This region contains 4 Sp1 elements and an IFN-gamma activated sequence ( GAS ) that overlaps with the second Sp1 element .

This region contains 4 <dna>Sp1 elements</dna> and an <dna>IFN-gamma activated sequence</dna> ( <dna>GAS</dna> ) that overlaps with the second <dna>Sp1 element</dna> .


In the monocytic cell line THP-1 cell , the combination of IFN-gamma and all-trans-retinoic acid ( a known differentiation agent ) enhances the ACAT-1 P1 promoter but not the P7 promoter .

In the <cell_type>monocytic cell line THP-1 cell</cell_type> , the combination of <protein>IFN-gamma</protein> and all-trans-retinoic acid ( a known differentiation agent ) enhances the <dna>ACAT-1 P1 promoter</dna> but not the <dna>P7 promoter</dna> .


Additional experiments showed that all-trans-retinoic acid causes large induction of the transcription factor STAT1 , while IFN-gamma causes activation of STAT1 such that it binds to the GAS/Sp1 site in the ACAT-1 P1 promoter .

Additional experiments showed that all-trans-retinoic acid causes large induction of the <protein>transcription factor STAT1</protein> , while <protein>IFN-gamma</protein> causes activation of <protein>STAT1</protein> such that it binds to the <dna>GAS/Sp1 site</dna> in the <dna>ACAT-1 P1 promoter</dna> .


Our work provides a molecular mechanism to account for the effect of IFN-gamma in causing transcriptional activation of ACAT-1 in macrophage-like cells .

Our work provides a molecular mechanism to account for the effect of <protein>IFN-gamma</protein> in causing transcriptional activation of <protein>ACAT-1</protein> in <cell_type>macrophage-like cells</cell_type> .


Inhaled nitric oxide down-regulates intrapulmonary nitric oxide production in lipopolysaccharide-induced acute lung injury .


OBJECTIVE : To examine whether inhaled nitric oxide ( NO ) affected the intrapulmonary production of NO , reactive oxygen species , and nuclear factor-kappaB in a lipopolysaccharide ( LPS ) -induced model of acute lung injury .

OBJECTIVE : To examine whether inhaled nitric oxide ( NO ) affected the intrapulmonary production of NO , reactive oxygen species , and <protein>nuclear factor-kappaB</protein> in a lipopolysaccharide ( LPS ) -induced model of acute lung injury .


DESIGN : Prospective , randomized , laboratory study .


SETTING : Experimental laboratory at a biomedical institute .


SUBJECTS : Twenty male rabbits weighing 2.5-3.5 kg .


INTERVENTIONS : Saline or LPS ( 5 mg/kg of body weight ) was administered intravenously with or without NO inhalation ( 10 ppm ) in each group of five rabbits .


MEASUREMENTS AND MAIN RESULTS : LPS increased the lung leak index , the neutrophils and NO levels in bronchoalveolar lavage fluid , and NO levels produced by resting and stimulated alveolar macrophages .

MEASUREMENTS AND MAIN RESULTS : LPS increased the lung leak index , the <cell_type>neutrophils</cell_type> and NO levels in bronchoalveolar lavage fluid , and NO levels produced by resting and stimulated <cell_type>alveolar macrophages</cell_type> .


Inhaled NO decreased the lung leak index , the neutrophils and NO levels as measured by nitrite levels in the lavage fluid , and NO produced by the resting and stimulated alveolar macrophages .

Inhaled NO decreased the lung leak index , the <cell_type>neutrophils</cell_type> and NO levels as measured by nitrite levels in the lavage fluid , and NO produced by the resting and stimulated <cell_type>alveolar macrophages</cell_type> .


Inhaled NO also blocked the activities of reactive oxygen species and nuclear factor-kappaB binding to DNA in lavage cells and in alveolar macrophages .

Inhaled NO also blocked the activities of reactive oxygen species and <protein>nuclear factor-kappaB</protein> binding to DNA in <cell_type>lavage cells</cell_type> and in <cell_type>alveolar macrophages</cell_type> .


CONCLUSION : Inhaled NO attenuates LPS-induced acute lung injury , possibly by decreasing NO production in the lungs .


The mechanism of reducing NO production resulting from inhaled NO may involve , in part , the activities of reactive oxygen species and/or nuclear factor-kappaB .

The mechanism of reducing NO production resulting from inhaled NO may involve , in part , the activities of reactive oxygen species and/or <protein>nuclear factor-kappaB</protein> .


Treatment of allergic airway inflammation and hyperresponsiveness by antisense-induced local blockade of GATA-3 expression .

Treatment of allergic airway inflammation and hyperresponsiveness by antisense-induced local blockade of <dna>GATA-3</dna> expression .


Recent studies in transgenic mice have revealed that expression of a dominant negative form of the transcription factor GATA-3 in T cells can prevent T helper cell type 2 ( Th2 ) -mediated allergic airway inflammation in mice .

Recent studies in transgenic mice have revealed that expression of a dominant negative form of the <protein>transcription factor GATA-3</protein> in <cell_type>T cells</cell_type> can prevent <cell_type>T helper cell type 2</cell_type> ( <cell_type>Th2</cell_type> ) -mediated allergic airway inflammation in mice .


However , it remains unclear whether GATA-3 plays a role in the effector phase of allergic airway inflammation and whether antagonizing the expression and/or function of GATA-3 can be used for the therapy of allergic airway inflammation and hyperresponsiveness .

However , it remains unclear whether <protein>GATA-3</protein> plays a role in the effector phase of allergic airway inflammation and whether antagonizing the expression and/or function of <dna>GATA-3</dna> can be used for the therapy of allergic airway inflammation and hyperresponsiveness .


Here , we analyzed the effects of locally antagonizing GATA-3 function in a murine model of asthma .

Here , we analyzed the effects of locally antagonizing <protein>GATA-3</protein> function in a murine model of asthma .


We could suppress GATA-3 expression in interleukin ( IL ) -4 -producing T cells in vitro and in vivo by an antisense phosphorothioate oligonucleotide overlapping the translation start site of GATA-3 , whereas nonsense control oligonucleotides were virtually inactive .

We could suppress GATA-3 expression in <protein>interleukin ( IL ) -4</protein> -producing <cell_type>T cells</cell_type> in vitro and in vivo by an antisense phosphorothioate oligonucleotide overlapping the <dna>translation start site</dna> of <dna>GATA-3</dna> , whereas nonsense control oligonucleotides were virtually inactive .


In a murine model of asthma associated with allergic pulmonary inflammation and hyperresponsiveness in ovalbumin ( OVA ) -sensitized mice , local intranasal administration of fluorescein isothiocyanate-labeled GATA-3 antisense oligonucleotides led to DNA uptake in lung cells associated with a reduction of intracellular GATA-3 expression .

In a murine model of asthma associated with allergic pulmonary inflammation and hyperresponsiveness in ovalbumin ( OVA ) -sensitized mice , local intranasal administration of fluorescein isothiocyanate-labeled GATA-3 antisense oligonucleotides led to DNA uptake in <cell_type>lung cells</cell_type> associated with a reduction of intracellular GATA-3 expression .


Such intrapulmonary blockade of GATA-3 expression caused an abrogation of signs of lung inflammation including infiltration of eosinophils and Th2 cytokine production .

Such intrapulmonary blockade of GATA-3 expression caused an abrogation of signs of lung inflammation including infiltration of <cell_type>eosinophils</cell_type> and <cell_type>Th2</cell_type> cytokine production .


Furthermore , treatment with antisense but not nonsense oligonucleotides induced a significant reduction of airway hyperresponsiveness in OVA-sensitized mice to levels comparable to saline-treated control mice , as assessed by both enhanced pause ( PenH ) responses and pulmonary resistance determined by body plethysmography .


These data indicate a critical role for GATA-3 in the effector phase of a murine asthma model and suggest that local delivery of GATA-3 antisense oligonucleotides may be a novel approach for the treatment of airway hyperresponsiveness such as in asthma .

These data indicate a critical role for <dna>GATA-3</dna> in the effector phase of a murine asthma model and suggest that local delivery of GATA-3 antisense oligonucleotides may be a novel approach for the treatment of airway hyperresponsiveness such as in asthma .


This approach has the potential advantage of suppressing the expression of various proinflammatory Th2 cytokines simultaneously rather than suppressing the activity of a single cytokine .

This approach has the potential advantage of suppressing the expression of various <protein>proinflammatory Th2 cytokines</protein> simultaneously rather than suppressing the activity of a single <protein>cytokine</protein> .


T helper-cell phenotype regulates atherosclerosis in mice under conditions of mild hypercholesterolemia .


BACKGROUND : T cells are implicated in atherosclerosis , but little is known about the genetic control or molecular pathways , especially under conditions of mild hypercholesterolemia .

BACKGROUND : <cell_type>T cells</cell_type> are implicated in atherosclerosis , but little is known about the genetic control or molecular pathways , especially under conditions of mild hypercholesterolemia .


METHODS AND RESULTS : BALB/c mice , making a CD4+ Th2 ( IL-4+ ) cell response , express both MHC class II antigens ( IA ( d ) , IE ( d ) ) and are atherosclerosis-resistant .

METHODS AND RESULTS : BALB/c mice , making a CD4+ Th2 ( IL-4+ ) cell response , express both <protein>MHC class II antigens</protein> ( <protein>IA ( d )</protein> , <protein>IE ( d )</protein> ) and are atherosclerosis-resistant .


C57Bl/6 mice produce a CD4+ Th1 ( interferon [ IFN ] gamma+ ) response , express IA ( b ) but no IE , and are atherosclerosis-prone .

C57Bl/6 mice produce a CD4+ Th1 ( interferon [ IFN ] gamma+ ) response , express <protein>IA ( b )</protein> but no <protein>IE</protein> , and are atherosclerosis-prone .


To evaluate T helper-cell phenotype in fatty streak formation , wild-type C57Bl/6 mice ( IA ( b ) +IE- ) and transgenic mice , either AB ( o ) , IA ( b ) -IE- ; ABEalpha , IA-IE ( k ) + ; or BL : TG : Ealpha , IA ( b ) +IE ( k ) + , were fed a high-cholesterol diet for 16 weeks and evaluated histomorphometrically for aortic lesions .


Lesion size in AB ( o ) , ABEalpha , and BL : TG : Ealpha strains was decreased by 54 % , 79 % , and 82 % , respectively , compared with wild-type , correlating with decreased Th1 and increased Th2 expression and suggesting that T helper-cell phenotype is important in fatty lesion development .

Lesion size in AB ( o ) , ABEalpha , and BL : TG : Ealpha strains was decreased by 54 % , 79 % , and 82 % , respectively , compared with wild-type , correlating with decreased <cell_type>Th1</cell_type> and increased <cell_type>Th2</cell_type> expression and suggesting that T helper-cell phenotype is important in fatty lesion development .


Decreasing Th1 cells by antibodies ( alpha-CD4 ) or cytokines ( IL-4 ) also caused > /=80 % reductions in lesion size .

Decreasing <cell_type>Th1 cells</cell_type> by <protein>antibodies</protein> ( <protein>alpha-CD4</protein> ) or <protein>cytokines</protein> ( <protein>IL-4</protein> ) also caused > /=80 % reductions in lesion size .


Immunohistology revealed IFN-gamma , but not IL-4 , colocalized with activated macrophages .

Immunohistology revealed <protein>IFN-gamma</protein> , but not <protein>IL-4</protein> , colocalized with activated <cell_type>macrophages</cell_type> .


Confirming these findings in a different mouse strain , BALB/c Stat 6 knockout mice ( Th2 cell-deficient ) developed aortic lesions comparable to C57Bl/6 mice on the same diet .


CONCLUSIONS : In mildly hypercholesterolemic C57Bl/6 mice , presence of IA ( b ) and absence of IE regulated CD4+ T helper-cell phenotype ; fatty lesions were proportional to IFNgamma+ Th1 cells in both C57Bl/6 and BALB/c strains .

