos SRE binding complexes .

Although the stimulation of <cell_type>T cells</cell_type> from young subjects resulted in coordinated increases of <protein>nuclear protein complexes</protein> binding the <dna>AP-1 TRE</dna> , <dna>c-jun TRE</dna> , and <dna>c-fos SRE DNA sequence motifs</dna> , age-related reductions in the activation of <protein>AP-1</protein> were accompanied by decreased levels of <protein>c-jun TRE and c-fos SRE binding complexes</protein> .


Furthermore , the nuclear protein complexes binding the SRE motif induced in activated T cells of young and elderly subjects contained serum response factor and Elk-1 pointing toward age-related defects in the activation of transcriptional regulatory proteins distinct from c-jun/AP-1 .

Furthermore , the <protein>nuclear protein complexes</protein> binding the <dna>SRE motif</dna> induced in <cell_type>activated T cells</cell_type> of young and elderly subjects contained <protein>serum response factor</protein> and <protein>Elk-1</protein> pointing toward age-related defects in the activation of <protein>transcriptional regulatory proteins</protein> distinct from <protein>c-jun/AP-1</protein> .


These results suggest that underlying aberrancies in the induction of c-fos/c-jun as well as their nuclear regulatory proteins may contribute to the age-related impairments of AP-1 activation in human T cells .

These results suggest that underlying aberrancies in the induction of <dna>c-fos/c-jun</dna> as well as their <protein>nuclear regulatory proteins</protein> may contribute to the age-related impairments of <protein>AP-1</protein> activation in <cell_type>human T cells</cell_type> .


Retinoic acid-induced modulation of IL-2 mRNA production and IL-2 receptor expression on T cells .

Retinoic acid-induced modulation of <rna>IL-2 mRNA</rna> production and <protein>IL-2</protein> receptor expression on <cell_type>T cells</cell_type> .


BACKGROUND : Retinoic acid ( RA ) has important immune-modulating effects on both T and B cell function .


Our laboratory has shown that RA can enhance in vitro polyclonal B cell immunoglobulin ( Ig ) response .

Our laboratory has shown that RA can enhance in vitro polyclonal B cell <protein>immunoglobulin</protein> ( Ig ) response .


Investigating cytokines known to affect B cell differentiation , we have recently shown that IL-6 production is augmented by RA .

Investigating <protein>cytokines</protein> known to affect B cell differentiation , we have recently shown that <protein>IL-6</protein> production is augmented by RA .


In the present study we have examined the immune modulating effects of RA on IL-2 mRNA , another important cytokine for B cell immunoglobulin production , the expression of IL-2 receptors on T cells , and the RA nuclear receptors .

In the present study we have examined the immune modulating effects of RA on <rna>IL-2 mRNA</rna> , another important <protein>cytokine</protein> for B cell <protein>immunoglobulin</protein> production , the expression of <protein>IL-2 receptors</protein> on <cell_type>T cells</cell_type> , and the <protein>RA nuclear receptors</protein> .


METHODS : Purified T cells were obtained from adenoidal tissues , and incubated with RA ( 10 ( -7 ) M ) or DMSO solvent/media control for 0 , 6-8 , and 24 h .

METHODS : Purified <cell_type>T cells</cell_type> were obtained from adenoidal tissues , and incubated with RA ( 10 ( -7 ) M ) or DMSO solvent/media control for 0 , 6-8 , and 24 h .


Total mRNA was extracted from T cells , and using RT-PCR , changes in the production of IL-2 and RA receptors ( RAR ) - alpha , beta , gamma mRNA were determined .

Total <rna>mRNA</rna> was extracted from <cell_type>T cells</cell_type> , and using RT-PCR , changes in the production of <protein>IL-2</protein> and <protein>RA receptors</protein> ( <protein>RAR</protein> ) - <rna>alpha</rna> , <rna>beta</rna> , <rna>gamma mRNA</rna> were determined .


The effects of RA on IL-2-alpha receptor expression was determined by flow cytometry on T cells .

The effects of RA on IL-2-alpha receptor expression was determined by flow cytometry on <cell_type>T cells</cell_type> .


CONCLUSION : These studies suggest that RA can augment IL-2 mRNA production by T cells with a possible paracrine effect on IL-2R-alpha expression .

CONCLUSION : These studies suggest that RA can augment <rna>IL-2 mRNA</rna> production by <cell_type>T cells</cell_type> with a possible paracrine effect on <protein>IL-2R-alpha</protein> expression .


These changes appear to be mediated by RAR-alpha .

These changes appear to be mediated by <protein>RAR-alpha</protein> .


Thus , IL-2 may be another important cytokine modulated by RA in the immune response .

