In T lymphocytes , the hematopoietic cytokine interleukin-2 ( IL-2 ) uses phosphatidylinositol 3-kinase ( PI 3-kinase ) -induced signaling pathways to regulate E2F transcriptional activity , a critical cell cycle checkpoint .

In <cell_type>T lymphocytes</cell_type> , the <protein>hematopoietic cytokine</protein> <protein>interleukin-2</protein> ( <protein>IL-2</protein> ) uses <protein>phosphatidylinositol 3-kinase</protein> ( <protein>PI 3-kinase</protein> ) -induced signaling pathways to regulate <protein>E2F</protein> transcriptional activity , a critical cell cycle checkpoint .


PI 3-kinase also regulates the activity of p70 ( s6k ) , the 40S ribosomal protein S6 kinase , a response that is abrogated by the macrolide rapamycin .

<protein>PI 3-kinase</protein> also regulates the activity of <protein>p70 ( s6k )</protein> , the <protein>40S ribosomal protein S6 kinase</protein> , a response that is abrogated by the macrolide rapamycin .


This immunosuppressive drug is known to prevent T-cell proliferation , but the precise point at which rapamycin regulates T-cell cycle progression has yet to be elucidated .


Moreover , the effects of rapamycin on , and the role of p70 ( s6k ) in , IL-2 and PI 3-kinase activation of E2Fs have not been characterized .

Moreover , the effects of rapamycin on , and the role of <protein>p70 ( s6k )</protein> in , <protein>IL-2</protein> and <protein>PI 3-kinase</protein> activation of <protein>E2Fs</protein> have not been characterized .


Our present results show that IL-2 -and PI 3-kinase -induced pathways for the regulation of E2F transcriptional activity include both rapamycin-resistant and rapamycin-sensitive components .

Our present results show that <protein>IL-2</protein> -and <protein>PI 3-kinase</protein> -induced pathways for the regulation of <protein>E2F</protein> transcriptional activity include both rapamycin-resistant and rapamycin-sensitive components .


Expression of a rapamycin-resistant mutant of p70 ( s6k ) in T cells could restore rapamycin-suppressed E2F responses .

Expression of a <protein>rapamycin-resistant mutant</protein> of <protein>p70 ( s6k )</protein> in <cell_type>T cells</cell_type> could restore rapamycin-suppressed <protein>E2F</protein> responses .


Thus , the rapamycin-controlled processes involved in E2F regulation appear to be mediated by p70 ( s6k ) .

Thus , the rapamycin-controlled processes involved in <protein>E2F</protein> regulation appear to be mediated by <protein>p70 ( s6k )</protein> .


However , the rapamycin-resistant p70 ( s6k ) could not rescue rapamycin inhibition of T-cell cycle entry , consistent with the involvement of additional , rapamycin-sensitive pathways in the control of T-cell cycle progression .

However , the <protein>rapamycin-resistant p70 ( s6k )</protein> could not rescue rapamycin inhibition of T-cell cycle entry , consistent with the involvement of additional , rapamycin-sensitive pathways in the control of T-cell cycle progression .


The present results thus show that p70 ( s6k ) is able to regulate E2F transcriptional activity and provide direct evidence for the first time for a link between IL-2 receptors , PI 3-kinase , and p70 ( s6k ) that regulates a crucial G1 checkpoint in T lymphocytes .

The present results thus show that <protein>p70 ( s6k )</protein> is able to regulate <protein>E2F</protein> transcriptional activity and provide direct evidence for the first time for a link between <protein>IL-2</protein> receptors , <protein>PI 3-kinase</protein> , and <protein>p70 ( s6k )</protein> that regulates a crucial G1 checkpoint in <cell_type>T lymphocytes</cell_type> .


Oxidative stress triggers STAT3 tyrosine phosphorylation and nuclear translocation in human lymphocytes .

Oxidative stress triggers <protein>STAT3</protein> tyrosine phosphorylation and nuclear translocation in <cell_type>human lymphocytes</cell_type> .


Oxidizing agents are powerful activators of factors responsible for the transcriptional activation of cytokine-encoding genes involved in tissue injury .

Oxidizing agents are powerful activators of factors responsible for the transcriptional activation of <dna>cytokine-encoding genes</dna> involved in tissue injury .


In this study we show evidence that STAT3 is a transcription factor whose activity is modulated by H2O2 in human lymphocytes , in which endogenous catalase had previously been inhibited .

In this study we show evidence that <protein>STAT3</protein> is a <protein>transcription factor</protein> whose activity is modulated by H2O2 in <cell_type>human lymphocytes</cell_type> , in which endogenous <protein>catalase</protein> had previously been inhibited .


H2O2-induced nuclear translocation of STAT3 to form sequence-specific DNA-bound complexes was evidenced by immunoblotting of nuclear fractions and electrophoretic mobility shift assays , and vanadate was found to strongly synergize with H2O2 .

H2O2-induced nuclear translocation of <protein>STAT3</protein> to form <protein>sequence-specific DNA-bound complexes</protein> was evidenced by immunoblotting of nuclear fractions and electrophoretic mobility shift assays , and vanadate was found to strongly synergize with H2O2 .


Moreover , anti- STAT3 antibodies specifically precipitated a protein of 92 kDa that becomes phosphorylated on tyrosine upon lymphocyte treatment with H2O2 .

Moreover , anti- <protein>STAT3 antibodies</protein> specifically precipitated a protein of <protein>92 kDa</protein> that becomes phosphorylated on tyrosine upon lymphocyte treatment with H2O2 .


Phenylarsine oxide , a tyrosine phosphatase inhibitor , and genistein , a tyrosine kinase inhibitor , cooperated and cancelled , respectively , the H2O2-promoted STAT3 nuclear translocation .

Phenylarsine oxide , a tyrosine phosphatase inhibitor , and genistein , a tyrosine kinase inhibitor , cooperated and cancelled , respectively , the H2O2-promoted <protein>STAT3</protein> nuclear translocation .


Evidence is also presented , using Fe2+/Cu2+ ions , that.OH generated from H2O2 through Fenton reactions could be a candidate oxygen reactive species to directly activate STAT3 .

Evidence is also presented , using Fe2+/Cu2+ ions , that.OH generated from H2O2 through Fenton reactions could be a candidate oxygen reactive species to directly activate <protein>STAT3</protein> .


Present data suggest that H2O2 and vanadate are likely to inhibit the activity of intracellular tyrosine phosphatase ( s ) , leading to enhanced STAT3 tyrosine phosphorylation and hence its translocation to the nucleus .

Present data suggest that H2O2 and vanadate are likely to inhibit the activity of <protein>intracellular tyrosine phosphatase</protein> ( s ) , leading to enhanced <protein>STAT3</protein> tyrosine phosphorylation and hence its translocation to the nucleus .


These results demonstrate that the DNA binding activity of STAT3 can be modulated by oxidizing agents and provide a framework to understand the effects of oxidative stress on the JAK -STAT signaling pathway .

These results demonstrate that the DNA binding activity of <protein>STAT3</protein> can be modulated by oxidizing agents and provide a framework to understand the effects of oxidative stress on the <protein>JAK</protein> <protein>-STAT</protein> signaling pathway .


Transcriptional regulation of T lymphocyte development and function .


The development and function of T lymphocytes are regulated tightly by signal transduction pathways that include specific cell-surface receptors , intracellular signaling molecules , and nuclear transcription factors .

The development and function of <cell_type>T lymphocytes</cell_type> are regulated tightly by signal transduction pathways that include specific <protein>cell-surface receptors</protein> , <protein>intracellular signaling molecules</protein> , and <protein>nuclear transcription factors</protein> .


Since 1988 , several families of functionally important T cell transcription factors have been identified .

Since 1988 , several families of functionally important <protein>T cell transcription factors</protein> have been identified .


These include the Ikaros , LKLF , and GATA3 zinc-finger proteins ; the Ets , CREB/ATF , and NF-kappa B/Rel/NFAT transcription factors ; the Stat proteins ; and HMG box transcription factors such as LEF1 , TCF1 , and Sox4 .

These include the <protein>Ikaros</protein> , <protein>LKLF</protein> , and <protein>GATA3 zinc-finger proteins</protein> ; the <protein>Ets</protein> , <protein>CREB/ATF</protein> , and <protein>NF-kappa B/Rel/NFAT transcription factors</protein> ; the <protein>Stat proteins</protein> ; and <protein>HMG box transcription factors</protein> such as <protein>LEF1</protein> , <protein>TCF1</protein> , and <protein>Sox4</protein> .


In this review , we summarize our current understanding of the transcriptional regulation of T cell development and function with particular emphasis on the results of recent gene targeting and transgenic experiments .


In addition to increasing our understanding of the molecular pathways that regulate T cell development and function , these results have suggested novel targets for genetic and pharmacological manipulation of T cell immunity .


In vivo inhibition of NF-kappa B in T-lineage cells leads to a dramatic decrease in cell proliferation and cytokine production and to increased cell apoptosis in response to mitogenic stimuli , but not to abnormal thymopoiesis .

In vivo inhibition of <protein>NF-kappa B</protein> in <cell_type>T-lineage cells</cell_type> leads to a dramatic decrease in cell proliferation and <protein>cytokine</protein> production and to increased cell apoptosis in response to mitogenic stimuli , but not to abnormal thymopoiesis .


To understand the role of NF-kappa B complexes in T cell development and activation , we have generated transgenic mice in which RelA and c-Rel complexes were selectively inhibited in the T-lineage cells by specific expression of a trans-dominant form of I kappa B alpha .

To understand the role of <protein>NF-kappa B</protein> complexes in T cell development and activation , we have generated transgenic mice in which <protein>RelA</protein> and <protein>c-Rel complexes</protein> were selectively inhibited in the <cell_type>T-lineage cells</cell_type> by specific expression of a trans-dominant form of <protein>I kappa B alpha</protein> .


Transgene expression did not affect the thymic development , but led to lowered numbers of splenic T cells and to a dramatic decrease in the ex vivo proliferative response of splenic T lymphocytes .

Transgene expression did not affect the thymic development , but led to lowered numbers of splenic <cell_type>T cells</cell_type> and to a dramatic decrease in the ex vivo proliferative response of <cell_type>splenic T lymphocytes</cell_type> .


Analysis of IL-2 and IL-2R alpha expression demonstrated that the perturbation of the proliferation response was not attributable to an abnormal expression of these genes .

Analysis of <protein>IL-2</protein> and <protein>IL-2R alpha</protein> expression demonstrated that the perturbation of the proliferation response was not attributable to an abnormal expression of these genes .


In contrast , expression of IL-4 , IL-10 , and IFN-gamma was strongly inhibited in the transgenic T cells .

In contrast , expression of <protein>IL-4</protein> , <protein>IL-10</protein> , and <protein>IFN-gamma</protein> was strongly inhibited in the <cell_type>transgenic T cells</cell_type> .


The proliferative deficiency of the transgenic T cells was associated with an increased apoptosis .

The proliferative deficiency of the <cell_type>transgenic T cells</cell_type> was associated with an increased apoptosis .


These results point out the involvement of NF-kappa B/Rel family proteins in growth signaling pathways by either regulating proteins involved in the IL-2 signaling or by functionally interfering with the cell cycle progression .

These results point out the involvement of <protein>NF-kappa B/Rel family proteins</protein> in growth signaling pathways by either <protein>regulating proteins</protein> involved in the <protein>IL-2</protein> signaling or by functionally interfering with the cell cycle progression .


Paradoxical priming effects of IL-10 on cytokine production .

Paradoxical priming effects of <protein>IL-10</protein> on <protein>cytokine</protein> production .


IL-10 is a well-known immunosuppressive and/or anti-inflammatory cytokine .