CONCLUSIONS : In mildly hypercholesterolemic C57Bl/6 mice , presence of <protein>IA ( b )</protein> and absence of <protein>IE</protein> regulated CD4+ T helper-cell phenotype ; fatty lesions were proportional to <cell_type>IFNgamma+ Th1 cells</cell_type> in both C57Bl/6 and BALB/c strains .


IFN-gamma may participate through macrophage activation , whereas IL-4 may act to limit Th1-cell response .

<protein>IFN-gamma</protein> may participate through macrophage activation , whereas <protein>IL-4</protein> may act to limit Th1-cell response .


Requirement for p38 and p44/p42 mitogen-activated protein kinases in RAGE -mediated nuclear factor-kappaB transcriptional activation and cytokine secretion .

Requirement for <protein>p38</protein> and <protein>p44/p42</protein> <protein>mitogen-activated protein kinases</protein> in <protein>RAGE</protein> -mediated <protein>nuclear factor-kappaB</protein> transcriptional activation and cytokine secretion .


Advanced glycation end product ( AGE ) activation of the signal-transducing receptor for AGE ( RAGE ) has been linked to a proinflammatory phenotypic change within cells .

Advanced glycation end product ( AGE ) activation of the <protein>signal-transducing receptor for AGE</protein> ( <protein>RAGE</protein> ) has been linked to a proinflammatory phenotypic change within cells .


However , the precise intracellular signaling pathways involved have not been elucidated .


We demonstrate here that human serum albumin modified with N ( varepsilon ) - ( carboxymethyl ) lysine ( CML ) , a major AGE adduct that progressively accumulates with aging , diabetes , and renal failure , induced nuclear factor ( NF ) -kappaB -driven reporter gene expression in human monocytic THP-1 cells .

We demonstrate here that human serum albumin modified with N ( varepsilon ) - ( carboxymethyl ) lysine ( CML ) , a major AGE adduct that progressively accumulates with aging , diabetes , and renal failure , induced <protein>nuclear factor ( NF ) -kappaB</protein> -driven reporter gene expression in <cell_type>human monocytic THP-1 cells</cell_type> .


The NF-kappaB response was blocked with a synthetic peptide corresponding to the putative ligand-binding domain of RAGE , with anti- RAGE antiserum , and by coexpression of truncated receptors lacking the intracellular domain .

The NF-kappaB response was blocked with a synthetic peptide corresponding to the <protein>putative ligand-binding domain</protein> of <protein>RAGE</protein> , with anti- <protein>RAGE</protein> antiserum , and by coexpression of <protein>truncated receptors</protein> lacking the <protein>intracellular domain</protein> .


Signal transduction from RAGE to NF-kappaB involved the generation of reactive oxygen species , since reporter gene expression was blocked with the antioxidant N-acetyl-L-cysteine .

Signal transduction from <protein>RAGE</protein> to <protein>NF-kappaB</protein> involved the generation of reactive oxygen species , since reporter gene expression was blocked with the antioxidant N-acetyl-L-cysteine .


CML-modified albumin produced rapid transient activation of tyrosine phosphorylation , extracellular signal-regulated kinase 1 and 2 , and p38 mitogen-activated protein kinase ( MAPK ) , but not c-Jun NH ( 2 ) -terminal kinase .

<protein>CML-modified albumin</protein> produced rapid transient activation of tyrosine phosphorylation , <protein>extracellular signal-regulated kinase 1 and 2</protein> , and <protein>p38 mitogen-activated protein kinase</protein> ( <protein>MAPK</protein> ) , but not <protein>c-Jun NH ( 2 ) -terminal kinase</protein> .


RAGE -mediated NF-kappaB activation was suppressed by the selective p38 MAPK inhibitor SB203580 and by coexpression of a kinase-dead p38 dominant-negative mutant .

<protein>RAGE</protein> -mediated NF-kappaB activation was suppressed by the selective <protein>p38 MAPK</protein> inhibitor SB203580 and by coexpression of a kinase-dead p38 dominant-negative mutant .


Activation of NF-kappaB by CML-modified albumin increased secretion of proinflammatory cytokines ( tumor necrosis factor-alpha , interleukin-1beta , and monocyte chemoattractant protein-1 ) severalfold , and inhibition of p38 MAPK blocked these increases .

Activation of <protein>NF-kappaB</protein> by <protein>CML-modified albumin</protein> increased secretion of <protein>proinflammatory cytokines</protein> ( <protein>tumor necrosis factor-alpha</protein> , <protein>interleukin-1beta</protein> , and <protein>monocyte chemoattractant protein-1</protein> ) severalfold , and inhibition of <protein>p38 MAPK</protein> blocked these increases .


These results indicate that p38 MAPK activation mediates RAGE -induced NF-kappaB -dependent secretion of proinflammatory cytokines and suggest that accelerated inflammation may be a consequence of cellular activation induced by this receptor .

These results indicate that <protein>p38 MAPK</protein> activation mediates <protein>RAGE</protein> -induced <protein>NF-kappaB</protein> -dependent secretion of <protein>proinflammatory cytokines</protein> and suggest that accelerated inflammation may be a consequence of cellular activation induced by this receptor .


Antigen-receptor cross-linking and lipopolysaccharide trigger distinct phosphoinositide 3-kinase -dependent pathways to NF-kappa B activation in primary B cells .

Antigen-receptor cross-linking and lipopolysaccharide trigger distinct <protein>phosphoinositide 3-kinase</protein> -dependent pathways to NF-kappa B activation in <cell_type>primary B cells</cell_type> .


The NF-kappaB/Rel transcription factors play an important role in the expression of genes involved in B cell development , differentiation and function .

The <protein>NF-kappaB/Rel transcription factors</protein> play an important role in the expression of genes involved in B cell development , differentiation and function .


Nuclear NF-kappaB is induced in B cells by engagement of either the BCR or CD40 or by stimulation with lipopolysaccharide ( LPS ) .

<protein>Nuclear NF-kappaB</protein> is induced in <cell_type>B cells</cell_type> by engagement of either the <protein>BCR</protein> or <protein>CD40</protein> or by stimulation with lipopolysaccharide ( LPS ) .


Despite the importance of NF-kappaB to B cell function , little is known about the signaling pathways leading to NF-kappaB activation .

Despite the importance of <protein>NF-kappaB</protein> to B cell function , little is known about the signaling pathways leading to NF-kappaB activation .


In this report we address the role of phosphoinositide 3'-kinase ( PI 3-kinase ) in BCR - and LPS-induced NF-kappaB activation using populations of primary murine resting B cells .

In this report we address the role of <protein>phosphoinositide 3'-kinase</protein> ( <protein>PI 3-kinase</protein> ) in <protein>BCR</protein> - and LPS-induced NF-kappaB activation using populations of <cell_type>primary murine resting B cells</cell_type> .


Using the specific pharmacological inhibitors of PI 3-kinase , Wortmannin and LY294002 , we demonstrate that PI 3-kinase activity is vital for BCR -induced NF-kappaB DNA-binding activity .

Using the specific pharmacological inhibitors of <protein>PI 3-kinase</protein> , Wortmannin and LY294002 , we demonstrate that <protein>PI 3-kinase</protein> activity is vital for <protein>BCR</protein> -induced <protein>NF-kappaB</protein> DNA-binding activity .


Furthermore , we show that this is achieved via protein kinase C -dependent degradation of IkappaBalpha .

Furthermore , we show that this is achieved via <protein>protein kinase C</protein> -dependent degradation of <protein>IkappaBalpha</protein> .


Similar analyses reveal that PI 3-kinase is also critical in triggering NF-kappaB DNA-binding activity and IkappaBalpha degradation following LPS stimulation .

Similar analyses reveal that <protein>PI 3-kinase</protein> is also critical in triggering <protein>NF-kappaB</protein> DNA-binding activity and <protein>IkappaBalpha</protein> degradation following LPS stimulation .


Interestingly , a PKC inhibitor which blocked the BCR -induced IkappaBalpha degradation had no effect on the degradation of IkappaBalpha after LPS stimulation .

Interestingly , a PKC inhibitor which blocked the <protein>BCR</protein> -induced <protein>IkappaBalpha</protein> degradation had no effect on the degradation of <protein>IkappaBalpha</protein> after LPS stimulation .


Taken together , our results indicate the involvement of PI 3-kinase in at least two distinct signaling pathways leading to activation of NF-kappaB in B cells .

Taken together , our results indicate the involvement of <protein>PI 3-kinase</protein> in at least two distinct signaling pathways leading to activation of <protein>NF-kappaB</protein> in <cell_type>B cells</cell_type> .


Tetramer-guided epitope mapping : rapid identification and characterization of immunodominant CD4+ T cell epitopes from complex antigens .

Tetramer-guided epitope mapping : rapid identification and characterization of <cell_type>immunodominant CD4+ T cell epitopes</cell_type> from complex <protein>antigens</protein> .


T cell responses to Ags involve recognition of selected peptide epitopes contained within the antigenic protein .

T cell responses to <protein>Ags</protein> involve recognition of selected <protein>peptide epitopes</protein> contained within the <protein>antigenic protein</protein> .


In this report , we describe a new approach for direct identification of CD4+ T cell epitopes of complex Ags that uses human class II tetramers to identify reactive cells .

In this report , we describe a new approach for direct identification of <cell_type>CD4+ T cell epitopes</cell_type> of complex <protein>Ags</protein> that uses <protein>human class II tetramers</protein> to identify reactive cells .


With a panel of 60 overlapping peptides covering the entire sequence of the VP16 protein , a major Ag for HSV-2 , we generated a panel of class II MHC tetramers loaded with peptide pools that were used to stain peripheral lymphocytes of an HSV-2 infected individual .

With a panel of 60 overlapping peptides covering the entire sequence of the <protein>VP16 protein</protein> , a major <protein>Ag</protein> for HSV-2 , we generated a panel of <protein>class II MHC tetramers</protein> loaded with peptide pools that were used to stain <cell_type>peripheral lymphocytes</cell_type> of an HSV-2 infected individual .


With this approach , we identified four new DRA1*0101/DRB1*0401- and two DRA1*0101/DRB1*0404-restricted , VP16-specific epitopes .

With this approach , we identified four new DRA1*0101/DRB1*0401- and two DRA1*0101/DRB1*0404-restricted , <protein>VP16-specific epitopes</protein> .


By using tetramers to sort individual cells , we easily obtained a large number of clones specific to these epitopes .

By using <protein>tetramers</protein> to sort individual cells , we easily obtained a large number of clones specific to these <protein>epitopes</protein> .


Although DRA1*0101/DRB1*0401 and DRA1*0101/DRB1*0404 are structurally very similar , nonoverlapping VP16 epitopes were identified , illustrating high selectivity of individual allele polymorphisms within common MHC variants .

Although DRA1*0101/DRB1*0401 and DRA1*0101/DRB1*0404 are structurally very similar , nonoverlapping <protein>VP16 epitopes</protein> were identified , illustrating high selectivity of individual allele polymorphisms within common MHC variants .


This rapid approach to detecting CD4+ T cell epitopes from complex Ags can be applied to any known Ag that gives a T cell response .

This rapid approach to detecting <cell_type>CD4+ T cell epitopes</cell_type> from complex <protein>Ags</protein> can be applied to any known <protein>Ag</protein> that gives a T cell response .


Localized pancreatic NF-kappaB activation and inflammatory response in taurocholate-induced pancreatitis .


Transcription factor nuclear factor-kappaB ( NF-kappaB ) is activated in cerulein pancreatitis and mediates cytokine expression .

<protein>Transcription factor nuclear factor-kappaB</protein> ( <protein>NF-kappaB</protein> ) is activated in cerulein pancreatitis and mediates cytokine expression .


The role of transcription factor activation in other models of pancreatitis has not been established .


Here we report upregulation of NF-kappaB and inflammatory molecules , and their correlation with local pancreatic injury , in a model of severe pancreatitis .

Here we report upregulation of <protein>NF-kappaB</protein> and <protein>inflammatory molecules</protein> , and their correlation with local pancreatic injury , in a model of severe pancreatitis .


Rats received intraductal infusion of taurocholate or saline , and the pancreatic head and tail were analyzed separately .