Thus , <protein>IL-2</protein> may be another important <protein>cytokine</protein> modulated by RA in the immune response .


Involvement of an SAF-like transcription factor in the activation of serum amyloid A gene in monocyte/macrophage cells by lipopolysaccharide .

Involvement of an <protein>SAF-like transcription factor</protein> in the activation of <dna>serum amyloid A gene</dna> in <cell_type>monocyte/macrophage cells</cell_type> by lipopolysaccharide .


Serum amyloid A ( SAA ) has been linked to atherosclerosis because of its ability to remodel high-density lipoprotein by the depletion of apolipoprotein A1 , its ability to bind cholesterol , and its presence in the atherosclerotic plaques of coronary and carotid arteries .

<protein>Serum amyloid A</protein> ( <protein>SAA</protein> ) has been linked to atherosclerosis because of its ability to remodel high-density lipoprotein by the depletion of apolipoprotein A1 , its ability to bind cholesterol , and its presence in the atherosclerotic plaques of coronary and carotid arteries .


In the present study , we investigated the induction mechanism of SAA gene in THP-1 monocyte/macrophage cells which play a critical role in the development of atherosclerotic fatty streak and plaque formation .

In the present study , we investigated the induction mechanism of <dna>SAA gene</dna> in <cell_line>THP-1 monocyte/macrophage cells</cell_line> which play a critical role in the development of atherosclerotic fatty streak and plaque formation .


We and others have shown that SAA gene is induced in monocyte/macrophage cells by lipopolysaccharide ( LPS ) .

We and others have shown that <protein>SAA</protein> gene is induced in <cell_type>monocyte/macrophage cells</cell_type> by lipopolysaccharide ( LPS ) .


By promoter function analysis , we show that the SAA promoter sequence between -280 and -226 can confer LPS responsiveness .

By promoter function analysis , we show that the <dna>SAA promoter sequence</dna> between -280 and -226 can confer LPS responsiveness .


Gel electrophoretic mobility shift assay detected an induced DNA-binding activity in these cells in response to LPS .


Characterization of the DNA-binding protein by UV cross-linking , Southwestern blot , and antibody ablation/supershift assays revealed that it is similar to a recently reported nuclear factor designated SAF .

Characterization of the <protein>DNA-binding protein</protein> by UV cross-linking , Southwestern blot , and antibody ablation/supershift assays revealed that it is similar to a recently reported <protein>nuclear factor</protein> designated <protein>SAF</protein> .


These results demonstrated that LPS-mediated SAA gene induction in monocyte/macrophage cells is primarily due to the induction of SAF activity .

These results demonstrated that LPS-mediated <protein>SAA</protein> gene induction in <cell_type>monocyte/macrophage cells</cell_type> is primarily due to the induction of <protein>SAF</protein> activity .


Cytomegalovirus immediate early genes upregulate interleukin-6 gene expression .

<dna>Cytomegalovirus immediate early genes</dna> upregulate <protein>interleukin-6</protein> gene expression .


BACKGROUND : The immediate early genes ( IE ) of human cytomegalovirus ( CMV ) can be expressed in monocytic cells and are known to regulate viral and cellular genes .

BACKGROUND : The <dna>immediate early genes</dna> ( <dna>IE</dna> ) of human cytomegalovirus ( CMV ) can be expressed in <cell_type>monocytic cells</cell_type> and are known to regulate <dna>viral and cellular genes</dna> .


Interleukin-6 ( IL-6 ) plays a central role in numerous inflammatory and immune processes .

<protein>Interleukin-6</protein> ( <protein>IL-6</protein> ) plays a central role in numerous inflammatory and immune processes .


Interleukin-6 levels are increased in lung transplant patients clinically diagnosed with CMV pneumonitis .

<protein>Interleukin-6</protein> levels are increased in lung transplant patients clinically diagnosed with CMV pneumonitis .


The regulation of IL-6 is dependent on various stimuli that include lipopolysaccharide ( LPS ) , viruses , and other cytokines .

The regulation of <protein>IL-6</protein> is dependent on various stimuli that include lipopolysaccharide ( LPS ) , viruses , and other <protein>cytokines</protein> .


These studies examined the ability of CMV IE gene products to modulate IL-6 production .

These studies examined the ability of CMV IE gene products to modulate <protein>IL-6</protein> production .


METHODS : THP-1 cells , a monocytic cell line , were transfected with the CMV IE genes .

METHODS : <cell_line>THP-1 cells</cell_line> , a <cell_line>monocytic cell line</cell_line> , were transfected with the <dna>CMV IE genes</dna> .


Interleukin-6 protein and IL-6 mRNA were measured in control and CMV immediate early transfected cells .

<protein>Interleukin-6</protein> protein and <rna>IL-6 mRNA</rna> were measured in control and <cell_type>CMV immediate early transfected cells</cell_type> .