<protein>IL-10</protein> is a well-known <protein>immunosuppressive and/or anti-inflammatory cytokine</protein> .


However , we report in vitro experimental studies in which IL-10 primed leukocytes and led to an enhanced production of tumor necrosis factor ( TNF ) upon further stimulation by lipopolysaccharide ( LPS ) .

However , we report in vitro experimental studies in which <cell_line>IL-10 primed leukocytes</cell_line> and led to an enhanced production of <protein>tumor necrosis factor</protein> ( <protein>TNF</protein> ) upon further stimulation by lipopolysaccharide ( LPS ) .


Monocytes and peripheral blood mononuclear cells ( PBMC ) prepared from whole blood maintained for 20 h at 37 degrees C in the presence of recombinant human IL-10 had an enhanced capacity to produce TNF in response to LPS .

<cell_type>Monocytes</cell_type> and <cell_type>peripheral blood mononuclear cells</cell_type> ( <cell_type>PBMC</cell_type> ) prepared from whole blood maintained for 20 h at 37 degrees C in the presence of <protein>recombinant human IL-10</protein> had an enhanced capacity to produce <protein>TNF</protein> in response to LPS .


In addition to TNF , LPS-induced IL-6 and spontaneous IL-1ra production were also enhanced .

In addition to <protein>TNF</protein> , LPS-induced IL-6 and spontaneous IL-1ra production were also enhanced .


When isolated PBMC were first cultured for 20 h in the presence of IL-10 on Teflon to prevent adherence , washed to remove IL-10 and then further cultured in plastic dishes for an additional 20 h in the presence of LPS or IL-1beta , an enhanced release of TNF was observed .

When isolated <cell_type>PBMC</cell_type> were first cultured for 20 h in the presence of <protein>IL-10</protein> on Teflon to prevent adherence , washed to remove <protein>IL-10</protein> and then further cultured in plastic dishes for an additional 20 h in the presence of LPS or <protein>IL-1beta</protein> , an enhanced release of <protein>TNF</protein> was observed .


This was not the case when PBMC were pre-cultured in plastic multidishes in the presence of IL-10 .

This was not the case when <cell_type>PBMC</cell_type> were pre-cultured in plastic multidishes in the presence of <protein>IL-10</protein> .


TNF mRNA expression induced by LPS was decreased when the pre-treatment of PBMC with IL-10 was performed on plastic , whereas this was not the case when cells were pre-cultured with IL-10 on Teflon .

<rna>TNF mRNA</rna> expression induced by LPS was decreased when the pre-treatment of <cell_type>PBMC</cell_type> with <protein>IL-10</protein> was performed on plastic , whereas this was not the case when cells were pre-cultured with <protein>IL-10</protein> on Teflon .


Furthermore , NFkappaB translocation following LPS activation was higher after IL-10 pre-treatment on Teflon than on plastic .

Furthermore , NFkappaB translocation following LPS activation was higher after <protein>IL-10</protein> pre-treatment on Teflon than on plastic .


Interestingly , an enhanced frequency of CD16 and CD68 ( + ) cells among the CD14 ( + ) cells was observed in the presence of IL-10 , independently of the pre-culture conditions of the PBMC .

Interestingly , an enhanced frequency of <protein>CD16</protein> and CD68 ( + ) cells among the CD14 ( + ) cells was observed in the presence of <protein>IL-10</protein> , independently of the pre-culture conditions of the <cell_type>PBMC</cell_type> .


Altogether , these results indicate that the IL-10 -induced up-regulation of cytokine production depends on the prevention of monocyte adherence by red cells in the whole blood assays or by cultures of PBMC on Teflon .

Altogether , these results indicate that the <protein>IL-10</protein> -induced up-regulation of <protein>cytokine</protein> production depends on the prevention of monocyte adherence by <cell_type>red cells</cell_type> in the whole blood assays or by cultures of <cell_type>PBMC</cell_type> on Teflon .


In contrast , the adherence parameter has no effect on the IL-10 -induced modulation of some monocyte surface markers .

In contrast , the adherence parameter has no effect on the <protein>IL-10</protein> -induced modulation of some <protein>monocyte surface markers</protein> .


Cell growth-regulated expression of mammalian MCM5 and MCM6 genes mediated by the transcription factor E2F .

Cell growth-regulated expression of <dna>mammalian MCM5 and MCM6 genes</dna> mediated by the <protein>transcription factor</protein> <protein>E2F</protein> .


Initiation of DNA replication requires the function of MCM gene products , which participate in ensuring that DNA replication occurs only once in the cell cycle .

Initiation of DNA replication requires the function of <protein>MCM gene products</protein> , which participate in ensuring that DNA replication occurs only once in the cell cycle .


Expression of all mammalian genes of the MCM family is induced by growth stimulation , unlike yeast , and the mRNA levels peak at G1/S boundary .

Expression of all <dna>mammalian genes</dna> of the <dna>MCM family</dna> is induced by growth stimulation , unlike yeast , and the mRNA levels peak at G1/S boundary .


In this study , we examined the transcriptional activities of isolated human MCM gene promoters .

In this study , we examined the transcriptional activities of <dna>isolated human MCM gene promoters</dna> .


Human MCM5 and MCM6 promoters with mutation in the E2F sites failed in promoter regulation following serum stimulation and exogenous E2F expression .

<dna>Human MCM5 and MCM6 promoters</dna> with mutation in the <dna>E2F sites</dna> failed in promoter regulation following serum stimulation and exogenous <protein>E2F</protein> expression .


In addition , we identified a novel E2F-like sequence in human MCM6 promoter which cooperates with the authentic E2F sites in E2F -dependent regulation .

In addition , we identified a novel <dna>E2F-like sequence</dna> in <dna>human MCM6 promoter</dna> which cooperates with the authentic <dna>E2F sites</dna> in <protein>E2F</protein> -dependent regulation .


Forced expression of E2F1 could induce expression of all members of the endogenous MCM genes in rat embryonal fibroblast REF52 cells .

Forced expression of <protein>E2F1</protein> could induce expression of all members of the <dna>endogenous MCM genes</dna> in <cell_line>rat embryonal fibroblast REF52 cells</cell_line> .


Our results demonstrated that the growth-regulated expression of mammalian MCM5 and MCM6 genes , and presumably other MCM members , is primarily regulated by E2F through binding to multiple E2F sites in the promoters .

Our results demonstrated that the growth-regulated expression of <dna>mammalian MCM5 and MCM6 genes</dna> , and presumably other MCM members , is primarily regulated by <protein>E2F</protein> through binding to <dna>multiple E2F sites</dna> in the <dna>promoters</dna> .


An essential role for NF-kappaB in human CD34 ( + ) bone marrow cell survival .

An essential role for <protein>NF-kappaB</protein> in <cell_type>human CD34 ( + ) bone marrow cell</cell_type> survival .


The transcription factor , NF-kappaB , is important for T-cell activation , B-cell maturation , and human immunodeficiency virus transcription and plays a role in alternatively mediating and protecting against apoptosis in a variety of cell types .

The <protein>transcription factor</protein> , <protein>NF-kappaB</protein> , is important for T-cell activation , B-cell maturation , and human immunodeficiency virus transcription and plays a role in alternatively mediating and protecting against apoptosis in a variety of cell types .


However , a role for NF-kappaB in human CD34 ( + ) bone marrow cells has not been described .

However , a role for <protein>NF-kappaB</protein> in <cell_type>human CD34 ( + ) bone marrow cells</cell_type> has not been described .


We provide evidence here that virtually all human CD34 ( + ) bone marrow cells express NF-kappaB that can be activated by exposure to phorbol 12-myristate 13-acetate and a variety of cytokines , eg , tumor necrosis factor alpha , interleukin-3 , and granulocyte-macrophage colony-stimulating factor .

We provide evidence here that virtually all <cell_type>human CD34 ( + ) bone marrow cells</cell_type> express <protein>NF-kappaB</protein> that can be activated by exposure to phorbol 12-myristate 13-acetate and a variety of <protein>cytokines</protein> , eg , <protein>tumor necrosis factor alpha</protein> , <protein>interleukin-3</protein> , and <protein>granulocyte-macrophage colony-stimulating factor</protein> .


In addition , we demonstrate that NF-kappaB may be required for human CD34 ( + ) bone marrow cell clonogenic function and survival .

In addition , we demonstrate that <protein>NF-kappaB</protein> may be required for human <cell_type>CD34 ( + ) bone marrow cell</cell_type> clonogenic function and survival .


These results offer insight into a new role for NF-kappaB in maintaining survival and function in hematopoietic stem and progenitor cells and suggest that proposed strategies involving inhibition of NF-kappaB activation as an adjunct to cancer chemotherapy should be approached with caution .

These results offer insight into a new role for <protein>NF-kappaB</protein> in maintaining survival and function in <cell_type>hematopoietic stem and progenitor cells</cell_type> and suggest that proposed strategies involving inhibition of <protein>NF-kappaB</protein> activation as an adjunct to cancer chemotherapy should be approached with caution .


High molecular weight dextran sulfate increases the activity of NF-kappaB-regulated promoter in monocyte-derived macrophages .

High molecular weight dextran sulfate increases the activity of <dna>NF-kappaB-regulated promoter</dna> in <cell_type>monocyte-derived macrophages</cell_type> .


It is known that sulfated polysaccharides can mimic the action of common T-cell mitogens .

It is known that sulfated polysaccharides can mimic the action of <protein>common T-cell mitogens</protein> .


To investigate the molecular basis of the mitogenic effect of high molecular weight dextran sulfate ( HMDS ) , monocyte-derived macrophages were transfected with recombinant plasmid containing chloramphenicol acetyl transferase ( CAT ) reporter gene under the control of the HIV-1 long terminal repeat ( LTR ) promoter , which is regulated by transcription factor NF-kappaB .

To investigate the molecular basis of the mitogenic effect of high molecular weight dextran sulfate ( HMDS ) , <cell_type>monocyte-derived macrophages</cell_type> were transfected with <dna>recombinant plasmid</dna> containing <dna>chloramphenicol acetyl transferase ( CAT ) reporter gene</dna> under the control of the <dna>HIV-1 long terminal repeat ( LTR ) promoter</dna> , which is regulated by <protein>transcription factor</protein> <protein>NF-kappaB</protein> .


We observed that HMDS , similar to bacterial lipopolysaccharide ( LPS ) , increases the expression of CAT reporter gene suggesting increased activity of NF-kappaB .

We observed that HMDS , similar to bacterial lipopolysaccharide ( LPS ) , increases the expression of <dna>CAT reporter gene</dna> suggesting increased activity of <protein>NF-kappaB</protein> .


The activation of NF-kappaB correlated with the increased expression of B7.1 molecules .

The activation of <protein>NF-kappaB</protein> correlated with the increased expression of <protein>B7.1 molecules</protein> .


It was postulated that this NF-kappaB-regulated promoter might play a role in the activation of the accessory cells as well as the rate of replication of HIV-1 in monocyte-derived macrophages .

It was postulated that this <dna>NF-kappaB-regulated promoter</dna> might play a role in the activation of the accessory cells as well as the rate of replication of HIV-1 in <cell_type>monocyte-derived macrophages</cell_type> .


Transcription factors Sp1 and AP-2 mediate induction of acid sphingomyelinase during monocytic differentiation .

<protein>Transcription factors</protein> <protein>Sp1</protein> and <protein>AP-2</protein> mediate induction of <protein>acid sphingomyelinase</protein> during monocytic differentiation .


Cells from the human monocytic leukemia cell line THP-1 differentiate towards a macrophage-like phenotype when stimulated with phorbol 12-myristate -13- acetate ( PMA ) , 1 , 25-dihydroxy-vitamin D3 , and various other agents .