NF-kappaB and activator protein-1 ( AP-1 ) activation were assessed by gel shift assay , and mRNA expression of interleukin-6 , tumor necrosis factor-alpha , KC , monocyte chemoattractant protein-1 , and inducible nitric oxide synthase was assessed by semiquantitative RT-PCR .

<protein>NF-kappaB</protein> and <protein>activator protein-1</protein> ( <protein>AP-1</protein> ) activation were assessed by gel shift assay , and mRNA expression of <rna>interleukin-6</rna> , <protein>tumor necrosis factor-alpha</protein> , KC , <protein>monocyte chemoattractant protein-1</protein> , and <protein>inducible nitric oxide synthase</protein> was assessed by semiquantitative RT-PCR .


Morphological damage and trypsin activation were much greater in the pancreatic head than tail , in parallel with a stronger activation of NF-kappaB and cytokine mRNA .

Morphological damage and trypsin activation were much greater in the pancreatic head than tail , in parallel with a stronger activation of <rna>NF-kappaB and cytokine mRNA</rna> .


Saline infusion mildly affected these parameters .


AP-1 was strongly activated in both pancreatic segments after either taurocholate or saline infusion .

<protein>AP-1</protein> was strongly activated in both pancreatic segments after either taurocholate or saline infusion .


NF-kappaB inhibition with N-acetylcysteine ameliorated the local inflammatory response .

<protein>NF-kappaB</protein> inhibition with N-acetylcysteine ameliorated the local inflammatory response .


Correlation between localized NF-kappaB activation , cytokine upregulation , and tissue damage suggests a key role for NF-kappaB in the development of the inflammatory response of acute pancreatitis .

Correlation between localized <protein>NF-kappaB</protein> activation , cytokine upregulation , and tissue damage suggests a key role for <protein>NF-kappaB</protein> in the development of the inflammatory response of acute pancreatitis .


CD45 tyrosine phosphatase controls common gamma-chain cytokine -mediated STAT and extracellular signal-related kinase phosphorylation in activated human lymphoblasts : inhibition of proliferation without induction of apoptosis .

<protein>CD45 tyrosine phosphatase</protein> controls common <protein>gamma-chain cytokine</protein> -mediated <protein>STAT</protein> and extracellular signal-related kinase phosphorylation in <cell_type>activated human lymphoblasts</cell_type> : inhibition of proliferation without induction of apoptosis .


The objective of this study was to test whether CD45 signals can influence signaling processes in activated human lymphoblasts .

The objective of this study was to test whether CD45 signals can influence signaling processes in <cell_type>activated human lymphoblasts</cell_type> .


To this end , we generated lymphoblasts which proliferate in response to common gamma-chain cytokines , but readily undergo apoptosis after cytokine withdrawal .

To this end , we generated <cell_type>lymphoblasts</cell_type> which proliferate in response to common <protein>gamma-chain cytokines</protein> , but readily undergo apoptosis after cytokine withdrawal .


In experiments with the CD45R0 mAb UCHL-1 , but not control CD45 mAbs , we found significant inhibition of proliferation .

In experiments with the <protein>CD45R0 mAb UCHL-1</protein> , but not <protein>control CD45 mAbs</protein> , we found significant inhibition of proliferation .


Interestingly , the pan-CD45 mAb GAP8.3 , which is most effective in inhibition of OKT-3-mediated proliferation in quiescent lymphocytes , was ineffective in lymphoblasts .

Interestingly , the <protein>pan-CD45 mAb GAP8.3</protein> , which is most effective in inhibition of OKT-3-mediated proliferation in <cell_type>quiescent lymphocytes</cell_type> , was ineffective in <cell_type>lymphoblasts</cell_type> .


Addition of CD3 mAb OKT-3 had no influence on IL-2-mediated proliferation ( with or without UCHL-1 ) .

Addition of <protein>CD3 mAb OKT-3</protein> had no influence on IL-2-mediated proliferation ( with or without <protein>UCHL-1</protein> ) .


In contrast , after addition of OKT-3 to IL-4 - and IL-7 -stimulated proliferation assays , UCHL-1 signals could not significantly alter cellular proliferation .

In contrast , after addition of <protein>OKT-3</protein> to <protein>IL-4</protein> - and <protein>IL-7</protein> -stimulated proliferation assays , <protein>UCHL-1</protein> signals could not significantly alter cellular proliferation .


We did not find induction of apoptosis following CD45R0 signaling .


In Western blots using mAbs detecting phosphorylated STAT-3 , STAT-5 , STAT-6 , or extracellular signal-related kinase 1/2 , we found that CD45R0 signaling could effectively diminish phosphorylation of these intracellular signaling components .

In Western blots using <protein>mAbs</protein> detecting <protein>phosphorylated STAT-3 , STAT-5 , STAT-6</protein> , or <protein>extracellular signal-related kinase 1/2</protein> , we found that CD45R0 signaling could effectively diminish phosphorylation of these intracellular signaling components .


Using RT-PCR , we found that CD45R0 signaling inhibited IL-2 mRNA production without major influence on IL-13 , IL-5 , or IFN-gamma mRNA levels .

Using RT-PCR , we found that CD45R0 signaling inhibited IL-2 mRNA production without major influence on <protein>IL-13</protein> , <protein>IL-5</protein> , or <rna>IFN-gamma mRNA</rna> levels .


Costimulation with OKT-3 and IL-2 optimally induced secretion of IFN-gamma , TNF-alpha , and IL-5 , which was not decreased by CD45 signals .

Costimulation with <protein>OKT-3</protein> and <protein>IL-2</protein> optimally induced secretion of <protein>IFN-gamma</protein> , <protein>TNF-alpha</protein> , and <protein>IL-5</protein> , which was not decreased by CD45 signals .


In conclusion , we illustrate that CD45R0 signals control early cytokine receptor-associated signaling processes and mRNA and DNA synthesis in activated human lymphoblasts .

In conclusion , we illustrate that CD45R0 signals control early cytokine receptor-associated signaling processes and mRNA and DNA synthesis in <cell_type>activated human lymphoblasts</cell_type> .


Furthermore , we show the existence of CD45 epitopes ( GAP8.3 ) , which are active and critical for signaling in quiescent lymphocytes , but are nonfunctional in activated human lymphoblasts .

Furthermore , we show the existence of <protein>CD45 epitopes</protein> ( <protein>GAP8.3</protein> ) , which are active and critical for signaling in <cell_type>quiescent lymphocytes</cell_type> , but are nonfunctional in <cell_type>activated human lymphoblasts</cell_type> .


Pax5 determines the identity of B cells from the beginning to the end of B-lymphopoiesis .

<dna>Pax5</dna> determines the identity of <cell_type>B cells</cell_type> from the beginning to the end of B-lymphopoiesis .


Despite being one of the most intensively studied cell types , the molecular basis of B cell specification is largely unknown .


The Pax5 gene encoding the transcription factor BSAP is required for progression of B-lymphopoiesis beyond the pro-B cell stage .

The <dna>Pax5 gene</dna> encoding the <protein>transcription factor BSAP</protein> is required for progression of B-lymphopoiesis beyond the pro-B cell stage .


Pax5-deficient pro-B cells are , however , not yet committed to the B-lymphoid lineage , but instead have a broad lymphomyeloid developmental potential .

<cell_type>Pax5-deficient pro-B cells</cell_type> are , however , not yet committed to the <cell_type>B-lymphoid lineage</cell_type> , but instead have a broad lymphomyeloid developmental potential .


Pax5 appears to mediate B-lineage commitment by repressing the transcription of non-B-lymphoid genes and by simultaneously activating the expression of B-lineage-specific genes .

<dna>Pax5</dna> appears to mediate B-lineage commitment by repressing the transcription of <dna>non-B-lymphoid genes</dna> and by simultaneously activating the expression of <dna>B-lineage-specific genes</dna> .


Pax5 thus functions both as a transcriptional repressor and activator , depending on its interactions with corepressors of the Groucho protein family or with positive regulators such as the TATA-binding protein .

<dna>Pax5</dna> thus functions both as a transcriptional repressor and activator , depending on its interactions with <protein>corepressors</protein> of the <protein>Groucho protein family</protein> or with positive regulators such as the <protein>TATA-binding protein</protein> .


Once committed to the B-lineage , B cells require Pax5 function to maintain their B-lymphoid identity throughout B cell development

Once committed to the <cell_type>B-lineage</cell_type> , <cell_type>B cells</cell_type> require <dna>Pax5</dna> function to maintain their B-lymphoid identity throughout B cell development


Partners in transcription : NFAT and AP-1 .

Partners in transcription : <protein>NFAT</protein> and <protein>AP-1</protein> .


Combinatorial regulation is a powerful mechanism that enables tight control of gene expression , via integration of multiple signaling pathways that induce different transcription factors required for enhanceosome assembly .

Combinatorial regulation is a powerful mechanism that enables tight control of gene expression , via integration of multiple signaling pathways that induce different <protein>transcription factors</protein> required for enhanceosome assembly .


The four calcium-regulated transcription factors of the NFAT family act synergistically with AP-1 ( Fos/Jun ) proteins on composite DNA elements which contain adjacent NFAT and AP-1 binding sites , where they form highly stable ternary complexes to regulate the expression of diverse inducible genes .

The four calcium-regulated <protein>transcription factors</protein> of the <protein>NFAT family</protein> act synergistically with <protein>AP-1 ( Fos/Jun ) proteins</protein> on composite DNA elements which contain adjacent <dna>NFAT and AP-1 binding sites</dna> , where they form highly stable ternary complexes to regulate the expression of diverse inducible genes .


Concomitant induction of NFAT and AP-1 requires concerted activation of two different signaling pathways : calcium/calcineurin , which promotes NFAT dephosphorylation , nuclear translocation and activation ; and protein kinase C ( PKC ) /Ras , which promotes the synthesis , phosphorylation and activation of members of the Fos and Jun families of transcription factors .

Concomitant induction of <protein>NFAT</protein> and <protein>AP-1</protein> requires concerted activation of two different signaling pathways : calcium/calcineurin , which promotes NFAT dephosphorylation , nuclear translocation and activation ; and <protein>protein kinase C ( PKC ) /Ras</protein> , which promotes the synthesis , phosphorylation and activation of members of the <protein>Fos and Jun families</protein> of <protein>transcription factors</protein> .


A fifth member of the NFAT family , NFAT5 , controls the cellular response to osmotic stress , by a mechanism that requires dimer formation and is independent of calcineurin or of interaction with AP-1 .

A fifth member of the <protein>NFAT family</protein> , <protein>NFAT5</protein> , controls the cellular response to osmotic stress , by a mechanism that requires dimer formation and is independent of <protein>calcineurin</protein> or of interaction with <protein>AP-1</protein> .


Pharmacological interference with theNFAT : AP-1 interaction may be useful in selective manipulation of the immune response .


Balanced activation of NFAT and AP-1 is known to be required for productive immune responses , but the role of NFAT : AP-1 interactions in other cell types and biological processes remains to be understood .

Balanced activation of <protein>NFAT</protein> and <protein>AP-1</protein> is known to be required for productive immune responses , but the role of NFAT : AP-1 interactions in other cell types and biological processes remains to be understood .


Cytokine production by Vgamma ( + ) -T-cell subsets is an important factor determining CD4 ( + ) -Th-cell phenotype and susceptibility of BALB/c mice to coxsackievirus B3-induced myocarditis .

Cytokine production by <cell_type>Vgamma ( + ) -T-cell subsets</cell_type> is an important factor determining CD4 ( + ) -Th-cell phenotype and susceptibility of BALB/c mice to coxsackievirus B3-induced myocarditis .


Two coxsackievirus B3 ( CVB3 ) variants ( H3 and H310A1 ) differ by a single amino acid mutation in the VP2 capsid protein .

Two coxsackievirus B3 ( CVB3 ) variants ( H3 and H310A1 ) differ by a single amino acid mutation in the <protein>VP2 capsid protein</protein> .


H3 induces severe myocarditis in BALB/c mice , but H310A1 is amyocarditic .