Cotransfection of CMV IE genes and IL-6 chloramphenicol acetyl transferase ( CAT ) or IL-6 luciferase constructs were used to study IL-6 promoter activity .

Cotransfection of <dna>CMV IE genes</dna> and <dna>IL-6 chloramphenicol acetyl transferase ( CAT ) or IL-6 luciferase constructs</dna> were used to study <protein>IL-6</protein> promoter activity .


RESULTS : Interleukin-6 protein and mRNA production were significantly increased in cells transfected with the CMV IE genes and stimulated with LPS compared to LPS-stimulated control cells .

RESULTS : <protein>Interleukin-6 protein</protein> and mRNA production were significantly increased in cells transfected with the <dna>CMV IE genes</dna> and stimulated with LPS compared to <cell_line>LPS-stimulated control cells</cell_line> .


Cytomegalovirus IE gene products significantly enhanced LPS stimulation of IL-6 promoter activity in both IL-6 CAT and IL-6 luciferase assays .

<protein>Cytomegalovirus IE gene products</protein> significantly enhanced LPS stimulation of <dna>IL-6 promoter</dna> activity in both <protein>IL-6</protein> CAT and IL-6 luciferase assays .


A deletion construct that contains a NF-kappa B site but is missing the multiple response region demonstrated a continued increase in IL-6 luciferase activity in LPS-stimulated CMV transfected cells .

A deletion construct that contains a <dna>NF-kappa B site</dna> but is missing the multiple response region demonstrated a continued increase in <protein>IL-6</protein> <protein>luciferase</protein> activity in <cell_line>LPS-stimulated CMV transfected cells</cell_line> .


CONCLUSION : Cytomegalovirus immediate early gene products significantly enhanced expression of IL-6 in LPS-stimulated cells .

CONCLUSION : <protein>Cytomegalovirus immediate early gene products</protein> significantly enhanced expression of <protein>IL-6</protein> in LPS-stimulated cells .


The increase in IL-6 luciferase activity occurs in the absence of the multiple response region , the area of the IL-6 promoter responsive to IL-1 , TNF alpha , cyclic amp , and phorbol 12-myristate 13-acetate .

The increase in <protein>IL-6</protein> <protein>luciferase</protein> activity occurs in the absence of the multiple response region , the area of the <dna>IL-6 promoter</dna> responsive to <protein>IL-1</protein> , <protein>TNF alpha</protein> , cyclic amp , and phorbol 12-myristate 13-acetate .


The ability of CMV IE gene products to enhance IL-6 production may play an important role in immune inflammatory states associated with CMV infection .

The ability of <protein>CMV IE gene products</protein> to enhance <protein>IL-6</protein> production may play an important role in immune inflammatory states associated with CMV infection .


Induction of relA ( p65 ) and I kappa B alpha subunit expression during differentiation of human peripheral blood monocytes to macrophages .

Induction of <protein>relA ( p65 )</protein> and <protein>I kappa B alpha subunit</protein> expression during differentiation of <cell_type>human peripheral blood monocytes</cell_type> to <cell_type>macrophages</cell_type> .


We evaluated the expression and DNA binding activity of nuclear factor ( NF ) -kappa B subunits in human peripheral blood monocytes and in monocyte-derived macrophages ( MDMs ) .

We evaluated the expression and DNA binding activity of <protein>nuclear factor ( NF ) -kappa B subunits</protein> in <cell_type>human peripheral blood monocytes</cell_type> and in <cell_type>monocyte-derived macrophages</cell_type> ( <cell_type>MDMs</cell_type> ) .


Constitutive DNA binding activity consisting of p50 homodimers was detected in nuclear extracts from both cell types .

Constitutive DNA binding activity consisting of <protein>p50 homodimers</protein> was detected in nuclear extracts from both cell types .


An additional complex composed of p50/RelA ( p65 ) heterodimers appeared only in nuclear extracts from 7-day MDMs .

An additional complex composed of <protein>p50/RelA ( p65 ) heterodimers</protein> appeared only in nuclear extracts from 7-day <cell_type>MDMs</cell_type> .


Immunoblot analysis showed that the p50 subunit was constitutively expressed in monocytes and MDMs .

Immunoblot analysis showed that the p50 subunit was constitutively expressed in <cell_type>monocytes</cell_type> and <cell_type>MDMs</cell_type> .


In contrast , the RelA ( p65 ) subunit was barely detectable in monocytes , but its level increased markedly in MDMs .

In contrast , the <protein>RelA ( p65 ) subunit</protein> was barely detectable in <cell_type>monocytes</cell_type> , but its level increased markedly in <cell_type>MDMs</cell_type> .