Cells from the <cell_line>human monocytic leukemia cell line</cell_line> <cell_line>THP-1</cell_line> differentiate towards a macrophage-like phenotype when stimulated with phorbol 12-myristate -13- acetate ( PMA ) , 1 , 25-dihydroxy-vitamin D3 , and various other agents .


We demonstrate here that the expression of the lysosomal enzyme acid sphingomyelinase ( ASM ; E.C.3.1.4.12 ) is induced during this process and is strongly elevated in differentiated THP-1 cells , as well as in differentiated human mononuclear phagocytes .

We demonstrate here that the expression of the <protein>lysosomal enzyme</protein> <protein>acid sphingomyelinase</protein> ( ASM ; E.C.3.1.4.12 ) is induced during this process and is strongly elevated in differentiated <cell_line>THP-1 cells</cell_line> , as well as in differentiated <cell_type>human mononuclear phagocytes</cell_type> .


Using Northern blotting , RNase protection assay , and nuclear run-on analyses , we show that the up-regulation of ASM expression is regulated mainly at the level of transcription and that new protein synthesis is required for enhanced ASM activity .

Using Northern blotting , <protein>RNase</protein> protection assay , and nuclear run-on analyses , we show that the up-regulation of ASM expression is regulated mainly at the level of transcription and that new protein synthesis is required for enhanced ASM activity .


This cell-type specific induction by PMA treatment was further investigated with respect to transcriptional control .


A series of 5 ' deletion derivatives of the upstream regulatory region were used in transient transfection assays to identify promoter elements required for basal and inducible gene expression .

A series of <dna>5 ' deletion derivatives</dna> of the <dna>upstream regulatory region</dna> were used in transient transfection assays to identify <dna>promoter elements</dna> required for basal and inducible gene expression .


A PMA responsive element was localized to a region between -319 and -219 bp upstream of the initiation codon and co-transfections with transcription factor expression plasmids for AP-2 and Sp1 resulted in augmented ASM promoter activity , which was abolished when the binding sites for these two factors were deleted .

A <dna>PMA responsive element</dna> was localized to a region between <dna>-319 and -219 bp upstream</dna> of the initiation codon and co-transfections with <dna>transcription factor expression plasmids</dna> for <protein>AP-2</protein> and <protein>Sp1</protein> resulted in augmented ASM promoter activity , which was abolished when the <dna>binding sites</dna> for these two factors were deleted .


Using electrophoretic mobility shift assays and supershift assays we demonstrate that this region is specifically bound by Sp1 and AP-2 .

Using electrophoretic mobility shift assays and supershift assays we demonstrate that this region is specifically bound by <protein>Sp1</protein> and <protein>AP-2</protein> .


These factors are present in nuclear extracts prepared from both induced and uninduced THP-1 cells .

These factors are present in nuclear extracts prepared from both induced and uninduced <cell_line>THP-1 cells</cell_line> .


However , the intensity of the complex formed appeared to increase when nuclear extracts from PMA-treated cells were used .

However , the intensity of the complex formed appeared to increase when nuclear extracts from <cell_line>PMA-treated cells</cell_line> were used .


From these studies , we conclude that a concerted action of the transcription factors AP-2 and Sp1 is essential for the up -regulation of ASM expression during the process of macrophage differentiation .

From these studies , we conclude that a concerted action of the <protein>transcription factors</protein> <protein>AP-2</protein> and <protein>Sp1</protein> is essential for the up -regulation of ASM expression during the process of macrophage differentiation .


LPS-Induced NF-kappaB activation and TNF-alpha release in human monocytes are protein tyrosine kinase dependent and protein kinase C independent .

LPS-Induced <protein>NF-kappaB</protein> activation and <protein>TNF-alpha</protein> release in <cell_type>human monocytes</cell_type> are <protein>protein tyrosine kinase</protein> dependent and <protein>protein kinase C</protein> independent .


BACKGROUND : Tumor necrosis factor alpha ( TNF-alpha ) is an important mediator of septic shock .

BACKGROUND : <protein>Tumor necrosis factor alpha</protein> ( <protein>TNF-alpha</protein> ) is an important mediator of septic shock .


Endotoxin ( LPS ) signal transduction in human monocytes leads to activation of nuclear factor-kappa B ( NF-kappaB ) and TNF-alpha release .

Endotoxin ( LPS ) signal transduction in <cell_type>human monocytes</cell_type> leads to activation of <protein>nuclear factor-kappa B</protein> ( <protein>NF-kappaB</protein> ) and <protein>TNF-alpha</protein> release .


Previous studies have implicated activation of both protein kinase C ( PKC ) and protein tyrosine kinases ( PTK ) in LPS-induced NF-kappaB activation and TNF-alpha production .

Previous studies have implicated activation of both <protein>protein kinase C</protein> ( <protein>PKC</protein> ) and <protein>protein tyrosine kinases</protein> ( <protein>PTK</protein> ) in LPS-induced <protein>NF-kappaB</protein> activation and <protein>TNF-alpha</protein> production .


We hypothesized that inhibition of either PKC or PTK would decrease LPS-induced NF-kappaB DNA binding and TNF-alpha release in human monocytes .

We hypothesized that inhibition of either <protein>PKC</protein> or <protein>PTK</protein> would decrease LPS-induced <protein>NF-kappaB</protein> DNA binding and <protein>TNF-alpha</protein> release in <cell_type>human monocytes</cell_type> .


MATERIALS AND METHODS : Human monocytes were stimulated with PMA ( 50 ng/ml ) alone or LPS ( 100 ng/ml ) with and without a nonspecific serine/threonine protein kinase inhibitor staurosporine ( Stauro ) , a specific pan- PKC inhibitor bisindolylmaleimide ( Bis ) , or an inhibitor of PTK genistein ( Gen ) .

MATERIALS AND METHODS : <cell_type>Human monocytes</cell_type> were stimulated with PMA ( 50 ng/ml ) alone or LPS ( 100 ng/ml ) with and without a nonspecific <protein>serine/threonine protein kinase</protein> inhibitor staurosporine ( Stauro ) , a specific pan- <protein>PKC</protein> inhibitor bisindolylmaleimide ( Bis ) , or an inhibitor of <protein>PTK</protein> genistein ( Gen ) .


TNF-alpha release in culture supernatants was measured by an ELISA .

<protein>TNF-alpha</protein> release in culture supernatants was measured by an ELISA .


NF-kappaB DNA binding was evaluated by electrophoretic mobility shift assay .

<protein>NF-kappaB</protein> DNA binding was evaluated by electrophoretic mobility shift assay .


RESULTS : LPS increased NF-kappaB DNA binding and TNF-alpha release in human monocytes .

RESULTS : LPS increased <protein>NF-kappaB</protein> DNA binding and <protein>TNF-alpha</protein> release in <cell_type>human monocytes</cell_type> .


Nonspecific protein kinase inhibition inhibited NF-kappaB activation and TNF-alpha release , while specific PKC inhibition with Bis had no effect on LPS-induced NF-kappaB DNA binding or TNF-alpha release .

Nonspecific <protein>protein kinase</protein> inhibition inhibited <protein>NF-kappaB</protein> activation and <protein>TNF-alpha</protein> release , while specific <protein>PKC</protein> inhibition with Bis had no effect on LPS-induced <protein>NF-kappaB</protein> DNA binding or <protein>TNF-alpha</protein> release .


PTK inhibition with Gen attenuated both LPS-induced NF-kappaB DNA binding and TNF-alpha production in human monocytes .

<protein>PTK</protein> inhibition with Gen attenuated both LPS-induced <protein>NF-kappaB</protein> DNA binding and <protein>TNF-alpha</protein> production in <cell_type>human monocytes</cell_type> .


Direct activation of PKC with PMA induced both NF-kappaB activation and TNF-alpha production by human monocytes .

Direct activation of <protein>PKC</protein> with PMA induced both <protein>NF-kappaB</protein> activation and <protein>TNF-alpha</protein> production by <cell_type>human monocytes</cell_type> .


CONCLUSIONS : These results suggest that LPS-induced NF-kappaB activation and TNF-alpha release in human monocytes are independent of PKC activity .

CONCLUSIONS : These results suggest that LPS-induced <protein>NF-kappaB</protein> activation and <protein>TNF-alpha</protein> release in <cell_type>human monocytes</cell_type> are independent of <protein>PKC</protein> activity .


Furthermore , our results provide evidence that PTK plays a role in LPS-induced NF-kappaB activation and TNF-alpha release in human monocytes and thus could be a potential therapeutic target in inflammatory states .

Furthermore , our results provide evidence that <protein>PTK</protein> plays a role in LPS-induced <protein>NF-kappaB</protein> activation and <protein>TNF-alpha</protein> release in <cell_type>human monocytes</cell_type> and thus could be a potential therapeutic target in inflammatory states .


Copyright 1999 Academic Press .


Amelioration of rat cerulein pancreatitis by guamerin-derived peptide , a novel elastase inhibitor .

Amelioration of rat cerulein pancreatitis by guamerin-derived peptide , a novel <protein>elastase</protein> inhibitor .


Increased activity of various proteases is observed in both human and experimental pancreatitis ; however , the information on the effects of specific protease inhibitors on the disease is limited .

Increased activity of various <protein>proteases</protein> is observed in both human and experimental pancreatitis ; however , the information on the effects of specific protease inhibitors on the disease is limited .


In this study we show that a novel elastase inhibitor , guamerin-derived synthetic peptide ( GDSP ) , improves the parameters of cerulein-induced acute pancreatitis in the rat .


The effects of GDSP on pancreatic weight , serum amylase and lipase , morphologic changes in the pancreas , neutrophil infiltration , and nuclear factor KB ( NF-KB ) activation were measured in rats infused with supramaximal dose of cerulein ( 5 ( g/kg/h ) for 6 h .

The effects of GDSP on pancreatic weight , <protein>serum amylase and lipase</protein> , morphologic changes in the pancreas , neutrophil infiltration , and <protein>nuclear factor KB</protein> ( <protein>NF-KB</protein> ) activation were measured in rats infused with supramaximal dose of cerulein ( 5 ( g/kg/h ) for 6 h .


The effects of GDSP were also measured on superoxide formation by activated human neutrophils .

The effects of GDSP were also measured on superoxide formation by <cell_type>activated human neutrophils</cell_type> .


The effects of GDSP were compared with those of another elastase inhibitor , elastatinal .


GDSP significantly inhibited edema formation , neutrophil infiltration , acinar cell damage , and plasma lipase and amylase increases caused by cerulein .

GDSP significantly inhibited edema formation , neutrophil infiltration , acinar cell damage , and <protein>plasma lipase</protein> and <protein>amylase</protein> increases caused by cerulein .


GDSP also completely inhibited superoxide formation in the human neutrophils stimulated by N-formyl-methionine-leucine-phenyl-alanine ( fMLP ) or 12-O-tetradecanoylphorbol-13-acetate ( TPA ) .

GDSP also completely inhibited superoxide formation in the <cell_type>human neutrophils</cell_type> stimulated by N-formyl-methionine-leucine-phenyl-alanine ( fMLP ) or 12-O-tetradecanoylphorbol-13-acetate ( TPA ) .


Elastatinal had some of the same effects as GDSP but was less potent and effective .


These results demonstrate a beneficial effect of GDSP , a novel specific elastase inhibitor , on the development of rat cerulein pancreatitis .


A novel lipopolysaccharide-induced transcription factor regulating tumor necrosis factor alpha gene expression : molecular cloning , sequencing , characterization , and chromosomal assignment .

A novel <protein>lipopolysaccharide-induced transcription factor</protein> regulating <protein>tumor necrosis factor alpha</protein> gene expression : molecular cloning , sequencing , characterization , and chromosomal assignment .