Infection with H3 , but not H310A1 , preferentially activates Vgamma4 Vdelta4 cells , which are strongly positive for gamma interferon ( IFN-gamma ) , whereas Vgamma1 Vdelta4 cells are increased in both H3 and H310A1 virus-infected animals .

Infection with H3 , but not H310A1 , preferentially activates <cell_type>Vgamma4 Vdelta4 cells</cell_type> , which are strongly positive for <protein>gamma interferon</protein> ( <protein>IFN-gamma</protein> ) , whereas <cell_type>Vgamma1 Vdelta4 cells</cell_type> are increased in both H3 and H310A1 virus-infected animals .


Depletion of Vgamma1 ( + ) cells using monoclonal anti-Vgamma1 antibody enhanced myocarditis and CD4 ( + ) - , IFN-gamma ( + ) -cell responses in both H3- and H310A1-infected mice yet decreased the CD4 ( + ) - , IL-4 ( + ) -cell response .

Depletion of <cell_type>Vgamma1 ( + ) cells</cell_type> using <protein>monoclonal anti-Vgamma1 antibody</protein> enhanced myocarditis and CD4 ( + ) - , <protein>IFN-gamma</protein> ( + ) -cell responses in both H3- and H310A1-infected mice yet decreased the CD4 ( + ) - , IL-4 ( + ) -cell response .


Depleting Vgamma4 ( + ) cells suppressed myocarditis and reduced CD4 ( + ) IFN-gamma ( + ) cells but increased CD4 ( + ) IL-4 ( + ) T cells .

Depleting <cell_type>Vgamma4 ( + ) cells</cell_type> suppressed myocarditis and reduced <cell_type>CD4 ( + ) IFN-gamma ( + ) cells</cell_type> but increased <cell_type>CD4 ( + ) IL-4 ( + ) T cells</cell_type> .


The role of cytokine production by Vgamma1 ( + ) and Vgamma4 ( + ) T cells was investigated by adoptively transferring these cells isolated from H3-infected BALB/c Stat4 knockout ( Stat4ko ) ( defective in IFN-gamma expression ) or BALB/c Stat6ko ( defective in IL-4 expression ) mice into H3 virus-infected wild-type BALB/c recipients .

The role of cytokine production by <cell_type>Vgamma1 ( + ) and Vgamma4 ( + ) T cells</cell_type> was investigated by adoptively transferring these cells isolated from H3-infected BALB/c Stat4 knockout ( Stat4ko ) ( defective in <protein>IFN-gamma</protein> expression ) or BALB/c Stat6ko ( defective in IL-4 expression ) mice into H3 virus-infected wild-type BALB/c recipients .


Vgamma4 and Vgamma1 ( + ) T cells from Stat4ko mice expressed IL-4 but no or minimal IFN-gamma , whereas these cell populations derived from Stat6ko mice expressed IFN-gamma but no IL-4 .

<cell_type>Vgamma4 and Vgamma1 ( + ) T cells</cell_type> from Stat4ko mice expressed <protein>IL-4</protein> but no or minimal <protein>IFN-gamma</protein> , whereas these cell populations derived from Stat6ko mice expressed <protein>IFN-gamma</protein> but no <protein>IL-4</protein> .


Stat4ko Vgamma1 ( + ) cells ( IL-4 ( + ) ) suppress myocarditis .

<cell_type>Stat4ko Vgamma1 ( + ) cells ( IL-4 ( + )</cell_type> ) suppress myocarditis .


Stat6ko Vgamma1 ( + ) cells ( IFN-gamma ( + ) ) were not inhibitory .

<cell_type>Stat6ko Vgamma1 ( + ) cells ( IFN-gamma ( + ) )</cell_type> were not inhibitory .


Stat6ko Vgamma4 ( + ) cells ( IFN-gamma ( + ) ) significantly enhanced myocarditis .

<cell_type>Stat6ko Vgamma4 ( + ) cells ( IFN-gamma ( + ) )</cell_type> significantly enhanced myocarditis .


Stat4ko Vgamma4 ( + ) cells ( IL-4 ( + ) ) neither inhibited nor enhanced disease .

<cell_type>Stat4ko Vgamma4 ( + ) cells ( IL-4 ( + ) )</cell_type> neither inhibited nor enhanced disease .


These results show that distinct gammadelta-T-cell subsets control myocarditis susceptibility and bias the CD4 ( + ) -Th-cell response .

These results show that distinct <cell_type>gammadelta-T-cell subsets</cell_type> control myocarditis susceptibility and bias the <cell_type>CD4 ( + ) -Th-cell</cell_type> response .


The cytokines produced by the Vgamma subpopulation have a significant influence on the CD4 ( + ) -Th-cell phenotype .

The <protein>cytokines</protein> produced by the <cell_type>Vgamma subpopulation</cell_type> have a significant influence on the <protein>CD4</protein> ( + ) -Th-cell phenotype .


Plasmin -induced expression of cytokines and tissue factor in human monocytes involves AP-1 and IKKbeta -mediated NF-kappaB activation .

<protein>Plasmin</protein> -induced expression of <protein>cytokines</protein> and <protein>tissue factor</protein> in <cell_type>human monocytes</cell_type> involves <protein>AP-1</protein> and <protein>IKKbeta</protein> -mediated NF-kappaB activation .


It was previously shown that plasmin activates human peripheral monocytes in terms of lipid mediator release and chemotactic migration .

It was previously shown that <protein>plasmin</protein> activates <cell_type>human peripheral monocytes</cell_type> in terms of lipid mediator release and chemotactic migration .


Here it is demonstrated that plasmin induces proinflammatory cytokine release and tissue factor ( TF ) expression by monocytes .

Here it is demonstrated that <protein>plasmin</protein> induces <protein>proinflammatory cytokine</protein> release and <protein>tissue factor</protein> ( <protein>TF</protein> ) expression by <cell_type>monocytes</cell_type> .


Plasmin 0.043 to 1.43 CTA U/mL , but not active site-blocked plasmin , triggered concentration-dependent expression of mRNA for interleukin-1alpha ( IL-1alpha ) , IL-1beta , tumor necrosis factor-alpha ( TNF-alpha ) , and TF with maximum responses after 4 hours .

<protein>Plasmin</protein> 0.043 to 1.43 CTA U/mL , but not <protein>active site-blocked plasmin</protein> , triggered concentration-dependent expression of mRNA for <protein>interleukin-1alpha</protein> ( <protein>IL-1alpha</protein> ) , <protein>IL-1beta</protein> , <protein>tumor necrosis factor-alpha</protein> ( <protein>TNF-alpha</protein> ) , and <protein>TF</protein> with maximum responses after 4 hours .


Plasmin -mediated mRNA expression was inhibited in a concentration-dependent manner by the lysine analogue trans-4- ( aminomethyl ) cyclohexane-1-carboxylic acid ( t-AMCA ) .

<protein>Plasmin</protein> -mediated mRNA expression was inhibited in a concentration-dependent manner by the lysine analogue trans-4- ( aminomethyl ) cyclohexane-1-carboxylic acid ( t-AMCA ) .


Increases in mRNA levels were followed by concentration- and time-dependent release of IL-1alpha , IL-1beta and TNF-alpha and by TF expression on monocyte surfaces .

Increases in mRNA levels were followed by concentration- and time-dependent release of <protein>IL-1alpha</protein> , <protein>IL-1beta</protein> and <protein>TNF-alpha</protein> and by <protein>TF</protein> expression on monocyte surfaces .


Neither cytokines nor TF could be detected when monocytes were preincubated with actinomycin D or cycloheximide .

Neither <protein>cytokines</protein> nor <protein>TF</protein> could be detected when <cell_type>monocytes</cell_type> were preincubated with actinomycin D or cycloheximide .


Electrophoretic mobility shift assays indicated plasmin -induced activation of NF-kappaB ; DNA-binding complexes were composed of p50 , p65 , and c-Rel , as shown by supershift experiments .

Electrophoretic mobility shift assays indicated <protein>plasmin</protein> -induced activation of <protein>NF-kappaB</protein> ; <protein>DNA-binding complexes</protein> were composed of <protein>p50</protein> , <protein>p65</protein> , and <protein>c-Rel</protein> , as shown by supershift experiments .


Nuclear translocation of NF-kappaB/Rel proteins coincided with IkappaBalpha degradation .

Nuclear translocation of <protein>NF-kappaB/Rel proteins</protein> coincided with IkappaBalpha degradation .


At variance with endotoxic lipopolysaccharide , plasmin elicited the rapid degradation of another cytoplasmic NF-kappaB inhibitor , p105 .

At variance with endotoxic lipopolysaccharide , <protein>plasmin</protein> elicited the rapid degradation of another <protein>cytoplasmic NF-kappaB inhibitor , p105</protein> .


Proteolysis of NF-kappaB inhibitors was apparently due to transient activation of IkappaB kinase ( IKK ) beta that reached maximum activity at 1 hour after plasmin stimulation .

Proteolysis of NF-kappaB inhibitors was apparently due to transient activation of <protein>IkappaB kinase ( IKK ) beta</protein> that reached maximum activity at 1 hour after <protein>plasmin</protein> stimulation .


In addition , AP-1 binding was increased in plasmin -treated monocytes , with most complexes composed of JunD , c-Fos , and FosB .

In addition , AP-1 binding was increased in <protein>plasmin</protein> -treated <cell_type>monocytes</cell_type> , with most complexes composed of <protein>JunD</protein> , <protein>c-Fos</protein> , and <protein>FosB</protein> .


These findings further substantiate the role of plasmin as a proinflammatory activator of human monocytes and reveal an important new link between the plasminogen-plasmin system and inflammation .

These findings further substantiate the role of <protein>plasmin</protein> as a proinflammatory activator of <cell_type>human monocytes</cell_type> and reveal an important new link between the plasminogen-plasmin system and inflammation .


( Blood. 2001 ; 97 : 3941-3950 )


STAT3 is constitutively active in some patients with Polycythemia rubra vera .

<protein>STAT3</protein> is constitutively active in some patients with Polycythemia rubra vera .


OBJECTIVE : Polycythemia vera is a clonal stem cell disorder characterized by hyperproliferation of the erythroid , myeloid , and megakaryocytic lineages .

OBJECTIVE : Polycythemia vera is a clonal stem cell disorder characterized by hyperproliferation of the <cell_type>erythroid , myeloid , and megakaryocytic lineages</cell_type> .


While it has been shown that progenitor cells of P. vera patients are hypersensitive to several growth factors including erythropoietin , insulin-like growth factor-1 , thrombopoietin , interleukin-3 , and granulocyte/monocyte colony-stimulating factor , the molecular pathogenesis of this disease remains unknown .

While it has been shown that <cell_type>progenitor cells</cell_type> of P. vera patients are hypersensitive to several growth factors including <protein>erythropoietin</protein> , <protein>insulin-like growth factor-1</protein> , <protein>thrombopoietin</protein> , <protein>interleukin-3</protein> , and <protein>granulocyte/monocyte colony-stimulating factor</protein> , the molecular pathogenesis of this disease remains unknown .


Growth factor hypersensitivity could be mediated by changes in signal transduction pathways .


We therefore investigated a common downstream effector of cytokines , the signal transducers and activators of transcription ( STATs ) .

We therefore investigated a common downstream effector of <protein>cytokines</protein> , the <protein>signal transducers and activators of transcription</protein> ( <protein>STATs</protein> ) .


A constitutive activation of STAT factors could explain the increased proliferation of P. vera cells even in the absence of growth factor stimulation .

A constitutive activation of <protein>STAT factors</protein> could explain the increased proliferation of <cell_type>P. vera cells</cell_type> even in the absence of growth factor stimulation .


METHODS : Peripheral granulocytes from patients with P. vera and from healthy volunteers were assayed for STAT1 , 3 , and 5 DNA binding by electrophoretic mobility shift assay .

METHODS : <cell_type>Peripheral granulocytes</cell_type> from patients with P. vera and from healthy volunteers were assayed for <protein>STAT1 , 3 , and 5</protein> DNA binding by electrophoretic mobility shift assay .