Lipopolysaccharide ( LPS ) is a potent stimulator of monocytes and macrophages , causing secretion of tumor necrosis factor alpha ( TNF-alpha ) and other inflammatory mediators .

Lipopolysaccharide ( LPS ) is a potent stimulator of <cell_type>monocytes</cell_type> and <cell_type>macrophages</cell_type> , causing secretion of <protein>tumor necrosis factor alpha</protein> ( <protein>TNF-alpha</protein> ) and other inflammatory mediators .


Given the deleterious effects to the host of TNF-alpha , it has been postulated that TNF-alpha gene expression must be tightly regulated .

Given the deleterious effects to the host of <protein>TNF-alpha</protein> , it has been postulated that <protein>TNF-alpha</protein> gene expression must be tightly regulated .


The nature of the nuclear factor ( s ) that control TNF-alpha gene transcription in humans remains obscure , although NF-kappaB has been suggested .

The nature of the <protein>nuclear factor</protein> ( s ) that control <protein>TNF-alpha</protein> gene transcription in humans remains obscure , although <protein>NF-kappaB</protein> has been suggested .


Our previous studies pertaining to macrophage response to LPS identified a novel DNA-binding domain located from -550 to -487 in the human TNF-alpha promoter that contains transcriptional activity , but lacks any known NF-kappaB -binding sites .

Our previous studies pertaining to macrophage response to LPS identified a novel <protein>DNA-binding domain</protein> located from -550 to -487 in the <dna>human TNF-alpha promoter</dna> that contains transcriptional activity , but lacks any known <protein>NF-kappaB</protein> <dna>-binding sites</dna> .


We have used this DNA fragment to isolate and purify a 60-kDa protein binding to this fragment and obtained its amino-terminal sequence , which was used to design degenerate probes to screen a cDNA library from THP-1 cells .

We have used this DNA fragment to isolate and purify a <protein>60-kDa protein</protein> binding to this fragment and obtained its <protein>amino-terminal sequence</protein> , which was used to design <dna>degenerate probes</dna> to screen a cDNA library from <cell_line>THP-1 cells</cell_line> .


A novel cDNA clone ( 1.8 kb ) was isolated and fully sequenced .

A novel <dna>cDNA clone</dna> ( 1.8 kb ) was isolated and fully sequenced .


Characterization of this cDNA clone revealed that its induction was dependent on LPS activation of THP-1 cells ; hence , the name LPS-induced TNF-alpha factor ( LITAF ) .

Characterization of this <dna>cDNA clone</dna> revealed that its induction was dependent on LPS activation of <cell_line>THP-1 cells</cell_line> ; hence , the name <protein>LPS-induced TNF-alpha factor</protein> ( <protein>LITAF</protein> ) .


Inhibition of LITAF mRNA expression in THP-1 cells resulted in a reduction of TNF-alpha transcripts .

Inhibition of <rna>LITAF mRNA</rna> expression in <cell_line>THP-1 cells</cell_line> resulted in a reduction of <protein>TNF-alpha</protein> transcripts .


In addition , high level of expression of LITAF mRNA was observed predominantly in the placenta , peripheral blood leukocytes , lymph nodes , and the spleen .

In addition , high level of expression of <rna>LITAF mRNA</rna> was observed predominantly in the placenta , <cell_type>peripheral blood leukocytes</cell_type> , lymph nodes , and the spleen .


Finally , chromosomal localization using fluorescence in situ hybridization revealed that LITAF mapped to chromosome 16p12-16p13.3 .

Finally , chromosomal localization using fluorescence in situ hybridization revealed that <protein>LITAF</protein> mapped to <dna>chromosome 16p12-16p13.3</dna> .


Together , these findings suggest that LITAF plays an important role in the activation of the human TNF-alpha gene and proposes a new mechanism to control TNF-alpha gene expression .

Together , these findings suggest that <protein>LITAF</protein> plays an important role in the activation of the <dna>human TNF-alpha gene</dna> and proposes a new mechanism to control <protein>TNF-alpha</protein> gene expression .


A human IFNGR1 small deletion hotspot associated with dominant susceptibility to mycobacterial infection [ see comments ]

A <dna>human IFNGR1 small deletion hotspot</dna> associated with dominant susceptibility to mycobacterial infection [ see comments ]


The immunogenetic basis of severe infections caused by bacille Calmette-Guerin vaccine and environmental mycobacteria in humans remains largely unknown .


We describe 18 patients from several generations of 12 unrelated families who were heterozygous for 1 to 5 overlapping IFNGR1 frameshift small deletions and a wild-type IFNGR1 allele .

We describe 18 patients from several generations of 12 unrelated families who were heterozygous for 1 to 5 <dna>overlapping IFNGR1 frameshift small deletions</dna> and a <dna>wild-type IFNGR1 allele</dna> .


There were 12 independent mutation events at a single mutation site , defining a small deletion hotspot .

There were 12 independent mutation events at a <dna>single mutation site</dna> , defining a <dna>small deletion hotspot</dna> .


Neighbouring sequence analysis favours a small deletion model of slipped mispairing events during replication .


The mutant alleles encode cell-surface IFNgamma receptors that lack the intra-cytoplasmic domain , which , through a combination of impaired recycling , abrogated signalling and normal binding to IFNgamma exert a dominant-negative effect .

The mutant alleles encode <protein>cell-surface IFNgamma receptors</protein> that lack the <protein>intra-cytoplasmic domain</protein> , which , through a combination of impaired recycling , abrogated signalling and normal binding to <protein>IFNgamma</protein> exert a dominant-negative effect .


We thus report a hotspot for human IFNGR1 small deletions that confer dominant susceptibility to infections caused by poorly virulent mycobacteria .

We thus report a hotspot for <dna>human IFNGR1 small deletions</dna> that confer dominant susceptibility to infections caused by poorly virulent mycobacteria .


Characterization of expression of the gene for human pterin carbinolamine dehydratase/dimerization cofactor of HNF1 .

Characterization of expression of the gene for <protein>human pterin carbinolamine dehydratase/dimerization cofactor</protein> of <protein>HNF1</protein> .


Pterin carbinolamine dehydratase/dimerization cofactor of HNF1 ( PCD/DCoH ) is a dual-function protein .

<protein>Pterin carbinolamine dehydratase/dimerization cofactor</protein> of <protein>HNF1</protein> ( <protein>PCD/DCoH</protein> ) is a <protein>dual-function protein</protein> .


In the cytoplasm it acts as a dehydratase in the regeneration of tetrahydrobiopterin , the cofactor for aromatic amino acid hydroxylases .

In the cytoplasm it acts as a <protein>dehydratase</protein> in the regeneration of <protein>tetrahydrobiopterin</protein> , the cofactor for <protein>aromatic amino acid hydroxylases</protein> .


In the nucleus , it functions as a dimerization cofactor of HNF1 and increases the transcriptional activity of HNF1 .

In the nucleus , it functions as a <protein>dimerization cofactor</protein> of <protein>HNF1</protein> and increases the transcriptional activity of <protein>HNF1</protein> .


To deepen our understanding of this protein , we characterized its expression in human tissues and cells .


Human PCD/DCoH was present predominantly in liver and kidney , with significant amounts in testis and ovary , trace amounts in lung , and undetectable levels in whole brain , heart , and spleen .

Human <protein>PCD/DCoH</protein> was present predominantly in liver and kidney , with significant amounts in testis and ovary , trace amounts in lung , and undetectable levels in whole brain , heart , and spleen .


It was expressed in all of the cells that were examined .


Importantly , it was also present in the nucleus of HeLa cells , which lack HNF1 , and in the cytoplasm of fibroblasts that have little or no tetrahydrobiopterin .

Importantly , it was also present in the nucleus of <cell_line>HeLa cells</cell_line> , which lack <protein>HNF1</protein> , and in the cytoplasm of <cell_type>fibroblasts</cell_type> that have little or no <protein>tetrahydrobiopterin</protein> .


The expression of human PCD/DCoH in the liver and nonhepatic cells was compared at both the mRNA and protein levels .

The expression of <protein>human PCD/DCoH</protein> in the liver and <cell_type>nonhepatic cells</cell_type> was compared at both the mRNA and protein levels .


Although the mRNA level in liver was only fourfold higher than that in keratinocytes and fibroblasts , the hepatic PCD/DCoH protein level was 20-fold higher than that in normal human epidermal keratinocytes and dermal fibroblasts .

Although the mRNA level in liver was only fourfold higher than that in <cell_type>keratinocytes</cell_type> and <cell_type>fibroblasts</cell_type> , the <protein>hepatic PCD/DCoH protein</protein> level was 20-fold higher than that in <cell_type>normal human epidermal keratinocytes</cell_type> and <cell_type>dermal fibroblasts</cell_type> .


Cloning of the 5 ' and 3 ' untranslated region ( UTR ) of human keratinocyte PCD/DCoH revealed that it has 53 bp more of GC-rich 5 ' untranslated sequence than the published liver PCD/DCoH .

Cloning of the <dna>5 ' and 3 ' untranslated region</dna> ( <dna>UTR</dna> ) of <protein>human keratinocyte PCD/DCoH</protein> revealed that it has 53 bp more of <dna>GC-rich 5 ' untranslated sequence</dna> than the published <protein>liver PCD/DCoH</protein> .


In vitro transcription and translation analysis showed that the longer 5 ' UTR resulted in about a 35 % decrease in translation efficiency .

In vitro transcription and translation analysis showed that the longer <dna>5 ' UTR</dna> resulted in about a 35 % decrease in translation efficiency .


These data show that human PCD/DCoH is not only present in cells where tetrahydrobiopterin is synthesized or HNF1 is present but is a widely distributed protein .

These data show that <protein>human PCD/DCoH</protein> is not only present in cells where <protein>tetrahydrobiopterin</protein> is synthesized or <protein>HNF1</protein> is present but is a widely distributed protein .


Its differential expression in different tissues and cells is regulated not only at the transcriptional level but also at the translational level .


NF-kappaB regulates Fas/APO-1/CD95- and TCR- mediated apoptosis of T lymphocytes .

<protein>NF-kappaB</protein> regulates Fas/APO-1/CD95- and TCR- mediated apoptosis of <cell_type>T lymphocytes</cell_type> .


The maintenance of lymphocyte homeostasis by apoptosis is a critical regulatory mechanism in the normal immune system .


The transcription factor NF-kappaB has been shown to play a role in protecting cells against death mediated by TNF We show here that NF-kappaB also has a role in regulating Fas / APO-1 / CD95 -mediated death , a major pathway of peripheral T cell death .

The <protein>transcription factor</protein> <protein>NF-kappaB</protein> has been shown to play a role in protecting cells against death mediated by <protein>TNF</protein> We show here that <protein>NF-kappaB</protein> also has a role in regulating <protein>Fas</protein> / <protein>APO-1</protein> / <protein>CD95</protein> -mediated death , a major pathway of <cell_type>peripheral T cell</cell_type> death .


Transfection of Jurkat cells with the NF-kappaB subunits p50 and p65 confers resistance against Fas-mediated apoptosis .

Transfection of <cell_line>Jurkat cells</cell_line> with the <protein>NF-kappaB subunits</protein> <protein>p50</protein> and <protein>p65</protein> confers resistance against Fas-mediated apoptosis .


Reciprocally , inhibition of NF-kappaB activation by a soluble peptide inhibitor or a dominant form of the NF-kappaB inhibitor , IkappaB , makes the cells more susceptible to Fas-mediated apoptosis .

Reciprocally , inhibition of <protein>NF-kappaB</protein> activation by a soluble peptide inhibitor or a dominant form of the <protein>NF-kappaB</protein> inhibitor , <protein>IkappaB</protein> , makes the cells more susceptible to Fas-mediated apoptosis .