RESULTS : Four of 14 P. vera patients analyzed showed constitutive STAT3 DNA binding in unstimulated peripheral granulocytes , while none of the 17 healthy volunteers tested did .

RESULTS : Four of 14 P. vera patients analyzed showed constitutive STAT3 DNA binding in <cell_type>unstimulated peripheral granulocytes</cell_type> , while none of the 17 healthy volunteers tested did .


of the subjects showed constitutive STAT1 or STAT5 activity .

of the subjects showed constitutive <protein>STAT1</protein> or <protein>STAT5</protein> activity .


Western blotting demonstrated that , in the three patients , STAT3 is constitutively phosphorylated on Tyr 705 , whereas it is unphosphorylated in the other patients and in controls .

Western blotting demonstrated that , in the three patients , <protein>STAT3</protein> is constitutively phosphorylated on Tyr 705 , whereas it is unphosphorylated in the other patients and in controls .


Interestingly , constitutive STAT3 activity did not correlate with the duration of disease or the treatment regimen .

Interestingly , constitutive <protein>STAT3</protein> activity did not correlate with the duration of disease or the treatment regimen .


It was observed in a recently diagnosed patient and in two patients treated only with phlebotomy .


CONCLUSION : Our data suggest that constitutive phosphorylation and activation of STAT3 is not a secondary event induced by mutagenizing agents or by prolonged hyperproliferation of hematopoietic cells , but rather represents a primary molecular aberration .

CONCLUSION : Our data suggest that constitutive phosphorylation and activation of <protein>STAT3</protein> is not a secondary event induced by mutagenizing agents or by prolonged hyperproliferation of <cell_type>hematopoietic cells</cell_type> , but rather represents a primary molecular aberration .


Constitutively active STAT3 may contribute to the growth factor hypersensitivity of P. vera cells .

<protein>Constitutively active STAT3</protein> may contribute to the growth factor hypersensitivity of <cell_type>P. vera cells</cell_type> .


Identification of phosphorylation sites for Bruton 's tyrosine kinase within the transcriptional regulator BAP/TFII-I .

Identification of <protein>phosphorylation sites</protein> for <protein>Bruton 's tyrosine kinase</protein> within the <protein>transcriptional regulator BAP/TFII-I</protein> .


Bruton 's tyrosine kinase ( Btk ) , a member of the Tec family of cytosolic kinases , is essential for B cell development and function .

<protein>Bruton 's tyrosine kinase</protein> ( <protein>Btk</protein> ) , a member of the <protein>Tec family of cytosolic kinases</protein> , is essential for <cell_type>B cell</cell_type> development and function .


BAP/TFII-I , a protein implicated in transcriptional regulation , is associated with Btk in B cells and is transiently phosphorylated on tyrosine following B cell receptor engagement .

<protein>BAP/TFII-I</protein> , a protein implicated in transcriptional regulation , is associated with <protein>Btk</protein> in <cell_type>B cells</cell_type> and is transiently phosphorylated on tyrosine following <cell_type>B cell</cell_type> receptor engagement .


BAP/TFII-I is a substrate for Btk in vitro and is hyperphosphorylated on tyrosine upon coexpression with Btk in mammalian cells .

<protein>BAP/TFII-I</protein> is a substrate for <protein>Btk</protein> in vitro and is hyperphosphorylated on tyrosine upon coexpression with <protein>Btk</protein> in <cell_type>mammalian cells</cell_type> .


In an effort to understand the physiologic consequences of BAP/TFII-I tyrosine phosphorylation following B cell receptor stimulation , site-directed mutagenesis and phosphopeptide mapping were used to locate the predominant sites of BAP/TFII-I phosphorylation by Btk in vitro .

In an effort to understand the physiologic consequences of <protein>BAP/TFII-I</protein> tyrosine phosphorylation following B cell receptor stimulation , site-directed mutagenesis and phosphopeptide mapping were used to locate the predominant sites of <protein>BAP/TFII-I</protein> phosphorylation by <protein>Btk</protein> in vitro .


These residues , Tyr248 , Tyr357 , and Tyr462 , were also found to be the major sites for Btk -dependent phosphorylation of BAP/TFII-I in vivo .

These residues , Tyr248 , Tyr357 , and Tyr462 , were also found to be the major sites for <protein>Btk</protein> -dependent phosphorylation of <protein>BAP/TFII-I</protein> in vivo .


Residues Tyr357 and Tyr462 are contained within the loop regions of adjacent helix-loop-helix-like repeats within BAP/TFII-I .

Residues Tyr357 and Tyr462 are contained within the <protein>loop regions</protein> of adjacent <protein>helix-loop-helix-like repeats</protein> within <protein>BAP/TFII-I</protein> .


Mutation of either Tyr248 , Tyr357 , or Tyr462 to phenylalanine reduced transcription from a c-fos promoter relative to wild-type BAP/TFII-I in transfected COS-7 cells , consistent with the interpretation that phosphorylation at these sites contributes to transcriptional activation .

Mutation of either Tyr248 , Tyr357 , or Tyr462 to phenylalanine reduced transcription from a <dna>c-fos promoter</dna> relative to <dna>wild-type BAP/TFII-I</dna> in transfected <cell_line>COS-7 cells</cell_line> , consistent with the interpretation that phosphorylation at these sites contributes to transcriptional activation .


Phosphorylation of BAP/TFII-I by Btk may link engagement of receptors such as surface immunoglobulin to modulation of gene expression .

Phosphorylation of <protein>BAP/TFII-I</protein> by <protein>Btk</protein> may link engagement of receptors such as surface <protein>immunoglobulin</protein> to modulation of gene expression .


Expression of interferon consensus sequence binding protein induces potent immunity against BCR/ABL-induced leukemia .

Expression of <protein>interferon consensus sequence binding protein</protein> induces potent immunity against BCR/ABL-induced leukemia .


Mice deficient in the interferon consensus sequence binding protein ( ICSBP ) develop a disease resembling chronic myeloid leukemia ( CML ) , which in humans is caused by the BCR/ABL oncoprotein .

Mice deficient in the <protein>interferon consensus sequence binding protein</protein> ( <protein>ICSBP</protein> ) develop a disease resembling chronic myeloid leukemia ( CML ) , which in humans is caused by the <protein>BCR/ABL oncoprotein</protein> .


Interferon-alpha ( IFN-alpha ) induces ICSBP expression and is an effective therapy for CML .

<protein>Interferon-alpha</protein> ( <protein>IFN-alpha</protein> ) induces <protein>ICSBP</protein> expression and is an effective therapy for CML .


This study examined whether enforced expression of ICSBP might antagonize BCR/ABL-induced leukemia ; results demonstrated that ICSBP-modified cells generated a protective CD8 ( + ) cytotoxic T-cell response against BCR/ABL-transformed BaF3 cells in a murine leukemia model .

This study examined whether enforced expression of <protein>ICSBP</protein> might antagonize BCR/ABL-induced leukemia ; results demonstrated that <cell_type>ICSBP-modified cells</cell_type> generated a protective CD8 ( + ) cytotoxic T-cell response against <cell_type>BCR/ABL-transformed BaF3 cells</cell_type> in a murine leukemia model .


ICSBP expression represents a novel means of stimulating a host immune response to BCR/ABL ( + ) leukemia cells and a potential strategy for immunotherapy of CML .

<protein>ICSBP</protein> expression represents a novel means of stimulating a host immune response to <cell_type>BCR/ABL ( + ) leukemia cells</cell_type> and a potential strategy for immunotherapy of CML .


( Blood. 2001 ; 97 : 3491-3497 )


A transcriptional block in the IL-2 promoter at the -150 AP-1 site in effector CD8+ T cells .

A transcriptional block in the <dna>IL-2 promoter</dna> at the <dna>-150 AP-1 site</dna> in <cell_type>effector CD8+ T cells</cell_type> .


Both CD4+ and CD8+ T cells that produce IL-2 in response to Ag recognition have been isolated .

Both <cell_type>CD4+ and CD8+ T cells</cell_type> that produce <protein>IL-2</protein> in response to Ag recognition have been isolated .


However , most effector CD8+ T cells recovered after exposure to Ag do not produce sufficient IL-2 to sustain growth , and depend on CD4+ T helper cells for this obligate growth factor .

However , most <cell_type>effector CD8+ T cells</cell_type> recovered after exposure to <protein>Ag</protein> do not produce sufficient <protein>IL-2</protein> to sustain growth , and depend on <cell_type>CD4+ T helper cells</cell_type> for this obligate growth factor .


IL-2 expression in CD4+ T cells is primarily controlled at the level of transcription , but mechanisms restricting IL-2 production in CD8+ T cells have not been elucidated .

IL-2 expression in <cell_type>CD4+ T cells</cell_type> is primarily controlled at the level of transcription , but mechanisms restricting IL-2 production in <cell_type>CD8+ T cells</cell_type> have not been elucidated .


To evaluate transcriptional regulation of the IL-2 gene in CD8+ T cells , we stably transfected reporter genes into Ag -specific CD8+ T cell clones .

To evaluate transcriptional regulation of the <dna>IL-2 gene</dna> in <cell_type>CD8+ T cells</cell_type> , we stably transfected <dna>reporter genes</dna> into <protein>Ag</protein> -specific <cell_line>CD8+ T cell clones</cell_line> .


CD28+ CD8 ( + ) T cells unable to transcribe the IL-2 gene in response to antigenic stimulation had a block in transactivation of the -150 CD28 response element ( CD28RE ) /AP-1 site of the IL-2 promoter , but did transactivate the composite NFAT/AP-1 and OCT/AP-1 sites , and a consensus AP-1 motif .

<cell_type>CD28+ CD8 ( + ) T cells</cell_type> unable to transcribe the <dna>IL-2 gene</dna> in response to antigenic stimulation had a block in transactivation of the <dna>-150 CD28 response element ( CD28RE ) /AP-1 site</dna> of the <dna>IL-2 promoter</dna> , but did transactivate the composite <dna>NFAT/AP-1 and OCT/AP-1 sites</dna> , and a <dna>consensus AP-1 motif</dna> .


Mutation of the nonconsensus -150 AP-1 site to a consensus AP-1 site , or insertion of a CD28RE/AP-1 consensus site upstream of the native -150 CD28RE/AP-1 site restored transactivation of the altered promoter .

Mutation of the <dna>nonconsensus -150 AP-1 site</dna> to a <dna>consensus AP-1 site</dna> , or insertion of a <dna>CD28RE/AP-1 consensus site</dna> upstream of the native <dna>-150 CD28RE/AP-1 site</dna> restored transactivation of the altered <dna>promoter</dna> .


These results suggest that the defect at the -150 site may reflect the absence or inactivity of a required factor rather than repression of the IL-2 promoter .

These results suggest that the defect at the <dna>-150 site</dna> may reflect the absence or inactivity of a required factor rather than repression of the <dna>IL-2 promoter</dna> .


Stem cell factor and interleukin-3 induce stepwise generation of erythroid precursor cells from a basic fibroblast growth factor -dependent hematopoietic stem cell line , A-6 .

<protein>Stem cell factor</protein> and <protein>interleukin-3</protein> induce stepwise generation of <cell_type>erythroid precursor cells</cell_type> from a basic <protein>fibroblast growth factor</protein> -dependent <cell_line>hematopoietic stem cell line , A-6</cell_line> .


A m ultipotent immature myeloid cell population was produced from a basic fibroblast growth factor ( bFGF ) -dependent hematopoietic stem cell line , A-6 , when cultured with stem cell factor ( SCF ) replacing bFGF .

A m <cell_type>ultipotent immature myeloid cell population</cell_type> was produced from a <protein>basic fibroblast growth factor</protein> ( <protein>bFGF</protein> ) -dependent <cell_line>hematopoietic stem cell line , A-6</cell_line> , when cultured with <protein>stem cell factor</protein> ( <protein>SCF</protein> ) replacing <protein>bFGF</protein> .


Those cells were positive for stem cell markers , c-kit and CD34 , and a myeloid cell marker , F4/80 .

Those cells were positive for <protein>stem cell markers</protein> , <protein>c-kit</protein> and <protein>CD34</protein> , and a <protein>myeloid cell marker , F4/80</protein> .