Furthermore , inhibition of NF-kappaB activation by a soluble peptide inhibitor rendered a T cell hybridoma more susceptible to TCR-mediated apoptosis .

Furthermore , inhibition of <protein>NF-kappaB</protein> activation by a soluble peptide inhibitor rendered a <cell_line>T cell hybridoma</cell_line> more susceptible to TCR-mediated apoptosis .


Correspondingly , transfection of p50 and p65 provided considerable protection from TCR-mediated apoptosis .

Correspondingly , transfection of <protein>p50</protein> and <protein>p65</protein> provided considerable protection from TCR-mediated apoptosis .


These observations were corroborated by studies on Fas -mediated death in primary T cells .

These observations were corroborated by studies on <protein>Fas</protein> -mediated death in <cell_type>primary T cells</cell_type> .


Concanavalin A-activated cycling T cell blasts from mice that are transgenic for the dominant IkappaB molecule have increased sensitivity to Fas-mediated apoptosis , associated with a down-regulation of NF-kappaB complexes in the nucleus .

Concanavalin A-activated <cell_line>cycling T cell blasts</cell_line> from mice that are transgenic for the <protein>dominant IkappaB molecule</protein> have increased sensitivity to Fas-mediated apoptosis , associated with a down-regulation of <protein>NF-kappaB complexes</protein> in the nucleus .


In addition , blocking TNF , itself a positive regulator of NF-kappaB , with neutralizing antibodies renders the cells more susceptible to anti-Fas-mediated apoptosis .

In addition , blocking <protein>TNF</protein> , itself a positive regulator of <protein>NF-kappaB</protein> , with <protein>neutralizing antibodies</protein> renders the cells more susceptible to anti-Fas-mediated apoptosis .


In summary , our results provide compelling evidence that NF-kappaB protects against Fas -mediated death and is likely to be an important regulator of T cell homeostasis and tolerance .

In summary , our results provide compelling evidence that <protein>NF-kappaB</protein> protects against <protein>Fas</protein> -mediated death and is likely to be an important regulator of T cell homeostasis and tolerance .


T helper differentiation proceeds through Stat1-dependent , Stat4-dependent and Stat4-independent phases .


Much of our focus in understanding Th1/Th2 development has been on the signals delivered by IL-12 and IL-4 as final determinants of terminal T cell differentiation .

Much of our focus in understanding Th1/Th2 development has been on the signals delivered by <protein>IL-12</protein> and <protein>IL-4</protein> as final determinants of terminal T cell differentiation .


Because extinction of IL-12 signaling in early Th2 development could potentially be important in imprinting a more permanent Th2 phenotype on a population of T cells , we have also examined various parameters regulating the IL-12 signaling pathway .

Because extinction of <protein>IL-12</protein> signaling in early Th2 development could potentially be important in imprinting a more permanent Th2 phenotype on a population of <cell_type>T cells</cell_type> , we have also examined various parameters regulating the <protein>IL-12</protein> signaling pathway .


Whereas IL-4 appears to repress functional IL-12 signaling through inhibition of IL-12R beta 2 expression , IFN-gamma in the mouse , and IFN-alpha in the human appear to induce IL-12R beta 2 expression and promote IL-12 responsiveness .

Whereas <protein>IL-4</protein> appears to repress functional <protein>IL-12</protein> signaling through inhibition of <protein>IL-12R beta 2</protein> expression , <protein>IFN-gamma</protein> in the mouse , and <protein>IFN-alpha</protein> in the human appear to induce <protein>IL-12R beta 2</protein> expression and promote <protein>IL-12</protein> responsiveness .


We propose that Th1 development can be considered in two stages , capacitance and development .


Capacitance would simply involve expression of IL-12R beta 1 and beta 2 subunits , regulated by TCR , IL-4 and IFNs .

Capacitance would simply involve expression of <protein>IL-12R beta 1 and beta 2 subunits</protein> , regulated by TCR , <protein>IL-4</protein> and <protein>IFNs</protein> .


The second stage , development , we propose is the true IL-12 induced developmental stage , involving expression of Stat4 inducible proteins .

The second stage , development , we propose is the true <protein>IL-12</protein> induced developmental stage , involving expression of <protein>Stat4 inducible proteins</protein> .


In the human , this may also occur via IFN-alpha , which is able to activate Stat4 .

In the human , this may also occur via <protein>IFN-alpha</protein> , which is able to activate <protein>Stat4</protein> .


It is perhaps possible that all of Stat4 actions on Th1 development may be exert directly by Stat4 at the IFN-gamma gene , however we suggest that , more likely , Stat4 may act to induce Th1 development through the induction of other non-cytokine genes , whose stable expression maintains the transcriptional state of a Th1 cell .

It is perhaps possible that all of <protein>Stat4</protein> actions on Th1 development may be exert directly by <protein>Stat4</protein> at the <dna>IFN-gamma gene</dna> , however we suggest that , more likely , <protein>Stat4</protein> may act to induce Th1 development through the induction of other <dna>non-cytokine genes</dna> , whose stable expression maintains the transcriptional state of a <cell_type>Th1 cell</cell_type> .


Lineage-specific activation of STAT3 by interferon-gamma in human neutrophils .

Lineage-specific activation of <protein>STAT3</protein> by <protein>interferon-gamma</protein> in <cell_type>human neutrophils</cell_type> .


Binding of interferon-gamma ( IFN-gamma ) to its heterodimeric receptor induces activation of the tyrosine kinases JAK1 and JAK2 followed by tyrosine phosphorylation of STAT1alpha .

Binding of <protein>interferon-gamma</protein> ( <protein>IFN-gamma</protein> ) to its heterodimeric receptor induces activation of the <protein>tyrosine kinases</protein> <protein>JAK1</protein> and <protein>JAK2</protein> followed by tyrosine phosphorylation of <protein>STAT1alpha</protein> .


Selective activation of STAT1alpha at the IFN-gamma receptor is achieved by specific interaction between a cytosolic tyrosine motif including Y440 in the IFN-gamma receptor alpha-chain and the SH2 domain of STAT1alpha .

Selective activation of <protein>STAT1alpha</protein> at the <protein>IFN-gamma receptor</protein> is achieved by specific interaction between a <protein>cytosolic tyrosine motif</protein> including <protein>Y440</protein> in the <protein>IFN-gamma receptor alpha-chain</protein> and the <protein>SH2 domain</protein> of <protein>STAT1alpha</protein> .


We demonstrate that , in addition to STAT1alpha , STAT3 is also activated by IFN-gamma in human neutrophils .

We demonstrate that , in addition to <protein>STAT1alpha</protein> , <protein>STAT3</protein> is also activated by <protein>IFN-gamma</protein> in <cell_type>human neutrophils</cell_type> .


The activation of STAT3 was not found in human eosinophils , monocytes , and HL-60 cells , although the STAT3 protein was expressed in these cells .

The activation of <protein>STAT3</protein> was not found in <cell_type>human eosinophils</cell_type> , <cell_type>monocytes</cell_type> , and <cell_line>HL-60 cells</cell_line> , although the <protein>STAT3 protein</protein> was expressed in these cells .


The cell type-specific activation of STAT3 by IFN-gamma was also observed in neutrophils that are differentiated in vitro from human CD34+ hematopoietic stem cells .

The cell type-specific activation of <protein>STAT3</protein> by <protein>IFN-gamma</protein> was also observed in <cell_type>neutrophils</cell_type> that are differentiated in vitro from <cell_type>human CD34+ hematopoietic stem cells</cell_type> .


These results indicate that a single cytokine receptor can activate different STAT family members in a cell-specific manner , which might result in cell-specific gene transcription .

These results indicate that a single <protein>cytokine receptor</protein> can activate different <protein>STAT family members</protein> in a cell-specific manner , which might result in cell-specific gene transcription .


Alternative polyadenylation events contribute to the induction of NF-ATc in effector T cells .

Alternative polyadenylation events contribute to the induction of <protein>NF-ATc</protein> in <cell_type>effector T cells</cell_type> .


The transcription factor NF-ATc is synthesized in three prominent isoforms .

The <protein>transcription factor</protein> <protein>NF-ATc</protein> is synthesized in three prominent <protein>isoforms</protein> .


These differ in the length of their C terminal peptides and mode of synthesis .


Due to a switch from the use of a 3 ' polyA site to a more proximal polyA site , NF-ATc expression switches from the synthesis of the two longer isoforms in naive T cells to that of short isoform A in T effector cells .

Due to a switch from the use of a <dna>3 ' polyA site</dna> to a more <dna>proximal polyA site</dna> , <protein>NF-ATc</protein> expression switches from the synthesis of the two longer isoforms in <cell_type>naive T cells</cell_type> to that of short <protein>isoform A</protein> in <cell_type>T effector cells</cell_type> .


The relative low binding affinity of cleavage stimulation factor CstF-64 to the proximal polyA site seems to contribute to its neglect in naive T cells .

The relative low binding affinity of <protein>cleavage stimulation factor</protein> <protein>CstF-64</protein> to the <dna>proximal polyA site</dna> seems to contribute to its neglect in <cell_type>naive T cells</cell_type> .


These alternative polyadenylation events ensure the rapid accumulation of high concentrations of NF-ATc necessary to exceed critical threshold levels of NF-ATc for gene induction in effector T cells .

These alternative polyadenylation events ensure the rapid accumulation of high concentrations of <protein>NF-ATc</protein> necessary to exceed critical threshold levels of <protein>NF-ATc</protein> for gene induction in <cell_type>effector T cells</cell_type> .


Estrogen and progesterone induction of survival of monoblastoid cells undergoing TNF-alpha -induced apoptosis .

Estrogen and progesterone induction of survival of <cell_type>monoblastoid cells</cell_type> undergoing <protein>TNF-alpha</protein> -induced apoptosis .


Induction of apoptosis of mononucleated cells is a physiological process for regulating the intensity of the immune response .

Induction of apoptosis of <cell_type>mononucleated cells</cell_type> is a physiological process for regulating the intensity of the immune response .


The female steroid hormones estrogen ( E2 ) and progesterone ( Prog ) are known to modulate the reactivity of the immune system ; recently it has been demonstrated that they can regulate induction of apoptosis of endothelial cells and osteoblasts .

The female steroid hormones estrogen ( E2 ) and progesterone ( Prog ) are known to modulate the reactivity of the immune system ; recently it has been demonstrated that they can regulate induction of apoptosis of <cell_type>endothelial cells</cell_type> and <cell_type>osteoblasts</cell_type> .


TNF-alpha -mediated induction of apoptosis has been well characterized in myeloid cells .

<protein>TNF-alpha</protein> -mediated induction of apoptosis has been well characterized in <cell_type>myeloid cells</cell_type> .


We investigated whether E2 and Prog could interfere with TNF-alpha -induced apoptosis of the monoblastoid U937 cell line .

We investigated whether E2 and Prog could interfere with <protein>TNF-alpha</protein> -induced apoptosis of the <cell_line>monoblastoid U937 cell line</cell_line> .


Treatment with E2 or Prog increased survival and prevented apoptosis induced by TNF-alpha in both undifferentiated and macrophage-like PMA-differentiated U937 cells , as assessed by trypan blue exclusion cell counting , thymidine incorporation , AnnexinV labeling , followed by flow cytometry and DNA fragmentation studies .

Treatment with E2 or Prog increased survival and prevented apoptosis induced by <protein>TNF-alpha</protein> in both undifferentiated and macrophage-like <cell_line>PMA-differentiated U937 cells</cell_line> , as assessed by trypan blue exclusion cell counting , thymidine incorporation , AnnexinV labeling , followed by flow cytometry and DNA fragmentation studies .


This effect can be associated with the activation of specific hormone receptors , since we observed the expression of the estrogen receptor alpha ( ER-alpha ) , ER-beta , and progesterone receptor ( PR ) mRNAs ; the ER-alpha protein expression was confirmed by immunocytochemical analysis .

This effect can be associated with the activation of specific hormone receptors , since we observed the expression of the <rna>estrogen receptor alpha ( ER-alpha ) , ER-beta , and progesterone receptor ( PR ) mRNAs</rna> ; the <protein>ER-alpha</protein> protein expression was confirmed by immunocytochemical analysis .


In addition , hormone-mediated survival against apoptosis was concentration dependent , reaching the half-maximal effect at 10 nM and blocked by the ER antagonist ICI 182 , 780 in undifferentiated cells , further supporting a receptor-mediated mechanism of cell survival .

In addition , hormone-mediated survival against apoptosis was concentration dependent , reaching the half-maximal effect at 10 nM and blocked by the ER antagonist ICI 182 , 780 in <cell_type>undifferentiated cells</cell_type> , further supporting a receptor-mediated mechanism of cell survival .


Other steroid receptor drugs such as Raloxifene , RU486 , or the ICI 182 , 780 in PMA-differentiated cells displayed agonist activity by preventing TNF-alpha -induced apoptosis as efficiently as the hormones alone , providing further evidence to the notion that steroid receptor drugs may manifest agonist or antagonist activities depending on the cellular context in which they are studied .

Other steroid receptor drugs such as Raloxifene , RU486 , or the ICI 182 , 780 in <cell_line>PMA-differentiated cells</cell_line> displayed agonist activity by preventing <protein>TNF-alpha</protein> -induced apoptosis as efficiently as the hormones alone , providing further evidence to the notion that steroid receptor drugs may manifest agonist or antagonist activities depending on the cellular context in which they are studied .


Treatment with E2 was also associated with a time-dependent decrease in the mRNA level of the proapoptotic Nip-2 protein , supporting the hypothesis that hormone responsiveness of U937 cells is mediated by target gene transcription .

Treatment with E2 was also associated with a time-dependent decrease in the mRNA level of the <protein>proapoptotic Nip-2 protein</protein> , supporting the hypothesis that hormone responsiveness of <cell_line>U937 cells</cell_line> is mediated by target gene transcription .


Together , these results demonstrate that ER and PR can be activated by endogenous or exogenous ligands to induce a genetic response that impairs TNF-alpha -induced apoptosis in U937 cells .

Together , these results demonstrate that <protein>ER</protein> and <protein>PR</protein> can be activated by endogenous or exogenous ligands to induce a genetic response that impairs <protein>TNF-alpha</protein> -induced apoptosis in <cell_line>U937 cells</cell_line> .


The data presented here suggest that the female steroid receptors play a role in regulation of the immune response by preventing apoptosis of monoblastoid cells ; this effect might have important consequences in the clinical use of steroid receptor drugs .

The data presented here suggest that the <protein>female steroid receptors</protein> play a role in regulation of the immune response by preventing apoptosis of <cell_type>monoblastoid cells</cell_type> ; this effect might have important consequences in the clinical use of steroid receptor drugs .


-- Vegeto , E. , Pollio , G. , Pellicciari , C. , Maggi , A .


Estrogen and progesterone induction of survival of monoblastoid cells undergoing TNF-alpha -inuced apoptosis .

Estrogen and progesterone induction of survival of <cell_type>monoblastoid cells</cell_type> undergoing <protein>TNF-alpha</protein> -inuced apoptosis .


Dicarba-closo-dodecaboranes as a pharmacophore .


Retinoidal antagonists and potential agonists .


Synthesis and biological evaluation of the first dicarba-closo-dodecaborane ( carborane ) derivatives of retinoids are described .


Their retinoidal activity were examined in terms of the differentiation-inducing ability toward human promyelocytic leukemia HL-60 cells .

Their retinoidal activity were examined in terms of the differentiation-inducing ability toward <cell_line>human promyelocytic leukemia HL-60 cells</cell_line> .


High retinoidal activity ( agonist or antagonist for retinoic acid receptor ( RAR ) requires a carboxylic acid moiety and an appropriate hydrophobic group located at a suitable position on the molecule .

High retinoidal activity ( agonist or antagonist for <protein>retinoic acid receptor</protein> ( <protein>RAR</protein> ) requires a carboxylic acid moiety and an appropriate hydrophobic group located at a suitable position on the molecule .


The 4-carboranyl-substituted compounds ( 7 , 11 ) showed antagonistic activity but no agonistic activity even in the presence of the potent synergist HX630 .


On the other hand , the 3-carboranyl-substituted compounds ( 8 , 12 ) showed potential agonistic activity , but no antagonistic activity .


The results indicates that carboranes are applicable as the hydrophobic moiety of biologically active molecules .


Inhibition of T cell signaling by mitogen-activated protein kinase-targeted hematopoietic tyrosine phosphatase ( HePTP ) .

Inhibition of T cell signaling by <protein>mitogen-activated protein kinase-targeted hematopoietic tyrosine phosphatase</protein> ( <protein>HePTP</protein> ) .


Activation of T lymphocytes to produce cytokines is regulated by the counterbalance of protein-tyrosine kinases and protein-tyrosine phosphatases , many of which have a high degree of substrate specificity because of physical association with their targets .

Activation of <cell_type>T lymphocytes</cell_type> to produce <protein>cytokines</protein> is regulated by the counterbalance of <protein>protein-tyrosine kinases</protein> and protein-tyrosine phosphatases , many of which have a high degree of substrate specificity because of physical association with their targets .


Overexpression of hematopoietic protein-tyrosine phosphatase ( HePTP ) results in suppression of T lymphocyte activation as measured by T cell antigen receptor -induced activation of transcription factors binding to the 5 ' promoter of the interleukin-2 gene .

Overexpression of <protein>hematopoietic protein-tyrosine phosphatase</protein> ( <protein>HePTP</protein> ) results in suppression of T lymphocyte activation as measured by <protein>T cell antigen receptor</protein> -induced activation of <protein>transcription factors</protein> binding to the <dna>5 ' promoter</dna> of the <dna>interleukin-2 gene</dna> .


Efforts to pinpoint the exact site of action and specificity of HePTP in the signaling cascade revealed that HePTP acts directly on the mitogen-activated protein ( MAP ) kinases Erk1 and 2 and consequently reduces the magnitude and duration of their catalytic activation in intact T cells .

Efforts to pinpoint the exact site of action and specificity of <protein>HePTP</protein> in the signaling cascade revealed that <protein>HePTP</protein> acts directly on the <protein>mitogen-activated protein ( MAP ) kinases</protein> <protein>Erk1 and 2</protein> and consequently reduces the magnitude and duration of their catalytic activation in <cell_type>intact T cells</cell_type> .


In contrast , HePTP had no effects on N-terminal c-Jun kinase or on events upstream of the MAP kinases .

In contrast , <protein>HePTP</protein> had no effects on <protein>N-terminal c-Jun kinase</protein> or on events upstream of the MAP kinases .


The specificity of HePTP correlated with its physical association through its noncatalytic N terminus with Erk and another MAP kinase , p38 , but not Jnk or other proteins .

The specificity of <protein>HePTP</protein> correlated with its physical association through its noncatalytic <protein>N terminus</protein> with <protein>Erk</protein> and another <protein>MAP kinase</protein> , <protein>p38</protein> , but not <protein>Jnk</protein> or other proteins .


We propose that HePTP plays a negative role in antigen receptor signaling by specifically regulating MAP kinases in the cytosol and at early time points of T cell activation before the activation-induced expression of nuclear dual-specific MAP kinase phosphatases .

We propose that <protein>HePTP</protein> plays a negative role in antigen receptor signaling by specifically regulating <protein>MAP kinases</protein> in the cytosol and at early time points of T cell activation before the activation-induced expression of <protein>nuclear dual-specific MAP kinase phosphatases</protein> .


p38 mitogen-activated protein kinase mediates signal integration of TCR /CD28 costimulation in primary murine T cells .

<protein>p38 mitogen-activated protein kinase</protein> mediates signal integration of <protein>TCR</protein> <protein>/CD28</protein> costimulation in <cell_type>primary murine T cells</cell_type> .


Optimal T cell activation requires two signals , one generated by TCR and another by the CD28 costimulatory receptor .

Optimal T cell activation requires two signals , one generated by <protein>TCR</protein> and another by the <protein>CD28 costimulatory receptor</protein> .


In this study , we investigated the regulation of costimulation-induced mitogen-activated protein kinase ( MAPK ) activation in primary mouse T cells .

In this study , we investigated the regulation of costimulation-induced <protein>mitogen-activated protein kinase</protein> ( <protein>MAPK</protein> ) activation in <cell_type>primary mouse T cells</cell_type> .


In contrast to that reported for human Jurkat T cells , we found that p38 MAPK , but not Jun NH2-terminal kinase ( JNK ) , is weakly activated upon stimulation with either anti-CD3 or anti-CD28 in murine thymocytes and splenic T cells .

In contrast to that reported for <cell_type>human Jurkat T cells</cell_type> , we found that <protein>p38 MAPK</protein> , but not <protein>Jun NH2-terminal kinase</protein> ( <protein>JNK</protein> ) , is weakly activated upon stimulation with either <protein>anti-CD3</protein> or <protein>anti-CD28</protein> in <cell_type>murine thymocytes</cell_type> and <cell_type>splenic T cells</cell_type> .


However , p38 MAPK is activated strongly and synergistically by either CD3/CD28 coligation or PMA/Ca2+ ionophore stimulation , which mimics TCR - CD3/CD28 -mediated signaling .

However , <protein>p38 MAPK</protein> is activated strongly and synergistically by either <protein>CD3/CD28</protein> coligation or PMA/Ca2+ ionophore stimulation , which mimics <protein>TCR</protein> - <protein>CD3/CD28</protein> -mediated signaling .


Activation of p38 MAPK correlates closely with the stimulation of T cell proliferation .

Activation of <protein>p38 MAPK</protein> correlates closely with the stimulation of T cell proliferation .


In contrast , PMA-induced JNK activation is inhibited by Ca2+ ionophore .

In contrast , PMA-induced <protein>JNK</protein> activation is inhibited by Ca2+ ionophore .


T cell proliferation and production of IL-2 , IL-4 , and IFN-gamma induced by both CD3 and CD3 /CD28 ligation and the nuclear expression of the c-Jun and ATF-2 proteins are each blocked by the p38 MAPK inhibitor SB203580 .

T cell proliferation and production of <protein>IL-2</protein> , <protein>IL-4</protein> , and <protein>IFN-gamma</protein> induced by both <protein>CD3</protein> and <protein>CD3</protein> <protein>/CD28</protein> ligation and the nuclear expression of the <protein>c-Jun and ATF-2 proteins</protein> are each blocked by the <protein>p38 MAPK</protein> inhibitor SB203580 .


Our findings demonstrate that p38 MAPK 1 ) plays an important role in signal integration during costimulation of primary mouse T cells , 2 ) may be involved in the induction of c-Jun activation and augmentation of AP-1 transcriptional activity , and 3 ) regulates whether T cells enter a state of functional unresponsiveness .

Our findings demonstrate that <protein>p38 MAPK</protein> 1 ) plays an important role in signal integration during costimulation of <cell_type>primary mouse T cells</cell_type> , 2 ) may be involved in the induction of <protein>c-Jun</protein> activation and augmentation of <protein>AP-1</protein> transcriptional activity , and 3 ) regulates whether <cell_type>T cells</cell_type> enter a state of functional unresponsiveness .


Erythroid gene expression is differentially regulated by erythropoietin , haemin and delta-aminolaevulinic acid in UT-7 cells .