Some cell fractions were also positive for Mac-1 , a macrophage marker or Gr-1 , a granulocytic maker , but negative for an erythroid marker TER119 .

Some cell fractions were also positive for <protein>Mac-1 , a macrophage marker</protein> or <protein>Gr-1 , a granulocytic maker</protein> , but negative for an <protein>erythroid marker TER119</protein> .


They also showed the expression of mRNA for the myeloid-specific PU.1 but did not that for the erythroid-specific GATA-1 .

They also showed the expression of mRNA for the <protein>myeloid-specific PU.1</protein> but did not that for the <protein>erythroid-specific GATA-1</protein> .


Among various cytokines , interleukin-3 ( IL-3 ) induced erythroid precursor cells that expressed the erythroid-specific GATA-1 and beta-major globin .

Among various <protein>cytokines</protein> , <protein>interleukin-3</protein> ( <protein>IL-3</protein> ) induced <cell_type>erythroid precursor cells</cell_type> that expressed the <protein>erythroid-specific GATA-1</protein> and <protein>beta-major globin</protein> .


The quantitative analysis showed that erythroid precursor cells were newly produced from the immature myeloid cells by cultivation with IL-3 .

The quantitative analysis showed that <cell_type>erythroid precursor cells</cell_type> were newly produced from the <cell_type>immature myeloid cells</cell_type> by cultivation with <protein>IL-3</protein> .


SCF and IL-3 induced stepwise generation of erythroid precursor cells from an A-6 hematopoietic stem cell line .

<protein>SCF</protein> and <protein>IL-3</protein> induced stepwise generation of <cell_type>erythroid precursor cells</cell_type> from an <cell_line>A-6 hematopoietic stem cell line</cell_line> .


Copyright 2001 Academic Press .


Distinct BMI-1 and EZH2 expression patterns in thymocytes and mature T cells suggest a role for Polycomb genes in human T cell differentiation .

Distinct <protein>BMI-1</protein> and <protein>EZH2</protein> expression patterns in <cell_type>thymocytes</cell_type> and <cell_type>mature T cells</cell_type> suggest a role for <dna>Polycomb genes</dna> in human T cell differentiation .


BMI-1 and EZH2 Polycomb-group ( PcG ) proteins belong to two distinct protein complexes involved in the regulation of hematopoiesis .

<protein>BMI-1</protein> and <protein>EZH2</protein> <protein>Polycomb-group ( PcG ) proteins</protein> belong to two distinct protein complexes involved in the regulation of hematopoiesis .


Using unique PcG-specific antisera and triple immunofluorescence , we found that mature resting peripheral T cells expressed BMI-1 , whereas dividing blasts were EZH2 ( + ) .

Using unique PcG-specific antisera and triple immunofluorescence , we found that <cell_type>mature resting peripheral T cells</cell_type> expressed <protein>BMI-1</protein> , whereas <cell_type>dividing blasts</cell_type> were EZH2 ( + ) .


By contrast , subcapsular immature double-negative ( DN ) ( CD4 ( - ) /CD8 ( - ) ) T cells in the thymus coexpressed BMI-1 and EZH2 or were BMI-1 single positive .

By contrast , <cell_type>subcapsular immature double-negative ( DN ) ( CD4 ( - ) /CD8 ( - ) ) T cells</cell_type> in the thymus coexpressed <protein>BMI-1</protein> and <protein>EZH2</protein> or were BMI-1 single positive .


Their descendants , double-positive ( DP ; CD4 ( + ) /CD8 ( + ) ) cortical thymocytes , expressed EZH2 without BMI-1 .

Their descendants , <cell_type>double-positive ( DP ; CD4 ( + ) /CD8 ( + ) ) cortical thymocytes</cell_type> , expressed <protein>EZH2</protein> without <protein>BMI-1</protein> .


Most EZH2 ( + ) DN and DP thymocytes were dividing , while DN BMI-1 ( + ) /EZH2 ( - ) thymocytes were resting and proliferation was occasionally noted in DN BMI-1 ( + ) /EZH2 ( + ) cells .

Most <cell_type>EZH2 ( + ) DN and DP thymocytes</cell_type> were dividing , while <cell_type>DN BMI-1 ( + ) /EZH2 ( - ) thymocytes</cell_type> were resting and proliferation was occasionally noted in <cell_type>DN BMI-1 ( + ) /EZH2 ( + ) cells</cell_type> .


Maturation of DP cortical thymocytes to single-positive ( CD4 ( + ) /CD8 ( - ) or CD8 ( + ) /CD4 ( - ) ) medullar thymocytes correlated with decreased detectability of EZH2 and continued relative absence of BMI-1 .

Maturation of <cell_type>DP cortical thymocytes</cell_type> to <cell_type>single-positive ( CD4 ( + ) /CD8 ( - )</cell_type> or <cell_type>CD8 ( + ) /CD4 ( - ) ) medullar thymocytes</cell_type> correlated with decreased detectability of <protein>EZH2</protein> and continued relative absence of <protein>BMI-1</protein> .


Our data show that BMI-1 and EZH2 expression in mature peripheral T cells is mutually exclusive and linked to proliferation status , and that this pattern is not yet established in thymocytes of the cortex and medulla .

Our data show that <protein>BMI-1</protein> and <protein>EZH2</protein> expression in <cell_type>mature peripheral T cells</cell_type> is mutually exclusive and linked to proliferation status , and that this pattern is not yet established in <cell_type>thymocytes</cell_type> of the cortex and medulla .


T cell stage-specific PcG expression profiles suggest that PcG genes contribute to regulation of T cell differentiation .

<cell_type>T cell</cell_type> stage-specific PcG expression profiles suggest that <dna>PcG genes</dna> contribute to regulation of T cell differentiation .


They probably reflect stabilization of cell type-specific gene expression and irreversibility of lineage choice .


The difference in PcG expression between medullar thymocytes and mature interfollicular T cells indicates that additional maturation processes occur after thymocyte transportation from the thymus .

The difference in PcG expression between <cell_type>medullar thymocytes</cell_type> and <cell_type>mature interfollicular T cells</cell_type> indicates that additional maturation processes occur after thymocyte transportation from the thymus .


Stepwise lineage restriction of progenitors in lympho-myelopoiesis .

Stepwise lineage restriction of <cell_type>progenitors</cell_type> in lympho-myelopoiesis .


It has long been controversial whether hematopoiesis progresses through ordered stages of determination as in embryonic development .


This is due to the absence of a methodology capable of exactly determining the developmental potential of hematopoietic stem/progenitor cells .

This is due to the absence of a methodology capable of exactly determining the developmental potential of <cell_type>hematopoietic stem/progenitor cells</cell_type> .


The multilineage progenitor ( MLP ) assay enabled us to discriminate among seven types of hematopoietic progenitors , which are multipotent progenitor p-MTB ( capable of generating myeloid , T and B cells ) , bipotent progenitors p-MT , p-MB and p-TB , and unipotent progenitors p-M , p-T and p-B .

The multilineage progenitor ( MLP ) assay enabled us to discriminate among seven types of hematopoietic progenitors , which are multipotent progenitor p-MTB ( capable of generating <cell_type>myeloid , T and B cells</cell_type> ) , bipotent progenitors p-MT , p-MB and p-TB , and unipotent progenitors p-M , p-T and p-B .


Among these seven types , the p-TB type progenitor was found to be absent .


These findings indicate that the process of lineage commitment proceeds through an ordered but not random process .


By extending the area of investigation to include the erythroid lineage , more convincing evidence for the ordered process was obtained .

By extending the area of investigation to include the <cell_type>erythroid lineage</cell_type> , more convincing evidence for the ordered process was obtained .


Detailed and exact illustration of the process of hematopoiesis will provide an opportunity to revive hematopoiesis as one of the most fascinating targets of research in developmental biology


Epstein-Barr Virus and its glycoprotein-350 upregulate IL-6 in human B-lymphocytes via CD21 , involving activation of NF-kappaB and different signaling pathways .

Epstein-Barr Virus and its <protein>glycoprotein-350</protein> upregulate <protein>IL-6</protein> in <cell_type>human B-lymphocytes</cell_type> via <protein>CD21</protein> , involving activation of <protein>NF-kappaB</protein> and different signaling pathways .


Epstein-Barr virus ( EBV ) is a ubiquitous and highly immunotropic gamma herpesvirus that infects more than 90 % of humans worldwide .


Its pathogenicity leads to a number of diseases including tumors that result from EBV 's ability to readily transform B-lymphocytes and , to a lesser extent , epithelial cells .

Its pathogenicity leads to a number of diseases including tumors that result from EBV 's ability to readily transform <cell_type>B-lymphocytes</cell_type> and , to a lesser extent , <cell_type>epithelial cells</cell_type> .


EBV utilizes CD21/CR2 as its receptor on B cells to initiate the infection process .

EBV utilizes <protein>CD21/CR2</protein> as its receptor on <cell_type>B cells</cell_type> to initiate the infection process .


EBV binds to CR2 through its major envelope glycoprotein-350 ( gp350 ) and is also a remarkable immunomodulating agent .

EBV binds to <protein>CR2</protein> through its <protein>major envelope glycoprotein-350</protein> ( <protein>gp350</protein> ) and is also a remarkable immunomodulating agent .


We had previously shown that EBV is capable of modulating the synthesis of a number of cytokines .

We had previously shown that EBV is capable of modulating the synthesis of a number of <protein>cytokines</protein> .


We now show that while both purified recombinant gp350 ( rgp350 ) and EBV upregulate IL-6 mRNA synthesis in B cells , EBV-induced IL-6 gene activation occurs for a significantly longer period of time ( i.e. 12 hours for EBV as compared to 6 hours for rgp350 ) .

We now show that while both purified <protein>recombinant gp350</protein> ( <protein>rgp350</protein> ) and EBV upregulate <rna>IL-6 mRNA</rna> synthesis in <cell_type>B cells</cell_type> , EBV-induced IL-6 gene activation occurs for a significantly longer period of time ( i.e. 12 hours for EBV as compared to 6 hours for <protein>rgp350</protein> ) .


Moreover , the half-life of EBV-induced IL-6 mRNA was also significantly longer ( 10 hours ) than that of mRNA induced by rgp350 ( about 6 hours ) .

Moreover , the half-life of EBV-induced <rna>IL-6 mRNA</rna> was also significantly longer ( 10 hours ) than that of mRNA induced by <protein>rgp350</protein> ( about 6 hours ) .


Both EBV and gp350 enhance the binding of the NF-kappaB transcription factor , as determined by band-shift and augment NF-kappaB -mediated activation of a CAT reporter plasmid .

Both EBV and <protein>gp350</protein> enhance the binding of the <protein>NF-kappaB transcription factor</protein> , as determined by band-shift and augment <protein>NF-kappaB</protein> -mediated activation of a <dna>CAT reporter plasmid</dna> .


Furthermore , we demonstrate that while the activation of IL-6 gene expression by gp350 is mediated primarily by the protein kinase C pathway , EBV can mediate its effects through multiple signaling pathways .

Furthermore , we demonstrate that while the activation of IL-6 gene expression by <protein>gp350</protein> is mediated primarily by the protein kinase C pathway , EBV can mediate its effects through multiple signaling pathways .


To our knowledge this is the first report showing that the binding of a herpesvirus envelope glycoprotein to CR2 on human B cells results in the activation of the NF-kappaB transcription factor leading to the upregulation of IL-6 gene expression in these lymphocytes .

To our knowledge this is the first report showing that the binding of a <protein>herpesvirus envelope glycoprotein</protein> to <protein>CR2</protein> on <cell_type>human B cells</cell_type> results in the activation of the <protein>NF-kappaB transcription factor</protein> leading to the upregulation of IL-6 gene expression in these <cell_type>lymphocytes</cell_type> .


Copyright 2001 Academic Press .


Gene- and tissue-specificity of mutation in Big Blue rats treated with the hepatocarcinogen N-hydroxy-2-acetylaminofluorene .