Erythroid gene expression is differentially regulated by <protein>erythropoietin</protein> , haemin and delta-aminolaevulinic acid in <cell_line>UT-7 cells</cell_line> .


Erythropoietin ( Epo ) is essential for the later stages of erythropoiesis , acting to promote cell survival and proliferation , but its role in differentiation remains to be defined .


The UT-7 cell line exhibits both erythroid and megakaryocytic characteristics and can be induced to differentiate along the erythroid pathway by Epo or the megakaryocytic pathway by phorbol myristic acetate .

The <cell_line>UT-7 cell line</cell_line> exhibits both erythroid and megakaryocytic characteristics and can be induced to differentiate along the erythroid pathway by <protein>Epo</protein> or the megakaryocytic pathway by phorbol myristic acetate .


We have compared the effects of Epo and the chemical inducers , delta-aminolaevulinic acid ( delta-ALA ) and haemin on the differentiation capacity of UT-7 cells .

We have compared the effects of <protein>Epo</protein> and the chemical inducers , delta-aminolaevulinic acid ( delta-ALA ) and haemin on the differentiation capacity of <cell_line>UT-7 cells</cell_line> .


Epo alone promoted relatively early events in erythroid maturation , without significant changes in haemoglobin production or morphology .

<protein>Epo</protein> alone promoted relatively early events in erythroid maturation , without significant changes in haemoglobin production or morphology .


GATA-2 and c-myb were down-regulated by Epo , and GATA-2 was further down-modulated by the inducers .

<protein>GATA-2</protein> and <protein>c-myb</protein> were down-regulated by <protein>Epo</protein> , and <protein>GATA-2</protein> was further down-modulated by the inducers .


Conversely , SCL expression was up-regulated by Epo and further increased by haemin and delta-ALA .

Conversely , <protein>SCL</protein> expression was up-regulated by <protein>Epo</protein> and further increased by haemin and delta-ALA .


Epo caused an increase in the proportion of cells expressing cell surface glycophorin A ( GPA ) and up-regulated beta- and gamma-globin by several fold .

<protein>Epo</protein> caused an increase in the proportion of cells expressing cell surface <protein>glycophorin A</protein> ( <protein>GPA</protein> ) and up-regulated <protein>beta- and gamma-globin</protein> by several fold .


Both haemin and delta-ALA caused a de novo increase in alpha-globin expression as well as enhancing Epo -induced beta-globin expression , leading to a marked increase in haemoglobin production .

Both haemin and delta-ALA caused a de novo increase in <protein>alpha-globin</protein> expression as well as enhancing <protein>Epo</protein> -induced <protein>beta-globin</protein> expression , leading to a marked increase in haemoglobin production .


These results suggest that haemoglobin production in UT-7 cells is limited by a deficiency of erythroid-specific aminolaevulinic acid synthase ( ALAS-E ) activity or globin synthesis as a consequence of their immaturity as a multipotential cell line .

These results suggest that haemoglobin production in <cell_line>UT-7 cells</cell_line> is limited by a deficiency of <protein>erythroid-specific aminolaevulinic acid synthase</protein> ( <protein>ALAS-E</protein> ) activity or globin synthesis as a consequence of their immaturity as a <cell_line>multipotential cell line</cell_line> .


Essential role of alveolar macrophages in intrapulmonary activation of NF-kappaB .

Essential role of <cell_type>alveolar macrophages</cell_type> in intrapulmonary activation of <protein>NF-kappaB</protein> .


Acute inflammatory injury in rat lung induced by deposition of immunoglobulin G immune complexes requires expression of cytokines and chemokines as well as activation of the transcription factor nuclear factor ( NF ) -kappaB .

Acute inflammatory injury in rat lung induced by deposition of <protein>immunoglobulin G immune complexes</protein> requires expression of <protein>cytokines</protein> and <protein>chemokines</protein> as well as activation of the <protein>transcription factor</protein> <protein>nuclear factor ( NF ) -kappaB</protein> .


There is little direct evidence regarding the role of alveolar macrophages in these activation events .

There is little direct evidence regarding the role of <cell_type>alveolar macrophages</cell_type> in these activation events .


In the present studies , rat lungs were depleted of alveolar macrophages by airway instillation of liposome-encapsulated dichloromethylene diphosphonate .

In the present studies , rat lungs were depleted of <cell_type>alveolar macrophages</cell_type> by airway instillation of liposome-encapsulated dichloromethylene diphosphonate .


These procedures , which greatly reduced the number of retrievable alveolar macrophages , suppressed activation of lung NF-kappaB in the inflammatory model .

These procedures , which greatly reduced the number of retrievable <cell_type>alveolar macrophages</cell_type> , suppressed activation of <protein>lung NF-kappaB</protein> in the inflammatory model .


In addition , bronchoalveolar lavage levels of tumor necrosis factor-alpha ( TNF-alpha ) and the CXC chemokine , macrophage inflammatory protein-2 , were substantially reduced .

In addition , bronchoalveolar lavage levels of <protein>tumor necrosis factor-alpha</protein> ( <protein>TNF-alpha</protein> ) and the <protein>CXC chemokine</protein> , <protein>macrophage inflammatory protein-2</protein> , were substantially reduced .


In parallel , upregulation of the lung vascular adhesion molecule , intercellular adhesion molecule-1 , was greatly reduced by intrapulmonary instillation of phosphonate-containing liposomes .

In parallel , upregulation of the <protein>lung vascular adhesion molecule</protein> , <protein>intercellular adhesion molecule-1</protein> , was greatly reduced by intrapulmonary instillation of phosphonate-containing liposomes .


Neutrophil accumulation and development of lung injury were also substantially diminished .


Lung instillation of TNF-alpha in alveolar macrophage-depleted rats restored the NF-kappaB activation response in whole lung .

Lung instillation of <protein>TNF-alpha</protein> in alveolar macrophage-depleted rats restored the <protein>NF-kappaB</protein> activation response in whole lung .


These data suggest that , in this inflammatory model , initial activation of NF-kappaB occurs in alveolar macrophages and the ensuing production of TNF-alpha may propagate NF-kappaB activation to other cell types in the lung .

These data suggest that , in this inflammatory model , initial activation of <protein>NF-kappaB</protein> occurs in <cell_type>alveolar macrophages</cell_type> and the ensuing production of <protein>TNF-alpha</protein> may propagate <protein>NF-kappaB</protein> activation to other cell types in the lung .


Interferon-beta mediates stromal cell rescue of T cells from apoptosis .

<protein>Interferon-beta</protein> mediates stromal cell rescue of <cell_type>T cells</cell_type> from apoptosis .


The resolution of immune responses is characterized by extensive apoptosis of activated T cells .

The resolution of immune responses is characterized by extensive apoptosis of activated <cell_type>T cells</cell_type> .


However , to generate and maintain immunological memory , some antigen-specific T cells must survive and revert to a resting G0/G1 state .

However , to generate and maintain immunological memory , some <cell_type>antigen-specific T cells</cell_type> must survive and revert to a resting G0/G1 state .


Cytokines that bind to the common gamma chain of the IL-2 receptor promote the survival of T cell blasts , but also induce proliferation .

<protein>Cytokines</protein> that bind to the common <protein>gamma chain</protein> of the <protein>IL-2</protein> receptor promote the survival of <cell_type>T cell blasts</cell_type> , but also induce proliferation .


In contrast , soluble factors secreted by stromal cells induce Tcell survival in a resting G0/G1 state .

In contrast , <protein>soluble factors</protein> secreted by <cell_type>stromal cells</cell_type> induce Tcell survival in a resting G0/G1 state .


We now report that interferon-beta is the principal mediator of stromal cell-mediated Tcell rescue from apoptosis .


Interferon-alpha and -beta promote the reversion of blast Tcells to a resting G0/G1 configuration with all the characteristic features of stromal cell rescue ; such as high Bcl-XL expression and low Bcl-2 .

<protein>Interferon-alpha and -beta</protein> promote the reversion of <cell_type>blast Tcells</cell_type> to a resting G0/G1 configuration with all the characteristic features of stromal cell rescue ; such as high <protein>Bcl-XL</protein> expression and low <protein>Bcl-2</protein> .


Type I interferons and stromal cells stimulate apparently identical signaling pathways , leading to STAT-1 activation .

<protein>Type I interferons</protein> and <cell_type>stromal cells</cell_type> stimulate apparently identical signaling pathways , leading to <protein>STAT-1</protein> activation .


We also show that this mechanism may play a fundamental role in the persistence of T cells at sites of chronic inflammation ; suggesting that chronic inflammation is an aberrant consequence of immunological memory .

We also show that this mechanism may play a fundamental role in the persistence of <cell_type>T cells</cell_type> at sites of chronic inflammation ; suggesting that chronic inflammation is an aberrant consequence of immunological memory .


Reduction of tumour necrosis factor alpha expression and signalling in peripheral blood mononuclear cells from patients with thalassaemia or sickle cell anaemia upon treatment with desferrioxamine .

Reduction of <protein>tumour necrosis factor alpha</protein> expression and signalling in <cell_type>peripheral blood mononuclear cells</cell_type> from patients with thalassaemia or sickle cell anaemia upon treatment with desferrioxamine .


Recent evidence indicates that the rate of progression of the HIV-1 disease is significantly reduced in thalassaemia major patients upon treatment with high doses of desferrioxamine ( DFX ) .


The authors have previously demonstrated that in vitro exposure of mononuclear cells to DFX decreases the bioavailability of tumour necrosis factor alpha ( TNF-alpha ) which has a stimulatory effect on HIV-1 replication .

The authors have previously demonstrated that in vitro exposure of <cell_type>mononuclear cells</cell_type> to DFX decreases the bioavailability of <protein>tumour necrosis factor alpha</protein> ( <protein>TNF-alpha</protein> ) which has a stimulatory effect on HIV-1 replication .


In this study , therefore , TNF-alpha bioavailability from mononuclear cells isolated from 10 patients with thalassaemia or sickle cell anaemia given DFX as compared to 10 untreated subjects has been evaluated .

In this study , therefore , <protein>TNF-alpha</protein> bioavailability from <cell_type>mononuclear cells</cell_type> isolated from 10 patients with thalassaemia or sickle cell anaemia given DFX as compared to 10 untreated subjects has been evaluated .


Evidence is presented showing that DFX treatment reduces TNF-alpha bioavailability ( P < 0.05 ) by inhibiting its steady state ( P < 0.05 ) and by enhancing its inactivation through binding to soluble TNF-alpha receptor type II ( P < 0.05 ) .

Evidence is presented showing that DFX treatment reduces <protein>TNF-alpha</protein> bioavailability ( P < 0.05 ) by inhibiting its steady state ( P < 0.05 ) and by enhancing its inactivation through binding to soluble <protein>TNF-alpha</protein> receptor type II ( P < 0.05 ) .


We also show that DFX treatment limits the in vivo activation of NF-kappaB , a transcription factor involved in both TNF-alpha gene transcription and TNF-alpha signalling ( P < 0.005 ) .

We also show that DFX treatment limits the in vivo activation of <protein>NF-kappaB</protein> , a <protein>transcription factor</protein> involved in both <protein>TNF-alpha</protein> gene transcription and <protein>TNF-alpha</protein> signalling ( P < 0.005 ) .


We conclude that TNF-alpha bioavailability and signalling are impaired in patients upon DFX treatment .

We conclude that <protein>TNF-alpha</protein> bioavailability and signalling are impaired in patients upon DFX treatment .


This mechanism may contribute to delayed progression of the HIV-1 infection in vivo .


Copyright 1999 Academic Press .


Involvement of NF-kappaB p50/p65 heterodimer in activation of the human pro-interleukin-1beta gene at two subregions of the upstream enhancer element .

Involvement of <protein>NF-kappaB p50/p65 heterodimer</protein> in activation of the <dna>human pro-interleukin-1beta gene</dna> at two subregions of the <dna>upstream enhancer element</dna> .


A region between-3134 and -2729 bp upstream from the transcription site of the human pro-interleukin 1beta ( proIL-1beta ) gene was identified as an LPS-responsive enhancer element .

A region between-3134 and -2729 bp upstream from the <dna>transcription site</dna> of the <dna>human pro-interleukin 1beta ( proIL-1beta ) gene</dna> was identified as an <dna>LPS-responsive enhancer element</dna> .


In this study , the influence of the sequences located between -3134 and -2987 on the transcriptional activity of the proIL-1beta gene in LPS-stimulated Raw 264.7 cells was examined in detail .

In this study , the influence of the sequences located <dna>between -3134 and -2987</dna> on the transcriptional activity of the <dna>proIL-1beta gene</dna> in <cell_line>LPS-stimulated Raw 264.7 cells</cell_line> was examined in detail .


The results obtained by transient transfection of fos -CAT constructs that contained serial 5'-deletion mutations showed that the region between -3134 and -3059 appears to be required for the induction of transcription by LPS .

The results obtained by transient transfection of fos <dna>-CAT constructs</dna> that contained <dna>serial 5'-deletion mutations</dna> showed that the region between -3134 and -3059 appears to be required for the induction of transcription by LPS .


Gel shift assay studies with synthetic oligonucleotides corresponding to partial sequences of the latter region and nuclear extracts from stimulated cells revealed specific protein binding sites between -3110 and -3090 and between -3079 and -3059 .

Gel shift assay studies with <dna>synthetic oligonucleotides</dna> corresponding to partial sequences of the latter region and nuclear extracts from stimulated cells revealed specific <dna>protein binding sites</dna> between -3110 and -3090 and between -3079 and -3059 .


These specific bindings were time and LPS dose dependent .


The results of supershift analysis using specific antibodies against transcription factors suggested that both binding complexes contained the NF-kappaB components p50 and p65 , and did not contain other NF-kappaB proteins ( p52 , c-Rel , Rel B ) , AP-1 proteins ( c-Fos , C-Jun ) , CREB or C/EBPbeta ( NF-IL6 ) .

The results of supershift analysis using specific antibodies against <protein>transcription factors</protein> suggested that both binding complexes contained the <protein>NF-kappaB components</protein> <protein>p50</protein> and <protein>p65</protein> , and did not contain other <protein>NF-kappaB proteins</protein> ( <protein>p52</protein> , <protein>c-Rel</protein> , <protein>Rel B</protein> ) , <protein>AP-1 proteins</protein> ( <protein>c-Fos</protein> , <protein>C-Jun</protein> ) , <protein>CREB</protein> or <protein>C/EBPbeta</protein> ( <protein>NF-IL6</protein> ) .


Mutation of either of the putative NF-kappaB-binding sites in the enhancer element decreased the LPS-stimulated transcriptional activity .

Mutation of either of the putative <dna>NF-kappaB-binding sites</dna> in the enhancer element decreased the LPS-stimulated transcriptional activity .


These data indicated that two NF-kappaB-binding sites , which are located between -3134 and -3059 , are critical for the activation of proIL-1beta gene transcription .

These data indicated that two <dna>NF-kappaB-binding sites</dna> , which are located between <dna>-3134 and -3059</dna> , are critical for the activation of <dna>proIL-1beta gene</dna> transcription .


N-acetyl-L-cysteine inhibits primary human T cell responses at the dendritic cell level : association with NF-kappaB inhibition .

N-acetyl-L-cysteine inhibits primary human T cell responses at the dendritic cell level : association with <protein>NF-kappaB</protein> inhibition .


N-acetyl-L-cysteine ( NAC ) is an antioxidant molecule endowed with immunomodulatory properties .


To investigate the effect of NAC on the induction phase of T cell responses , we analyzed its action on human dendritic cells ( DC ) derived from adherent PBMC cultured with IL-4 and granulocyte-macrophage CSF .

To investigate the effect of NAC on the induction phase of T cell responses , we analyzed its action on human dendritic cells ( DC ) derived from adherent PBMC cultured with <protein>IL-4</protein> and <protein>granulocyte-macrophage CSF</protein> .


We first found that NAC inhibited the constitutive as well as the LPS-induced activity of the transcription factor NF-kappaB .

We first found that NAC inhibited the constitutive as well as the LPS-induced activity of the <protein>transcription factor</protein> <protein>NF-kappaB</protein> .


In parallel , NAC was shown to down-regulate the production of cytokines by DC as well as their surface expression of HLA-DR , CD86 ( B7-2 ) , and CD40 molecules both at the basal state and upon LPS activation .

In parallel , NAC was shown to down-regulate the production of <protein>cytokines</protein> by <cell_type>DC</cell_type> as well as their surface expression of <protein>HLA-DR</protein> , <protein>CD86</protein> ( <protein>B7-2</protein> ) , and <protein>CD40 molecules</protein> both at the basal state and upon LPS activation .


NAC also inhibited DC responses induced by CD40 engagement .

NAC also inhibited DC responses induced by <protein>CD40</protein> engagement .


The inhibitory effects of NAC were not due to nonspecific toxicity as neither the viability of DC nor their mannose receptor-mediated endocytosis were modified by NAC .

The inhibitory effects of NAC were not due to nonspecific toxicity as neither the viability of <cell_type>DC</cell_type> nor their mannose receptor-mediated endocytosis were modified by NAC .


Finally , we found that the addition of NAC to MLR between naive T cells and allogeneic DC resulted in a profound inhibition of alloreactive responses , which could be attributed to a defect of DC as APC-independent T cell responses were not inhibited by NAC .

Finally , we found that the addition of NAC to MLR between <cell_type>naive T cells</cell_type> and allogeneic <cell_type>DC</cell_type> resulted in a profound inhibition of alloreactive responses , which could be attributed to a defect of <cell_type>DC</cell_type> as APC-independent T cell responses were not inhibited by NAC .


Altogether , our results suggest that NAC might impair the generation of primary immune responses in humans through its inhibitory action on DC .

Altogether , our results suggest that NAC might impair the generation of primary immune responses in humans through its inhibitory action on <cell_type>DC</cell_type> .


Thrombin-induced p65 homodimer binding to downstream NF-kappa B site of the promoter mediates endothelial ICAM-1 expression and neutrophil adhesion .

<protein>Thrombin-induced p65 homodimer</protein> binding to downstream <dna>NF-kappa B site</dna> of the promoter mediates endothelial <protein>ICAM-1</protein> expression and <cell_type>neutrophil</cell_type> adhesion .


We investigated the mechanisms by which proinflammatory mediator , thrombin , released during intravascular coagulation and tissue injury , induces ICAM-1 ( CD54 ) expression in endothelial cells .

We investigated the mechanisms by which <protein>proinflammatory mediator</protein> , <protein>thrombin</protein> , released during intravascular coagulation and tissue injury , induces <protein>ICAM-1</protein> ( <protein>CD54</protein> ) expression in <cell_type>endothelial cells</cell_type> .


Stimulation of HUVEC with thrombin resulted in dose- and time-dependent increases in ICAM-1 mRNA and cell surface expression and in ICAM-1 -dependent endothelial adhesivity toward polymorphonuclear leukocytes .

Stimulation of <cell_line>HUVEC</cell_line> with <protein>thrombin</protein> resulted in dose- and time-dependent increases in <rna>ICAM-1 mRNA</rna> and cell surface expression and in <protein>ICAM-1</protein> -dependent endothelial adhesivity toward <cell_type>polymorphonuclear leukocytes</cell_type> .


Transient transfection of endothelial cells with ICAM-1 promoter luciferase reporter gene ( ICAM-1LUC ) constructs indicated that deletion of upstream NF-kappa B site ( -533 bases from translation start site ) had no effect on thrombin responsiveness , whereas mutation/deletion of downstream NF-kappa B site ( -223 bases from the translation start site ) prevented the activation of ICAM-1 promoter , indicating that the downstream NF-kappa B site is critical for thrombin inducibility .

Transient transfection of <cell_type>endothelial cells</cell_type> with <dna>ICAM-1 promoter luciferase reporter gene ( ICAM-1LUC ) constructs</dna> indicated that deletion of upstream <dna>NF-kappa B site</dna> ( <dna>-533 bases</dna> from <dna>translation start site</dna> ) had no effect on <protein>thrombin</protein> responsiveness , whereas mutation/deletion of downstream <dna>NF-kappa B site</dna> ( <dna>-223 bases</dna> from the <dna>translation start site</dna> ) prevented the activation of <dna>ICAM-1 promoter</dna> , indicating that the downstream <dna>NF-kappa B site</dna> is critical for <protein>thrombin</protein> inducibility .


NF-kappa B -directed luciferase activity increased approximately 3-fold when cells transfected with the plasmid pNF-kappa BLUC containing five copies of consensus NF-kappa B site linked to a minimal adenovirus E1B promoter-luciferase gene were exposed to thrombin , indicating that activation of NF-kappa B was essential for thrombin response .

<protein>NF-kappa B</protein> -directed <protein>luciferase</protein> activity increased approximately 3-fold when cells transfected with the <dna>plasmid pNF-kappa BLUC</dna> containing five copies of <dna>consensus NF-kappa B site</dna> linked to a minimal <dna>adenovirus E1B promoter-luciferase gene</dna> were exposed to <protein>thrombin</protein> , indicating that activation of <protein>NF-kappa B</protein> was essential for <protein>thrombin</protein> response .


Gel supershift assays demonstrated that thrombin induced binding of NF-kappa Bp65 ( Rel A ) to downstream NF-kappa B site of the ICAM-1 promoter .

Gel supershift assays demonstrated that <protein>thrombin</protein> induced binding of <protein>NF-kappa Bp65</protein> ( <protein>Rel A</protein> ) to downstream <dna>NF-kappa B site</dna> of the <dna>ICAM-1 promoter</dna> .


Thrombin receptor activation peptide , a 14-amino-acid peptide representing the new NH2 terminus of proteolytically activated receptor-1 , mimicked thrombin 's action in inducing ICAM-1 expression .

<protein>Thrombin</protein> receptor activation peptide , a 14-amino-acid peptide representing the new <protein>NH2 terminus</protein> of proteolytically activated <protein>receptor-1</protein> , mimicked <protein>thrombin</protein> 's action in inducing <protein>ICAM-1</protein> expression .


These data indicate that thrombin activates endothelial ICAM-1 expression and polymorphonuclear leukocyte adhesion by NF-kappa Bp65 binding to the downstream NF-kappa B site of ICAM-1 promoter after proteolytically activated receptor-1 activation .

These data indicate that <protein>thrombin</protein> activates endothelial <protein>ICAM-1</protein> expression and <cell_type>polymorphonuclear leukocyte</cell_type> adhesion by <protein>NF-kappa Bp65</protein> binding to the downstream <dna>NF-kappa B site</dna> of <dna>ICAM-1 promoter</dna> after proteolytically activated receptor-1 activation .


Differential inhibition of Smad6 and Smad7 on bone morphogenetic protein- and activin-mediated growth arrest and apoptosis in B cells .

Differential inhibition of <protein>Smad6</protein> and <protein>Smad7</protein> on bone morphogenetic protein- and activin-mediated growth arrest and apoptosis in <cell_type>B cells</cell_type> .


Smad6 and Smad7 prevent ligand-induced activation of signal-transducing Smad proteins in the transforming growth factor-beta family .

<protein>Smad6</protein> and <protein>Smad7</protein> prevent ligand-induced activation of <protein>signal-transducing Smad proteins</protein> in the <protein>transforming growth factor-beta family</protein> .