In a previous study , we found that treating transgenic Big Blue rats with the hepatocarcinogen N-hydroxy-2-acetylaminofluorene ( N-OH-AAF ) produced the same major DNA adduct in the target liver and the nontarget spleen lymphocytes and bone marrow cells , induced lacI mutants in the liver , and induced much lower frequencies of l acI and hprt mutants in spleen lymphocytes .

In a previous study , we found that treating transgenic Big Blue rats with the hepatocarcinogen N-hydroxy-2-acetylaminofluorene ( N-OH-AAF ) produced the same major DNA adduct in the target liver and the <cell_type>nontarget spleen lymphocytes</cell_type> and <cell_type>bone marrow cells</cell_type> , induced <cell_type>lacI mutants</cell_type> in the liver , and induced much lower frequencies of l <cell_type>acI and hprt mutants</cell_type> in <cell_type>spleen lymphocytes</cell_type> .


In the present study , sequence analysis was conducted on lacI DNA and hprt cDNA from the mutants , to determine the mutational specificity of N-OH-AAF in the rat .

In the present study , sequence analysis was conducted on <dna>lacI DNA</dna> and <dna>hprt cDNA</dna> from the mutants , to determine the mutational specificity of N-OH-AAF in the rat .


All the mutation spectra from N-OH-AAF-treated rats differed significantly from corresponding mutation profiles from untreated animals ( P = 0.02 to P < 0.0001 ) .


Although there were similarities among the mutational patterns derived from N-OH-AAF-treated rats ( e.g. , G : C -- > T : A transversion was the most common mutation in all mutation sets ) , there were significant differences in the patterns of basepair substitution and frameshift mutation between the liver and spleen lymphocyte lacI mutants ( P = 0.02 ) and between the spleen lymphocyte lacI and hprt mutants ( P = 0.04 ) .

Although there were similarities among the mutational patterns derived from N-OH-AAF-treated rats ( e.g. , G : C -- > T : A transversion was the most common mutation in all mutation sets ) , there were significant differences in the patterns of basepair substitution and frameshift mutation between the liver and <cell_type>spleen lymphocyte lacI mutants</cell_type> ( P = 0.02 ) and between the <cell_type>spleen lymphocyte</cell_type> <cell_type>lacI and hprt mutants</cell_type> ( P = 0.04 ) .


Also , multiplex PCR analysis of genomic DNA from the hprt mutants indicated that 12 % of mutants from treated rats had major deletions in the hprt gene ; no corresponding incidence of large deletions was evident among lacI mutations .

Also , multiplex PCR analysis of <dna>genomic DNA</dna> from the <cell_type>hprt mutants</cell_type> indicated that 12 % of mutants from treated rats had major deletions in the <dna>hprt gene</dna> ; no corresponding incidence of large deletions was evident among lacI mutations .


All the mutation profiles reflect the general mutational specificity of the major DNA adduct formed by N-OH-AAF .


The differences between N-OH-AAF mutation in the endogenous gene and transgene can be partially explained by the structures of the two genes .

The differences between N-OH-AAF mutation in the <dna>endogenous gene</dna> and <dna>transgene</dna> can be partially explained by the structures of the two genes .


The tissue-specificity of the mutation spectra may contribute to targeting tumor formation to the liver .


Environ. Mol. Mutagen. 37 : 203-214 , 2001 .


Published 2001 Wiley-Liss , Inc .


Caspase -dependent cleavage of the hematopoietic specific adaptor protein Gads alters signalling from the T cell receptor .

<protein>Caspase</protein> -dependent cleavage of the <protein>hematopoietic specific adaptor protein Gads</protein> alters signalling from the <protein>T cell receptor</protein> .


Gads is a SH2 and SH3 domain -containing , hematopoietic-specific adaptor protein that functions in signalling from the T cell receptor .

<protein>Gads</protein> is a <protein>SH2 and SH3 domain</protein> -containing , <protein>hematopoietic-specific adaptor protein</protein> that functions in signalling from the <protein>T cell receptor</protein> .


Gads acts by linking SLP-76 , bound by the carboxy-terminal Gads SH3 domain , to tyrosine phosphorylated LAT which contains binding sites for the Gads SH2 domain .

<protein>Gads</protein> acts by linking <protein>SLP-76</protein> , bound by the <protein>carboxy-terminal Gads SH3 domain</protein> , to <protein>tyrosine phosphorylated LAT</protein> which contains binding sites for the <protein>Gads SH2 domain</protein> .


Gads is distinguished from Grb2 and the closely related Grap protein by the presence of a 120 amino acid unique region between the SH2 domain and the carboxy terminal SH3 domain .

<protein>Gads</protein> is distinguished from <protein>Grb2</protein> and the closely related <protein>Grap protein</protein> by the presence of a <protein>120 amino acid unique region</protein> between the <protein>SH2 domain</protein> and the <protein>carboxy terminal SH3 domain</protein> .


Here we demonstrate that the unique region of Gads contains a capase cleavage site .

Here we demonstrate that the unique region of <protein>Gads</protein> contains a <protein>capase cleavage site</protein> .


Induction of apoptosis in lymphocytes results in detectable Gads cleavage by 60 min .

Induction of apoptosis in <cell_type>lymphocytes</cell_type> results in detectable <protein>Gads</protein> cleavage by 60 min .


Gads cleavage is blocked in vivo by treating cells with a caspase 3 inhibitor .

<protein>Gads</protein> cleavage is blocked in vivo by treating cells with a caspase 3 inhibitor .


A putative caspase 3 cleavage site was identified within the unique region and mutation of this site prevented Gads cleavage in vitro , and in vivo .

A <protein>putative caspase 3 cleavage site</protein> was identified within the unique region and mutation of this site prevented <protein>Gads</protein> cleavage in vitro , and in vivo .


The Gads cleavage products retained the predicted binding specificity for SLP-76 and LAT .

The <protein>Gads</protein> cleavage products retained the predicted binding specificity for <protein>SLP-76</protein> and <protein>LAT</protein> .


Expression of the Gads cleavage products in Jurkat T cells inhibited NFAT activation following TCR cross linking .

Expression of the <protein>Gads</protein> cleavage products in <cell_line>Jurkat T cells</cell_line> inhibited NFAT activation following TCR cross linking .


These findings indicate that cleavage of Gads in vivo could function to alter signalling downstream of the T cell receptor by disrupting cross talk between SLP-76 and LAT .

These findings indicate that cleavage of <protein>Gads</protein> in vivo could function to alter signalling downstream of the <protein>T cell receptor</protein> by disrupting cross talk between <protein>SLP-76</protein> and <protein>LAT</protein> .


Targeting of p300 to the interleukin-2 promoter via CREB-Rel cross-talk during mitogen and oncogenic molecular signaling in activated T-cells .

Targeting of <protein>p300</protein> to the <dna>interleukin-2 promoter</dna> via CREB-Rel cross-talk during <protein>mitogen</protein> and oncogenic molecular signaling in <cell_type>activated T-cells</cell_type> .


In this report , we explore the mechanisms of targeting of p300 to the interleukin-2 ( IL-2 ) promoter in response to mitogenic and oncogenic molecular signals .

In this report , we explore the mechanisms of targeting of <protein>p300</protein> to the <dna>interleukin-2 ( IL-2 ) promoter</dna> in response to mitogenic and oncogenic molecular signals .


Recruitment of p300 by cAMP-responsive element-binding protein-Rel cross-talk at the composite CD28 response element ( CD28RE ) - TRE element of the IL-2 promoter is essential for promoter inducibility during T-cell activation , and CD28RE-TRE is the exclusive target of the human T-cell lymphotropic virus type I oncoprotein Tax .

Recruitment of <protein>p300</protein> by <protein>cAMP-responsive element-binding protein-Rel</protein> cross-talk at the <protein>composite CD28 response element</protein> ( <protein>CD28RE</protein> ) - <dna>TRE element</dna> of the <dna>IL-2 promoter</dna> is essential for promoter inducibility during T-cell activation , and <dna>CD28RE-TRE</dna> is the exclusive target of the <cell_type>human T-cell</cell_type> lymphotropic virus type I <protein>oncoprotein Tax</protein> .


The intrinsic histone acetyltransferase activity of p300 is dispensable for activation of the IL-2 promoter , and the N-terminal 743 residues contain the minimal structural requirements for synergistic transactivation of the CD28RE-TRE , the IL-2 promoter , and endogenous IL-2 gene expression .

The intrinsic histone acetyltransferase activity of <protein>p300</protein> is dispensable for activation of the <dna>IL-2 promoter</dna> , and the <protein>N-terminal 743 residues</protein> contain the minimal structural requirements for synergistic transactivation of the <dna>CD28RE-TRE</dna> , the <dna>IL-2 promoter</dna> , and endogenous <dna>IL-2 gene</dna> expression .


Mutational analysis of p300 reveals differential structural requirements for the N-terminal p300 module by individual cis-elements within the IL-2 promoter .

Mutational analysis of <protein>p300</protein> reveals differential structural requirements for the N-terminal p300 module by individual <dna>cis-elements</dna> within the <dna>IL-2 promoter</dna> .


These findings provide evidence that p300 assembles at the IL-2 promoter to form an enhanceosome-like signal transduction target that is centrally integrated at the CD28RE-TRE element of the IL-2 promoter through specific protein module-targeted associations in activated T-cells .

These findings provide evidence that <protein>p300</protein> assembles at the <dna>IL-2 promoter</dna> to form an enhanceosome-like signal transduction target that is centrally integrated at the <dna>CD28RE-TRE element</dna> of the <dna>IL-2 promoter</dna> through specific protein module-targeted associations in <cell_type>activated T-cells</cell_type> .


Regulation of cytokine production in T-cell responses to inhalant allergen : GATA-3 expression distinguishes between Th1- and Th2-polarized immunity .

Regulation of cytokine production in <cell_type>T-cell</cell_type> responses to inhalant allergen : <protein>GATA-3</protein> expression distinguishes between Th1- and Th2-polarized immunity .


BACKGROUND : The precise nature of allergen-specific cytokine responses in atopics versus non-atopics , in particular the 'Th1 polarity ' of responses in non-atopics , remains controversial .


This is due in part to the relative insensitivity of cytokine detection systems , and associated variations in kinetics of cytokine production and catabolism in in vitro culture systems .


As an alternative to cytokine measurement , this study focuses on expression of the transcription factor GATA-3 for analysis of allergen-specific Th cell responses .

As an alternative to cytokine measurement , this study focuses on expression of the <protein>transcription factor GATA-3</protein> for analysis of allergen-specific Th cell responses .


METHODS : Cord blood mononuclear cells were Th1- or Th2-polarized by culture in IL-12- or IL-4-employing established methods ; PBMC from house dust mite ( HDM ) -sensitive atopics and controls were stimulated overnight with HDM ; cytokine production was measured by ELISA and GATA-3 mRNA expression by PCR .

METHODS : <cell_type>Cord blood mononuclear cells</cell_type> were Th1- or Th2-polarized by culture in IL-12- or IL-4-employing established methods ; <cell_type>PBMC</cell_type> from house dust mite ( HDM ) -sensitive atopics and controls were stimulated overnight with HDM ; cytokine production was measured by ELISA and <rna>GATA-3 mRNA</rna> expression by PCR .


RESULTS : Cytokine -driven Th2 polarization of naive T cells is associated with marked upregulation of GATA-3 expression , whereas a reciprocal expression pattern accompanies differentiation towards the Th1 cytokine phenotype .

RESULTS : <protein>Cytokine</protein> -driven Th2 polarization of <cell_type>naive T cells</cell_type> is associated with marked upregulation of <protein>GATA-3</protein> expression , whereas a reciprocal expression pattern accompanies differentiation towards the Th1 cytokine phenotype .


In T cells from HDM skin prick test-positive ( HDM-SPT+/HDM-IgE+ ) volunteers , overnight stimulation results in marked upregulation of GATA-3 expression , compared to an equally marked downregulation of expression in T cells from SPT-/IgE- subjects .

In <cell_type>T cells</cell_type> from HDM skin prick test-positive ( HDM-SPT+/HDM-IgE+ ) volunteers , overnight stimulation results in marked upregulation of <protein>GATA-3</protein> expression , compared to an equally marked downregulation of expression in <cell_type>T cells</cell_type> from SPT-/IgE- subjects .


In subjects who are HDM-SPT+ but IgE- , GATA-3 expression levels remained relatively stable during culture with HDM .

In subjects who are HDM-SPT+ but IgE- , <protein>GATA-3</protein> expression levels remained relatively stable during culture with HDM .


CONCLUSIONS : Upregulation of GATA-3 expression in PBMC is a hallmark of the early phase of Th2 recall responses to specific allergen in atopics .

CONCLUSIONS : Upregulation of <protein>GATA-3</protein> expression in <cell_type>PBMC</cell_type> is a hallmark of the early phase of Th2 recall responses to specific allergen in atopics .


The reciprocal expression pattern observed in HDM-specific recall responses of non-atopics provides independent confirmation of the presence of underlying Th1-like immunity in these subjects .


The parallel findings in neonatal T cells suggest that the same approach may be utilized for monitoring the progress of allergen-specific Th1/Th2 memory development during early childhood , and hence in assessment of risk for future allergic disease .

The parallel findings in <cell_type>neonatal T cells</cell_type> suggest that the same approach may be utilized for monitoring the progress of allergen-specific Th1/Th2 memory development during early childhood , and hence in assessment of risk for future allergic disease .


Copyright 2001 S. Karger AG , Basel


The heat shock response reduces myelin oligodendrocyte glycoprotein -induced experimental autoimmune encephalomyelitis in mice .

The heat shock response reduces <protein>myelin oligodendrocyte glycoprotein</protein> -induced experimental autoimmune encephalomyelitis in mice .


The stress response ( SR ) can block inflammatory gene expression by preventing activation of transcription factor nuclear factor-kappa B ( NF-kappaB ) .

The stress response ( SR ) can block inflammatory gene expression by preventing activation of <protein>transcription factor</protein> <protein>nuclear factor-kappa B</protein> ( <protein>NF-kappaB</protein> ) .


As inflammatory gene expression contributes to the pathogenesis of demyelinating diseases , we tested the effects of the SR on the progression of the demyelinating disease experimental autoimmune encephalomyelitis ( EAE ) .


EAE was actively induced in C57BL/6 mice using an encephalitogenic myelin oligodendrocyte glycoprotein ( MOG ( 35-55 ) ) peptide .

EAE was actively induced in C57BL/6 mice using an encephalitogenic <protein>myelin oligodendrocyte glycoprotein</protein> ( <protein>MOG ( 35-55 )</protein> ) peptide .


Whole body hyperthermia was used to induce a heat shock response ( HSR ) in immunized mice 2 days after the booster MOG ( 35-55 ) peptide injection .


The HSR reduced the incidence of EAE by 70 % , delayed disease onset by 6 days , and attenuated disease severity .


The HSR attenuated leukocyte infiltration into CNS assessed by quantitation of perivascular infiltrates , and by reduced staining for CD4 and CD25 immunopositive T-cells .

The HSR attenuated <cell_type>leukocyte</cell_type> infiltration into CNS assessed by quantitation of perivascular infiltrates , and by reduced staining for <protein>CD4</protein> and <protein>CD25</protein> immunopositive <cell_type>T-cells</cell_type> .


T-cell activation , assessed by the production of interferon gamma ( IFNgamma ) in response to MOG ( 35-55 ) , was also decreased by the HSR .

T-cell activation , assessed by the production of <protein>interferon gamma</protein> ( <protein>IFNgamma</protein> ) in response to <protein>MOG ( 35-55 )</protein> , was also decreased by the HSR .


The HSR reduced inflammatory gene expression in the brain that normally occurs during EAE , including the early increase in RANTES ( regulated on activation of normal T-cell expressed and secreted ) expression , and the later expression of the inducible form of nitric oxide synthase .

The HSR reduced inflammatory gene expression in the brain that normally occurs during EAE , including the early increase in <protein>RANTES</protein> ( <protein>regulated on activation of normal T-cell expressed and secreted</protein> ) expression , and the later expression of the inducible form of <protein>nitric oxide synthase</protein> .


The early activation of transcription factor NF-kappaB was also blocked by the HSR .

The early activation of <protein>transcription factor</protein> <protein>NF-kappaB</protein> was also blocked by the HSR .


The finding that the SR reduces inflammation in the brain and the clinical severity of EAE opens a novel therapeutic approach for prevention of autoimmune diseases .


In vivo detection of intracellular signaling pathways in developing thymocytes .

In vivo detection of intracellular signaling pathways in developing <cell_type>thymocytes</cell_type> .


Information regarding the intracellular signaling processes that occur during the development of T cells has largely been obtained with the use of transgenic mouse models , which although providing invaluable information are time consuming and costly .

Information regarding the intracellular signaling processes that occur during the development of <cell_type>T cells</cell_type> has largely been obtained with the use of transgenic mouse models , which although providing invaluable information are time consuming and costly .


To this end , we have developed a novel system that facilitates the in vivo analysis of signal transduction pathways during T-lymphocyte development .

To this end , we have developed a novel system that facilitates the in vivo analysis of signal transduction pathways during <cell_type>T-lymphocyte</cell_type> development .


This approach uses reporter-plasmids for the detection of intracellular signals mediated by the mitogen-activated protein kinase or cyclic AMP-dependent protein kinase .

This approach uses <dna>reporter-plasmids</dna> for the detection of intracellular signals mediated by the <protein>mitogen-activated protein kinase</protein> or <protein>cyclic AMP-dependent protein kinase</protein> .


Reporter-plasmids are transfected into thymocytes in fetal thymic organ culture by accelerated DNA/particle bombardment ( gene gun ) , and the activation of a signaling pathway is determined in the form of a standard luciferase assay .

<dna>Reporter-plasmids</dna> are transfected into <cell_type>thymocytes</cell_type> in fetal thymic organ culture by accelerated DNA/particle bombardment ( gene gun ) , and the activation of a signaling pathway is determined in the form of a standard luciferase assay .


Importantly , this powerful technique preserves the structural integrity of the thymus , and will provide an invaluable tool to study how thymocytes respond to normal environmental stimuli encountered during differentiation within the thymic milieu .

Importantly , this powerful technique preserves the structural integrity of the thymus , and will provide an invaluable tool to study how <cell_type>thymocytes</cell_type> respond to normal environmental stimuli encountered during differentiation within the thymic milieu .


Thus , this method allows for the monitoring of signals that occur in a biological time frame , such as during differentiation , and within the natural environment of differentiating cells .

Thus , this method allows for the monitoring of signals that occur in a biological time frame , such as during differentiation , and within the natural environment of <cell_type>differentiating cells</cell_type> .


Core-binding factor beta ( CBFbeta ) , but not CBFbeta-smooth muscle myosin heavy chain , rescues definitive hematopoiesis in CBFbeta -deficient embryonic stem cells .

<protein>Core-binding factor beta</protein> ( <protein>CBFbeta</protein> ) , but not <protein>CBFbeta-smooth muscle myosin heavy chain</protein> , rescues definitive hematopoiesis in <protein>CBFbeta</protein> -deficient <cell_type>embryonic stem cells</cell_type> .


Core-binding factor beta ( CBFbeta ) is the non-DNA-binding subunit of the heterodimeric CBFs .

<protein>Core-binding factor beta</protein> ( <protein>CBFbeta</protein> ) is the <protein>non-DNA-binding subunit</protein> of the <protein>heterodimeric CBFs</protein> .


Genes encoding CBFbeta ( CBFB ) , and one of the DNA-binding CBFalpha subunits , Runx1 ( also known as CBFalpha2 , AML1 , and PEBP2alphaB ) , are required for normal hematopoiesis and are also frequent targets of chromosomal translocations in acute leukemias in humans .

<dna>Genes encoding CBFbeta</dna> ( <dna>CBFB</dna> ) , and one of the <protein>DNA-binding CBFalpha subunits , Runx1</protein> ( also known as <protein>CBFalpha2</protein> , <protein>AML1</protein> , and <protein>PEBP2alphaB</protein> ) , are required for normal hematopoiesis and are also frequent targets of chromosomal translocations in acute leukemias in humans .


Homozygous disruption of either the Runx1 or Cbfb gene in mice results in embryonic lethality at midgestation due to hemorrhaging in the central nervous system , and severely impairs fetal liver hematopoiesis .

Homozygous disruption of either the <dna>Runx1 or Cbfb gene</dna> in mice results in embryonic lethality at midgestation due to hemorrhaging in the central nervous system , and severely impairs fetal liver hematopoiesis .


Results of this study show that Cbfb-deficient mouse embryonic stem ( ES ) cells can differentiate into primitive erythroid colonies in vitro , but are impaired in their ability to produce definitive erythroid and myeloid colonies , mimicking the in vivo defect .

Results of this study show that Cbfb-deficient mouse <cell_type>embryonic stem ( ES ) cells</cell_type> can differentiate into <cell_type>primitive erythroid colonies</cell_type> in vitro , but are impaired in their ability to produce definitive <cell_type>erythroid and myeloid colonies</cell_type> , mimicking the in vivo defect .


Definitive hematopoiesis is restored by ectopic expression of full-length Cbfb transgenes , as well as by a transgene encoding only the heterodimerization domain of CBFbeta .

Definitive hematopoiesis is restored by ectopic expression of <dna>full-length Cbfb transgenes</dna> , as well as by a transgene encoding only the <protein>heterodimerization domain</protein> of <protein>CBFbeta</protein> .


In contrast , the CBFbeta - smooth muscle myosin heavy chain ( SMMHC ) fusion protein generated by the inv ( 16 ) associated with acute myeloid leukemias ( M4Eo ) can not rescue definitive hematopoiesis by Cbfb-deficient ES cells .

In contrast , the <protein>CBFbeta</protein> - <protein>smooth muscle myosin heavy chain ( SMMHC ) fusion protein</protein> generated by the <protein>inv ( 16 )</protein> associated with <cell_line>acute myeloid leukemias</cell_line> ( <cell_line>M4Eo</cell_line> ) can not rescue definitive hematopoiesis by <cell_type>Cbfb-deficient ES cells</cell_type> .


Sequences responsible for the inability of CBFbeta -SMMHC to rescue definitive hematopoiesis reside in the SMMHC portion of the fusion protein .

Sequences responsible for the inability of <protein>CBFbeta</protein> -SMMHC to rescue definitive hematopoiesis reside in the <protein>SMMHC portion</protein> of the <protein>fusion protein</protein> .


Results also show that the CBFbeta-SMMHC fusion protein transdominantly inhibits definitive hematopoiesis , but not to the same extent as homozygous loss of Runx1 or Cbfb .

Results also show that the <protein>CBFbeta-SMMHC fusion protein</protein> transdominantly inhibits definitive hematopoiesis , but not to the same extent as homozygous loss of <protein>Runx1</protein> or <protein>Cbfb</protein> .


CBFbeta-SMMHC preferentially inhibits the differentiation of myeloid lineage cells , while increasing the number of blastlike cells in culture .

<protein>CBFbeta-SMMHC</protein> preferentially inhibits the differentiation of <cell_type>myeloid lineage cells</cell_type> , while increasing the number of <cell_type>blastlike cells</cell_type> in culture .


The latency pattern of Epstein-Barr virus infection and viral IL-10 expression in cutaneous natural killer/T-cell lymphomas .

The latency pattern of Epstein-Barr virus infection and viral <protein>IL-10</protein> expression in <cell_type>cutaneous natural killer/T-cell lymphomas</cell_type> .


The nasal type , extranodal natural killer or T ( NK/T ) -cell lymphoma is usually associated with latent Epstein-Barr virus ( EBV ) infection .

The <cell_type>nasal type , extranodal natural killer or T ( NK/T ) -cell lymphoma</cell_type> is usually associated with latent Epstein-Barr virus ( EBV ) infection .


In order to elucidate the EBV gene expression patterns in vivo , we examined eight patients with cutaneous EBV-related NK/T-cell lymphomas , including six patients with a NK-cell phenotype and two patients with a T-cell phenotype .